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WO2013026587A1 - 1,4 disubstituted pyrrolidine - 3 - yl -amine derivatives and their use for the treatment of metabolic disorders - Google Patents

1,4 disubstituted pyrrolidine - 3 - yl -amine derivatives and their use for the treatment of metabolic disorders Download PDF

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Publication number
WO2013026587A1
WO2013026587A1 PCT/EP2012/057964 EP2012057964W WO2013026587A1 WO 2013026587 A1 WO2013026587 A1 WO 2013026587A1 EP 2012057964 W EP2012057964 W EP 2012057964W WO 2013026587 A1 WO2013026587 A1 WO 2013026587A1
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WO
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Prior art keywords
preparation
phenyl
pyrrolidin
methyl
piperidin
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Application number
PCT/EP2012/057964
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French (fr)
Inventor
Oscar Barba
Lisa Sarah Bertram
Emma Louise Carswell
Susan Helen Davis
Peter Timothy Fry
Robert James Gleave
Revathy Perpetua Jeevaratnam
Craig Johnstone
John Keily
Martin James Procter
Karen Lesley Schofield
Alan John William Stewart
Simon Andrew Swain
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Prosidion Limited
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Publication of WO2013026587A1 publication Critical patent/WO2013026587A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P3/00—Drugs for disorders of the metabolism
    • A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
    • A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings

Definitions

  • the present invention is directed to therapeutic compounds useful for the treatment of metabolic disorders including type II diabetes.
  • the present invention is 5 directed to compounds which have activity as agonists of GPR1 19.
  • Drugs aimed at the pathophysiology associated with non-insulin dependent type II diabetes have many potential side effects and do not adequately address the dyslipidaemia and hyperglycaemia in a high proportion of patients. Treatment is often focused at individual patient needs using diet, exercise, hypoglycaemic agents and insulin, but there is a0 continuing need for novel antidiabetic agents, particularly ones that may be better tolerated with fewer adverse effects.
  • metabolic syndrome places people at high risk of coronary artery disease, and is characterized by a cluster of risk factors including central obesity (excessive fat tissue in the abdominal region), glucose intolerance, high triglycerides and low5 HDL cholesterol, and high blood pressure.
  • central obesity excessive fat tissue in the abdominal region
  • glucose intolerance high triglycerides
  • low5 HDL cholesterol low5 HDL cholesterol
  • Obesity is characterized by an excessive adipose tissue mass relative to body size.
  • body fat mass is estimated by the body mass index (BMI; weight (kg)/height0 (m) 2 ), or waist circumference.
  • BMI body mass index
  • Individuals are considered obese when the BMI is greater than 30 and there are established medical consequences of being overweight. It has been an accepted medical view for some time that an increased body weight, especially as a result of abdominal body fat, is associated with an increased risk for diabetes, hypertension, heart disease, and numerous other health complications, such as arthritis, stroke, gallbladder5 disease, muscular and respiratory problems, back pain and even certain cancers.
  • GPR119 (previously referred to as GPR116) is a GPCR identified as SNORF25 in0 WO00/50562 which discloses both the human and rat receptors, US 6,468,756 also discloses the mouse receptor (accession numbers: AAN95194 (human), AAN95195 (rat) and A 95196 (mouse)).
  • GPR1 19 is expressed in the pancreas, small intestine, colon and adipose tissue. The expression profile of the human GPR1 19 receptor indicates its potential utility as a target for the treatment of diabetes.
  • GPR119 agonists have been shown to stimulate the release of GLP-1 from the GI tract. In doing so, GPR119 agonists (1) enhance glucose-dependent insulin release from the pancreas leading to improvements in oral glucose tolerance; (2) attenuate disease progression by increasing ⁇ -cell cAMP concentrations; and (3) induce weight loss possibly through GLP-1 's ability to reduce food intake.
  • DPP-IV Dipeptidyl peptidase IV
  • GLP- 1 Dipeptidyl peptidase IV
  • DPP-IV inhibitors are of use for the treatment of type II diabetes, examples of DPP-IV inhibitors include vildagliptin, sitagliptin, alogliptin and saxagliptin.
  • WO 2009/034388, WO 2010/103333, WO 2010/103334 and WO 2010/103333 disclose dual GPR119 agonists and DPP-IV inhibitors.
  • the compounds of the invention preferably have dual activity as agonists of GPR1 19 and inhibitors of DPP-IV
  • the present invention provides compounds of formula (I) and pharmaceutically acceptable salts thereof:
  • A is a para-linked phenyl, pyridinyl, pyrimidinyl, pyrazinyl or triazinyl;
  • R1 is hydrogen, halo, cyano, Ci _4alkyl, C ⁇ ⁇ haloalkyl, Ci _4alkoxy or
  • N-pyridonyl optionally substituted by one or more groups independently selected from halo, methyl or halomethyl groups
  • R3 is C3_5cycloalkyl, 4 to 6 membered saturated heterocyclyl (comprising 1 or 2 ring heteroatoms selected from ⁇ , O and S) or Ci _4alkyl wherein the C3_ cycloalkyl and Ci _ 4alkyl are optionally substituted by one to three groups independently selected from halo, cyano, hydroxy or Ci ⁇ alkoxy;
  • p and q are each 0, 1 or 2, provided that 0 ⁇ p+q ⁇ 2;
  • Z is selected from the group consisting of:
  • heteroaryl ring or fused bicyclic system optionally substituted by one or two groups independently selected from halo, cyano, C1 .4 alkyl, Ci _4haloalkyl, Ci _4alkoxy, Ci _4hydroxyalkyl, Ci _4alkoxyCi _4alkyl, heterocyclyl (a 4 to 6 membered saturated heterocycle comprising 1 or 2 ring heteroatoms selected from N , O and S), heterocyclyl Ci .
  • heterocyclyl is a 4 to 6 membered saturated heterocycle comprising 1 or 2 ring heteroatoms selected from N, O and S
  • C3_ cycloalkyl optionally substituted by Ci _4 alkyl, C i .4 alkoxy or halo
  • R4 is selected from the group consisting of:
  • R7 is hydrogen, halo, C ⁇ .2 alkyl, C ⁇ ⁇ haloalkyl or C ⁇ .3 alkoxy;
  • (la) p and q are independently 0, 1 or 2 provided that p+q does not exceed 2, i.e., forming a 4-, 5- or 6-membered ring.
  • p and q may be the same, i.e., forming a 4- or 6-membered ring.
  • p and q are both 1, therefore the compounds of the invention may have the formula:
  • A is suitably pyridine, pyrimidine or pyrazine typically pyridine or pyrimidine, e.g., 2- or 3-pyridyl or 2- or 5-pyrimidinyl, where the 2-, 3- or 5- refers to the point of attachment of the pyrrolidine ring (X being attached in the para position and R 1 being attached at any suitable position).
  • R1 is suitably hydrogen, halogen or cyano.
  • R ⁇ is preferably hydrogen.
  • R ⁇ may optionally be substituted by 1, 2 or 3 substituents.
  • R ⁇ is optionally substituted by 1 or 2 substituents and in further embodiments it is substituted by 2 or 3 substituents.
  • R ⁇ is preferably optionally substituted by 1, 2, or 3 substituents independently selected from halo and methyl.
  • R ⁇ is preferably optionally substituted by 1 or 2 substituents independently selected from halo and methyl.
  • R ⁇ is suitably phenyl substituted by 1, 2 or 3 groups selected from halo and methyl groups.
  • R ⁇ is suitably phenyl substituted by 2 or 3 halo groups.
  • Halo is suitably fluoro.
  • R ⁇ may be an optionally substituted phenyl or pyridyl, e.g.
  • R ⁇ examples include phenyl, 2-pyridinyl, 4-fluoro-2-pyridinyl, 5-fluoro-2-pyridinyl, 4- methyl-2-pyridinyl, 5-methyl-2-pyridinyl, 2-fluorophenyl, 3 -fluorophenyl, 4-fluorophenyl, 2,3-difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl, 2,3,4- trifluorophenyl, 2,3,5-trifluorophenyl, 2,3,6-trifluorophenyl, 2,4,5-trifluorophenyl, 2,4,6- trifluorophenyl, 2-fluoro-5-methylphenyl and 2,4-difluoro-5-methylphenyl.
  • R3 may suitably be C3_ cycloalkyl or Ci _4 alkyl wherein the Ci _4 alkyl is optionally substituted by one to three groups independently selected from halo, cyano, hydroxy or Ci _2alkoxy.
  • Suitable examples of include methyl, ethyl, n-propyl, i-propyl, cyclopropyl, CH 2 CH 2 OCH 3 and CH 2 CF 3 .
  • Z is -C(0)OR ⁇ . In further embodiments Z is C(0)R ⁇ . In still further embodiments it is S(0)2R ⁇ .
  • R4 may suitably be C ⁇ alkyl, C ⁇ _ alkoxyC2-6alkyl, C3_ cycloalkyl or C3.
  • R ⁇ examples include methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl or t-butyl, cyclopropyl and 1-methylcyclopropyl.
  • R ⁇ is propyl, especially isopropyl.
  • Z may be a heteroaryl group optionally substituted by one or two groups selected from halo, C 1.4 alkyl, Ci _4haloalkyl, Ci _4alkoxy, Ci _4alkoxyCi _4alkyl, Ci ⁇ hydroxyalkyl,
  • Ci _4haloalkyls are perfluoroalkyls, e.g. CF 3 , CF2Me and CHF 2
  • suitable heteroaryl groups include oxadiazole, pyrimidine, pyridazine, thiazole, tetrazole, benzothiazole and thiadiazole, e.g., oxadiazole and pyrimidine.
  • Z may comprise l,2,4-oxadiazol-3-yl, l,2,4-oxadiazol-5- yl, pyrimidin-2-yl or a 2H-tetrazol-5-yl, which may be substituted by any of the aforementioned substituents.
  • Z is a substituted heteroaryl
  • suitable substituents include ethyl, i-propyl, t- butyl, cyclopropyl, chloro, 1-methylcyclopropyl, CF 3 , CH 2 F, CF 2 H, CH(OMe)Me, C(Me) 2 OH, C(Me) 2 F, CF 2 Me, CH 2 OMe and 2-tetrahydrofuranyl.
  • R ⁇ is preferably hydrogen.
  • the molecular weight of the compounds of the invention is suitably less than about 800, typically less than about 600.
  • the invention also includes isotopically labeled compounds, which are similar to those recited in formulae (I), (la) and (lb) and following, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number most commonly found in nature.
  • isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, fluorine, such as 3 ⁇ 4, ⁇ C, and ⁇ F.
  • Isotopically labeled compounds of the present invention for example those into which radioactive isotopes such as 3 ⁇ 4, l ⁇ C are incorporated, are useful in drug and/or substrate tissue distribution assays. Tritiated, i.e., 3 ⁇ 4, and carbon-14, i.e., ⁇ C, isotopes are particularly preferred for their ease of preparation and detectability. and isotopes are particularly useful in PET (positron emission tomography). PET is useful in brain imaging.
  • isotopically labeled compounds of formula (I), (la) and (lb) and following of this invention can generally be prepared by carrying out the procedures disclosed in the Schemes and/or in the Examples below, by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent.
  • the compounds of formula (I), (la) and (lb) or salts thereof are not isotopically labelled.
  • alkyl means carbon chains which may be linear or branched. Examples of alkyl groups include ethyl, propyl, isopropyl, butyl, sec- and tert-butyl. Such alkyl groups may in some embodiments be substituted with one or more halo groups, particularly fluoro.
  • cycloalkyl refers to a 3 to 6 C saturated carbocycle moiety e.g. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.
  • heterocyclyl refers to a 4 to 6 membered saturated heterocycle comprising 1 or 2 ring heteroatoms selected from N, O and S.
  • heterocycles include tetrahydrofuran, pyrrolidine, tetrahydropyran, piperidine, oxetane and azetidine.
  • heteroaryl refers to a 5- or 6-membered aromatic heteroaryl ring optionally containing one or more, e.g. 1 , 2 or 3 heteroatoms selected from N, O and S, or a 8 to 10 membered fused aromatic bicyclic system optionally containing one or more, e.g.
  • heteroaryl rings 1, 2 or 3 heteroatoms selected from N, O and S.
  • heteroaryl rings include pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, tetrazolyl and benzothiazolyl.
  • Reference to para-linked in relation to ring A refers to the relative positions of the carbonyl group and the pyrrolidine group.
  • Compounds described herein may contain one or more asymmetric centers and may thus give rise to diastereomers and optical isomers.
  • the present invention includes all such possible diastereomers as well as their racemic mixtures, their substantially pure resolved enantiomers, all possible geometric isomers, and pharmaceutically acceptable salts thereof.
  • the present invention includes all stereoisomers of the compounds of the invention and pharmaceutically acceptable salts thereof. Further, mixtures of stereoisomers as well as isolated specific stereoisomers are also included. During the course of the synthetic procedures used to prepare such compounds, or in using racemization or epimerization procedures known to those skilled in the art, the products of such procedures can be a mixture of stereoisomers.
  • the present invention includes any possible tautomers and pharmaceutically acceptable salts thereof, and mixtures thereof, except where specifically drawn or stated otherwise.
  • the present invention includes any possible solvates and polymorphic forms.
  • a type of a solvent that forms the solvate is not particularly limited so long as the solvent is pharmacologically acceptable.
  • water, ethanol, propanol, acetone or the like can be used.
  • salts refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids.
  • pharmaceutically acceptable non-toxic bases including inorganic bases and organic bases.
  • Salts derived from such inorganic bases include aluminum, ammonium, calcium, copper (ic and ous), ferric, ferrous, lithium, magnesium, potassium, sodium, zinc and the like salts. Particularly preferred are the ammonium, calcium, magnesium, potassium and sodium salts.
  • Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, as well as cyclic amines and substituted amines such as naturally occurring and synthesized substituted amines.
  • organic non-toxic bases from which salts can be formed include arginine, betaine, caffeine, choline, N',N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2- dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N- ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, trimethamine and the like.
  • the compound of the invention When the compound of the invention is basic, its corresponding salt can be conveniently prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids.
  • acids include, for example, acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic, trifluoroacetic acid and the like.
  • the compounds of the invention are intended for pharmaceutical use they are preferably provided in substantially pure form, for example at least 60% pure, more suitably at least 75% pure, especially at least 98% pure (% are on a weight for weight basis).
  • R 3 , R 4 , R 5 , R 6 , R 7 , A, Z, p, q and r are as defined for formula (I).
  • PG is a protecting group
  • LG is a leaving group
  • Hal is halogen
  • Het is heteroaryl.
  • Esters of formula (V) can be prepared by S A T displacement of suitable haloaromatic compounds of formula (III) with amines of formula (IV) under standard conditions, for example, DBU and DMSO at 80 - 100°C.
  • Acids of formula (VI) can be prepared from esters of formula (V) under standard conditions, for example, LiOH, water and methanol at room temperature.
  • Amides of formula (II) can be prepared from acids of formula (VI) with amines of formula (VII) under standard amide coupling conditions, for example, HOBT and EDCI, in a suitable solvent, such as DCM.
  • Sulfonamides of formula (XIV) can be prepared from amines of formula (VIII) and sulfonyl chlorides of formula (XIII) under standard conditions, for example, triethylamine in a suitable solvent, such as DCM. Removal of the protecting group PG from the amine functionality in compounds of formula (XIV) can be achieved using standard conditions well known to those skilled in the art.
  • Compounds of formula (VII) where Z is heteroaryl can be prepared as outlined in Scheme 6.
  • Compounds of formula (XVI) can be prepared by S displacement of suitable haloaromatic compounds of formula (XV) with amines of formula (VIII) under standard conditions, for example, DBU and DMSO at 80 - 100°C.
  • compounds of formula (XVI) can be prepared by reaction of suitable haloaromatic compounds of formula (XV) with amines of formula (VIII) under Buchwald-Hartwig conditions, such as, Pd2(dba and ⁇ in a suitable solvent, such as toluene at 1 10°C. Removal of the protecting group PG from the amine functionality in compounds of formula (XVI) can be achieved using standard conditions well known to those skilled in the art.
  • VIM XV XVI VII Compounds of formula (IV) where R 2 is N-pyridonyl can be prepared as outlined in Scheme 7.
  • Compounds of formula (XVIII) can be prepared from alcohols of formula (XVII) under standard conditions, for example, methanesulfonyl chloride and triethylamine in a suitable solvent, such as, THF at 0°C.
  • Diazabicyclo compounds of formula (XIX) can be prepared from compounds of formula (XVIII) by treatment with sodium hydride in a suitable solvent, such as, THF at 0°C.
  • Compounds of formula (XX) can be prepared from compounds of formula (XIX), pyrid-2-one and potassium tert-butoxide in a suitable solvent, such as, ⁇ in a microwave reactor at 100°C. Removal of the protecting group PG' from the amine functionality in compounds of formula (XX) can be achieved using standard conditions well known to those skilled in the art.
  • the compounds of formula (I) may be prepared singly or as compound libraries comprising at least 2, for example 5 to 1 ,000, compounds and more preferably 10 to 100 compounds of formula (I).
  • Compound libraries may be prepared by a combinatorial "split and mix” approach or by multiple parallel synthesis using either solution or solid phase chemistry, using procedures known to those skilled in the art.
  • labile functional groups in the intermediate compounds e.g. hydroxy, carboxy and amino groups
  • the protecting groups may be removed at any stage in the synthesis of the compounds of formula (I) or may be present on the final compound of formula (I).
  • a comprehensive discussion of the ways in which various labile functional groups may be protected and methods for cleaving the resulting protected derivatives is given in, for example, Protective Groups in Organic Chemistry, T.W. Greene and P.G.M. Wuts, (1991) Wiley-Interscience, New York, 2 nd edition.
  • the compounds of the invention are useful as GPR1 19 agonists, e.g. for the treatment and/or prophylaxis of diabetes.
  • the compounds of the invention will generally be administered in the form of a pharmaceutical composition.
  • the compounds of the invention may also be useful as dual GPR1 19 agonists/DPP- IV inhibitors, e.g. for the treatment and/or prophylaxis of diabetes.
  • the compounds of the invention will generally be administered in the form of a pharmaceutical composition.
  • the invention also provides a compound of the invention, or a pharmaceutically acceptable salt thereof, for use as a pharmaceutical.
  • the invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of the invention, in combination with a pharmaceutically acceptable carrier.
  • composition is comprised of a pharmaceutically acceptable carrier and a non-toxic therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof.
  • the invention also provides a pharmaceutical composition for the treatment of disease by modulating GPR1 19 and optionally DPP-IV, resulting in the prophylactic or therapeutic treatment of diabetes, comprising a pharmaceutically acceptable carrier and a non-toxic therapeutically effective amount of compound of the invention, or a pharmaceutically acceptable salt thereof.
  • compositions may optionally comprise other therapeutic ingredients or adjuvants.
  • the compositions include compositions suitable for oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most suitable route in any given case will depend on the particular host, and nature and severity of the conditions for which the active ingredient is being administered.
  • the pharmaceutical compositions may be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.
  • the compounds of the invention can be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques.
  • a pharmaceutical carrier may take a wide variety of forms depending on the form of preparation desired for administration, e.g. oral or parenteral (including intravenous).
  • compositions can be presented as discrete units suitable for oral administration such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient. Further, the compositions can be presented as a powder, as granules, as a solution, as a suspension in an aqueous liquid, as a non-aqueous liquid, as an oil-in-water emulsion, or as a water-in-oil liquid emulsion.
  • the compound of the invention, or a pharmaceutically acceptable salt thereof may also be administered by controlled release means and/or delivery devices.
  • the compositions may be prepared by any of the methods of pharmacy.
  • such methods include a step of bringing into association the active ingredient with the carrier that constitutes one or more necessary ingredients.
  • the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both. The product can then be conveniently shaped into the desired presentation.
  • the compounds of the invention can also be included in pharmaceutical compositions in combination with one or more other therapeutically active compounds.
  • the pharmaceutical carrier employed can be, for example, a solid, liquid, or gas.
  • solid carriers include lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid.
  • liquid carriers are sugar syrup, peanut oil, olive oil, and water.
  • gaseous carriers include carbon dioxide and nitrogen.
  • any convenient pharmaceutical media may be employed.
  • water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and the like may be used to form oral liquid preparations such as suspensions, elixirs and solutions; while carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like may be used to form oral solid preparations such as powders, capsules and tablets. Because of their ease of administration, tablets and capsules are the preferred oral dosage units whereby solid pharmaceutical carriers are employed.
  • tablets may be coated by standard aqueous or nonaqueous techniques.
  • a tablet containing the composition of this invention may be prepared by compression or molding, optionally with one or more accessory ingredients or adjuvants.
  • Compressed tablets may be prepared by compressing, in a suitable machine, the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine, a mixture of the powdered compound moistened with an inert liquid diluent.
  • Each tablet preferably contains from about 0.05mg to about 5g of the active ingredient and each cachet or capsule preferably containing from about 0.05mg to about 5g of the active ingredient.
  • a formulation intended for the oral administration to humans may contain from about 0.5mg to about 5g of active agent, compounded with an appropriate and convenient amount of carrier material which may vary from about 5 to about 95 percent of the total composition.
  • Unit dosage forms will generally contain between from about lmg to about 2g of the active ingredient, typically 25mg, 50mg, lOOmg, 200mg, 300mg, 400mg, 500mg, 600mg, 800mg, or lOOOmg.
  • compositions of the present invention suitable for parenteral administration may be prepared as solutions or suspensions of the active compounds in water.
  • a suitable surfactant can be included such as, for example, hydroxypropylcellulose.
  • Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof in oils. Further, a preservative can be included to prevent the detrimental growth of microorganisms.
  • compositions of the present invention suitable for injectable use include sterile aqueous solutions or dispersions.
  • the compositions can be in the form of sterile powders for the extemporaneous preparation of such sterile injectable solutions or dispersions.
  • the final injectable form must be sterile and must be effectively fluid for easy syringability.
  • the pharmaceutical compositions must be stable under the conditions of manufacture and storage; thus, preferably should be preserved against the contaminating action of microorganisms such as bacteria and fungi.
  • the carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g. glycerol, propylene glycol and liquid polyethylene glycol), vegetable oils, and suitable mixtures thereof.
  • compositions of the present invention can be in a form suitable for topical use such as, for example, an aerosol, cream, ointment, lotion, dusting powder, or the like. Further, the compositions can be in a form suitable for use in transdermal devices. These formulations may be prepared, using a compound of the invention, or a pharmaceutically acceptable salt thereof, via conventional processing methods. As an example, a cream or ointment is prepared by admixing hydrophilic material and water, together with about 5wt% to about 10wt% of the compound, to produce a cream or ointment having a desired consistency.
  • compositions of this invention can be in a form suitable for rectal administration wherein the carrier is a solid. It is preferable that the mixture forms unit dose suppositories. Suitable carriers include cocoa butter and other materials commonly used in the art. The suppositories may be conveniently formed by first admixing the composition with the softened or melted carrier(s) followed by chilling and shaping in molds.
  • the pharmaceutical formulations described above may include, as appropriate, one or more additional carrier ingredients such as diluents, buffers, flavoring agents, binders, surface-active agents, thickeners, lubricants, preservatives (including anti-oxidants) and the like.
  • additional carrier ingredients such as diluents, buffers, flavoring agents, binders, surface-active agents, thickeners, lubricants, preservatives (including anti-oxidants) and the like.
  • additional carrier ingredients such as diluents, buffers, flavoring agents, binders, surface-active agents, thickeners, lubricants, preservatives (including anti-oxidants) and the like.
  • additional carrier ingredients such as diluents, buffers, flavoring agents, binders, surface-active agents, thickeners, lubricants, preservatives (including anti-oxidants) and the like.
  • other adjuvants can be included to render the formulation isotonic with the blood of the intended recipient
  • dosage levels on the order of O.Olmg/kg to about 150mg/kg of body weight per day are useful in the treatment of the above-indicated conditions, or alternatively about 0.5mg to about 7g per patient per day.
  • obesity may be effectively treated by the administration of from about 0.01 to 50mg of the compound per kilogram of body weight per day, or alternatively about 0.5mg to about 3.5g per patient per day. It is understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination and the severity of the particular disease undergoing therapy.
  • the compounds of the invention may be used in the treatment of diseases or conditions in which GPRl 19 and optionally DPP-IV play a role.
  • the invention also provides a method for the treatment of a disease or condition in which GPRl 19 and optionally DPP-IV play a role comprising a step of administering to a subject in need thereof an effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof.
  • diseases or conditions diabetes, obesity, impaired glucose tolerance, insulin resistance and diabetic complications such as neuropathy, nephropathy, retinopathy, cataracts, cardiovascular complications and dyslipidaemia).
  • the compounds of the invention may also be used for treating metabolic diseases such as metabolic syndrome (syndrome X), impaired glucose tolerance, hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, low HDL levels and hypertension.
  • the invention also provides a method for the treatment of type II diabetes, comprising a step of administering to a patient in need thereof an effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof.
  • the invention also provides a method for the treatment of obesity, metabolic syndrome (syndrome X), impaired glucose tolerance, hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, low HDL levels or hypertension comprising a step of administering to a patient in need thereof an effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof.
  • the invention also provides a compound of the invention, or a pharmaceutically acceptable salt thereof, for use in the treatment of a condition as defined above.
  • the invention also provides the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a condition as defined above.
  • treatment includes both therapeutic and prophylactic treatment.
  • the compounds of the invention may exhibit advantageous properties compared to known compounds or combination therapies for the treatment of diabetes.
  • the compounds of the invention, or pharmaceutically acceptable salts thereof, may be administered alone or in combination with one or more other therapeutically active compounds.
  • the other therapeutically active compounds may be for the treatment of the same disease or condition as the compounds of the invention or a different disease or condition.
  • the therapeutically active compounds may be administered simultaneously, sequentially or separately.
  • the compounds of the invention may be administered with other active compounds for the treatment of obesity and/or diabetes, for example insulin and insulin analogs, gastric lipase inhibitors, pancreatic lipase inhibitors, sulfonyl ureas and analogs, biguanides, a2 agonists, glitazones, PPAR- ⁇ agonists, GPR40 agonists, mixed PPAR- ⁇ / ⁇ agonists, RXR agonists, fatty acid oxidation inhibitors, a-glucosidase inhibitors, ⁇ -agonists, phosphodiesterase inhibitors, lipid lowering agents, glycogen phosphorylase inhibitors, antiobesity agents e.g.
  • pancreatic lipase inhibitors MCH-1 antagonists and CB-1 antagonists (or inverse agonists), amylin antagonists, lipoxygenase inhibitors, somostatin analogs, glucokinase activators, glucagon antagonists, insulin signalling agonists, PTP1B inhibitors, gluconeogenesis inhibitors, antilypolitic agents, GSK inhibitors, galanin receptor agonists, anorectic agents, CCK receptor agonists, leptin, serotonergic/dopaminergic antiobesity drugs, reuptake inhibitors e.g.
  • sibutramine CRF antagonists, CRF binding proteins, thyromimetic compounds, aldose reductase inhibitors, glucocorticoid receptor antagonists, NHE-1 inhibitors or sorbitol dehydrogenase inhibitors.
  • Combination therapy comprising the administration of a compound of the invention, or a pharmaceutically acceptable salt thereof, and at least one other agent represents a further aspect of the invention.
  • the present invention also provides a method for the treatment of diabetes in a mammal, such as a human, which method comprises administering an effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, and another agent, to a mammal in need thereof.
  • the invention also provides the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, and another agent for the treatment of diabetes.
  • the invention also provides the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for use in combination with another agent, for the treatment of diabetes.
  • the compound of the invention, or a pharmaceutically acceptable salt thereof, and the other agent(s) may be co-administered or administered sequentially or separately.
  • Co-administration includes administration of a formulation which includes both the compound of the invention, or a pharmaceutically acceptable salt thereof, and the other agent(s), or the simultaneous or separate administration of different formulations of each agent. Where the pharmacological profiles of the compound of the invention, or a pharmaceutically acceptable salt thereof, and the other agent(s) allow it, coadministration of the two agents may be preferred.
  • the invention also provides the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, and another agent in the manufacture of a medicament for the treatment of diabetes.
  • the invention also provides a pharmaceutical composition
  • a pharmaceutical composition comprising a compound of the invention, or a pharmaceutically acceptable salt thereof, and another antiobesity agent, and a pharmaceutically acceptable carrier.
  • the invention also encompasses the use of such compositions in the methods described above.
  • Gradient information 0.0-0.3 min 100% H 2 0; 0.3 ⁇ 1.25 min: Ramp up to 10% H 2 O-90% MeCN; 4.25-4.4 min: Ramp up to 100% MeCN; 4.4-4.9 min: Hold at 100% MeCN; 4.9-6.0 min: Return to 100% H 2 O.
  • the mass spectra were obtained using an electrospray ionisation source in either the positive (ES + ) or negative (ES ⁇ ) ion modes.
  • LCMS-method 2 Phenomenex Kinetex C18 column (3.0x30mm, 2.6 ⁇ , flow rate l .OmL/min) eluting with a H 2 0-MeCN solution containing 0.1% HCO 2 H over 2 min with UV detection at 220 nm.
  • Gradient information 0.0-0.1 min 2% MeCN 98% H 2 0 to 5% MeCN 95% H 2 0; 0.1-1.50 min: Ramp up to 100% MeCN; 1.5-1.75min: Hold at 100% MeCN; 1.75-1.8min: 100% MeCN to 2% MeCN 98% H 2 0 ; 1.8-2.0 min: Hold at 2% MeCN 98% H 2 0.
  • the mass spectra were obtained using an electrospray ionisation source in both the positive (ES + ) or negative (ES ⁇ ) ion modes.
  • LCMS-method 3 data were obtained as follows: Xbridge C18 column (3.0 x 150mm, 5 ⁇ , flow rate l .OmL/min) eluting with an MeCN-lOmM NH 4 HCO 3 solution over 5 min with UV detection at 215 - 350nm. Gradient information: 0-0.1 min: hold at 5% MeCN 95% NH4HCO3; 0.1 -3.0 min: 5% MeCN 95%NH4HC0 3 to 5% NH4HCO3 95% MeCN; 3.0- 3.9min: hold at 5% NH4HCO3 95% MeCN.
  • the mass spectra were obtained using an electrospray ionisation source in the positive (ES + ) mode.
  • LCMS -method 4 data were obtained as follows: Xbridge Ci 8 column (2.1 x 50mm, 2.5 ⁇ , flow rate 0.8mL/min) eluting with an MeCN-lOmM NH 4 HCO 3 solution over 1.5 min with UV detection at 215 - 350nm. Gradient information: 0-0.8 min: 98% MeCN 2% NH4HCO3 to 98% NH4HCO3 2% MeCN; 0.8-1.2min: hold at 98% NH4HCO3 2% MeCN. The mass spectra were obtained using an electrospray ionisation source in the positive (ES ⁇ ) mode.
  • Preparative HPLC purification was carried out using either a standard or basic method.
  • Standard method Gemini-NX Ci 8 column (21.2 x 100mm, 5 ⁇ , flow rate 20mL/min) eluting with a H 2 0-MeCN solution containing 0.1% HC0 2 H using a 10 minute gradient with UV detection at 220 nm.
  • Preparation 31 [l-(3-Ethyl-[l,2,4] eridin-4-yl]-methyl-amine
  • Preparation 171 [(3 ⁇ ,45)-1- ⁇ 5-[ ⁇ 1-[3-(1,1- ⁇ - ⁇ 1)-[1,2,4] ⁇ 3 ⁇ 3 ⁇ 1-5 ⁇ 1]- piperidin-4-yl ⁇ -(2,2,2-trifluoro-ethyl)-carbamoyl]-pyrimidin-2-yl ⁇ -4-(2,5-difluoro- phenyl)-pyrrolidin-3-yl]-carbamic acid teri-butyl ester
  • the title compound was prepared by reacting 2-[(3R,45')-3-iert- butoxycarbonylamino-4-(2,5-difluoro-phenyl) ⁇ yrrolidin-l-yl] ⁇ yrimidine-5-carboxylic acid (Preparation 80) with ?ra «s- ⁇ l-[3-(l ,l-difluoro-ethyl)-[l,2,4]oxadiazol-5-yl]-3-fluoro- piperidin-4-yl ⁇ -methyl-amine (Procedure 30) employing a procedure similar to that outlined in Preparation 95 to afford the title compound as a mixture of diastereoisomers.
  • Preparation 176 [(3/?,45)-4-(2,4-Difluoro-5-methyl-phenyl)-l-(5- ⁇ [l-(3-isopropyl- [ 1 ,2 ,4] oxa diazol-5-yl)-piperidin-4-yl] -methyl-carbamoyl ⁇ -pyrimidin-2 -yl)-pyr r olidin-3 - yl]-carbamic acid teri-b
  • Preparation 180 [(3/?,45)-l-(5- ⁇ [l-(5-Chloro-pyrimidin-2-yl)-piperidin-4-yl]-ethyl- carbamoyl ⁇ -pyrimidin-2-yl)-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
  • Acetic anhydride (665mL, 7.05mol) was added in one portion to l-(2,5-difluoro- phenyl)-2-nitro-ethanol (Preparation 187, 672g, 3.31mol) at 0 °C under argon.
  • DMAP 28.3g, 0.23mol was added and the solution darkened in colour.
  • the reaction was warmed to r.t. over 18 h.
  • the reaction mixture was cautiously poured into NaHC(3 ⁇ 4 (sat. aq., 3.5L) and stirred to form a yellow solid.
  • the slurry was stirred for 30 min at r.t. before filtering.
  • the solid was washed with NaHC(3 ⁇ 4 (sat.
  • the resulting paler yellow solution was stirred at -10 °C for 1 h before quenching cautiously with aHC0 3 (sat. aq., 1 L). The two batches were combined and the layers partitioned. The DCM layer was washed with water (2L) then brine (2L), dried over MgS0 4 and evaporated to give 209g of an orange oil. The material was purified by dry flash chromatography eluting with DCM to afford the title compound.
  • Triethylamine (244mL, 1.75mol) was added to a solution of l-benzyl-4-(2,5- difluoro-phenyl)-pyrrolidin-3-ylamine (Preparation 190, 252g, 0.87mol) in THF (3.8L) and the cloudy light brown solution was cooled to 0 °C.
  • Di-tert-butyldicarbonate (229g, 1.05mol) was added in one portion and the suspension warmed to room temperature over 18 h. The THF was removed by evaporation and the off-white slurry taken up into EtOAc (2.5L).
  • Nitromethane (274mL, 5.10mol) was added in one portion to a solution of 2,4,5- trifluorobenzaldehyde (680g, 4.25mol) in MeOH (7.2L) in a 10 L jacketed vessel.
  • the solution was cooled to 0 °C (jacket -10 °C) and a solution of NaOH (204g, 5.10mol) in water (680mL) was added over 30 min. A 5 °C exotherm was observed.
  • the solution was stirred for 30 min (jacket 0 °C) after which time a white precipitate had formed.
  • Half of the material was transferred to a second vessel and both slurries stirred for a further 30 min becoming very thick.
  • Acetic anhydride (849mL, 8.99mol) was added to 2-nitro-l -(2,4,5-trifluoro-phenyl)- ethanol (Preparation 194, 935g, 4.22mol) at 0 °C under argon.
  • DMAP 36g, 0.30mol was added in one portion and the solution darkened in colour. The temperature reached 50 °C over 20 min before cooling back to 0 °C. The solution was allowed to warm to r.t. over 18 h. The reaction mixture was cautiously poured into aHC0 3 (sat. aq., 6L) and stirred to form a yellow solid. The slurry was stirred for 30 min at r.t. before filtering and washing with NaHCCh (sat.
  • Triethylamine (210mL, 1.51mol) was added to l-benzyl-4-(2,4,5-trifluoro-phenyl)- pyrrolidin-3-ylamine (Preparation 197, 23 lg, 0.75mol) in THF (1.8L) and the light brown cloudy solution was cooled to 0 °C.
  • Di-tert-butyldicarbonate (198g, 0.91mol) was added in one portion and the suspension warmed to room temperature over 18 h. The suspension was concentrated and the off-white slurry dissolved into EtOAc (2.5L). The solution was washed with water (3 x 1L) and brine (1L) then dried over MgS0 4 and evaporated to give a pale yellow oil.
  • Preparation 205 [(S ⁇ -l-IS-CCyclopropyl-il-IS-Cl-methoxy-l-methyl-ethyl)- [l,2,4]oxadiazol-3-yl]-piperidin-4-yl ⁇ -carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro- phenyl)-pyrrolidin-3-yl -carbamic acid tert-but l ester
  • reaction mixture was diluted with EtOAc (50mL) and water (20mL); aqueous layer was back-extracted with more EtOAc, then the organics were combined, washed with brine (20mL), dried (MgS0 4 ), filtered and concentrated in vacuo.
  • Example 1 2-[(3/?,45)-3-Amino-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine- 5-carboxylic acid cyclopropyl-[l-(3-isopropyl-[l,2,4]oxadiazol-5-yl)-piperidin-4-yl]- amide hydrochloride
  • Example 2 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid [l-(3-iso ropyl-[l,2,4]oxadiazol-5-yl)-piperidin-4-yl]-methyl-amide
  • Example 3 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid ethyl-[l-(3-isopropyl-[l,2,4]oxadiazol-5-yl)-piperidin-4-yl]-amide hydrochloride
  • Example 4 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid ⁇ l-[3-(l-fluoro-l-methyl-ethyl)-[l,2,4]oxadiazol-5-yl]-piperidin-4-yl ⁇ - methyl-amide hydrochloride
  • Example 84 2- [(3/?,45)-3-Amino-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-l -yl] - pyrimidine-5-carboxylic acid [l-(3-ethyl-[l,2,4]oxadiazol-5-yl)-piperidin-4-yl]-methyl- amide hydrochloride
  • Example 85 2-[(3/?,4S)-3-Amino-4-(2-fluoro-5-methyl-phenyl)-pyrrolidin-l-yl]- pyrimidine-5-carboxylic acid [l-(3-ethyl-[l,2,4]oxadiazol-5-yl)-piperidin-4-yl]-methyl- amide hydrochloride
  • Example 86 2-[(3/?,4S)-3-Amino-4-(2-fluoro-5-methyl-phenyl)-pyrrolidin-l-yl]- pyrimidine-5-carboxylic acid [l-(3-isopropyl-[l,2,4]oxadiazol-5-yl)-piperidin-4-yl]- methyl-amide hydrochloride
  • Example 87 2-[(3/?,4S)-3-Amino-4-(2-fluoro-5-methyl-phenyl)-pyrrolidin-l-yl]- pyrimidine-5-carboxylic acid ethyl-[l-(3-isopropyl-[l,2,4]oxadiazol-5-yl)-piperidin-4- yl]-amide hydrochloride
  • Example 89 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl- ⁇ l-[5-(l-methoxy-l-methyl-ethyl)-[l,2,4]oxadiazol-3-yl]- piperidin-4-yl ⁇ -amide hydrochloride
  • Example 90 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl- ⁇ l-[5-(l,l-difluoro-ethyl)-[l,2,4]oxadiazol-3-yl]-piperidin- 4-yl ⁇ -amide hydrochloride
  • Example 91 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl- ⁇ l-[(/?)-5-(tetrahydro-furan-3-yl)-[l,2,4]oxadiazol-3-yl]- piperidin-4-yl ⁇ -amide h drochloride
  • Example 92 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl- ⁇ l-[(5)-3-(tetrahydro-furan-3-yl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl ⁇ -amide
  • Example 94 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl-[l-(2-ethyl-2H-tetrazol-5-yl)-piperidin-4-yl]-amide hydrochloride
  • Example 95 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl- ⁇ l-[3-(tetrahydro-pyran-4-yl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl ⁇ -amide
  • Example 96 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl- ⁇ l-[3-(l-methoxy-cyclopropyl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl ⁇ -amide
  • Example 97 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl- ⁇ l-[(5)-3-(tetrahydro-furan-2-yl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl ⁇ -amide hydrochloride
  • Example 98 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl- ⁇ l-[(/?)-3-(tetrahydro-furan-2-yl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl ⁇ -amide h drochloride
  • Example 99 5-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrazine-2- carboxylic acid ⁇ l-[3-(l,l-difluoro-ethyl)-[l,2,4]oxadiazol-5-yl]-piperidin-4-yl ⁇ -methyl- amide hydrochloride
  • Example 100 6-[(3 J R,45)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-7V- ⁇ l-[3-(l,l- difluoro-ethyl)-[l,2,4]oxadiazol-5-yl]-piperidin-4-yl ⁇ -7V-methyl-nicotinamide hydrochloride
  • Example 101 6-[(3/?,45)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-5-cyano-iV-
  • the biological activity of the compounds of the invention may be tested in the following assay systems:
  • GPR119 cAMP Assay A stable cell line expressing recombinant human GPR1 19 was established and this cell line was used to investigate the effect of compounds of the invention on intracellular levels of cyclic AMP (cAMP).
  • the cell monolayers were washed with phosphate buffered saline and stimulated at 37°C for 30 min with various concentrations of compound in stimulation buffer plus 1% DMSO. Cells were then lysed and cAMP content determined using the Perkin Elmer AlphaScreenTM (Amplified Luminescent Proximity Homogeneous Assay) cAMP kit. Buffers and assay conditions were as described in the manufacturer's protocol.
  • DPP-IV activity was measured by monitoring the cleavage of the fluorogenic peptide substrate, H-Gly-Pro-7-amino-4-methylcoumarin (GP-AMC) whereby the product 7-amino-
  • 4-methylcoumarin is quantified by fluorescence at excitation 380 nm and emission 460 nm.
  • Asn29-Pro766 was purchased from BioMol.
  • HIT-T15 cells (passage 60) were obtained from ATCC, and were cultured in
  • HIT-T15 cells were plated in standard culture medium in 96-well plates at 100,000 cells/ 0.1 mL/ well and cultured for 24 h and the medium was then discarded. Cells were incubated for 15min at room temperature with ⁇ ⁇ stimulation buffer (Hanks buffered salt solution, 5mM HEPES, 0.5mM IBMX, 0.1% BSA, pH 7.4). This was discarded and replaced with compound dilutions over the range 0.001, 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3, 10, 30 ⁇ in stimulation buffer in the presence of 0.5% DMSO. Cells were incubated at room temperature for 30 min.
  • ⁇ ⁇ stimulation buffer Hors buffered salt solution, 5mM HEPES, 0.5mM IBMX, 0.1% BSA, pH 7.4
  • 75 uL lysis buffer (5mM HEPES, 0.3% Tween-20, 0.1% BSA, pH 7.4) was added per well and the plate was shaken at 900 rpm for 20 min. Particulate matter was removed by centrifugation at 3000rpm for 5 min, then the samples were transferred in duplicate to 384-well plates, and processed following the Perkin Elmer AlphaScreen cAMP assay kit instructions. Briefly 25 ⁇ reactions were set up containing 8 ⁇ sample, 5 ⁇ acceptor bead mix and 12 ⁇ detection mix, such that the concentration of the final reaction components is the same as stated in the kit instructions. Reactions were incubated at room temperature for 150 min, and the plate was read using a Packard Fusion instrument.
  • Measurements for cAMP were compared to a standard curve of known cAMP amounts (0.01, 0.03, 0.1, 0.3, 1, 3, 10, 30, 100, 300, 1000 nM) to convert the readings to absolute cAMP amounts. Data was analysed using XLfit 3 software. Representative compounds of the invention were found to increase cAMP at an EC5 0 of less than 10 ⁇ . Compounds showing an EC5 0 of less than 1 ⁇ in the cAMP assay may be preferred.
  • HIT-T15 cells are plated in standard culture medium in 12-well plates at 106 cells/ 1 ml/ well and cultured for 3 days and the medium then discarded. Cells are washed x 2 with supplemented Krebs-Ringer buffer (KRB) containing 1 19 mM NaCl, 4.74 mM KC1, 2.54 mM CaCl 2 , 1.19 mM MgS0 4 , 1.19 mM KH 2 P0 4 , 25 mM NaHC0 3 , 10 mM HEPES at pH 7.4 and 0.1% bovine serum albumin. Cells are incubated with 1ml KRB at 37°C for 30 min which is then discarded.
  • KRB Krebs-Ringer buffer
  • Compounds of the invention preferably increase insulin secretion at an EC5 0 of less than 10 ⁇ .
  • the effects of compounds of the invention on oral glucose (Glc) tolerance may be evaluated in male Sprague-Dawley rats. Food is withdrawn 16 h before administration of Glc and remains withdrawn throughout the study. Rats have free access to water during the study. A cut is made to the animals' tails, then blood (1 drop) is removed for measurement of basal Glc levels 60 min before administration of the Glc load. Then, the rats are weighed and dosed orally with test compound or vehicle (20% aqueous hydroxypropyl-yS- cyclodextrin) 45 min before the removal of an additional blood sample and treatment with the Glc load (2 g kg 1 p.o.).
  • Blood samples are taken from the cut tip of the tail 5, 15, 30, 60, 120, and 180 min after Glc administration. Blood glucose levels are measured just after collection using a commercially available glucose-meter (OneTouch® UltraTM from Lifescan). Compounds of the invention preferably statistically reduce the Glc excursion at doses ⁇ 100 mg kg -1 .
  • the effects of compounds of the invention on oral glucose (Glc) tolerance were evaluated in male C57B1/6 or male ob/ob mice. Food was withdrawn 5 h before
  • mice had free access to water during the study. A cut was made to the animals' tails, then blood (20 ⁇ ,) was removed for measurement of basal Glc levels 45 min before administration of the Glc load. Then, the mice were weighed and dosed orally with test compound or vehicle (20% aqueous hydroxypropyl-yS-cyclodextrin or 25% aqueous Gelucire 44/14) 30 min before the removal of an additional blood sample (20 ⁇ ) and treatment with the Glc load (2-5 g kg 1 p.o.). Blood samples (20 ⁇ ) were then taken 25, 50, 80, 120, and 180 min after Glc administration.
  • the 20 blood samples for measurement of Glc levels were taken from the cut tip of the tail into disposable micro-pipettes (Dade Diagnostics Inc., Puerto Rico) and the sample added to 480 ⁇ , of haemolysis reagent. Duplicate 20 ⁇ , aliquots of the diluted haemolysed blood were then added to 180 ⁇ , of Trinders glucose reagent (Sigma enzymatic (Trinder) colorimetric method) in a 96-well assay plate. After mixing, the samples were left at room temperature for 30 min before being read against Glc standards (Sigma glucose/urea nitrogen combined standard set). Compounds of the invention statistically reduced the Glc excursion at doses ⁇ 100 mg kg -1 , for example at a dose of 30 mg kg 1 the compound of Example 6 showed a > 40% reduction in the Glc excursion.

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Abstract

Therapeutic compounds are disclosed having the general formula (I) that are useful for the treatment of metabolic disorders, including type II diabetes. The compounds have activity as agonists of GPR1 19. Compounds having stereochemistry of formula (la) may also demonstrate DPP-IV inhibitory activity.

Description

1,4 DISUBS ITUTED PYRROLIDINE - 3 - YL -AMINE DERIVATIVES AND THEIR USE FOR THE TREATMENT OF METABOLIC DISORDERS
The present invention is directed to therapeutic compounds useful for the treatment of metabolic disorders including type II diabetes. In particular, the present invention is 5 directed to compounds which have activity as agonists of GPR1 19.
Drugs aimed at the pathophysiology associated with non-insulin dependent type II diabetes have many potential side effects and do not adequately address the dyslipidaemia and hyperglycaemia in a high proportion of patients. Treatment is often focused at individual patient needs using diet, exercise, hypoglycaemic agents and insulin, but there is a0 continuing need for novel antidiabetic agents, particularly ones that may be better tolerated with fewer adverse effects.
Similarly, metabolic syndrome (syndrome X) places people at high risk of coronary artery disease, and is characterized by a cluster of risk factors including central obesity (excessive fat tissue in the abdominal region), glucose intolerance, high triglycerides and low5 HDL cholesterol, and high blood pressure. Myocardial ischemia and microvascular disease is an established morbidity associated with untreated or poorly controlled metabolic syndrome.
Obesity is characterized by an excessive adipose tissue mass relative to body size. Clinically, body fat mass is estimated by the body mass index (BMI; weight (kg)/height0 (m)2), or waist circumference. Individuals are considered obese when the BMI is greater than 30 and there are established medical consequences of being overweight. It has been an accepted medical view for some time that an increased body weight, especially as a result of abdominal body fat, is associated with an increased risk for diabetes, hypertension, heart disease, and numerous other health complications, such as arthritis, stroke, gallbladder5 disease, muscular and respiratory problems, back pain and even certain cancers.
There is a continuing need for novel antidiabetic agents, particularly ones that are well tolerated with few adverse effects and in particular for agents which are weight neutral or preferably cause weight loss.
GPR119 (previously referred to as GPR116) is a GPCR identified as SNORF25 in0 WO00/50562 which discloses both the human and rat receptors, US 6,468,756 also discloses the mouse receptor (accession numbers: AAN95194 (human), AAN95195 (rat) and A 95196 (mouse)). In humans, GPR1 19 is expressed in the pancreas, small intestine, colon and adipose tissue. The expression profile of the human GPR1 19 receptor indicates its potential utility as a target for the treatment of diabetes.
GPR119 agonists have been shown to stimulate the release of GLP-1 from the GI tract. In doing so, GPR119 agonists (1) enhance glucose-dependent insulin release from the pancreas leading to improvements in oral glucose tolerance; (2) attenuate disease progression by increasing β-cell cAMP concentrations; and (3) induce weight loss possibly through GLP-1 's ability to reduce food intake.
International Patent Applications WO 2005/061489, WO 2006/070208, WO 2006/067532, WO 2006/067531, WO 2007/003960, WO 2007/003961, WO 2007/003962, WO 2007/003964, WO 2007/1 16229, WO 2007/1 16230, WO 2008/081204, WO 2008/081205, WO 2008/081206, WO 2008/081207, WO 2008/081208, WO 2009/050522, WO 2009/050523, WO 2010/001 166, WO 2010/004343, WO 2010/004344, WO 2010/004345, WO 2010/004346, WO 2010/004347 and WO 2010/004348 disclose GPR1 19 receptor agonists.
Dipeptidyl peptidase IV (DPP-IV) is a ubiquitous, yet highly specific, serine protease that cleaves N-terminal dipeptides from polypeptides with L-proline or L-alanine at the penultimate position. Studies with DPP-IV inhibitors show the principle role of DPP-IV is in the inactivation GLP- 1. By extending the duration of action of GLP- 1 , insulin secretion is stimulated, glucagon release inhibited, and gastric emptying slowed. DPP-IV inhibitors are of use for the treatment of type II diabetes, examples of DPP-IV inhibitors include vildagliptin, sitagliptin, alogliptin and saxagliptin.
The possibility of using a combination of a GPR1 19 agonist and a DPP-IV has been suggested, however this requires the administration of two separately formulated products to the patient or the co-formulation of two active ingredients with the inherent problems of achieving compatibility in the physicochemical and pharmacokinetic and pharmacodynamic properties of the two active ingredients. WO 2009/034388, WO 2010/103333, WO 2010/103334 and WO 2010/103333 disclose dual GPR119 agonists and DPP-IV inhibitors. The compounds of the invention preferably have dual activity as agonists of GPR1 19 and inhibitors of DPP-IV
The present invention provides compounds of formula (I) and pharmaceutically acceptable salts thereof:
Figure imgf000004_0001
wherein A is a para-linked phenyl, pyridinyl, pyrimidinyl, pyrazinyl or triazinyl; R1 is hydrogen, halo, cyano, Ci _4alkyl, C\ ^haloalkyl, Ci _4alkoxy or
C 1 _5alkoxyC 1 _4alkyl;
is selected from the group consisting of:
(a) phenyl optionally substituted by one or more groups independently
selected from halo, methyl or halomethyl groups,
(b) pyridyl optionally substituted by one or more groups independently selected from halo, methyl or halomethyl groups and
(c) N-pyridonyl optionally substituted by one or more groups independently selected from halo, methyl or halomethyl groups
R3 is C3_5cycloalkyl, 4 to 6 membered saturated heterocyclyl (comprising 1 or 2 ring heteroatoms selected from Ν, O and S) or Ci _4alkyl wherein the C3_ cycloalkyl and Ci _ 4alkyl are optionally substituted by one to three groups independently selected from halo, cyano, hydroxy or Ci ^alkoxy;
p and q are each 0, 1 or 2, provided that 0 < p+q < 2;
Z is selected from the group consisting of:
(a) C(0)OR4,
(b) C(0)R4
(c) S(0)2R4,
(d) a 5- or 6-membered N-containing heteroaryl ring optionally containing 1 to 3 additional heteroatoms selected from N, O and S, or a 8 to 10 membered fused bicyclic system optionally containing 1 to 3 heteroatoms selected from N, O and S, wherein the heteroaryl ring or fused bicyclic system is optionally substituted by one or two groups independently selected from halo, cyano, C1 .4 alkyl, Ci _4haloalkyl, Ci _4alkoxy, Ci _4hydroxyalkyl, Ci _4alkoxyCi _4alkyl, heterocyclyl (a 4 to 6 membered saturated heterocycle comprising 1 or 2 ring heteroatoms selected from N , O and S), heterocyclyl Ci . 4alkyl (wherein the heterocyclyl is a 4 to 6 membered saturated heterocycle comprising 1 or 2 ring heteroatoms selected from N, O and S) and C3_ cycloalkyl optionally substituted by Ci _4 alkyl, C i .4 alkoxy or halo;
R4 is selected from the group consisting of:
a) Chalky!,
b) phenyl,
c) Ci _galkoxyC2-6alkyl,
d) C3.gcycloalkyl optionally substituted by C 1 _4alkyl
e) C3 _5cycloalkylC 1 _4alkyl optionally substituted by C 1 .4 alkyl, and f) 4 to 6 membered saturated heterocycle comprising 1 or 2 ring heteroatoms selected from N, O and S, and
g) 5 or 6 membered heteroaryl ring containing 1 to 4 heteroatoms selected from N, O and S optionally substituted by one or two groups selected from halo, cyano, Ci _4alkyl, Ci _4halo alkyl, Ci _4alkoxy, C2-4alkoxyCi _
4alkyl, 4 to 6 membered saturated heterocycle comprising 1 or 2 ring heteroatoms selected from N, O and S and C3_ cycloalkyl wherein the cycloalkyl is optionally substituted by Ci .4 alkyl or halo: each of and is independently hydrogen, halo, Ci _2alkyl, C\ _2haloalkyl or Ci _3 alkoxy;
R7 is hydrogen, halo, C\ .2 alkyl, C\ ^haloalkyl or C\ .3 alkoxy;
and pharmaceutically acceptable salts thereof.
In particular the invention provides compounds of formula I having the stereochemistry defined below; such compounds may demonstrate DPP-IV inhibitory activity:
Figure imgf000005_0001
(la) p and q are independently 0, 1 or 2 provided that p+q does not exceed 2, i.e., forming a 4-, 5- or 6-membered ring. In some embodiments, p and q may be the same, i.e., forming a 4- or 6-membered ring. Suitably p and q are both 1, therefore the compounds of the invention may have the formula:
Figure imgf000006_0001
(lb)
A is suitably pyridine, pyrimidine or pyrazine typically pyridine or pyrimidine, e.g., 2- or 3-pyridyl or 2- or 5-pyrimidinyl, where the 2-, 3- or 5- refers to the point of attachment of the pyrrolidine ring (X being attached in the para position and R1 being attached at any suitable position).
R1 is suitably hydrogen, halogen or cyano. R^ is preferably hydrogen.
The ring of R^ may optionally be substituted by 1, 2 or 3 substituents. In certain embodiments R^ is optionally substituted by 1 or 2 substituents and in further embodiments it is substituted by 2 or 3 substituents. R^ is preferably optionally substituted by 1, 2, or 3 substituents independently selected from halo and methyl. R^ is preferably optionally substituted by 1 or 2 substituents independently selected from halo and methyl. R^ is suitably phenyl substituted by 1, 2 or 3 groups selected from halo and methyl groups. R^ is suitably phenyl substituted by 2 or 3 halo groups. Halo is suitably fluoro. R^ may be an optionally substituted phenyl or pyridyl, e.g. an optionally substituted phenyl or 2-pyridyl. Examples of R^ include phenyl, 2-pyridinyl, 4-fluoro-2-pyridinyl, 5-fluoro-2-pyridinyl, 4- methyl-2-pyridinyl, 5-methyl-2-pyridinyl, 2-fluorophenyl, 3 -fluorophenyl, 4-fluorophenyl, 2,3-difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl, 2,3,4- trifluorophenyl, 2,3,5-trifluorophenyl, 2,3,6-trifluorophenyl, 2,4,5-trifluorophenyl, 2,4,6- trifluorophenyl, 2-fluoro-5-methylphenyl and 2,4-difluoro-5-methylphenyl.
R3 may suitably be C3_ cycloalkyl or Ci _4 alkyl wherein the Ci _4 alkyl is optionally substituted by one to three groups independently selected from halo, cyano, hydroxy or Ci _2alkoxy. Suitable examples of include methyl, ethyl, n-propyl, i-propyl, cyclopropyl, CH2CH2OCH3 and CH2CF3.
In certain embodiments Z is -C(0)OR^. In further embodiments Z is C(0)R^. In still further embodiments it is S(0)2R^.
R4 may suitably be C^alkyl, C\ _ alkoxyC2-6alkyl, C3_ cycloalkyl or C3.
5cycloalkylCi _4alkyl, wherein said C3_ cycloalkyl or C3_5cycloalkylCi _4alkyl is optionally substituted by Ci .4 alkyl.
Examples of R^ include methyl, ethyl, n-propyl, i-propyl, n-butyl, s-butyl, i-butyl or t-butyl, cyclopropyl and 1-methylcyclopropyl. In certain embodiments R^ is propyl, especially isopropyl.
Z may be a heteroaryl group optionally substituted by one or two groups selected from halo, C 1.4 alkyl, Ci _4haloalkyl, Ci _4alkoxy, Ci _4alkoxyCi _4alkyl, Ci ^hydroxyalkyl,
4 to 6 membered saturated heterocyclyl comprising 1 or 2 ring heteroatoms selected from N, O and S, C4_5heterocyclylCi _4alkyl (wherein the heterocyclyl is a 4 to 6 membered saturated heterocycle comprising 1 or 2 ring heteroatoms selected from N, O and S) and C3.
5cycloalkyl wherein the cycloalkyl is optionally substituted by Ci .4 alkyl or halo. Preferred
Ci _4haloalkyls are perfluoroalkyls, e.g. CF3, CF2Me and CHF2
When Z is heteroaryl, suitable heteroaryl groups include oxadiazole, pyrimidine, pyridazine, thiazole, tetrazole, benzothiazole and thiadiazole, e.g., oxadiazole and pyrimidine. In some embodiments Z may comprise l,2,4-oxadiazol-3-yl, l,2,4-oxadiazol-5- yl, pyrimidin-2-yl or a 2H-tetrazol-5-yl, which may be substituted by any of the aforementioned substituents.
When Z is a substituted heteroaryl suitable substituents include ethyl, i-propyl, t- butyl, cyclopropyl, chloro, 1-methylcyclopropyl, CF3, CH2F, CF2H, CH(OMe)Me, C(Me)2OH, C(Me)2F, CF2Me, CH2OMe and 2-tetrahydrofuranyl.
R^ is preferably hydrogen.
The molecular weight of the compounds of the invention is suitably less than about 800, typically less than about 600.
The invention also includes isotopically labeled compounds, which are similar to those recited in formulae (I), (la) and (lb) and following, but for the fact that one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number most commonly found in nature. Examples of isotopes that can be incorporated into compounds of the invention include isotopes of hydrogen, carbon, nitrogen, fluorine, such as ¾, ^C, and ^F.
Compounds of the present invention and salts of said compounds that contain the aforementioned isotopes and/or other isotopes of other atoms are within the scope of the present invention. Isotopically labeled compounds of the present invention, for example those into which radioactive isotopes such as ¾, l^C are incorporated, are useful in drug and/or substrate tissue distribution assays. Tritiated, i.e., ¾, and carbon-14, i.e., ^C, isotopes are particularly preferred for their ease of preparation and detectability. and isotopes are particularly useful in PET (positron emission tomography). PET is useful in brain imaging. Further, substitution with heavier isotopes such as deuterium, i.e., ¾, can afford certain therapeutic advantages resulting from greater metabolic stability, for example increased in vivo half-life or reduced dosage requirements and, hence, may be preferred in some circumstances, isotopically labeled compounds of formula (I), (la) and (lb) and following of this invention can generally be prepared by carrying out the procedures disclosed in the Schemes and/or in the Examples below, by substituting a readily available isotopically labeled reagent for a non-isotopically labeled reagent. In one embodiment, the compounds of formula (I), (la) and (lb) or salts thereof are not isotopically labelled.
As used herein, unless stated otherwise, "alkyl" means carbon chains which may be linear or branched. Examples of alkyl groups include ethyl, propyl, isopropyl, butyl, sec- and tert-butyl. Such alkyl groups may in some embodiments be substituted with one or more halo groups, particularly fluoro. When used herein the term "cycloalkyl" refers to a 3 to 6 C saturated carbocycle moiety e.g. cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl. When used herein the term 'heterocyclyl' refers to a 4 to 6 membered saturated heterocycle comprising 1 or 2 ring heteroatoms selected from N, O and S. Examples of heterocycles include tetrahydrofuran, pyrrolidine, tetrahydropyran, piperidine, oxetane and azetidine. When used herein the term "heteroaryl" refers to a 5- or 6-membered aromatic heteroaryl ring optionally containing one or more, e.g. 1 , 2 or 3 heteroatoms selected from N, O and S, or a 8 to 10 membered fused aromatic bicyclic system optionally containing one or more, e.g. 1, 2 or 3 heteroatoms selected from N, O and S. Examples of heteroaryl rings include pyrrolyl, pyrazolyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, triazinyl, tetrazolyl and benzothiazolyl.
Reference to para-linked in relation to ring A refers to the relative positions of the carbonyl group and the pyrrolidine group.
Compounds described herein may contain one or more asymmetric centers and may thus give rise to diastereomers and optical isomers. The present invention includes all such possible diastereomers as well as their racemic mixtures, their substantially pure resolved enantiomers, all possible geometric isomers, and pharmaceutically acceptable salts thereof. The present invention includes all stereoisomers of the compounds of the invention and pharmaceutically acceptable salts thereof. Further, mixtures of stereoisomers as well as isolated specific stereoisomers are also included. During the course of the synthetic procedures used to prepare such compounds, or in using racemization or epimerization procedures known to those skilled in the art, the products of such procedures can be a mixture of stereoisomers.
When a tautomer of the compound of the invention exists, the present invention includes any possible tautomers and pharmaceutically acceptable salts thereof, and mixtures thereof, except where specifically drawn or stated otherwise.
When the compound of the invention and pharmaceutically acceptable salts thereof exist in the form of solvates or polymorphic forms, the present invention includes any possible solvates and polymorphic forms. A type of a solvent that forms the solvate is not particularly limited so long as the solvent is pharmacologically acceptable. For example, water, ethanol, propanol, acetone or the like can be used.
The term "pharmaceutically acceptable salts" refers to salts prepared from pharmaceutically acceptable non-toxic bases or acids. When the compound of the present invention is acidic, its corresponding salt can be conveniently prepared from pharmaceutically acceptable non-toxic bases, including inorganic bases and organic bases. Salts derived from such inorganic bases include aluminum, ammonium, calcium, copper (ic and ous), ferric, ferrous, lithium, magnesium, potassium, sodium, zinc and the like salts. Particularly preferred are the ammonium, calcium, magnesium, potassium and sodium salts. Salts derived from pharmaceutically acceptable organic non-toxic bases include salts of primary, secondary, and tertiary amines, as well as cyclic amines and substituted amines such as naturally occurring and synthesized substituted amines. Other pharmaceutically acceptable organic non-toxic bases from which salts can be formed include arginine, betaine, caffeine, choline, N',N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2- dimethylaminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N- ethylpiperidine, glucamine, glucosamine, histidine, hydrabamine, isopropylamine, lysine, methylglucamine, morpholine, piperazine, piperidine, polyamine resins, procaine, purines, theobromine, triethylamine, trimethylamine, tripropylamine, trimethamine and the like.
When the compound of the invention is basic, its corresponding salt can be conveniently prepared from pharmaceutically acceptable non-toxic acids, including inorganic and organic acids. Such acids include, for example, acetic, benzenesulfonic, benzoic, camphorsulfonic, citric, ethanesulfonic, fumaric, gluconic, glutamic, hydrobromic, hydrochloric, isethionic, lactic, maleic, malic, mandelic, methanesulfonic, mucic, nitric, pamoic, pantothenic, phosphoric, succinic, sulfuric, tartaric, p-toluenesulfonic, trifluoroacetic acid and the like.
Since the compounds of the invention are intended for pharmaceutical use they are preferably provided in substantially pure form, for example at least 60% pure, more suitably at least 75% pure, especially at least 98% pure (% are on a weight for weight basis).
The compounds of formula (I) can be prepared as described below, wherein R.1, R^,
R3, R4, R5, R6, R7, A, Z, p, q and r are as defined for formula (I). PG is a protecting group, LG is a leaving group, Hal is halogen, Het is heteroaryl.
Compounds of formula (I) can be prepared as outlined in Scheme 1. Removal of the protecting group PG from the amine functionality in compounds of formula (II) can be achieved using standard conditions well known to those skilled in the art.
Figure imgf000010_0001
Compounds of formula (II) can be prepared as outlined in Scheme 2. Esters of formula (V) can be prepared by S AT displacement of suitable haloaromatic compounds of formula (III) with amines of formula (IV) under standard conditions, for example, DBU and DMSO at 80 - 100°C. Acids of formula (VI) can be prepared from esters of formula (V) under standard conditions, for example, LiOH, water and methanol at room temperature. Amides of formula (II) can be prepared from acids of formula (VI) with amines of formula (VII) under standard amide coupling conditions, for example, HOBT and EDCI, in a suitable solvent, such as DCM.
Figure imgf000011_0001
Compounds of formula (VII) where Z is C(0)OR4 can be prepared as outlined in Scheme 3. Carbamates of formula (X) can be prepared from amines of formula (VIII) and chloro formates of formula (IX) under standard conditions, for example, triethylamine in a suitable solvent, such as DCM. Removal of the protecting group PG from the amine functionality in compounds of formula (X) can be achieved using standard conditions well known to those skilled in the art.
Scheme 3
Figure imgf000011_0002
Compounds of formula (VII) where Z is C(0)R4 can be prepared as outlined in Scheme 4. Amides of formula (XII) can be prepared from amines of formula (VIII) and acid chlorides of formula (XI) under standard conditions, for example, triethylamine in a suitable solvent, such as DCM. Removal of the protecting group PG from the amine functionality in compounds of formula (XII) can be achieved using standard conditions well known to those skilled in the art. Scheme 4
Figure imgf000012_0001
VIII XI XII
Compounds of formula (VII) where Z is S(0)2 4 can be prepared as outlined in Scheme 5. Sulfonamides of formula (XIV) can be prepared from amines of formula (VIII) and sulfonyl chlorides of formula (XIII) under standard conditions, for example, triethylamine in a suitable solvent, such as DCM. Removal of the protecting group PG from the amine functionality in compounds of formula (XIV) can be achieved using standard conditions well known to those skilled in the art.
Scheme 5
Figure imgf000012_0002
VIII XIII XIV VII
Compounds of formula (VII) where Z is heteroaryl can be prepared as outlined in Scheme 6. Compounds of formula (XVI) can be prepared by S displacement of suitable haloaromatic compounds of formula (XV) with amines of formula (VIII) under standard conditions, for example, DBU and DMSO at 80 - 100°C. Alternatively, compounds of formula (XVI) can be prepared by reaction of suitable haloaromatic compounds of formula (XV) with amines of formula (VIII) under Buchwald-Hartwig conditions, such as, Pd2(dba and ΒΓΝΑΡ in a suitable solvent, such as toluene at 1 10°C. Removal of the protecting group PG from the amine functionality in compounds of formula (XVI) can be achieved using standard conditions well known to those skilled in the art.
Scheme 6
Figure imgf000012_0003
VIM XV XVI VII Compounds of formula (IV) where R2 is N-pyridonyl can be prepared as outlined in Scheme 7. Compounds of formula (XVIII) can be prepared from alcohols of formula (XVII) under standard conditions, for example, methanesulfonyl chloride and triethylamine in a suitable solvent, such as, THF at 0°C. Diazabicyclo compounds of formula (XIX) can be prepared from compounds of formula (XVIII) by treatment with sodium hydride in a suitable solvent, such as, THF at 0°C. Compounds of formula (XX) can be prepared from compounds of formula (XIX), pyrid-2-one and potassium tert-butoxide in a suitable solvent, such as, ΝΜΡ in a microwave reactor at 100°C. Removal of the protecting group PG' from the amine functionality in compounds of formula (XX) can be achieved using standard conditions well known to those skilled in the art.
Scheme 7
Figure imgf000013_0001
XX IV
Examples and syntheses of building blocks of formula (IV) have been described elsewhere: Benbow et.al, WO2007148185; Backes et.al, Bioorg. Med. Chem. Lett., 2007, 17 2005-2012; Pei et.al, J. Med. Chem., 2007, 50 (8), 1983-1987; Cox et.al, Bioorg. Med. Chem. Lett, 2007, 17 4579-4583; Wright et.al, Bioorg. Med. Chem. Lett, 2007, 17 5638- 5642.
Other compounds of formula (I) may be prepared by methods analogous to those described above or by methods known per se. Further details for the preparation of the compounds of formula (I) are found in the examples.
The compounds of formula (I) may be prepared singly or as compound libraries comprising at least 2, for example 5 to 1 ,000, compounds and more preferably 10 to 100 compounds of formula (I). Compound libraries may be prepared by a combinatorial "split and mix" approach or by multiple parallel synthesis using either solution or solid phase chemistry, using procedures known to those skilled in the art.
During the synthesis of the compounds of formula (I), labile functional groups in the intermediate compounds, e.g. hydroxy, carboxy and amino groups, may be protected. The protecting groups may be removed at any stage in the synthesis of the compounds of formula (I) or may be present on the final compound of formula (I). A comprehensive discussion of the ways in which various labile functional groups may be protected and methods for cleaving the resulting protected derivatives is given in, for example, Protective Groups in Organic Chemistry, T.W. Greene and P.G.M. Wuts, (1991) Wiley-Interscience, New York, 2nd edition.
The processes for the production of the compounds of formula (I) and intermediates thereto as described above are also included as further aspects of the present invention. Any novel intermediates as defined in the Schemes above or in the Examples, are also included within the scope of the invention. Therefore according to a further aspect of the invention there is provided a compound of any one of formulae (II), (VI), (VII), (XIV) and (XVI) as defined above. The preferred groups for variables recited above in relation to the compounds of formula (I) also apply to the intermediate compounds.
As indicated above the compounds of the invention are useful as GPR1 19 agonists, e.g. for the treatment and/or prophylaxis of diabetes. For such use the compounds of the invention will generally be administered in the form of a pharmaceutical composition.
The compounds of the invention may also be useful as dual GPR1 19 agonists/DPP- IV inhibitors, e.g. for the treatment and/or prophylaxis of diabetes. For such use the compounds of the invention will generally be administered in the form of a pharmaceutical composition.
The invention also provides a compound of the invention, or a pharmaceutically acceptable salt thereof, for use as a pharmaceutical.
The invention also provides a pharmaceutical composition comprising a compound of the invention, in combination with a pharmaceutically acceptable carrier.
Preferably the composition is comprised of a pharmaceutically acceptable carrier and a non-toxic therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof.
Moreover, the invention also provides a pharmaceutical composition for the treatment of disease by modulating GPR1 19 and optionally DPP-IV, resulting in the prophylactic or therapeutic treatment of diabetes, comprising a pharmaceutically acceptable carrier and a non-toxic therapeutically effective amount of compound of the invention, or a pharmaceutically acceptable salt thereof.
The pharmaceutical compositions may optionally comprise other therapeutic ingredients or adjuvants. The compositions include compositions suitable for oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most suitable route in any given case will depend on the particular host, and nature and severity of the conditions for which the active ingredient is being administered. The pharmaceutical compositions may be conveniently presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy.
In practice, the compounds of the invention, or pharmaceutically acceptable salts thereof, can be combined as the active ingredient in intimate admixture with a pharmaceutical carrier according to conventional pharmaceutical compounding techniques. The carrier may take a wide variety of forms depending on the form of preparation desired for administration, e.g. oral or parenteral (including intravenous).
Thus, the pharmaceutical compositions can be presented as discrete units suitable for oral administration such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient. Further, the compositions can be presented as a powder, as granules, as a solution, as a suspension in an aqueous liquid, as a non-aqueous liquid, as an oil-in-water emulsion, or as a water-in-oil liquid emulsion. In addition to the common dosage forms set out above, the compound of the invention, or a pharmaceutically acceptable salt thereof, may also be administered by controlled release means and/or delivery devices. The compositions may be prepared by any of the methods of pharmacy. In general, such methods include a step of bringing into association the active ingredient with the carrier that constitutes one or more necessary ingredients. In general, the compositions are prepared by uniformly and intimately admixing the active ingredient with liquid carriers or finely divided solid carriers or both. The product can then be conveniently shaped into the desired presentation.
The compounds of the invention, or pharmaceutically acceptable salts thereof, can also be included in pharmaceutical compositions in combination with one or more other therapeutically active compounds.
The pharmaceutical carrier employed can be, for example, a solid, liquid, or gas. Examples of solid carriers include lactose, terra alba, sucrose, talc, gelatin, agar, pectin, acacia, magnesium stearate, and stearic acid. Examples of liquid carriers are sugar syrup, peanut oil, olive oil, and water. Examples of gaseous carriers include carbon dioxide and nitrogen.
In preparing the compositions for oral dosage form, any convenient pharmaceutical media may be employed. For example, water, glycols, oils, alcohols, flavoring agents, preservatives, coloring agents, and the like may be used to form oral liquid preparations such as suspensions, elixirs and solutions; while carriers such as starches, sugars, microcrystalline cellulose, diluents, granulating agents, lubricants, binders, disintegrating agents, and the like may be used to form oral solid preparations such as powders, capsules and tablets. Because of their ease of administration, tablets and capsules are the preferred oral dosage units whereby solid pharmaceutical carriers are employed. Optionally, tablets may be coated by standard aqueous or nonaqueous techniques.
A tablet containing the composition of this invention may be prepared by compression or molding, optionally with one or more accessory ingredients or adjuvants. Compressed tablets may be prepared by compressing, in a suitable machine, the active ingredient in a free-flowing form such as powder or granules, optionally mixed with a binder, lubricant, inert diluent, surface active or dispersing agent. Molded tablets may be made by molding in a suitable machine, a mixture of the powdered compound moistened with an inert liquid diluent. Each tablet preferably contains from about 0.05mg to about 5g of the active ingredient and each cachet or capsule preferably containing from about 0.05mg to about 5g of the active ingredient.
For example, a formulation intended for the oral administration to humans may contain from about 0.5mg to about 5g of active agent, compounded with an appropriate and convenient amount of carrier material which may vary from about 5 to about 95 percent of the total composition. Unit dosage forms will generally contain between from about lmg to about 2g of the active ingredient, typically 25mg, 50mg, lOOmg, 200mg, 300mg, 400mg, 500mg, 600mg, 800mg, or lOOOmg.
Pharmaceutical compositions of the present invention suitable for parenteral administration may be prepared as solutions or suspensions of the active compounds in water. A suitable surfactant can be included such as, for example, hydroxypropylcellulose. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, and mixtures thereof in oils. Further, a preservative can be included to prevent the detrimental growth of microorganisms.
Pharmaceutical compositions of the present invention suitable for injectable use include sterile aqueous solutions or dispersions. Furthermore, the compositions can be in the form of sterile powders for the extemporaneous preparation of such sterile injectable solutions or dispersions. In all cases, the final injectable form must be sterile and must be effectively fluid for easy syringability. The pharmaceutical compositions must be stable under the conditions of manufacture and storage; thus, preferably should be preserved against the contaminating action of microorganisms such as bacteria and fungi. The carrier can be a solvent or dispersion medium containing, for example, water, ethanol, polyol (e.g. glycerol, propylene glycol and liquid polyethylene glycol), vegetable oils, and suitable mixtures thereof.
Pharmaceutical compositions of the present invention can be in a form suitable for topical use such as, for example, an aerosol, cream, ointment, lotion, dusting powder, or the like. Further, the compositions can be in a form suitable for use in transdermal devices. These formulations may be prepared, using a compound of the invention, or a pharmaceutically acceptable salt thereof, via conventional processing methods. As an example, a cream or ointment is prepared by admixing hydrophilic material and water, together with about 5wt% to about 10wt% of the compound, to produce a cream or ointment having a desired consistency.
Pharmaceutical compositions of this invention can be in a form suitable for rectal administration wherein the carrier is a solid. It is preferable that the mixture forms unit dose suppositories. Suitable carriers include cocoa butter and other materials commonly used in the art. The suppositories may be conveniently formed by first admixing the composition with the softened or melted carrier(s) followed by chilling and shaping in molds.
In addition to the aforementioned carrier ingredients, the pharmaceutical formulations described above may include, as appropriate, one or more additional carrier ingredients such as diluents, buffers, flavoring agents, binders, surface-active agents, thickeners, lubricants, preservatives (including anti-oxidants) and the like. Furthermore, other adjuvants can be included to render the formulation isotonic with the blood of the intended recipient. Compositions containing a compound of the invention, or pharmaceutically acceptable salts thereof, may also be prepared in powder or liquid concentrate form.
Generally, dosage levels on the order of O.Olmg/kg to about 150mg/kg of body weight per day are useful in the treatment of the above-indicated conditions, or alternatively about 0.5mg to about 7g per patient per day. For example, obesity may be effectively treated by the administration of from about 0.01 to 50mg of the compound per kilogram of body weight per day, or alternatively about 0.5mg to about 3.5g per patient per day. It is understood, however, that the specific dose level for any particular patient will depend upon a variety of factors including the age, body weight, general health, sex, diet, time of administration, route of administration, rate of excretion, drug combination and the severity of the particular disease undergoing therapy.
The compounds of the invention may be used in the treatment of diseases or conditions in which GPRl 19 and optionally DPP-IV play a role.
Thus the invention also provides a method for the treatment of a disease or condition in which GPRl 19 and optionally DPP-IV play a role comprising a step of administering to a subject in need thereof an effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof. Such diseases or conditions diabetes, obesity, impaired glucose tolerance, insulin resistance and diabetic complications such as neuropathy, nephropathy, retinopathy, cataracts, cardiovascular complications and dyslipidaemia). And the treatment of patients who have an abnormal sensitivity to ingested fats leading to functional dyspepsia. The compounds of the invention may also be used for treating metabolic diseases such as metabolic syndrome (syndrome X), impaired glucose tolerance, hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, low HDL levels and hypertension.
The invention also provides a method for the treatment of type II diabetes, comprising a step of administering to a patient in need thereof an effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof.
The invention also provides a method for the treatment of obesity, metabolic syndrome (syndrome X), impaired glucose tolerance, hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, low HDL levels or hypertension comprising a step of administering to a patient in need thereof an effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof.
The invention also provides a compound of the invention, or a pharmaceutically acceptable salt thereof, for use in the treatment of a condition as defined above.
The invention also provides the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for the treatment of a condition as defined above.
In the methods of the invention the term "treatment" includes both therapeutic and prophylactic treatment.
The compounds of the invention may exhibit advantageous properties compared to known compounds or combination therapies for the treatment of diabetes. The compounds of the invention, or pharmaceutically acceptable salts thereof, may be administered alone or in combination with one or more other therapeutically active compounds. The other therapeutically active compounds may be for the treatment of the same disease or condition as the compounds of the invention or a different disease or condition. The therapeutically active compounds may be administered simultaneously, sequentially or separately.
The compounds of the invention may be administered with other active compounds for the treatment of obesity and/or diabetes, for example insulin and insulin analogs, gastric lipase inhibitors, pancreatic lipase inhibitors, sulfonyl ureas and analogs, biguanides, a2 agonists, glitazones, PPAR-γ agonists, GPR40 agonists, mixed PPAR-α/γ agonists, RXR agonists, fatty acid oxidation inhibitors, a-glucosidase inhibitors, β-agonists, phosphodiesterase inhibitors, lipid lowering agents, glycogen phosphorylase inhibitors, antiobesity agents e.g. pancreatic lipase inhibitors, MCH-1 antagonists and CB-1 antagonists (or inverse agonists), amylin antagonists, lipoxygenase inhibitors, somostatin analogs, glucokinase activators, glucagon antagonists, insulin signalling agonists, PTP1B inhibitors, gluconeogenesis inhibitors, antilypolitic agents, GSK inhibitors, galanin receptor agonists, anorectic agents, CCK receptor agonists, leptin, serotonergic/dopaminergic antiobesity drugs, reuptake inhibitors e.g. sibutramine, CRF antagonists, CRF binding proteins, thyromimetic compounds, aldose reductase inhibitors, glucocorticoid receptor antagonists, NHE-1 inhibitors or sorbitol dehydrogenase inhibitors.
Combination therapy comprising the administration of a compound of the invention, or a pharmaceutically acceptable salt thereof, and at least one other agent represents a further aspect of the invention.
The present invention also provides a method for the treatment of diabetes in a mammal, such as a human, which method comprises administering an effective amount of a compound of the invention, or a pharmaceutically acceptable salt thereof, and another agent, to a mammal in need thereof.
The invention also provides the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, and another agent for the treatment of diabetes.
The invention also provides the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, in the manufacture of a medicament for use in combination with another agent, for the treatment of diabetes. The compound of the invention, or a pharmaceutically acceptable salt thereof, and the other agent(s) may be co-administered or administered sequentially or separately.
Co-administration includes administration of a formulation which includes both the compound of the invention, or a pharmaceutically acceptable salt thereof, and the other agent(s), or the simultaneous or separate administration of different formulations of each agent. Where the pharmacological profiles of the compound of the invention, or a pharmaceutically acceptable salt thereof, and the other agent(s) allow it, coadministration of the two agents may be preferred.
The invention also provides the use of a compound of the invention, or a pharmaceutically acceptable salt thereof, and another agent in the manufacture of a medicament for the treatment of diabetes.
The invention also provides a pharmaceutical composition comprising a compound of the invention, or a pharmaceutically acceptable salt thereof, and another antiobesity agent, and a pharmaceutically acceptable carrier. The invention also encompasses the use of such compositions in the methods described above.
All publications, including, but not limited to, patents and patent application cited in this specification, are herein incorporated by reference as if each individual publication were specifically and individually indicated to be incorporated by reference herein as fully set forth.
The invention will now be described by reference to the following examples which are for illustrative purposes and are not to be construed as a limitation of the scope of the present invention.
EXAMPLES
Materials and methods
Column chromatography was carried out on S1O2 (40-63 mesh) unless specified otherwise. LCMS data were obtained as follows:
LCMS-method 1 : Atlantis 3μ Ci8 column (3.0 x 20.0mm, flow rate = 0.85mL/min) eluting with a EkO-MeC solution containing 0.1% HCO2H over 6 min with UV detection at 220 nm. Gradient information: 0.0-0.3 min 100% H20; 0.3^1.25 min: Ramp up to 10% H2O-90% MeCN; 4.25-4.4 min: Ramp up to 100% MeCN; 4.4-4.9 min: Hold at 100% MeCN; 4.9-6.0 min: Return to 100% H2O. The mass spectra were obtained using an electrospray ionisation source in either the positive (ES+) or negative (ES~) ion modes. LCMS-method 2: Phenomenex Kinetex C18 column (3.0x30mm, 2.6μΜ, flow rate l .OmL/min) eluting with a H20-MeCN solution containing 0.1% HCO2H over 2 min with UV detection at 220 nm. Gradient information: 0.0-0.1 min 2% MeCN 98% H20 to 5% MeCN 95% H20; 0.1-1.50 min: Ramp up to 100% MeCN; 1.5-1.75min: Hold at 100% MeCN; 1.75-1.8min: 100% MeCN to 2% MeCN 98% H20 ; 1.8-2.0 min: Hold at 2% MeCN 98% H20. The mass spectra were obtained using an electrospray ionisation source in both the positive (ES+) or negative (ES~) ion modes.
LCMS-method 3 data were obtained as follows: Xbridge C18 column (3.0 x 150mm, 5μΜ, flow rate l .OmL/min) eluting with an MeCN-lOmM NH4HCO3 solution over 5 min with UV detection at 215 - 350nm. Gradient information: 0-0.1 min: hold at 5% MeCN 95% NH4HCO3; 0.1 -3.0 min: 5% MeCN 95%NH4HC03 to 5% NH4HCO3 95% MeCN; 3.0- 3.9min: hold at 5% NH4HCO3 95% MeCN. The mass spectra were obtained using an electrospray ionisation source in the positive (ES+) mode.
LCMS -method 4 data were obtained as follows: Xbridge Ci8 column (2.1 x 50mm, 2.5μΜ, flow rate 0.8mL/min) eluting with an MeCN-lOmM NH4HCO3 solution over 1.5 min with UV detection at 215 - 350nm. Gradient information: 0-0.8 min: 98% MeCN 2% NH4HCO3 to 98% NH4HCO3 2% MeCN; 0.8-1.2min: hold at 98% NH4HCO3 2% MeCN. The mass spectra were obtained using an electrospray ionisation source in the positive (ES^) mode.
LCMS-method 5 data were obtained as follows: Xbridge Ci8 column (2.1 x 5.0mm,
2.55μΜ, flow rate 0.8mL/min) eluting with an MeCN-lOmM NH4HCO3 solution over 5 min with UV detection at 215 - 350nm. Gradient information: 0-4 min: 98% MeCN 2% NH4HCO3 to 98% NH4HCO3 2% MeCN; 4-4.6min: hold at 98% NH4HCO3 2% MeCN. The mass spectra were obtained using an electrospray ionisation source in the positive (ES+) mode.
Unless otherwise stated LCMS-method 1 was employed for analysis.
Preparative HPLC purification was carried out using either a standard or basic method. Standard method: Gemini-NX Ci8 column (21.2 x 100mm, 5μΜ, flow rate 20mL/min) eluting with a H20-MeCN solution containing 0.1% HC02H using a 10 minute gradient with UV detection at 220 nm.
Basic method: XBridge Prep Ci8 column (19 x 100mm, 5μΜ, flow rate 20mL/min) eluting with a H20-MeCN solution containing 0.2% NH4OH using a 10 minute gradient with UV detection at 220 nm. Unless otherwise stated, the standard method was employed for purification.
Chiral-HPLC was performed on a Daicel chiralpak IA 250 x 20 mm, 5 μΜ column. Abbreviations and acronyms: AcOH: Acetic acid; Aq: Aqueous; BOP: (Benzotriazol-1- yloxy)tris(dimethylamino)phosphonium hexafluorophosphate; DBU: 1 ,8-
Diazabicyclo[5.4.0]undec-7-ene; DCE: 1, 2 -Dichloro ethane; DCM: Dichloromethane; DEA: Diethylamine; DIPEA: Diisopropylethylamine; DMAP: Dimethylpyridin-4-ylamine; EDCI: (3-Dimethylaminopropyl)ethylcarbodiimide hydrochloride; Et20: Diethyl ether; EtOH: Ethanol; EtOAc: Ethyl acetate; h: hour(s); HC1: Hydrochloric acid; HCO2H: Formic acid; H2O: Water; HOBt: l -Hydroxybenzotriazole monohydrate; HPLC: High performance liquid chromatography; IH: Isohexane; IPA: Isopropyl alcohol; IMS: Industrial methylated spirits; M: Molar; MeCN: Acetonitrile; MeOH: Methanol; MgS04: Magnesium sulfate; min: minute/s; PPA: 1 -Propanephosphonic acid cyclic anhydride; aHCOs: Sodium hydrogen carbonate; NaOH: Sodium hydroxide; Na2S04: Sodium sulfate; NH3: Ammonia; NH4CI: Ammonium chloride; NH4HCO3: Ammonium carbamate; NH4OH: Ammonium hydroxide; RT: Retention time; r.t.: Room temperature; Sat: saturated; SCX: Strong Cation Exchange resin; S1O2: Silica gel; THF: Tetrahydrofuran; TFA: Trifluoroacetic acid; TMSC1: Trimethylsilyl Chloride. The syntheses of the following compounds have been described elsewhere: 2,2-Difluoro-N- hydroxy-acetamidine and 2,2-Difluoro-N-hydroxy-propionamidine: Bertram, L., et ah, WO2010004348; 2-Fluoro-N-hydroxy-2-methyl-propionamidine: Azimioara, M. et ah, WO201 1044001 ; (3-Benzyl-3-aza-bicyclo[3.2.1]oct-8-yl)-carbamic acid tert-butyl ester, Aranyi, P. et al., WO2005021536. All other compounds were available from commercial sources.
Preparation 1: (l-Benzyl-4-methyl-piperidin-4-yl)-methyl-carbamic acid teri-butyl ester
Figure imgf000022_0001
To an argon purged flask containing anhydrous THF (80mL) was added lithium aluminum hydride and the suspension cooled to 0 °C. A solution of (l-benzyl-4-methyl- piperidin-4-yl)-carbamic acid tert-butyl ester in THF was added dropwise and the mixture allowed to warm to room temperature before heating to reflux for 4 h. The mixture was then cooled to r.t. and water slowly added followed by aqueous sodium hydroxide solution (2M) then water. The mixture was filtered through celite and di-tert-butyldicarbonate added to the filtrate. The mixture was allowed to stir at room temperature under argon for 18 h. The mixture was concentrated in vacuo, the residue dissolved in DCM and washed with 1M NaOH solution then brine. The organics were dried (MgSC^), filtered and concentrated. Purification by chromatography (DCM to DCM:7M ammonia in methanol 95:5) afforded the title compound: H NMR DH (301MHz, CDC13): 7.36 - 7.15 (m, 5H), 3.47 (s, 2H), 2.83 (s, 3H), 2.51 - 2.16 (m, 8H), 1.85 - 1.68 (m, 2H), 1.44 (s, 9H), 1.24 (s, 3H).
Preparation 2: Methyl-(4-methyl-piperidin-4- l)-carbamic acid tert-butyl ester
Figure imgf000023_0001
To a suspension of 10% Pd/C (700mg) in MeOH (500mL) in an autoclave was added (l-benzyl-4-methyl-piperidin-4-yl)-methyl-carbamic acid tert-butyl ester (Preparation 1, 9.0g, 28.26mmol) and the mixture hydrogenated (40 psi, 75 °C) for 4 h. The mixture was filtered through celite and the filtrate concentrated in vacuo to afford the title compound: H NMR DH (301MHZ, CDCI3): 2.88 - 2.69 (m, 5H), 2.28 - 2.07 (m, 2H), 1.76 - 1.57 (m, 4H), 1.44 (s, 9H), 1.26 (s, 3H).
Preparation 3: 3-Fluoro-4-oxo-pip c acid benzyl ester
Figure imgf000023_0002
To a solution of 4-oxo-piperidine-l-carboxylic acid benzyl ester (15g, 64mmol) in anhydrous DMF (80mL) was added anhydrous triethylamine (22mL, 150mmol) and the reaction heated to 80 °C for 18 h. TMSC1 (9.8mL, 77mmol) was added followed by triethylamine (22mL) and the reaction heated to 80 °C for 18 h. The reaction was cooled to room temperature, diluted with hexane and washed with water then brine solution. The organics were dried (sodium sulfate), filtered and concentrated in vacuo. To a solution of the residue in MeCN (300 mL) at 0 °C under argon was added l-chloromethyl-4-fluoro-l,4- diazaoniabicyclo[2.2.2]octane bis(tetrafluoroborate) (25g, 71mmol) portion wise and the mixture allowed to warm to room temperature and stirred for 18 h. The mixture was concentrated in vacuo and the residue partitioned between water and ethyl acetate. The organics were then washed with brine, dried (sodium sulfate), filtered and concentrated in vacuo. Purification by flash chromatography (hexane:EtOAc 7:3 - 1 : 1) afforded the title compound: 'H NMR D h (400MHZ, CDC13): 7.41-7.39 (m, 5H), 5.35 (s, 2H), 4.91-4.80 (m, 1H), 4.60-4.49 (m, 1H), 4.29-4.23 (m, 1H), 3.41-3.36 (m, 2H), 2.69-2.55 (m, 2H).
Preparation 4: c s-3-Fluoro-4-met e-l-carboxylic acid benzyl ester
Figure imgf000024_0001
To a solution of 3-fluoro-4-oxo-piperidine-l-carboxylic acid benzyl ester (Preparation 3, 4.00g, 15.9mmol) in anhydrous 1 ,2 -dichloro ethane (40mL) was added a solution of methylamine in MeOH (2M, 24mL, 48mmol) and the mixture stirred for 2 h at r.t. before the addition of sodium triacetoxyborohydride (5.1 g, 24mmol). The reaction was stirred at r.t. for 18 h before the addition of saturated potassium carbonate solution and the organics extracted with DCM. The organics were then dried (sodium sulfate), filtered and concentrated in vacuo. Purification by flash chromatography (1 :2:97 7N ammonia in MeOH:MeOH:DCM) afforded the title compound: RT = 0.56 min, m/z (ES+) = 267.3 [M + H]+ (LCMS Method - 2).
Preparation 5: c s-4-(teri-Butoxycarbonyl-methyl-amino)-3-fluoro-piperidine-l- carboxylic acid benzyl ester
Figure imgf000024_0002
To a solution of c«-3-fluoro-4-methylamino-piperidine-l-carboxylic acid benzyl ester (Preparation 4, 542mg, 2.04mmol) in DCM (10 mL) was added triethylamine (0.851mL, 6.10mmol) followed by di-tert-butyldicarbonate (444mg, 2.04mmol) and the mixture stirred at r.t. for 18 h. The mixture was diluted with water and the organics dried (sodium sulfate), filtered and concentrated in vacuo. Purification by flash chromatography (hexane - 7:3 hexane:ethyl acetate) afforded the title compound. RT = 1.28 min, m/z (ES ) = 389.3 [M +Na]+ (LCMS Method - 2).
Preparation 6: c s-3-Fluoro-piperidin-4- l)-methyl-carbamic acid tert-butyl ester
Figure imgf000025_0001
To a solution of c«-4-(?ert-butoxycarbonyl-methyl-amino)-3-fluoro-piperidine-l- carboxylic acid benzyl ester (Preparation 5, 634mg, 1.73mmol) in EtOH (20 mL) and 1 ,4- cyclohexadiene (4.05mL, 43.2mmol) was added 10% Pd/C (150mg, 4.2mmol) as a slurry in EtOH under argon. The reaction was stirred at r.t. for 18 h then filtered through celite and concentrated in vacuo. The crude residue was loaded onto a SCX cartridge, washed with methanol, eluted with 7N ammonia in MeOH and concentrated in vacuo to afford the title compound: ¾ NMR DH (400MHz, CDC13): 4.79-4.66 (m, 1H), 4.27-4.18 (m, 1H), 3.31- 3.20 (m, 2H), 2.92 (s, 3H), 2.78-2.71 (m, 2H), 2.09-1.85 (dq, J = 14, 6 Hz, 1H), 1.57-1.50 (m, 10H).
Preparation 7: ir««s-3-Fluoro-4- ine-l-carboxylic acid benzyl ester
Figure imgf000025_0002
To a solution of 3-fluoro-4-oxo-piperidine-l-carboxylic acid benzyl ester (Preparation 3, l .OOg, 3.98mmol) in anhydrous MeOH (lOmL) was added AcOH (0.5mL, 9mmol) followed by a solution of methylamine in MeOH (2 M, 6mL). The mixture was stirred at r.t. for 1 h before the addition of sodium cyanoborohydride (0.38g, 6.0mmol) portion wise over 5 min. After 3 h the reaction was quenched with a saturated solution of sodium bicarbonate and the organics extracted with ethyl acetate. The organics were then dried (sodium sulfate), filtered and concentrated in vacuo. Purification by flash chromatography (97:3 DCM:MeOH then 1 :5:94 7N ammonia in MeOH:MeOH:DCM) afforded the title compound. RT = 0.56 min, m/z (ES+) = 267.3 [M +Na]+ (Method -2). Preparation 8: ir««s-4-(teri-Butoxycarbonyl-methyl-amino)-3-fluoro-piperidine-l- carboxylic acid benzyl ester
Figure imgf000026_0001
The title compound was synthesized from ?ra«s-3-fluoro-4-methylamino-piperidine- 1 -carboxylic acid benzyl ester (Preparation 7) employing a procedure similar to that outlined in Preparation 5: 1H NMR DH (400MHz, CDC13): 7.48 - 7.32 (m, 5H), 5.17 (s, 2H), 4.67 - 4.36 (m, 2H), 4.36 - 4.17 (m, 2H), 2.91 - 2.77 (m, 5H), 1.85 - 1.69 (m, 2H), 1.50 (s, 9H). Preparation 9: (ir««s-3-Fluoro-piperidin-4- l)-methyl-carbamic acid teri-butyl ester
Figure imgf000026_0002
The title compound was synthesized from £raws-4-(tert-butoxycarbonyl-methyl- amino)-3-fluoro-piperidine- 1-carboxylic acid benzyl ester (Preparation 8) employing a procedure similar to that outlined in Preparation 6: !H NMR DH (400MHz, CDC13): 4.64 - 4.39 (m, 1H), 4.31 - 3.89 (m, 1H), 3.52 - 3.39 (m, 1H), 3.12 - 3.01 (m, 1H), 2.86 (br. s., 3H), 2.74 - 2.56 (m, 2H), 1.83 - 1.62 (m, 2H), 1.50 (s, 9H).
Preparation 10: 4-Cyclopropylamino-piperidine-l-carboxylic acid benzyl ester
Figure imgf000026_0003
To a solution of 4-oxo-piperidine- 1-carboxylic acid benzyl ester (lOg, 43.1mmol) in
DCM (lOOmL) was added acetic acid (3.7mL, 64.6mmol) and cyclopropylamine (3.7g, 64.6mmol). The mixture was cooled to 10 °C, sodium triacetoxyborohydride (13.7g, 64.6mmol) added and the mixture was stirred at room temperature for 18 h. The reaction mixture was quenched with water and washed with NaHC03. The aqueous layer was extracted with DCM (3x), the organics dried (MgS04), filtered and concentrated in vacuo. Purification by column chromatography (DCM:7M ammonia in MeOH, 95 :5) afforded the title compound: RT = 0.78 min; mlz (ES+) = 275.5 [M + H]+ (LCMS Method - 4).
Preparation 11: 4-(2-Methoxy-et -l-carboxylic acid benzyl ester
Figure imgf000027_0001
The title compound was synthesized from 4-oxo-piperidine-l -carboxylic acid benzyl ester and 2-methoxy-ethylamine employing a procedure similar to that outlined in Preparation 10: ¾ NMR D H (301MHz, CDC13): 7.40 - 7.27 (m, 5H), 5.12 (s, 2H), 4.24 - 4.01 (m, 2H), 3.49 (t, J=5.0 Hz, 2H), 3.35 (s, 3H), 2.74 - 2.98 (m, 4H), 2.71 - 2.55 (m, 1H), 1.96 - 1.76 (m, 2H), 1.39 - 1.19 (m, 2H).
Preparation 12 : 4-(teri-Butoxycarbonyl-cyclopropyl-amino)-piperidine-l-carboxylic acid benzyl ester
Figure imgf000027_0002
To a solution of 4-cyclopropylamino-piperidine-l-carboxylic acid benzyl ester
(Preparation 10, 1 1.79g, 43.13mmol) in THF (1 18mL) was added aqueous sodium carbonate solution (2N, 151mL) followed by di-tert-butyldicarbonate (11.3g, 51.75mmol) and the mixture stirred at r.t. for 18 h. The mixture was diluted with DCM, washed with water and the organics dried (MgS04), filtered and concentrated in vacuo. Purification by column chromatography (heptane :EtO Ac, 50:50) afforded the title compound: H NMR ¾ (301MHz, CDCI3): 7.43 - 7.28 (m, 5H), 5.12 (s, 2H), 4.39 -4.14 (m, 2H), 3.88 -3.70 (m, 1H), 2.89 -2.66 (m, 2H), 2.38 -2.25 (m, 1H), 2.03 -1.18 (m, 2H), 1.79 -1.63 (m, 2H), 1.45 (s, 9H), 0.79-0.69 (m, 2H), 0.67-0.58 (m, 2H). Preparation 13: 4-[teri-Butoxycarbonyl-(2-methoxy-ethyl)-amino]-piperidine-l- carboxylic acid benzyl ester
Figure imgf000028_0001
The title compound was synthesized from 4-(2-methoxy-ethylamino)-piperidine- 1 - carboxylic acid benzyl ester (Preparation 11) employing a procedure similar to that outlined in Preparation 12: JH NMR DH (301MHz, CDC13): 7.41 - 7.28 (m, 5H), 5.12 (s, 2H), 4.39 - 4.14 (m, 2H), 3.47 - 3.36 (m, 2H), 3.32 (s, 3H), 3.29 - 3.14 (m, 2H), 2.91 - 2.69 (m, 3H), 1.80 - 1.59 (m, 4H), 1.46 (s, 9H).
Preparation 14: Cyclopropyl-piperidin-4- l-carbamic acid teri-butyl ester
Figure imgf000028_0002
To a solution of 5% Pd/C (0.50g) in IMS (lOOmL) was added -(tert- butoxycarbonyl-cyclopropyl-amino)-piperidine-l -carboxylic acid benzyl ester (Preparation 12, 14.92g, 39.9mmol) and the mixture hydrogenated at 40 bar for 18 h at room temperature. The mixture was then filtered through celite and the filtrate concentrated in vacuo to afford the title compound: 1H NMR DH (301MHz , CDC13): 3.81 -3.66 (m, 1H), 3.19 - 3.07 (m, 2H), 2.71 - 2.56 (m, 2H), 2.38 - 2.27 (m, 1H), 2.08 -1.98 (m, 1H), 1.98 - 1.81 (m, 2H), 1.79 - 1.67 (m, 2H), 1.46 (s, 9H), 0.80 - 0.70 (m, 2H), 0.70 - 0.61 (m, 2H).
Preparation 15: (2-Methoxy-ethyl)- iperidin-4-yl-carbamic acid teri-butyl ester
Figure imgf000028_0003
The title compound was synthesized from 4-[?ert-butoxycarbonyl-(2-methoxy-ethyl)- amino]-piperidine-l -carboxylic acid benzyl ester employing a procedure similar to that outlined in Preparation 13: RT = 0.63 min; mlz (ES+) = 259.49 [M + H]+ (LCMS Method - 4)· Preparation 16: (l-Cyano-piperidin-4- l)-meth l-carbamic acid tert-butyl ester
Figure imgf000029_0001
To a solution of methyl-piperidin-4-yl-carbamic acid tert-butyl ester (2.08g, 9.68mmol) in DCM (25mL) was added a slurry of NaHC03 (2.44g, 29.1mmol) in water (6mL). The resulting reaction mixture was cooled to 0 °C and a solution of cyanogen bromide (1.23g, 1 1.6mmol) in DCM (3mL) was added dropwise over 1 min, before warming to r.t. and stirring for 2.5 h. The reaction mixture was diluted with water, the layers separated and the aqueous layer extracted with DCM. The combined organic extracts were washed with brine, dried (MgS04), filtered and concentrated in vacuo to afford the title compound: RT = 3.12 min; mlz (ES+) = 240.17 [M + H]+.
Preparation 17: (l-Cyano-4-methyl-piperidin-4-yl)-methyl-carbamic acid tert-butyl ester
Figure imgf000029_0002
The title compound was synthesized from methyl-(4-methyl-piperidin-4-yl)-carbamic acid tert-butyl ester (Preparation 2) employing a procedure similar to that outlined in Preparation 16: *Η NMR δΗ (301MHz, CDC13): 3.29 - 3.1 1 (m, 4H), 2.80 (s, 3H), 2.56 - 2.42 (m, 2H), 1.78 - 1.62 (m, 2H), 1.44 (s, 9H), 1.25 (s, 3H).
Preparation 18: (c/s-l-Cyano-3-fluoro-piperidin-4-yl)-methyl-carbamic acid tert-butyl ester
Figure imgf000029_0003
The title compound was synthesized from (cz's-3-fluoro-piperidin-4-yl)-methyl- carbamic acid tert-butyl ester (Preparation 6) employing a procedure similar to that outlined in Preparation 16: RT = 0.97 min; mlz (ES+) = 280.29 [M + H]+ (LCMS Method - 2)· Preparation 19: (l-Cyano-piperidin-4-yl)-ethyl-carbamic acid tert-butyl ester
N=— —
The title compound was synthesized from ethyl-piperidin-4-yl-carbamic acid tert- butyl ester employing a procedure similar to that outlined in Preparation 16: RT = 3.43 min; mlz (ES+) = 254.16 [M + H]+.
Preparation 20: (l-Cyano-piperidin-4- l)-cyclo ropyl-carbamic acid tert-butyl ester
Figure imgf000030_0001
The title compound was synthesized from cyclopropyl-piperidin-4-yl-carbamic acid tert-butyl ester (Preparation 14) employing a procedure similar to that outlined in Preparation 16: RT = 1.07 min; mlz (ES+) = 288.32 [M+ H]+ (LCMS Method - 2).
Preparation 21: (l-Cyano-piperidin-4- l)-isopropyl-carbamic acid tert-butyl ester
Figure imgf000030_0002
The title compound was synthesized from isopropyl-piperidin-4-yl-carbamic acid tert-butyl ester employing a procedure similar to that outlined in Preparation 16: XH NMR □ H (301MHZ, CDC13): 3.56 - 3.38 (m, 3H), 3.17 - 2.96 (m, 3H), 2.26 - 1.92 (m, 2H), 1.69 - 1.57 (m, 2H), 1.48 (s, 9H), 1.20 (d, J=6.4 Hz, 6H). Preparation 22: (l-Cyano-piperidin-4- l)-pro l-carbamic acid tert-butyl ester
Figure imgf000030_0003
The title compound was synthesized from piperidin-4-yl-propyl-carbamic acid tert- butyl ester employing a procedure similar to that outlined in Preparation 16: H NMR 8H (301MHz, CDC13): 4.14 - 3.88 (m, 1H), 3.56 - 3.37 (m, 2H), 3.17 - 2.89 (m, 4H), 1.99 - 1.63 (m, 4H), 1.60 - 1.48 (m, 2H), 1.45 (s, 9H), 0.93 - 0.83 (m, 3H).
Preparation 23: (l-Cyano-piperidin-4-yl)-(2-methoxy-ethyl)-carbamic acid tert-butyl ester
Figure imgf000031_0001
The title compound was synthesized from (2-methoxy-ethyl)-piperidin-4-yl-carbamic acid tert-butyl ester (Preparation 15) employing a procedure similar to that outlined in Preparation 16: 1H NMR δΗ (301MHz, CDC13): 3.56 - 3.38 (m, 5H), 3.33 (s, 3H), 3.31 - 3.20 (m, 2H), 3.15 - 2.98 (m, 2H), 2.03 - 3.15 (m, 2H), 1.77 - 1.64 (m, 2H), 1.46 (s, 9H).
Preparation 24: 4-(2,2,2-Trifluoro-eth lamino)-piperidine-l-carbonitrile
Figure imgf000031_0002
The title compound was synthesized from piperidin-4-yl-(2,2,2-trifluoro-ethyl)-amine employing a procedure similar to that outlined in Preparation 16: H NMR 8H (301MHZ, CDCI3): 3.46 (dt, J=13.2, 4.0 Hz, 2H), 3.29 - 3.12 (m, 2H), 3.11 - 2.94 (m, 2H), 2.84 - 2.68 (m, 1H), 1.91 (dd, J=13.1, 3.4 Hz, 2H), 1.57 - 1.39 (m, 2H), 1.34 - 1.08 (m, 1H).
Preparation 25: (5)-iV-Hydroxy-2-methoxypropionamidine
Figure imgf000031_0003
To a solution of (S)-2-methoxypropionitrile (910mg, 10.69mmol) in IMS (lOmL) was added hydro xylamine (50% in water, 1.55mL, 23.52mmol) and the reaction was heated to 80 °C for 16 h. On cooling, the solvent was removed in vacuo and the product recrystallised from heptane:EtOAc (1 : 1). The mother liquor was concentrated in vacuo and crystallised for a second time. Both crops of material were combined and dried under vacuum to afford the title compound: RT = 0.33 min; mlz (ES ) = 119.0 [M + H] (LCMS Method - 2).
Preparation 26: (5)-7V-Hydroxytetra -carboxamidine
Figure imgf000032_0001
To a solution of (5)-tetrahydrofuran-2-carbonitrile (lOOOmg, 10.3mmol) in IMS (7mL) was added hydroxylamine (50% in water, 750μΕ, 11.3mmol) and the reaction was heated to 70 °C for 18 h. On cooling, the solvent was removed in vacuo, azeotroping with toluene. The residue was purified by column chromatography (EtOAc) to afford the title compound. H NMR δΗ (300MHz, CDC13): 4.85 (bs, 2H), 4.44 - 4.32 (m, 1H), 4.00 - 3.74 (m, 2H), 2.20 - 1.80 (m, 4H).
Preparation 27: [l-(3-Isopropyl-[l 2,4]oxadiazol-5-yl)-piperidin-4-yl]-methyl-amine
Figure imgf000032_0002
To a solution of (l-cyano-piperidin-4-yl)-methyl-carbamic acid tert-butyl ester (Preparation 16, l .OOg, 4.18mmol) and N-hydroxy-isobutyramidine (512mg, 5.01mmol) in EtOH (20mL) was added a solution of zinc dichloride (683mg, 5.01mmol) in EtOH (7mL) and the resulting reaction mixture was stirred at r.t. for 2 h. Aqueous HC1 solution (11.7 M, 1.07mL, 12.5mmol) was added and the reaction mixture stirred at 50 °C for 16 h and at r.t. for 120 h. The solvent was removed in vacuo and the residue triturated with MeCN. The solid was removed by filtration and the filtrate concentrated in vacuo, dissolved in water and washed with EtOAc. The aqueous was basified to pH12 with 2M aqueous NaOH solution, extracted with EtOAc and the resulting emulsion was filtered through celite. The layers of the filtrate were separated and the aqueous was extracted with EtOAc (3 x). The combined organic extracts were washed with brine, dried (MgS04), filtered and concentrated in vacuo to afford the title compound: RT = 1.68 min; mlz (ES+) = 225.13 [M+ H]+.
Preparation 28: [l-(3-Isopropyl-[l,2,4]oxadiazol-5-yl)-4-methyl-piperidin-4-yl]-methyl- amine
Figure imgf000033_0001
The title compound was synthesized from (l-cyano-4-methyl-piperidin-4-yl)-methyl- carbamic acid tert-butyl ester (Preparation 17) employing a procedure similar to that outlined in Preparation 27: RT = 0.62 min; mlz (ES+) = 239.4 [M + H]+ (LCMS Method - 4).
Preparation 29: c s-{l-[3-(l,l-Difluoro-ethyl)-[l,2,4]oxadiazol-5-yl]-3-fluoro-piperidin- 4-yl}-methyl-amine
Figure imgf000033_0002
The title compound was synthesized from c«-l-cyano-3-fluoro-piperidin-4-yl)- methyl-carbamic acid tert-butyl ester (Preparation 18) employing a procedure similar to that outlined in Preparation 27: RT = 0.42 min; mlz (ES+) = 265.3 [M + H]+ (LCMS Method - 2). Preparation 30: ir««s-{l-[3-(l,l-Difluoro-ethyl)-[l,2,4]oxadiazol-5-yl]-3-fluoro- piperidin-4-yl}-methyl-amine
Figure imgf000033_0003
To a cooled solution of ?ra«s-3-fluoro-piperidin-4-yl)-methyl-carbamic acid tert- butyl ester (Preparation 9, 95mg, 0.41mmol) in DCM (5mL) was added a solution of sodium bicarbonate (93.7mg, 1.12mmol) in water (lmL) and a solution of cyanogen bromide (43mg, 0.41mmol) in DCM (2mL) dropwise over 5 min. The mixture was stirred at 0 °C for 1 h then at r.t. for 10 min. The organics were separated using a phase separator and concentrated in vacuo. The residue was then dissolved in EtOH and the title compound synthesized employing a procedure similar to that outlined in Preparation 27: RT = 0.46 min; mlz (ES+) = 265.2 [M + H]+ (LCMS Method - 2).
Preparation 31: [l-(3-Ethyl-[l,2,4] eridin-4-yl]-methyl-amine
Figure imgf000033_0004
The title compound was synthesized from (l-cyano-piperidin-4-yl)-methyl-carbamic acid tert-butyl ester (Preparation 16) and N-hydroxy-propionamidine employing a procedure similar to that outlined in Preparation 27: RT = 0.43 min; mlz (ES ) = 21 1.24 [M+ H]+ (LCMS Method - 2).
The following compounds were prepared by reacting (l -cyano-piperidin-4-yl)-methyl- carbamic acid tert-butyl ester (Preparation 16) with the appropriate amidoxime intermediate employing a procedure similar to that outlined in Preparation 27:
Figure imgf000034_0001
Prep. Structure Name Spectral Data piperidin-4-yl]-amine
{ 1 -[3 -((S 1 -Methoxy-ethyl)- [ 1 ,2,4]oxadiazol-5-yl]- RT = 1.82 min; mlz (ES^) = 241.2
38
piperidin-4-yl} -methyl- [M + H]+ (LCMS Method - 3). amine
Methyl- { l -[(S)-3 - (tetrahydro-furan-2-yl)- RT = 1.86 min; mlz (ES^) = 253.2
39
[ 1 ,2,4]oxadiazol-5-yl]- [M + H]+ (LCMS Method - 3). piperidin-4-yl} -amine
The following compounds were prepared by reacting (l-cyano-piperidin-4-yl)-ethyl- carbamic acid tert-butyl ester (Preparation 19) with the appropriate amidoxime intermediate employing a procedure similar to that outlined in Preparation 27:
Figure imgf000035_0001
Prep. Structure Name Spectral Data
Ethyl- { 1 - [3 -((S 1 -methoxy- RT = 1.16 min; mlz (ES+) =
45 ethyl)-[l,2,4]oxadiazol-5-yl]- 255.19 [M+ H]+ (LCMS Method - piperidin-4-yl} -amine 5).
Ethyl- { 1 -[(S 3 -(tetrahydro-
RT = 2.04 min; mlz (ES ) = 267.2
46 furan-2-yl)-[ 1 ,2,4]oxadiazol- [M + H]+ (LCMS Method - 3). o 5-yl]-piperidin-4-yl}-amine
The following compounds were prepared by reacting the appropriate cyano-piperidinyl intermediate with the appropriate amidoxime intermediate employing a procedure similar to that outlined in Preparation 27:
5
Figure imgf000036_0001
Prep. Structure Name Spectral Data
trifluoro-ethyl)-amine
{ l-[3-(l,l -Difluoro-ethyl)- [ 1 ,2,4]oxadiazol-5-yl]- RT = 1.49 min; mlz (ES^) = 291.4
51
piperidin-4-yl} -(2-methoxy- [M + H]+ (LCMS Method - 5). ethyl)-amine
Cyclopropyl- { l-[3-((5)-l - methoxy-ethyl)- RT = 1.40 min; mlz (ES^) = 267.5
52
[ 1 ,2,4]oxadiazol-5-yl]- [M + H]+ (LCMS Method - 5). piperidin-4-yl} -amine
Cyclopropyl- [ 1 -(3 -
RT = 0.42 min; mlz (ES+) = methoxymethyl-
53 253.26 [M+ H]+ (LCMS Method -
[ 1 ,2,4]oxadiazol-5-yl)- 2)·
piperidin-4-yl]-amine
Preparation 54: 5-Trichloromethyl- -carboxylic acid ethyl ester
Figure imgf000037_0001
To a suspension of ethyl-2-oximinooxamate (1.26g, 9.54mmol) in toluene (13mL, 120mmol) was added perchloroacetic anhydride (1.74mL, 9.54mmol) and the resulting reaction mixture was heated at 1 10 °C for 17 h, prior to removal of the solvent in vacuo. The residue was dissolved in EtOAc (80mL), washed with saturated aqueous NaHC(¾ solution (2x) and concentrated in vacuo to afford the title compound: XH NMR 8H (400 MHz, CDC13): 4.58 (q, J=7.29 Hz, 2 H), 1.50 (t, J=7.22 Hz, 3 H).
Preparation 55: 5-[4-(teri-Butoxycarbonyl-cyclopropyl-amino)-piperidin-l-yl]- [l,2,4]oxadiazole-3-carboxylic acid eth l ester
Figure imgf000037_0002
To a solution of cyclopropyl-piperidin-4-yl-carbamic acid tert-butyl ester5 (Preparation 14: l.Og, 4.2mmol) in DMF (5.0mL) under an argon atmosphere was added trichloromethyl-[l,2,4]oxadiazole-3-carboxylic acid ethyl ester (Preparation 54, 0.7g, 3mmol) and the reaction mixture heated to 50 °C for 2.5 h. The reaction was diluted with EtOAc before washing with water (2x) and finally brine. The EtOAc layer was dried over magnesium sulfate before filtering and concentrating in vacuo. Purification by column chromatography (2: 1 - 3 :2 heptane :EtOAc) afforded the title compound: RT = 1.23 min; mlz (ES+) = 381.36 [M + H]+ (LCMS Method - 2).
Preparation 56: Cyclopropyl-{l-[3-(l-hydroxy-l-methyl-ethyl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl}-carbamic acid tert-but l ester
Figure imgf000038_0001
To a solution of 5-[4-(tert-butoxycarbonyl-cyclopropyl-amino)-piperidin-l -yl]- [l,2,4]oxadiazole-3-carboxylic acid ethyl ester (Preparation 55, 380mg, l .Ommol) in anhydrous THF (8mL) at -20°C was added a solution of methylmagnesium bromide in diethyl ether (3M, 1.6mL, 5.0mmol). The reaction was allowed to stir at -20 °C for 1.5 h before the addition of a saturated aqueous solution of ammonium chloride. The mixture was concentrated in vacuo and the residue partitioned between EtOAc and water. The organics were washed with water and brine then dried (MgS04), filtered and concentrated in vacuo to afford the title compound: RT = 1.04 min; mlz (ES ) = 367.37 [M + H]+ (LCMS Method - 2). Preparation 57: 2-[5-(4-Cyclopropylamino-piperidin-l-yl)-[l,2,4]oxadiazol-3-yl]- propan-2-ol
Figure imgf000038_0002
To a solution of cyclopropyl- { l-[3-(l-hydroxy- l -methyl-ethyl)-[l,2,4]oxadiazol-5- yl]-piperidin-4-yl} -carbamic acid tert-butyl ester (Preparation 56, 20mg, 0.05mmol) in DCM (400μΕ) at 0 °C was added TFA (ΙΟΟμΕ, lmmol). The reaction mixture was allowed to stir for 1 h before concentrating in vacuo. The residue was dissolved in MeOH and loaded onto a SCX cartridge. The product was eluted with a 7M ammonia in MeOH solution and concentrated in vacuo to afford the title compound: RT = 0.44 min; mlz (ES+) = 267.30 [M + H] (LCMS Method - 2).
Preparation 58: l-(2-Fluoro-5-methyl-phenyl)-2-nitro-ethanol
Figure imgf000039_0001
To a solution of 2 -fluoro-5 -methyl -benzaldehyde (5.0g, 36.2mmol) and nitromethane
(2.35mL, 43.5mmol) in MeOH (90mL) at 0 °C was added a solution of NaOH (1.52g, 38.0mmol) in ¾0 (15mL) dropwise over 10 min and the resulting reaction mixture was stirred at r.t. for 50 min. The reaction mixture was poured into saturated aqueous NH4CI solution and extracted with DCM (3 x). The combined organics were washed with brine, dried (MgSC^), filtered and concentrated in vacuo to afford the title compound: H NMR 8H (300MHz, CDCI3): 7.34-7.30 (m, 1H), 7.15-7.10 (m, 1H), 7.0-6.9 (m, 1H), 5.71-5.70 (m, 1H), 4.63-4.57 (m, 2H), 2.93 (br. s, 1H), 2.33 (s, 3H).
Preparation 59: l-Fluoro-4-methyl-2- -2-nitro-vinyl)-benzene
Figure imgf000039_0002
solution of l-(2-fluoro-5-methyl-phenyl)-2-nitro-ethanol (Preparation 58,
7.18g, 36.1mmol) in acetic anhydride (6.8mL, 22.2mmol) was added DMAP (300mg, 2.5mmol) and the resulting reaction mixture was stirred at r.t. for 19 h. The reaction mixture was slowly poured into saturated aqueous aHC03 solution and stirred vigorously for 1 h. The solid was collected by filtration, washing with saturated aqueous NaHCCh solution, then H20 to afford the title compound: 1H NMR δΗ (300MHz, CDC13): 8.04-7.99 (d, J=13.76 Hz, 1H), 7.74-7.69 (d, J=13.76 Hz, 1H), 7.30-7.26 (m, 2H), 7.1-7.0 (m, 1H), 2.36 (s, 3H).
Preparation 60: (ir««s)-l-Benzyl-3- 2-fluoro-5-methyl-phenyl)-4-nitro-pyrrolidine
Figure imgf000039_0003
To a solution of l-fluoro-4-methyl-2-((£)-2-nitro-vinyl)-benzene (Preparation 59, 6.38g, 35.3mmol) in DCM (70mL) at 10 °C was added TFA (0.27mL, 3.52mmol), followed by dropwise addition of N-(methoxymethyl)-N-(trimethylsilylmethyl)benzylamine (9.52mL, 38.8mmol) in DCM (30mL). The resulting reaction mixture was stirred at r.t. for 1.5 h, poured into saturated aqueous aHC03 solution and extracted with DCM (3 x). The combined organic extracts were washed with brine, dried (MgS04), filtered and concentrated in vacuo to afford the title compound: *Η NMR δΗ (300MHz, CDC13): 7.35-7.26 (m, 5H), 7.1 -7.0 (m, 2H), 7.0-6.9 (m, 1H), 5.04-5.02 (m, 1H), 4.17-4.15 (m, 1H), 3.77-3.65 (m, 2H), 3.5-3.46 (m, 1H), 3.3-3.26 (m, 1H), 3.1-3.0 (m, 1H), 2.65-2.60 (m, 1H), 2.30 (s, 3H). Preparation 61: (ir««s)-Benzyl-4-(2-fluoro-5-meth l-phenyl)-pyrrolidin-3-ylamine
Figure imgf000040_0001
To a solution of (?ra«s)-l-benzyl-3-(2-fluoro-5-methyl-phenyl)-4-nitro-pyrrolidine (Preparation 60, 1 1.4g, 35.3mmol) in AcOH (90mL) at 30 °C was added zinc (13.8g, 212mmol) portion wise over 30 min. The resulting reaction mixture was stirred at 30 °C for 1 h, then filtered through celite washing with AcOH. The filtrate was concentrated in vacuo, the remainder dissolved in DCM (30mL) and poured slowly into saturated aqueous NaHC(¾ solution (900mL). The resulting mixture was stirred at r.t. for 16 h, the layers separated and the aqueous extracted with DCM (2 x). The combined organic extracts were dried (MgS04), filtered and concentrated in vacuo. Purification by column chromatography (DCM:MeOH, 19: 1 to 9: 1 to 17:3 to 4: 1 to 7:3 to 3 :2) afforded the title compound: RT = 3.06 min; mlz (ES+) = 285.1 [M + H]+ (LCMS Method - 3).
Preparation 62: [(ir««s)-l-Benzyl-4-(2-fluoro-5-methyl-phenyl)-pyrrolidin-3-yl]- carbamic acid tert-butyl ester
Figure imgf000040_0002
To a solution of (?ra«s)-benzyl-4-(2-fluoro-5-methyl-phenyl)-pyrrolidin-3-ylamine (Preparation 61, 5.6g, 19.7mmol) in THF (80mL) at 0 °C was added triethylamine (5.65mL, 40mmol) and di-tert-butyldicarbonate (5.15 g, 23.6mmol) in THF (30mL) and the mixture was stirred at r.t. for 21 h. The mixture was poured into water, extracted with EtOAc (3 x) and the combined organic extracts washed with brine, dried (MgSC^), filtered and concentrated. The resulting oil was dissolved in IH, allowed to stand at r.t. and the solid collected by filtration, washing with IH, to afford the title compound: RT = 3.36 min; mlz (ES+) = 385.1 [M + H]+ (LCMS Method - 3).
Preparation 63: [(3/?,4S)-l-Benzyl-4-(2-fluoro-5-methyl-phenyl)-pyrrolidin-3-yl]- carbamic acid tert-butyl ester
Figure imgf000041_0001
The title compound was afforded via chiral HPLC separation of [(/raws)-l-benzyl-4-(2- fluoro-5-methyl-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 62): IH:IPA:DEA 96:4:0.1, 15ml/min, 270nm, RT = 8.2 min. Preparation 64: [(3/?,4S)-4-(2-Fluoro-5-methyl-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000041_0002
A solution of [(3R,45')-l-benzyl-4-(2-fluoro-5-methyl-phenyl)-pyrrolidin-3-yl]- carbamic acid tert-butyl ester (Preparation 63, 2.4g, 6.2mmol) in MeOH (124mL) was passed through an H-Cube apparatus (10% Pd/C Catcart 70, 100 bar, 50 °C) at a flow rate of lmL per min. The solvent was removed in vacuo to afford the title compound: RT = 2.42 min; mlz (ES+) = 295.32 [M + H]+.
Preparation 65: l-(2,4-Difluoro-5-methyl-phenyl)-2-nitro-ethanol
Figure imgf000042_0001
To a solution of 2,4-difluoro-5-methyl-benzaldehyde (15.0g, 96.1mmol) and nitromethane (7.33g, 120mmol) in IPA (150mL) at 0°C was added a solution of NaOH (5.22g, 131mmol) in H2O (20mL) dropwise over 10 min and the resulting reaction mixture was stirred at r.t. for 1 h. Water (90mL) and saturated aqueous NH4CI solution (105mL) were added and extracted with DCM (3 x 250mL). The combined organic extracts were washed with brine, dried ( a2S04), filtered and concentrated in vacuo. Purification by column chromatography (IH:EtOAc, 9: 1 to 7: 1 to 4: 1 to 3 : 1) afforded the title compound: H NMR δΗ (300MHz, CDC13): 7.38-7.34 (m, 1H), 6.82-6.76 (m, 1H), 5.69-5.66 (m, 1H), 4.61- 4.51 (m, 2H), 2.95 (s, 1H), 2.28 (s, 3H).
Preparation 66: l,5-Difluoro-2-meth -4-((is)-2-nitro-vinyl)-benzene
Figure imgf000042_0002
The title compound was synthesized from l-(2,4-difluoro-5-methyl-phenyl)-2-nitro- ethanol (Preparation 65, 14.5g, 66.9mmol) employing a procedure similar to that outlined in Preparation 59: 1H NMR δΗ (400MHz, CDC13): 7.99-7.96 (d, J=13.73 Hz, 1H), 7.68- 7.65 (d, J=13.73 Hz, 1H), 7.34-7.3 (m, 1H), 6.91-6.89 (m, 1H), 2.27 (s, 3H).
Preparation 67: (ir««s)-l-Benzyl-3-(2,4-difluoro-5-methyl-phenyl)-4-nitro-pyrrolidine
Figure imgf000042_0003
The title compound was synthesized from l,5-difluoro-2-methyl-4-((£)-2-nitro- vinyl)-benzene (Preparation 66, 12. Og, 60.3mmol) employing a procedure similar to that outlined in Preparation 60: RT= l .OOmin; mlz (ES+) = 333.2 [M+ H]+ (LCMS Method - 4).
Preparation 68: (ir««s)-l-Benzyl-4-(2,4-difluoro-5-methyl-phenyl)-pyrrolidin-3-ylamine
Figure imgf000043_0001
The title compound was synthesized from (ira«s)-l-benzyl-3-(2,4-difluoro-5-methyl- phenyl)-4-nitro-pyrrolidine (Preparation 67, 21. Og, 63.3mmol) employing a procedure similar to that outlined in Preparation 61: RT = 0.93 min; mlz (ES+) = 303.2 [M + H]+ (LCMS Method - 4).
Preparation 69: [(ir««s)-l-Benzyl-4-(2,4-difluoro-5-methyl-phenyl)-pyrrolidin-3-yl]- carbamic acid teri-butyl ester
Figure imgf000043_0002
The title compound was synthesised from (?ra«s)-l-benzyl-4-(2,4-difluoro-5-methyl- phenyl)-pyrrolidin-3-ylamine (Preparation 68, 9.5g, 31.5mmol) employing a procedure similar to that outlined in Preparation 62: RT = 3.25 min; mlz (ES ) = 403.3 [M + H]+ (LCMS Method - 5). Preparation 70: [(3/?,4S)-l-Benzyl-4-(2,4-difluoro-5-methyl-phenyl)-pyrrolidin-3-yl]- carbamic acid teri-butyl ester
Figure imgf000043_0003
The title compound was afforded via chiral HPLC separation of [(?ra«s)-l-benzyl-4- (2,4-difluoro-5-methyl-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 69): IH:IPA:DEA 96:4:0.1, 15ml/min, 270nm, RT = 9.0 min.
Preparation 71: [(3/?,4S)-4-(2,4-Difluoro-5-methyl-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000044_0001
A solution of [(3R,45')-l-benzyl-4-(2,4-difluoro-5-methyl-phenyl)-pyrrolidin-3-yl]- carbamic acid tert-butyl ester (Preparation 70, 1.97g, 4.9mmol) in MeOH (100 mL) was passed through an H-Cube apparatus (10% Pd/C Catcart 70, 80 bar, 50 °C) at a flow rate of lmL per min. The solvent was removed in vacuo to afford the title compound: RT = 0.68 min; mlz (ES+) = 313.32 [M + H]+ (LCMS Method - 2).
Preparation 72: l-(2-Fluoro-phenyl)- -nitro-ethanol
Figure imgf000044_0002
The title compound was synthesized from 2-fluoro-benzaldehyde (41g, 330mmol) employing a procedure similar to that outlined in Preparation 58: H NMR 8H (300MHZ, CDC13): 7.60-7.50 (m, IH), 7.40-7.29 (m, 1H), 7.23-7.13 (1H, m), 7.12-7.01 (m, 1H), 5.8- 5.68 (m, IH), 4.69-4.52 (m, 2H), 3.15-3.0 (m, IH).
Preparation 73: l-Fluoro-2-((is)-2-nit -vinyl)-benzene
Figure imgf000044_0003
The title compound was synthesized from l-(2-fluoro-phenyl)-2-nitro-ethanol (Preparation 72, 59g, 319mmol) employing a procedure similar to that outlined in Preparation 59: H NMR δΗ (400MHz, CDC13): 8.05 (d, J=13.79 Hz, IH), 7.72 (d, J=13.79 Hz, IH), 7.55-7.42 (m, 2H), 7.28-7.12 (m, 2H).
Preparation 74: (ir««s)-l-Benzyl-3-(2-fluoro-phenyl)-4-nitro-pyrrolidine
Figure imgf000045_0001
The title compound was synthesized from l-fluoro-2-((£)-2-nitro-vinyl)-benzene (Preparation 73, 26.5g, 159mmol) employing a procedure similar to that outlined in Preparation 60: RT = 0.90 min; mlz (ES+) = 301.2 [M+ H]+ (LCMS Method - 4).
Preparation 75: (ir««s)-l-Benzyl-4- 2-fluoro-phenyl)-pyrrolidin-3-ylamine
Figure imgf000045_0002
The title compound was synthesized from (?ra«s)-l-benzyl-3-(2-fluoro-phenyl)-4- nitro-pyrrolidine (Preparation 74, 54.5 g, 181mmol) employing a procedure similar to that outlined in Preparation 61: RT = 0.75 min; mlz (ES+) = 271.2 [M + H]+ (LCMS Method - 4)·
Preparation 76: [(ir««s)-l-Benzyl-4-(2-fluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid teri-butyl ester
Figure imgf000045_0003
The title compound was synthesised from (?ra«s)-l-benzyl-4-(2-fluoro-phenyl)- pyrrolidin-3-ylamine (Preparation 75, 11.7g, 43.3mmol) employing a procedure similar to that outlined in Preparation 62: RT = 3.00 min; mlz (ES+) = 371.3 [M + H]+ (LCMS Method - 5).
Preparation 77: [(3/?,4S)-l-Benzyl-4-(2-fluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid teri-butyl ester
Figure imgf000046_0001
The title compound was afforded via chiral HPLC separation of [(7ra«s)-l-benzyl-4-(2- fluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 76): IH:IPA:n- butylamine 96:4:0.1, 15ml/min, 270nm, RT = 10.6 min.
Preparation 78: [(3i?,4S)-4-(2-Fluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000046_0002
The title compound was synthesised from [(3R,45)-l-benzyl-4-(2-fluoro-phenyl)- pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 77, 880mg, 2.37mmol) employing a procedure similar to that outlined in Preparation 64: RT = 0.63 min; m/z (ES ) = 281.3 [M + H]+ (LCMS Method - 2).
Preparation 79: 2-[(3i?,45)-3-feri-Butoxycarbonylamino-4-(2,5-difluoro-phenyl)- pyrrolidin-l-yl]-pyrimidine-5-carbox lic acid methyl ester
Figure imgf000046_0003
To a solution of 2-chloro-pyrimidine-5-carboxylic acid methyl ester (2.1 g, 12mmol) and [(3R,45')-4-(2,5-difluorophenyl)pyrrolidin-3-yl]carbamic acid tert-butyl ester (Preparation 193, 4.0g, 13mmol) in DCE (l OOmL) was added triethylamine (3.3mL, 23mmol) and the resulting reaction mixture was stirred at r.t. for 16 h. The reaction mixture was diluted with DCM (200mL), washed with water (200mL) and brine (400mL), dried (MgS04), filtered and concentrated in vacuo. The remainder was triturated with MeOH and the solid collected by filtration to afford the title compound: RT = 3.95 min; mlz (ES ) = 435.18 [M+ H]+.
Preparation 80: 2-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2,5-difluoro-phenyl)- pyrrolidin-l-yl]-pyrimidine-5-carbox lic acid
Figure imgf000047_0001
To a solution of 2-[(3R,45')-3-iert-butoxycarbonylamino-4-(2,5-difluoro-phenyl)- pyrrolidin-l -yl]-pyrimidine-5-carboxylic acid methyl ester (Preparation 79, 3.89g, 8.95mmol) in THF (l OOmL) was added aqueous NaOH solution (1M, 50mL, 50mmol) and the resulting reaction mixture was stirred at r.t. for 16 h. The reaction mixture was acidified to pH 6 using 2M aqueous HC1 solution and extracted with EtOAc (3 x 200mL). The combined organic extracts were dried (MgS04), filtered and concentrated in vacuo. The remainder was triturated with Et20 and the solid collected by filtration to afford the title compound: RT = 3.50 min; mlz (ES+) = 421.19 [M+ H]+.
Preparation 81: 2-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2,4,5-trifluoro-phenyl)- pyrrolidin-l-yl]-pyrimidine-5- er
Figure imgf000047_0002
The title compound was synthesized from 2-chloro-pyrimidine-5-carboxylic acid methyl ester (350mg, 3.85mmol) and [(3R,45,)-4-(2,4,5-trifluorophenyl)pyrrolidin-3- yljcarbamic acid tert-butyl ester (Preparation 199, 784mg, 2.48mmol) employing a procedure similar to that outlined in Preparation 79: RT = 1.29 min; mlz (ES+) = 453.00 [M + H]+ (LCMS Method - 2). Preparation 82: 2-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2,4,5-trifluoro-phenyl)- pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid
Figure imgf000048_0001
The title compound was synthesized from 2-[(3R,45')-3-tert-butoxycarbonylamino-4- (2,4,5-trifluoro-phenyl)-pyrrolidin-l -yl]-pyrimidine-5-carboxylic acid methyl ester (Preparation 81, 820mg, 1.8mmol) employing a procedure similar to that outlined in Preparation 80: RT = 1.1 1 min; mlz (ES+) = 439.3 [M+ H]+ (LCMS Method - 2).
Preparation 83: 5-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2,4,5-trifluoro-phenyl)- pyrrolidin-l-yl]-pyrazine-2-carboxylic acid methyl ester
Figure imgf000048_0002
The title compound was synthesized from 2,5-chloro-pyrazine-2-carboxylic acid methyl ester (292mg, 1.69mmol) and [(3R,45,)-4-(2,4,5-trifluorophenyl)pyrrolidin-3- yljcarbamic acid tert-butyl ester (Preparation 199, 1.07g, 3.38mmol) employing a procedure similar to that outlined in Preparation 79: RT = 1.18 min; mlz (ES+) = 453.38 [M + H]+ (LCMS Method - 2).
Preparation 84: 5-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2,4,5-trifluoro-phenyl)- pyrrolidin-l-yl]-pyrazine-2-carboxylic acid
Figure imgf000048_0003
The title compound was synthesized from 5-[(3R,45')-3-tert-butoxycarbonylamino-4- (2,4,5-trifluoro-phenyl)-pyrrolidin-l -yl]-pyrazine-2-carboxylic acid methyl ester (Preparation 83, 400mg, 0.884mmol) employing a procedure similar to that outlined in Preparation 80: RT = 1.05 min; mlz (ES ) = 439.33 [M+ H] (LCMS Method - 2).
Preparation 85: 2-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2,4-difluoro-5-methyl- phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid methyl ester
Figure imgf000049_0001
The title compound was synthesized from 2-chloro-pyrimidine-5-carboxylic acid methyl ester (295mg, 1.71mmol) and [(3R,45')-4-(2,4-difluoro-5-methyl-phenyl)-pyrrolidin- 3-yl]-carbamic acid tert-butyl ester (Preparation 71, 640mg, 2.05mmol) employing a procedure similar to that outlined in Preparation 79: RT = 1.33 min; mlz (ES+) = 449.38 [M + H]+ (LCMS Method - 2).
Preparation 86: 2-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2,4-difluoro-5-methyl- phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid
Figure imgf000049_0002
To a solution of 2-[(3R,45')-3-iert-butoxycarbonylamino-4-(2,4-difluoro-5-methyl- phenyl)-pyrrolidin-l -yl]-pyrimidine-5-carboxylic acid methyl ester (Preparation 85, 767mg, 1.71mmol) in THF (5.5mL), MeOH (2.8mL) and water (2.5mL) was added lithium hydroxide monohydrate (86.1mg, 2.05mmol) and the resulting reaction mixture was heated at 30 °C for 5.5 h. The solvent was removed in vacuo, water was added and the mixture acidified to pH 1 with aqueous HCl solution (1M) prior to extraction with EtOAc. The organic extract was dried (MgS04), filtered and concentrated in vacuo to afford the title compound: RT = 1.14 min; mlz (ES+) = 435.39 [M+ H]+ (LCMS Method - 2).
Preparation 87: 2-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2-fluoro-5-methyl-phenyl)- pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid methyl ester
Figure imgf000050_0001
The title compound was synthesized from 2-chloro-pyrimidine-5-carboxylic acid methyl ester (375mg, 2.17mmol) and [(3R,45')-4-(2-fluoro-5-methyl-phenyl)-pyrrolidin-3- yl]-carbamic acid tert-butyl ester (Preparation 64, 704 mg, 2.39mmol), employing a procedure similar to that outlined in Preparation 79: RT = 1.32 min; mlz (ES ) = 431.39 [M + H]+ (LCMS Method - 2).
Preparation 88: 2-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2-fluoro-5-methyl-phenyl)- pyrrolidin-l-yl]-pyrimidine-5-carbox lic acid
Figure imgf000050_0002
The title compound was synthesized from 2-[(3R,45')-3-iert-butoxycarbonylamino-4- (2-fluoro-5-methyl-phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid methyl ester (Preparation 87, 850mg, 2.0mmol), employing a procedure similar to that outlined in Preparation 80: RT = 1.13 min; mlz (ES+) = 417.38 [M+ H]+ (LCMS Method - 2).
Preparation 89: 2-[(3/?,45)-3-feri-Butoxycarbonylamino-4-(2-fluoro-phenyl)-pyrrolidin- l-yl]-pyrimidine-5-carboxylic acid meth l ester
Figure imgf000050_0003
The compound was synthesized from 2-chloro-pyrimidine-5-carboxylic acid methyl ester (271mg, 1.57mmol) and [(3R,45')-4-(2-fluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 78, 440mg, 1.57mmol), employing a procedure similar to that outlined in Preparation 79: RT=1.24 min; mlz (ES+) = 417.33 [M+ H]+ (LCMS Method 2). Preparation 90: 2-[(3i?,45)-3-feri-Butoxycarbonylamino-4-(2-fluoro-phenyl)-pyrrolidin- l-yl]-pyrimidine-5-carboxylic acid
Figure imgf000051_0001
The title compound was synthesized from 2-[(3R,45')-3-iert-butoxycarbonylamino-4-
(2-fluoro-phenyl)-pyrrolidin-l -yl]-pyrimidine-5-carboxylic acid methyl ester (Preparation 89, 460mg, l . lmmol), employing a procedure similar to that outlined in Preparation 80: RT = 1.06 min; mlz (ES+) = 403.30 [M+ H]+ (LCMS Method - 2). Preparation 91: [(3i?,4S)-l-{5-[(l-Benzyl-piperidin-4-yl)-cyclopropyl-carbamoyl]- pyrimidin-2-yl}-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000051_0002
To a solution of 2-[(3R,45,)-3-iert-butoxycarbonylamino-4-(2,4,5-trifluoro-phenyl)- pyrrolidin-l -yl]-pyrimidine-5-carboxylic acid (Preparation 82, 400mg, 900μmol) in THF (12mL) was added EDCI (289mg, 1.50mmol), HOBt (2.30mg, 1.50mmol) and DIPEA (421 μΕ, 2.42mmol). The resulting reaction mixture was stirred at r.t. for 20 min prior to the addition of (l-benzyl-piperidin-4-yl)-cyclopropyl-amine (320mg, 1.4mmol) in THF (3mL) and the resulting reaction mixture was stirred at r.t. for 16 h. The solvent was removed in vacuo, then the residue was dissolved in EtOAc, washed with aqueous NaOH solution (2M, 2 x) and brine, dried (MgS04), filtered and concentrated in vacuo. Purification by column chromatography (EtOAc) afforded a crude product which was dissolved in DCM and added to an SCX cartridge. The SCX cartridge was washed with MeOH then eluted with 7M ¾ in MeOH and the basic fraction was concentrated in vacuo to afford the title compound: RT = 0.93 min; mlz (ES+) = 651.57 [M+ H]+ (LCMS Method - 2). Preparation 92: [(3i?,4S)-l-[5-(Cyclopropyl-piperidin-4-yl-carbamoyl)-pyrimidin-2-yl]- 4-(2,4,5-trifluoro-phenyl)- butyl ester
Figure imgf000052_0001
A solution of [(3R,45,)-l- {5-[(l-benzyl-piperidin-4-yl)-cyclopropyl-carbamoyl]- p rimidin-2-yl}-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 91, 162mg, 249μηιο1) in EtOH (5mL) was reacted on the H-cube cube (10% Pd/C Catcart 70, 50 bar, 70 °C) at a flow rate of 1 mL/min. The resulting solution was concentrated in vacuo and the remainder was re-dissolved in EtOH (25mL) and reacted on the H-cube (10% Pd/C Catcart 70, 100 bar, 75 °C) at a flow rate of 1 mL/min. The resulting solution was concentrated in vacuo and the remainder was re-dissolved in EtOH (35mL) and reacted on the H-cube (10% Pd/C Catcart 70, 100 bar, 80 °C) at a flow rate of 0.7 mL/min. The resulting solution was concentrated in vacuo to afford the title compound: RT = 0.81 min; mlz (ES+) = 561.57 [M + H]+ (LCMS Method - 2).
Preparation 93: [(3i?,4S)-l-{5-[(l-Cyano-piperidin-4-yl)-cyclopropyl-carbamoyl]- pyrimidin-2-yl}-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000052_0002
To a solution of [(3R,45)-l -[5-(cyclopropyl-piperidin-4-yl-carbamoyl)-pyrimidin-2- yl]-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 92, 78.2mg, 139μιηο1) in DCM (500μΓ) was added a slurry of NaHC03 (35mg, 420μιηο1) in water (400μί), followed by cyanogen bromide (18mg, ΠΟμιηοΙ) in DCM (500μΕ). The resulting reaction mixture was stirred at r.t. for 1.25 h, then diluted with DCM and water. The layers were separated, the aqueous layer extracted with DCM (3 x) and the combined organic extracts were washed with brine, dried (MgSC^), filtered and concentrated in vacuo. Purification by column chromatography (EtOAc:IH, 4: 1) afforded the title compound: RT = 1.16 min; mlz (ES+) = 586.52 [M + H]+ (LCMS Method - 2). Preparation 94: [(3/?,45)-4-(2,5-Difluoro-phenyl)-l-(5-{[l-(3-isopropyl-[l,2,4]oxadiazol- 5-yl)-piperidin-4-yl]-methyl-carbamoyl}-pyrimidin-2-yl)-pyrrolidin-3-yl]-carbamic ester tert-butyl ester
Figure imgf000053_0001
To a solution of 2-[(3R,45')-3-iert-butoxycarbonylamino-4-(2,5-difluoro-phenyl)- pyrrolidin-l -yl]-pyrimidine-5-carboxylic acid (Preparation 80, 120mg, 280μmol) in THF (4mL) was added EDCI (71.1mg, 371 μιηο1), HOBt (56.8mg, 371 μιηο1) and DIPEA (99.4μΕ, 571 μιηοΐ). The resulting reaction mixture was stirred at r.t. for 20 min prior to the addition of [l-(3-isopropyl-[l ,2,4]oxadiazol-5-yl)-piperidin-4-yl]-methyl-amine (Preparation 27, 96.0mg, 428μιηο1), then stirred at r.t. for 16 h. The solvent was removed in vacuo and the remainder dissolved in EtOAc, washed with aqueous NaOH solution (2M, 2 x), aqueous citric acid solution (10%, 2 x) and brine, dried (MgSC^), filtered and concentrated in vacuo. Purification by column chromatography (EtOAcTH, 4: 1 to 9: 1) afforded the title compound: RT = 3.87 min; mlz (ES+) = 627.33 [M + H]+. Preparation 95: {(S^SH^^-Difluoro-phen ^-l-IS-Cil-IS-Cl-fluoro-l-meth l-eth l)- [l,2,4]oxadiazol-5-yl]-piperidin-4-yl}-methyl-carbamoyl)-pyrimidin-2-yl]-pyrrolidin-3- yl}-carbamic acid tert-but l ester
Figure imgf000053_0002
To a solution of 2-[(3R,45)-3-?ert-butoxycarbonylamino-4-(2,5-difluoro-phenyl)- pyrrolidin-l -yl]-pyrimidine-5-carboxylic acid (Preparation 80, 70mg, 166μιηο1) in DCM (2mL) was added { l-[3-(l-fluoro-l-methyl-ethyl)-[l,2,4]oxadiazol-5-yl]-piperidin-4-yl}- methyl-amine (Preparation 36, 40mg, 166μιηο1) and DIPEA (87μί, 499μιηο1) followed by PPA (50% w/w in EtOAc, 149μΕ, 250μιηο1). The resulting reaction mixture was stirred at r.t. under argon for 3 h. After this time the mixture was diluted with water and filtered through a hydrophobic frit and concentrated in vacuo. Purification by column 5 chromatography (DCM to 2:98 MeOH:DCM) afforded the title compound: RT = 1.23 min; mlz (ES+) = 645.45 [M+ H]+ ((LCMS Method - 2 ).
The following compounds were prepared by reacting 2-[(3R,4S)-3-tert- butoxycarbonylamino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid 10 (Preparation 80) with the appropriate amine intermediate employing a procedure similar to that outlined in Preparation 94:
Figure imgf000054_0001
Figure imgf000055_0001
ac ferf- ty ester
Figure imgf000056_0001
acid tert-butyl ester
The following compounds were prepared by reacting 2-[(3R,4S)-3-tert- butoxycarbonylamino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid (Preparation 80) with the appropriate amine intermediate employing a procedure similar to that outlined in Preparation 95:
Figure imgf000057_0001
Figure imgf000058_0001
ester
The following compounds were prepared by reacting 2-[(3R,45')-3-iert- butoxycarbonylamino-4-(2,4,54rifluoro^henyl)^yrrolidin-l-yl]-pyrimidine-5-carboxylic acid (Preparation 82) with the appropriate amine intermediate employing a procedure 5 similar to that outlined in Preparation 94:
Figure imgf000058_0002
Figure imgf000059_0001
Figure imgf000060_0001
acid tert-butyl ester 2)·
Figure imgf000061_0001
butyl ester Method -
Figure imgf000062_0001
The following compounds were prepared by reacting the appropriate acid intermediate with the appropriate amine intermediate employing a procedure similar to that outlined in Preparation 95:
Figure imgf000063_0001
Figure imgf000064_0001
The following compounds were prepared by reacting 2-[(3R,45')-3-iert- butoxycarbonylamino-4-(2-fluoro-5-methyl-phenyl)-pyrrolidin- l -yl]-pyrimidine-5- carboxylic acid (Preparation 88) with the appropriate amine intermediate employing a 5 procedure similar to that outlined in Preparation 94:
Figure imgf000064_0002
Figure imgf000065_0001
Figure imgf000066_0001
Figure imgf000067_0001
carbamic acid tert-butyl ester
The following compounds were prepared by reacting 2-[(3 ?,45)-3-tert- butoxycarbonylamino-4-(2-fluoro-5-methyl-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid (Preparation 88) with the appropriate amine intermediate employing a 5 procedure similar to that outlined in Preparation 95:
Figure imgf000067_0002
Figure imgf000068_0001
butyl ester
The following compounds were prepared by reacting 2-[(3R,4S -3-tert- butoxycarbonylamino-4-(2-fluoro-phenyl)-pyrrolidin- 1 -yl]-pyrimidine-5-carboxylic acid (Preparation 90) with the appropriate amine intermediate employing a procedure similar to 5 that outlined in Preparation 95:
Figure imgf000068_0002
Figure imgf000069_0001
ester
The following compounds were prepared by reacting 2-[(3 ?,45)-3-tert- butoxycarbonylamino-4-(2,4-difluoro-5-methyl-phenyl)-pyrrolidin-l -yl]-pyrimidine-5- carboxylic acid (Preparation 86) and the appropriate amine intermediate employing a 5 procedure similar to that outlined in Preparation 94:
Prep. Structure Name LCMS Data
[(3R,45)-l-(5- {[l -(3- Difluoromethyl- RT = 4.07 min; m/z [l,2,4]oxadiazol-5-yl)- (ES+) = 649.07 [M
159 piperidin-4-yl]-methyl- carbamoyl} -pyrimidin-2-yl)- + H]+ (LCMS 4-(2,4-difluoro-5-methyl- Method - 1). phenyl)-pyrrolidin-3-yl]- carbamic acid tert-butyl ester
[(3R,45)-l-[5-({l -[3-(l,l- Difluoro-ethyl)- RT = 4.20 min; m/z [l,2,4]oxadiazol-5-yl]- (ES+) = 663.10 [M
160 piperidin-4-yl} -methyl- carbamoyl)-pyr imidin-2 -yl]- + H]+ (LCMS 4-(2,4-difluoro-5-methyl-
- Method - 1).
phenyl)-pyrrolidin-3-yl]- carbamic acid tert-butyl ester
[(3R,45)-l-(5- {[l -(3- Cyclopropyl- RT = 4.02 min; m/z [l,2,4]oxadiazol-5-yl)- (ES+) = 639.13 [M
161 piperidin-4-yl]-methyl- carbamoyl} -pyrimidin-2-yl)- + H]+ (LCMS 4-(2,4-difluoro-5-methyl- Method - 1). phenyl)-pyrrolidin-3-yl]- carbamic acid tert-butyl ester
Figure imgf000070_0001
Figure imgf000071_0001
ester
Preparation 171 : [(3Λ,45)-1-{5-[{1-[3-(1,1-ϋίηυοΓθ-β^1)-[1,2,4]οχ3ΰΪ3ζο1-5^1]- piperidin-4-yl}-(2,2,2-trifluoro-ethyl)-carbamoyl]-pyrimidin-2-yl}-4-(2,5-difluoro- phenyl)-pyrrolidin-3-yl]-carbamic acid teri-butyl ester
Figure imgf000072_0001
To a suspension of 2-[(3R,45')-3-tert-butoxycarbonylamino-4-(2,5-difluoro-phenyl)- pyrrolidin-l -yl]-pyrimidine-5-carboxylic acid (Preparation 80, 150mg, 360μηιο1) in DCM (3mL) was added pyridine (38μί, 460μηιο1), DMF (20μ∑, 200μηιο1) followed by the dropwise addition of oxalyl chloride (36μΙ,, 430μηιο1) under argon. The reaction mixture was stirred at r.t. for 1 h. To this was added a solution of ( l-[3-(l, l-difluoro-ethyl)- [l,2,4]oxadiazol-5-yl]-piperidin-4-yl} -(2,2,2 -trifluoro-ethyl)-amine (Preparation 50, 120mg, 390μιηο1) and pyridine (38μΚ) in DCM (2mL) and the mixture stirred at r.t. for 18 h. The reaction mixture was diluted with EtOAc and washed with water (15mL) then a saturated solution of sodium carbonate (15mL). The organic extracts were dried (MgSC^), filtered and concentrated in vacuo. The residue was purified by preparative HPLC (standard method) to afford the title compound: RT = 1.37 min; mlz (ES+) = 717.43 [M + H]+ (LCMS Method - 2). Preparation 172: [(S^SJ-l-IS-Cil- -Cia-Difluoro-eth ^-Il^^loxadiazol-S- ll-S- fluoro-piperidin-4-yl}-methyl-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro-phenyl)- pyrrolidin-3-yl]-carbamic acid tert-but l ester
Figure imgf000072_0002
The title compound was prepared by reacting 2-[(3R,4S)-3-tert- butoxycarbonylamino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid (Preparation 80) with c«- { l -[3-(l ,l-difluoro-ethyl)-[l,2,4]oxadiazol-5-yl]-3-fluoro- piperidin-4-yl}-methyl-amine (Procedure 29) employing a procedure similar to that outlined in Preparation 95 to afford the title compound as a mixture of diastereoisomers: RT = 1.30 min; mlz (ES+) = 667.44 [M + H]+ (LCMS Method - 2). Preparation 173: [(3i?,45)-l-[5-({(35,4i?)-l-[3-(l,l-Difluoro-ethyl)-[l,2,4]oxadiazol-5- yl]-3-fluoro-piperidin-4-yl}-methyl-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro-phenyl)- pyrrolidin-3-yl]-carbamic acid tert-but l ester
Figure imgf000073_0001
[(3R,4S 1 -[5-( { 1 -[3 -(1 , 1 -Difluoro-ethyl)-[ 1 ,2,4]oxadiazol-5 -yl]-3 -fluoro-piperidin-
4-yl}-methyl-carbamoyl)-pyrimidin-2-yl]-4-(2,^^
acid tert-butyl ester (Preparation 172) was separated via chiral HPLC: MTBE:MeOH:IH 35:35:30, 15ml/min, 270nm, RT = 12.1 min to afford the title single diastereomer with arbritrarily assigned stereochemistry: RT = 1.30 min; m/z (ES+) = 667.44 [M + H]+ (LCMS Method - 2).
Preparation 174: [(3i?,45)-l-[5-({(3i?,4S)-l-[3-(l,l-Difluoro-ethyl)-[l,2,4]oxadiazol-5- yl]-3-fluoro-piperidin-4-yl}-methyl-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro-phenyl)- pyrrolidin-3-yl]-carbamic acid tert-but l ester
Figure imgf000073_0002
[(3R,4S 1 -[5-( { 1 -[3 -(1 , 1 -Difluoro-ethyl)-[ 1 ,2,4]oxadiazol-5 -yl]-3 -fluoro-piperidin- 4-yl}-methyl-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 172) was separated via chiral HPLC: MTBE:MeOH:IH 35:35:30, 15ml/min, 270nm, RT = 9.7 min to afford the title single diastereomer with arbritrarily assigned stereochemistry. RT = 1.30 min; m/z (ES+) = 667.44 [M + H]+ (LCMS Method - 2).
Preparation 175: [(3i?,45)-l-[5-({(3i?,4i?)-l-[3-(l,l-Difluoro-ethyl)-[l,2,4]oxadiazol-5- yl]-3-fluoro-piperidin-4-yl}-methyl-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro-phenyl)- pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000074_0001
The title compound was prepared by reacting 2-[(3R,45')-3-iert- butoxycarbonylamino-4-(2,5-difluoro-phenyl)^yrrolidin-l-yl]^yrimidine-5-carboxylic acid (Preparation 80) with ?ra«s- { l-[3-(l ,l-difluoro-ethyl)-[l,2,4]oxadiazol-5-yl]-3-fluoro- piperidin-4-yl}-methyl-amine (Procedure 30) employing a procedure similar to that outlined in Preparation 95 to afford the title compound as a mixture of diastereoisomers. The diastereomers were separated via chiral HPLC: MTBE:MeOH 80:20, 15ml/min, 270nm, RT = 9.7 min to afford the title single diastereomer with arbitrarily assigned stereochemistry: RT = 1.26 min; mlz (ES+) = 667.5 [M + H]+ (LCMS Method - 2).
Preparation 176: [(3/?,45)-4-(2,4-Difluoro-5-methyl-phenyl)-l-(5-{[l-(3-isopropyl- [ 1 ,2 ,4] oxa diazol-5-yl)-piperidin-4-yl] -methyl-carbamoyl} -pyrimidin-2 -yl)-pyr r olidin-3 - yl]-carbamic acid teri-b
Figure imgf000074_0002
The title compound was synthesised from 2-[(3R,45')-3-iert-butoxycarbonylamino-4- (2,4-difluoro-5-methyl-phenyl)-pyrrolidin- 1 -yl]-pyrimidine-5-carboxylic acid (Preparation 86) and [l-(3-isopropyl-[l ,2,4]oxadiazol-5-yl)-piperidin-4-yl]-methyl-amine (Preparation 27) employing a procedure similar to that outlined in Preparation 94: RT = 1.28 min; mlz (ES+) = 641.61 [M + H]+ (LCMS Method - 2).
Preparation 177: (3/?,45)-l-(5-{Ethyl-[l-(3-isopropyl-[l,2,4]oxadiazol-5-yl)-piperidin-4- yl]-carbamoyl}-pyrazin-2-yl)-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid teri-butyl ester
Figure imgf000075_0001
The title compound was synthesised from 5-[(3R,45')-3-tert-butoxycarbonylamino-4- (2,4,5-trifluoro-phenyl)-pyrrolidin-l -yl]-pyrazine-2-carboxylic acid (Preparation 84) and ethyl-[l -(3-isopropyl-[l ,2,4]oxadiazol-5-yl)-piperidin-4-yl]-amine (Preparation 42) employing a procedure similar to that outlined in Preparation 94: RT = 4.15 min; mlz (ES ) = 659.19 [M + H]+ (LCMS Method - 1).
Preparation 178: 4-({2-[(3/?,45)-3-feri-Butoxycarbonylamino-4-(2,4,5-trifluoro-phi pyrrolidin-l-yl]-pyrimidine-5-carbonyl}-ethyl-amino)-piperidine-l-carboxylic benzyl ester
Figure imgf000075_0002
The title compound was prepared by reacting 2-[(3R,45')-3-iert- butoxycarbonylamino-4-(2,4,54rifluoro^henyl)^yrrolidin-l -yl]-pyrimidine-5-carboxylic acid (Preparation 82) with 4-ethylamino-piperidine-l-carboxylic acid benzyl ester employing a procedure similar to that outlined in Preparation 104: RT = 4.35 min; mlz (ES+) = 683.13 [M + H]+ (LCMS Method - 1).
Preparation 179: [(3/?,45)-l-[5-(Ethyl-piperidin-4-yl-carbamoyl)-pyrimidin-2-yl]-4- (2,4,5-trifluoro-phenyl)-p utyl ester
Figure imgf000075_0003
To a solution of 10% Pd/C (24mg, 0.022mmol) in EtOH (15mL) was added 4-({2- [(3R,4S)-3 -tert-butoxycarbonylamino-4-(2,4,5 -trifluoro-phenyl)-pyrrolidin- 1 -yl]-pyrimidine- 5-carbonyl} -ethyl-amino)-piperidine-l -carboxylic acid benzyl ester (Preparation 178, 240mg, 0.35mmol) and the mixture placed under hydrogen for 1 h at room temperature. The mixture was then filtered through celite and the filtrate concentrated in vacuo, azeotroping with DCM, to afford the title compound: RT = 2.75 min; mlz (ES+) = 549.16 [M + H]+ (LCMS Method - 1).
Preparation 180: [(3/?,45)-l-(5-{[l-(5-Chloro-pyrimidin-2-yl)-piperidin-4-yl]-ethyl- carbamoyl}-pyrimidin-2-yl)-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000076_0001
To a solution of [(3R,45,)-l-[5-(ethyl-piperidin-4-yl-carbamoyl)-pyrimidin-2-yl]-4- (2,4,5-trifluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 179, 40mg, 0.07mmol) in DMSO (lmL) was added DIPEA (19μΕ, O. l lmmol) and 2,5- dichloropyrimidine (16mg, O. l lmmol). The mixture was heated in the microwave at 150 °C for 1 h. The mixture was purified by preparative HPLC and the compound containing fractions concentrated in vacuo to afford the title compound: RT = 4.55 min; mlz (ES ) = 661.13 [M+ H]+ (LCMS Method - 1).
Preparation 181 : 4-({2-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2,5-difluoro-phenyl)- pyrrolidin-l-yl]-pyrimidine-5-carbonyl}-ethyl-amino)-piperidine-l-carboxylic acid benzyl ester
Figure imgf000076_0002
The title compound was prepared by reacting 2-[(3R,4S)-3-tert- butoxycarbonylamino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid (Preparation 80) with 4-ethylamino-piperidine-l -carboxylic acid benzyl ester employing a procedure similar to that outlined in Preparation 95: RT = 1.33 min; mlz (ES ) = 665.51 [M + H]+ (LCMS Method - 2).
Preparation 182: {(3/?,45)-4-(2,5-Difluoro-phenyl)-l-[5-(ethyl-piperidin-4-yl- carbamoyl)-pyrimidin-2-yl]- rrolidin-3-yl}-carbamic acid tert-butyl ester
Figure imgf000077_0001
The title compound was prepared from 4-({2-[(3R,45')-3-tert-butoxycarbonylamino- 4-(2,5-difluoro-phenyl)-pyrrolidin- 1 -yl]-pyrimidine-5-carbonyl} -ethyl-amino)-piperidine- 1 - carboxylic acid benzyl ester (Preparation 181) employing a procedure similar to that outlined in Preparation 179: RT = 0.81 min; mlz (ES+) = 531.51 [M + H]+ (LCMS Method - 2).
Preparation 183: 4-({2-[(3/?,45)-3-tert-Butoxycarbonylamino-4-(2,5-difluoro-phenyl)- pyrrolidin-l-yl]-pyrimidine-5-carbonyl}-ethyl-amino)-piperidine-l-carboxylic acid 1- methyl-cyclopropyl ester
Figure imgf000077_0002
To a stirred solution of {(3R,45)-4-(2,5-difluoro-phenyl)-l -[5-(ethyl-piperidin-4-yl- carbamoyl)-pyrimidin-2-yl]-pyrrolidin-3-yl} -carbamic acid tert-butyl ester (Preparation 182, 200mg, 0.3mmol) and triethylamine (72.5μΙ., 0.520mmol) in DCM (7mL) was added 1- methylcyclopropyl 4-nitrophenyl carbonate (82.3mg, 0.347mmol) followed by 4- dimethylaminopyridine (4.24mg, 0.0347mmol) and the solution stirred at r.t. for 6 h. The reaction mixture was diluted with EtOAc, washed with saturated aqueous sodium carbonate solution, 1M HC1 solution, water and finally brine. The organics were dried (magnesium sulfate), filtered and concentrated in vacuo. Purification by column chromatography (97:3 DCM:MeOH) afforded the title compound: RT = 1.25 min; mlz (ES+) = 629.51 [M + H]+ (LCMS Method - 2).
Preparation 184 : [(3R,4S)-1 -{5- [(1 -Cyano-piperidin-4-yl)-ethyl-carbamoyl] -pyrimidin- 2-yl}-4-(2,5-difluoro-phen l)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000078_0001
The title compound was prepared from {(3R,45)-4-(2,5-difluoro-phenyl)-l -[5-(ethyl- piperidin-4-yl-carbamoyl)-pyrimidin-2-yl]-pyrrolidin-3-yl}-carbamic acid tert-butyl ester (Preparation 182) employing a procedure similar to that outline in Preparation 16: RT = 1.12 min; mlz (ES+) = 556.49 [M + H]+ (LCMS Method - 2). Preparation 185: [(3/?,45)-4-(2,5-Difluoro-phenyl)-l-(5-{ethyl-[l-(2H-tetrazol-5-yl)- piperidin-4-yl]-carbamoyl}-pyrimidin-2-yl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000078_0002
To a solution of [(3R,45)-l- {5-[(l -cyano-piperidin-4-yl)-ethyl-carbamoyl]- pyrimidin-2-yl}-4-(2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 184, 0.180g, 0.324mmol) in DMF (2.5mL) was added ammonium chloride (26mg, 0.486mmol) and sodium azide (31.6mg, 0.486mmol). The mixture was heated at 100 °C for 18 h. The reaction mixture was diluted with water and extracted with EtOAc, then EtOAc:MeOH (95:5). The combined organic extracts were washed with brine, filtered through a hydrophobic frit and concentrated in vacuo. Purification by column chromatography (95 :5 DCM:MeOH) afforded the title compound: RT = 1.01 min; mlz (ES ) = 599.46 [M + H]+ (LCMS Method - 2).
Preparation 186: [(3R,45)-4-(2,5-Difluoro-phenyl)-l-(5-{ethyl-[l-(2-ethyl-2H-tetrazol-5- yl)-piperidin-4-yl]-carbamoyl}-pyrimidin-2-yl)-pyrrolidin-3-yl]-carbamic acid tert- butyl ester
Figure imgf000079_0001
To a solution of [(3R,45")-4-(2,5-difluoro-phenyl)-l-(5- {ethyl-[l -(2H-tetrazol-5-yl)- piperidin-4-yl]-carbamoyl} -pyrimidin-2-yl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 185, 60mg, O. lOmmol) in acetone (3mL) was added potassium carbonate (42mg, 0.30mmol) followed by the dropwise addition of iodoethane (ΙΟμΙ,, 0.13mmol). The mixture was then heated at 45 °C for 2.5 h then concentrated in vacuo. The residue was partitioned between EtOAc and water, the organic phase washed with brine, dried (magnesium sulfate), filtered and concentrated in vacuo. Purification by preparative HPLC (Standard Method) afforded the title compound: RT = 1.20 min; mlz (ES+) = 627.54 [M + H]+ (LCMS Method - 2).
Preparation 187: l-(2,5-Difluoro-phe hanol
Figure imgf000079_0002
Nitromethane (227mL, 4.22mol) was added in one portion to a solution of
2,5-difluorobenzaldehyde (500g, 3.52mol) in MeOH (5L) in a 10 L jacketed vessel The solution was cooled to 4 °C (jacket -10 °C ) and a solution of NaOH (169g, 4.22mol) in water (500mL) was added over 40 min causing a 6 °C exotherm. The solution was stirred for 60 min (jacket 0 °C) after which time a thick white precipitate formed. Ice water (3L) was added (jacket 5 °C) followed by NH4CI (sat. aq., 3.5 L) and the solution extracted with DCM (5 x 2 L). The combined organics were evaporated then taken up into DCM (1L), dried over MgS04 and evaporated to dryness to afford the title compound. ¾ NMR 8H (300MHZ, CDCI3): 7.35 (m, 1H), 7.05 (m, 2H), 5.70 (m, 1H), 4.60 (m, 2H), 3.10 (m, 1H).
Preparation 188: l,4-Difluoro-2-((£)- -nitro-vinyl)-benzene
Figure imgf000080_0001
Acetic anhydride (665mL, 7.05mol) was added in one portion to l-(2,5-difluoro- phenyl)-2-nitro-ethanol (Preparation 187, 672g, 3.31mol) at 0 °C under argon. DMAP (28.3g, 0.23mol) was added and the solution darkened in colour. The reaction was warmed to r.t. over 18 h. The reaction mixture was cautiously poured into NaHC(¾ (sat. aq., 3.5L) and stirred to form a yellow solid. The slurry was stirred for 30 min at r.t. before filtering. The solid was washed with NaHC(¾ (sat. aq., 2L) then water (2L). The solid was dissolved in DCM (1.5L) and washed with saturated NaHCCh (sat. aq., 4 x 1L), dried over MgS04 and evaporated to afford a yellow / orange oil which solidified to give a yellow solid. This was purified by dry flash chromatography eluting with DCM to afford the title compound. ¾ NMR δΗ (300MHz, CDC13): 8.05 (d, 1H), 7.70 (d, 1H), 7.20 (m, 3H).
Preparation 189: l-Benzyl-3-(2,5-difluoro- henyl)-4-nitro-pyrrolidine
Figure imgf000080_0002
l,4-Difluoro-2-((£)-2-nitro-vinyl)-benzene (Preparation 188, 120g, 0.65mol) was dissolved in analytical grade DCM (1.2L) and dried over MgS04. The solution was filtered and split into two portions (60g, 0.32mol). N-(Methoxymethyl)-N- (trimethylsilylmethyl)benzylamine (lOOmL, 0.39mol) was added to each portion under argon. The yellow solutions were then cooled to -5 °C and TFA (2.7mL, 0.04mol) added. After 5 min the temperature rapidly increased to 30 °C then cooled back to -5 °C. The resulting paler yellow solution was stirred at -10 °C for 1 h before quenching cautiously with aHC03 (sat. aq., 1 L). The two batches were combined and the layers partitioned. The DCM layer was washed with water (2L) then brine (2L), dried over MgS04 and evaporated to give 209g of an orange oil. The material was purified by dry flash chromatography eluting with DCM to afford the title compound. H NMR δΗ (300MHz, CDC13): 7.40 - 7.20 (m, 5H), 7.00 (m, 3H), 4.95 (m, 1H), 4.20 (m, 1H), 3.70 (m, 2H), 3.40 (m, 1H), 3.20 (m, 2H), 2.70 (m, 1H).
Preparation 190: l-Benzyl-4-(2,5-difluoro- henyl)-pyrrolidin-3-ylamine
Figure imgf000081_0001
(+/-)
l -Benzyl-3-(2,5-difluoro-phenyl)-4-nitro-pyrrolidine (Preparation 189, 340g, 1.07mol) was dissolved in IMS (5L) and charged into a 7.8L autoclave. RaNi (20g) was added (without washing). The autoclave was sealed and charged with ¾ (50 bar) and stirred at r.t. for 18 h (10 °C exotherm). The reaction mixture was filtered through celite, the pad washed with IMS and the filtrate evaporated to dryness. The brown oil was dissolved in DCM (1.5L), extracted with HC1 (1M, aq. 2.5L) and washed with DCM (3 x 1L) to remove the colour from the aqueous layer. The aqueous layer was basified to pH 14 with NaOH (2M, aq. 3L) and extracted into DCM (5 x 1L) then the organics were combined, dried over MgSC-4 and evaporated to afford the title compound. 1H NMR δΗ (300MHz, CDC13): 7.40 - 7.20 (m, 5H), 7.10 (m, 1H), 6.95 (m, 1H), 6.85 (m, 1H), 3.70 (m, 2H), 3.50 (m, 1H), 3.25 (m, 1H), 3.05 (m, 2H), 2.65 (m, 1H), 2.45 (m, 1H), 1.50 (br s, 2H).
Preparation 191: l-Benzyl-4-(2,5-difluoro-phenyl)-pyrrolidin-3-yl-carbamic acid tert- butyl ester
Figure imgf000081_0002
Triethylamine (244mL, 1.75mol) was added to a solution of l-benzyl-4-(2,5- difluoro-phenyl)-pyrrolidin-3-ylamine (Preparation 190, 252g, 0.87mol) in THF (3.8L) and the cloudy light brown solution was cooled to 0 °C. Di-tert-butyldicarbonate (229g, 1.05mol) was added in one portion and the suspension warmed to room temperature over 18 h. The THF was removed by evaporation and the off-white slurry taken up into EtOAc (2.5L). The solution was washed with water (3 x 1L), brine (1L), dried over MgS04 then evaporated to give a pale yellow oil. Heptane (0.5L) was added and evaporated to give an off white solid, which was triturated with heptane. This was filtered and the solid washed with heptane then dried at 40 °C for 18 h to afford the title compound. *Η NMR δΗ (300MHz, CDC13): 7.40 - 7.20 (m, 5H), 7.05 (m, 1H), 6.95 (m, 1H), 6.85 (m, 1H), 4.95 (m, 1H), 4.20 (m, 1H), 3.60 (s, 2H), 3.30 (m, 1H), 3.10 (m, 1H), 2.95 (m, 1H), 2.65 (m, 1H), 2.45 (m, 1H), 1.40 (s, 9H).
Preparation 192: [(3/?,4S)-l-Benzyl-4-(2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000082_0001
The title compound was afforded via chiral HPLC separation of l -benzyl-4-(2,5- difluoro-phenyl)-pyrrolidin-3-yl-carbamic acid tert-butyl ester (Preparation 191): IH:IPA:DEA 96:4:0.1, 15ml/min, 270nm, RT = 10.9 min. Preparation 193: [(3/?,4S)-4-(2,5-Difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert- butyl ester
Figure imgf000082_0002
10% Pd/C (20g) was added as a paste in toluene to a solution of [(3R,45)-l -benzyl-4- (2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 192, 212g, 0.55mol) in IMS (6L) in a 7.8L autoclave. The autoclave was sealed, charged with ¾ (50 bar) and heated to 60 °C for 6 h then r.t. for 72 h. The Pd/C was removed by filtration through celite, the pad washed with IMS and the filtrate evaporated to dryness. The reaction was repeated (200g, 0.52mol) and the two batches combined to give 288g of an off-white solid. This was triturated with heptane, filtered, washed with heptane and dried at 40 °C for 18 h to afford the title compound. Η NMR δΗ (300MHz, CDC13): 7.00 (m, 2H), 6.85 (m, 1H) 4.75 (br s, 1H), 4.15 (m, 1H), 3.45 (m, 2H), 3.25 (m, 1H), 2.90 (m, 2H), 1.90 (s, 1H), 1.35 (s, 9H). Preparation 194: 2-Nitro-l-(2,4,5-tr hanol
Figure imgf000083_0001
Nitromethane (274mL, 5.10mol) was added in one portion to a solution of 2,4,5- trifluorobenzaldehyde (680g, 4.25mol) in MeOH (7.2L) in a 10 L jacketed vessel. The solution was cooled to 0 °C (jacket -10 °C) and a solution of NaOH (204g, 5.10mol) in water (680mL) was added over 30 min. A 5 °C exotherm was observed. The solution was stirred for 30 min (jacket 0 °C) after which time a white precipitate had formed. Half of the material was transferred to a second vessel and both slurries stirred for a further 30 min becoming very thick. Ice water (2L) and NH4CI (sat. aq., 2.5L) was added to each to afford a solution. The solutions were extracted with DCM (5 x 1.5L) and the organics from both batches combined and evaporated then dissolved in DCM (1L), dried over MgS04 and evaporated to dryness to afford the title compound. 'H NMR δΗ (300MHz, CDC13): 7.45 (m, 1H), 6.95 (m, 1H), 5.70 (m, 1H), 4.55 (m, 2H), 3.10 (d, 1H).
Preparation 195: l,2,4-Trifluoro-5- -2-nitro-vinyl)-benzene
Figure imgf000083_0002
Acetic anhydride (849mL, 8.99mol) was added to 2-nitro-l -(2,4,5-trifluoro-phenyl)- ethanol (Preparation 194, 935g, 4.22mol) at 0 °C under argon. DMAP (36g, 0.30mol) was added in one portion and the solution darkened in colour. The temperature reached 50 °C over 20 min before cooling back to 0 °C. The solution was allowed to warm to r.t. over 18 h. The reaction mixture was cautiously poured into aHC03 (sat. aq., 6L) and stirred to form a yellow solid. The slurry was stirred for 30 min at r.t. before filtering and washing with NaHCCh (sat. aq., 3L) then water (3L). The solid was dissolved in DCM (2L) and washed with saturated NaHCCh (sat. aq., 2 x 1.5L) then the organics dried over MgS04 and evaporated to give 795g of an orange / brown oil which solidified to give an orange solid. Purification by dry flash chromatography eluting with DCM afforded the title compound. NMR δΗ (300MHz, CDC13): 7.95 (d, 1H), 7.65 (d, 1H), 7.35 (m, 1H), 7.05 (m, 1H).
Preparation 196: l-Benzyl-3-nitro-4-(2,4,5-trifluoro-phenyl)-pyrrolidine
Figure imgf000084_0001
l,2,4-Trifluoro-5-((£)-2-nitro-vinyl)-benzene (Preparation 195, 375g, 1.85mol) was dissolved into analytical grade DCM (3.75L) and dried over MgS04. The solution was filtered and split into five batches (75 g, 0.37mol). N-(Methoxymethyl)-N- (trimethylsilylmethyl)benzylamine (114mL, 0.44mol) was added to each under argon and the orange solutions were then cooled to -10 °C. TFA (3mL, 40.6mmol) was added to each in one portion and after 5 min the temperature rapidly increased to 30 °C then cooled back to - 10 °C. The resulting paler yellow / orange solutions were stirred at -10 °C for 1 h before quenching cautiously with aHC03 (sat. aq., IL). The layers were partitioned and the DCM layer washed with water (IL), brine (IL), dried over MgS04 and evaporated. The crude batches were combined and purified by dry flash chromatography eluting with DCM to afford the title compound. 1H NMR δΗ (300MHz, CDC13): 7.40 - 7.20 (m, 6H), 6.90 (m, 1H), 4.95 (m, 1H), 4.20 (m, 1H), 3.70 (m, 2H), 3.35 (m, 1H), 3.20 (m, 2H), 2.70 (m, 1H).
Preparation 197: l-Benzyl-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-3-ylamine
Figure imgf000084_0002
l -Benzyl-3-nitro-4-(2,4,5-trifluoro-phenyl)-pyrrolidine (Preparation 196, 62 l g, 923mol) was run as two batches, each dissolved into IMS (5L) and charged into a 7.8L autoclave. RaNi (20g) was added (without washing) and the autoclave charged with H2 (50 bar) and stirred at r.t. for 18 h (10 °C exotherm). The mixtures were filtered through celite and both batches combined and evaporated to dryness. The resulting brown oil was dissolved in DCM (1.5L) and extracted with HC1 (1M, aq. 2.5L) and washed with DCM (3 x IL) to remove the colour from the aqueous layer. The aqueous layer was basified to pH 14 with NaOH (2M, aq. 3L) and extracted with DCM (5 x 1L), combined, dried over MgS04 and evaporated to afford the title compound. *Η NMR δΗ (300MHz, CDC13): 7.40 - 7.20 (m, 6H), 6.85 (m, 1H), 3.70 (m, 2H), 3.45 (m, 1H), 3.20 (m, 1H), 3.10 (m, 2H), 2.65 (m, 1H), 2.45 (m, 1H), 2.20 (br s, 2H).
Preparation 198: l-Benzyl-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-3-yl-carbamic acid tert- butyl ester
Figure imgf000085_0001
Triethylamine (210mL, 1.51mol) was added to l-benzyl-4-(2,4,5-trifluoro-phenyl)- pyrrolidin-3-ylamine (Preparation 197, 23 lg, 0.75mol) in THF (1.8L) and the light brown cloudy solution was cooled to 0 °C. Di-tert-butyldicarbonate (198g, 0.91mol) was added in one portion and the suspension warmed to room temperature over 18 h. The suspension was concentrated and the off-white slurry dissolved into EtOAc (2.5L). The solution was washed with water (3 x 1L) and brine (1L) then dried over MgS04 and evaporated to give a pale yellow oil. Heptane (0.5L) was added and evaporated to give an off white solid. This was combined with a second batch (from 218g, 0.71mol starting material) and triturated with heptane, filtered, washed with heptane and dried at 40 °C for 18 h to afford the title compound. ¾ NMR δΗ (300MHz, CDC13): 7.40 - 7.20 (m, 6H), 6.85 (m, 1H), 4.85 (m, 1H), 4.20 (m, 1H), 3.60 (s, 2H), 3.30 (m, 1H), 3.00 (m, 2H), 2.60 (m, 1H), 2.45 (m, 1H), 1.40 (s, 9H).
Preparation 199: 4-(2,4,5-Trifluoro-phenyl)-pyrrolidin-3-yl-carbamic acid tert-butyl ester
Figure imgf000085_0002
l -Benzyl-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-3-yl-carbamic acid tert-butyl ester (Preparation 198, 612g, 1.51mol) was run in two batches each being suspended in EtOAc (0.5L) and IMS (4L) and charged into a 7.8L autoclave. 10% Pd/C (20g) was added to each as a paste in toluene. The autoclave was sealed and charged with ¾ (50 bar) then heated to 55 °C for 6 h, 20 °C for 18 h then 55 °C for 6 h. The reaction mixtures were filtered through celite and the pad washed with IMS before evaporating to dryness. The resulting off-white solids were triturated with heptane, filtered, washed with heptane then dried at 40 °C for 18 h before the two batches were combined to afford the title compound. H NMR 8H (300MHz, CDC13): 7.10 (m, lH), 6.90 (m, 1H) 4.70 (br s, 1H), 4.15 (m, 1H), 3.45 (m, 2H), 3.20 (m, lH), 2.85 (m, 2H), 1.75 (br s, 1H), 1.40 (s, 9H).
Preparation 199: [(3/?,45)-4-(2,4,5-Trifluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000086_0001
4-(2,4,5-Trifluorophenyl)pyrrolidin-3-yl]carbamic acid tert-butyl ester (Preparation
199, 30.0g, 95mmol) was suspended in EtOH (lOOmL) and heated to 70°C. To the suspension was added a warm solution of (5*)-(+)-naproxen (10.9g, 47mmol) in EtOH (lOOmL) and the mixture was heated to reflux. The heat was removed and the mixture was slowly allowed to cool to r.t, with stirring, for 16 h. The resulting precipitate was filtered, washing with EtOH, and the solid product was partitioned between DCM (2400mL) and IM NaOH (600mL). The organic phase was separated, washed with IM NaOH, brine, then dried (MgS04), and the solvent was removed in vacuo.
A second portion of [(ira«s)-4-(2,4,5-trifluorophenyl)pyrrolidin-3-yl]carbamic acid tert-butyl ester (Preparation 199, 29.5g, 93mmol), suspended in EtOH (lOOmL), was heated to 70°C. Treatment with a warm solution of («S)-(+)-naproxen (10.6g, 46mmol) in EtOH (lOOmL), employing the procedure outlined above, afforded a second batch of the enantiomerically enriched product.
The two products were combined, and suspended in EtOH (200mL) before being treated with a hot solution of («S)-(+)-naproxen (16g, 69mmol) in EtOH (150mL). The mixture was heated to reflux, employing the procedure outlined above, and work-up was repeated to afford the title compound: 'H NMR δΗ (400MHz , CD3OD): 7.38 - 7.25 (m, 1H), 7.14 - 7.01 (m, 1H), 4.20 - 4.09 (m,l H), 3.30 - 3.21 (m, 3H), 2.90 - 2.81 (m, 1H), 2.77 - 2.68 (m, 1H), 1.34 (br. s., 9H). Preparation 200: {l-[5-(l,l-Difluoro-ethyl)-[l,2,4]oxadiazol-3-yl]-piperidin-4-yl}- methyl-carbamic acid teri-butyl ester
Figure imgf000087_0001
To a solution of (l-cyano-piperidin-4-yl)-methyl-carbamic acid tert-butyl ester (Preparation 16, l .OOg, 4.18mmol) in EtOH (l OmL) was added 50% hydroxylamine in water (0.26mL, 4.2mmol) and the mixture heated at 70°C for 1 h. The reaction was cooled to r.t. and concentrated under reduced pressure. The resulting solid was dissolved in DMF (6.47mL, 83.6mmol) and HOBt (0.0640g, 0.418mmol), EDCI (0.961 g, 5.01mmol) and 2,2- difluoropropionic acid (0.552g, 5.01mmol) added. The reaction mixture was stirred at r.t. over 60 h, after which time it was diluted with EtOAc (150mL) and washed with a saturated solution of sodium bicarbonate (2 x lOOmL), 1 M citric acid solution (lOOmL) then brine (lOOmL). The organics were concentrated under reduced pressure, and the residue dissolved in toluene (5mL) then heated at reflux for 13 h. The reaction was cooled to r.t. and concentrated under reduced pressure. Purification by column chromatography (7:3 EtOAcTH) afforded the title compound: RT = 1.33 min; m/z (ES+) = 347.4 [M + H]+ (LCMS Method - 2).
Preparation 201: {l-[5-(l,l-Difluoro-ethyl)-[l,2,4]oxadiazol-3-yl]-piperidin-4-yl}- methyl-amine
Figure imgf000087_0002
{ 1 -[5-(l, 1 -Difluoro-ethyl)-[ 1 ,2,4]oxadiazol-3 -yl]-piperidin-4-yl} -methyl-carbamic acid tert-butyl ester (Preparation 200, l . lg, 3.2mmol) was dissolved in DCM (lOmL, 200mmol) and TFA (2mL, 20mmol) added. The reaction was stirred at r.t. for 1 h before being loaded directly onto an SCX cartridge. Elution with MeOH followed by 7N ammonia in MeOH solution and concentration in vacuo afforded the title compound: RT = 0.52 min, m/z (ES+) = 247.3 [M + H]+ (LCMS Method - 2). Preparation 202: [(3i?,4S)-l-[5-({l-[5-(l,l-Difluoro-ethyl)-[l,2,4]oxadiazol-3-yl]- piperidin-4-yl}-methyl-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro-phenyl)-pyrrolidin- 3-yl]-carbamic acid tert- butyl ester
Figure imgf000088_0001
The title compound was synthesized from 2-[(3R,4S)-3-tert-butoxycarbonylamino-4- (2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid (Preparation 80, 430mg, l .Ommol) and {l -[5-(l ,l -difluoro-ethyl)-[l,2,4]oxadiazol-3-yl]-piperidin-4-yl}- methyl-amine (Preparation 201, 250mg, 1.015mmol) employing a procedure similar to that outlined in Preparation 95: RT = 1.31 min; mlz (ES+) = 649.6 [M+ H]+.
Preparation 203: Cyclopropyl-{l-[5-(l-methoxy-l-methyl-ethyl)-[l,2,4]oxadiazol-3-yl]- piperidin-4-yl}-carbamic acid tert-but l ester
Figure imgf000088_0002
The title compound was synthesized from (l-cyano-piperidin-4-yl)-cyclopropyl- carbamic acid tert-butyl ester (Preparation 20, 621mg, 2.34mmol) and 2-methoxy-2- methyl-propionic acid (351mL, 2.82mmol) employing a procedure similar to that outlined in Preparation 200: RT = 1.34 min; mlz (ES+) = 381.5 [M+ H]+.
Preparation 204: Cyclopropyl-{l-[5-(l-methoxy-l-methyl-ethyl)-[l,2,4]oxadiazol-3-yl]- piperidin-4-yl}-amine
Figure imgf000088_0003
The title compound was synthesized from cyclopropyl- { l -[5-(l-methoxy-l -methyl- ethyl)-[l,2,4]oxadiazol-3-yl]-piperidin-4-yl} -carbamic acid tert-butyl ester (Preparation 203, 590mg, 1.55mmol) employing a procedure similar to that outlined in Preparation RT = 0.51 min; mlz (ES+) = 281.4 [M + H]+.
Preparation 205: [(S^^-l-IS-CCyclopropyl-il-IS-Cl-methoxy-l-methyl-ethyl)- [l,2,4]oxadiazol-3-yl]-piperidin-4-yl}-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro- phenyl)-pyrrolidin-3-yl -carbamic acid tert-but l ester
Figure imgf000089_0001
The title compound was synthesized from 2-[(3R,45')-3-iert-butoxycarbonylamino-4- (2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid (Preparation 80, 277mg, 0.66mmol) and cyclopropyl- { l -[5-(l -methoxy-l -methyl-ethyl)-[l,2,4]oxadiazol-3- yl]-piperidin-4-yl} -amine (Preparation 204, 185mg, 0.66mmol) employing a procedure similar to that outlined in Preparation 95: RT = 1.33 min; mlz (ES ) = 683.7 [M+ H]+.
Preparation 206: [(3/?,4S)-l-{5-[(l-Carbamoyl-piperidin-4-yl)-cyclopropyl-carbamoyl]- pyrimidin-2-yl}-4-(2,5-difluoro- henyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000089_0002
To a stirred solution of (l-cyano-piperidin-4-yl)-cyclopropyl-carbamic acid tert-butyl ester (Preparation 20, 1.09g, 4.12mmol) in DCM (20mL) at 0°C was added TFA (4mL, 50mmol). The solution was allowed to warm to r.t. and stirred for 4 h. The reaction mixture was passed through a SCX cartridge and the free base eluted with a 3.5M solution of ammonia in MeOH. The mixture was concentrated in vacuo and the residue dissolved in DCM (60mL). To this was added 2-[(3R,45')-3-tert-butoxycarbonylamino-4-(2,5-difluoro- phenyl)-pyrrolidin-l -yl]-pyrimidine-5-carboxylic acid (Preparation 80, 1.73g, 4.12mmol), triethylamine (2.29mL, 16.5mmol) and PPA (50% w/w in EtOAc, 4.90mL, 8.23mmol) and the reaction stirred at r.t. for 18 h. The mixture was quenched with MeOH and concentrated under reduced pressure. The residue was dissolved in MeOH and loaded directly onto a SCX cartridge and eluted with MeOH, then 7N ammonia in MeOH solution and concentrated in vacuo. Purification of the residue by column chromatography (7:3 EtOAcTH) afforded the title compound: RT = 0.99 min, m/z (ES+) = 586.6 [M + H]+ (LCMS Method - 2).
Preparation 207: [(3/?,4S)-l-{5-[(l-Cyano-piperidin-4-yl)-cyclopropyl-carbamoyl]- pyrimidin-2-yl}-4-(2,5-difluoro- henyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000090_0001
[(3R,45,)-l- {5-[(l-Carbamoyl-piperidin-4-yl)-cyclopropyl-carbamoyl]-pyrimidin-2- yl} -4-(2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 206, 0.700g, 1.20mmol) and MeCN (20mL) were combined under an argon atmosphere. Methanesulfonyl chloride (0.139mL, 1.79mmol) followed by pyridine (0.242mL, 2.99mmol) were added via syringe. The resulting suspension was stirred for 5 min prior to being heated to 50 °C for 18 h. The solvent was removed under reduced pressure and water (50mL) added to the resulting residue. The mixture was extracted with EtOAc (3 x 150mL), the organics combined and dried (magnesium sulfate) before concentration under reduced pressure afford the title compound: RT = 1.15 min, m/z (ES+) = 568.8 [M+ H]+ (LCMS Method -2). Preparation 208: [(3/?,4S)-l-(5-{Cyclopropyl-[l-(iV-hydroxycarbamimidoyl)-piperidin- 4-yl]-carbamoyl}-pyrimidin-2-yl)-4-(2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000090_0002
[(3R,45,)-l- {5-[(l-Cyano-piperidin-4-yl)-cyclopropyl-carbamoyl]-pyrimidin-2-yl} -4- (2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 207, 500mg, 0.881mmol) was dissolved in EtOH (lOmL) and 50% hydroxylamine in water (0.1 lmL, 1.8mmol) added via syringe. The reaction mixture was stirred overnight at r.t. The mixture was then concentrated under reduced pressure and further azeotroped with toluene to afford the title compound: RT = 0.84 min, m/z (ES ) = 601.6 [M+ H]+. Preparation 209: [(S^^-l-IS-CC clo ro l-il-IS-Cl^-difluoro-eth ^-Il^^loxadiazol- 3-yl]-piperidin-4-yl}-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro-phenyl)-pyrrolidin-3- yl]-carbamic acid tert-but l ester
Figure imgf000091_0001
[(3R,4S)-l -(5- {Cyclopropyl-[ 1 -(N-hydroxycarbamimidoyl)-piperidin-4-yl]- carbamoyl}-pyrimidin-2-yl)-4-(2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert- butyl ester (Preparation 208, 230mg, 0.383mmol), 2,2-difluoropropionic acid (50.6mg, 0.459mmol), EDCI (88.1mg, 0.459mmol) and triethylamine (0.117mL, 0.842mmol) were combined in DCM (20mL) and HOBt (0.8mg, 0.005mmol) added. The reaction was stirred at r.t. for 18 h before quenching with MeOH and concentrating under reduced pressure, azeotroping with toluene. Purification of the residue by preparative HPLC afforded the title compound: RT = 4.32 min, m/z (ES+) = 675.9 [M+ H]+ (LCMS Method -1).
Preparation 210: [(S.R^-l-IS-CC cloprop l-il-IW-S-Ctetrah dro-furan-S- l)- [l,2,4]oxadiazol-3-yl]-piperidin-4-yl}-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro- phenyl)-pyrrolidin-3-yl -carbamic acid tert-butyl ester
Figure imgf000091_0002
The title compound was synthesized from [(SR^^-l-iS- jcyclopropyl-f l-iN- hydroxycarbamimidoyl)-piperidin-4-yl]-carbamoyl}-pyrimidin-2-yl)-4-(2,5-difluoro- phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 208) and (R)- tetrahydro-furan-3-carboxylic acid employing a procedure similar to that outlined in Preparation 209: RT = 1.22 min, m/z (ES+) = 681.6 [M+ H]+ (LCMS Method - 2). Preparation 211: (i?)-Tetrahydro-fura -3-carboxylic acid amide
Figure imgf000092_0001
To a solution of (R)-tetrahydro-furan-3-carboxylic acid (500mg, 4.31mmol) in DCM (5mL) at 0 °C under inert atmosphere was added oxalyl chloride (444μί, 5.17mmol) followed by DMF (50μί, 0.65mmol). The reaction mixture was stirred for 1 h then allowed to warm to room temperature and stirred for further 2 h. After cooling to 0 °C, H3 in MeOH solution (7N, 50mL) was added and the mixture stirred overnight. The solvents were then removed in vacuo, azeotroping with DCM to dryness to afford the title compound: ¾ NMR δΗ (300MHz, CDC13): 5.80-5.30 (bs, 2H), 3.90 (m, 4H), 2.90 (m, 1H), 2.20 (m, 2H).
Preparation 212: (5)-Tetrahydro-furan- -carbonitrile
Figure imgf000092_0002
To a solution of (R)-tetrahydro-furan-3-carboxylic acid amide (Preparation 211, 435mg, 3.78mmol) in THF (lOmL) was added pyridine (2.31mL, 28.61mmol) and the reaction cooled to 0 °C. Trifluoroacetic anhydride (2.70mL, 28.61mmol) was added and the mixture stirred for 1 h. The mixture was partitioned between saturated aqueous sodium bicarbonate solution and DCM; the aqueous layer was back-extracted with further DCM. The combined organics were washed with brine and dried with magnesium sulfate before concentrating in vacuo. Purification of the residue by column chromatography (EtOAc) afforded the title compound: 'H NMR δΗ (300MHz, CDC13): 4.05 (t, 1H), 3.90 (m, 3H), 3.05 (m, 1H), 2.3 (m, 2H).
Preparation 213: (5)-7V-Hydroxy-tetrahydro-furan-3-carboxamidine
Figure imgf000092_0003
To a solution of (S)-tetrahydrofuran-3-carbonitrile (Preparation 212, 280mg, 2.89mmol) in IMS (2mL) was added hydro xylamine (50% in water, 229μΙ., 3.46mmol) and the reaction was heated to 70 °C for 18 h. On cooling, the solvent was removed in vacuo, azeotroping with toluene. The residue was triturated with toluene then DCM. Purification by column chromatography (DCM:MeOH, 90: 10) afforded the title compound: !H NMR δΗ (300MHz, CDC13): 4.70 (bs, 2H), 4.05 (m, 2H), 3.70 (m, 2H), 2.9 (m, 1H), 2.25 (m, 1H), 2.00 (m, 1H). Preparation 214: Cyclopropyl-{l-[(5)-3-(tetrahydro-furan-3-yl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl}-amine
Figure imgf000093_0001
The title compound was synthesized from (l-cyano-piperidin-4-yl)-cyclopropyl-carbamic acid tert-butyl ester (Preparation 20) and (5)-N-hydroxy-tetrahydro-furan-3-carboxamidine (Preparation 213) employing a procedure similar to that outlined in Preparation 27: RT = 0.46 min; mlz (ES+) = 279.37 [M + H]+ (LCMS Method - 2).
Preparation 215: [(S.R^-l-IS-CC cloprop l-il-IC^-S-Ctetrah dro-furan-S- l)- [l,2,4]oxadiazol-5-yl]-piperidin-4-yl}-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro- phenyl)-pyrrolidin-3-y -carbamic acid teri-butyl ester
Figure imgf000093_0002
The title compound was synthesized from 2-[(3R,45')-3-iert-butoxycarbonylamino-4- (2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid (Preparation 80) and cyclopropyl- { 1 -[(S)-3-(tetrahydro-furan-3 -yl)-[ l,2,4]oxadiazol-5-yl]-piperidin-4-yl} -amine (Preparation 214) employing a procedure similar to that outlined in Preparation 95: RT = 1.17 min; mlz (ES+) = 681.56 [M + H]+.
Preparation 216: (l-Cyclopropanesulfon l-piperidin-4-yl)-cyclopropyl-amine
Figure imgf000093_0003
Cyclopropyl-piperidin-4-yl-carbamic acid tert-bu yl ester (Preparation 14, 0.200g, 0.832mmol) was dissolved in DCM (lO.OmL, 156mmol) and DIPEA (0.362mL, 2.08mmol) added followed by cyclopropylsulfonylchloride (0.102mL, 0.998mmol). The reaction mixture was stirred at r.t. for 18 h. The mixture was diluted with DCM and washed with water followed by brine, filtered through a hydrophobic frit and then concentrated in vacuo. The residue was passed through a SCX cartridge and eluted with MeOH before concentrating in vacuo. The residue was dissolved in DCM (lOmL) and cooled to 0 °C before the dropwise addition of TFA (l .OmL, 13mmol). The reaction mixture was stirred at room temperature for 1 h. The mixture was then passed through a SCX cartridge and eluted with 10% ammonia in MeOH and concentrated in vacuo. Purification by column chromatography (8:92 MeOH:DCM) afforded the title compound: ¾ NMR δΗ (301MHz , CDC13): 3.44 - 3.37 (m, 2H), 2.67 - 2.56 (m, 2H), 2.38 - 2.27 (m, 1H), 1.99 -1.89 (m, 1H), 1.84 - 1.78 (m, 1H), 1.73 - 1.64 (m, 2H), 1.29 - 1.10(m, 4H), 0.89 - 0.81 (m, 2H), 0.68 - 0.61 (m, 2H), 0.19 - 0.12 (m, 2H), 0.09 - 0.00 (m, 2H). Preparation 217: [(3/?,45)-l-{5-[(l-Cyclopropanesulfonyl-piperidin-4-yl)-cyclopropyl- carbamoyl]-pyrimidin-2-yl}-4-(2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert- butyl ester
Figure imgf000094_0001
The title compound was synthesized from 2-[(3R,45')-3-tert-butoxycarbonylamino-4- (2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid (Preparation 80) and (l-cyclopropanesulfonyl-piperidin-4-yl)-cyclopropyl-amine (Preparation 216) employing a procedure similar to that outlined in Preparation 95: RT = 1.21 min; m/z (ES+) = 647.61 [M + H]+ (LCMS Method - 2). Preparation 218: Cyclopropyl-[l-(2H-tetrazol-5-yl)-piperidin-4-yl]-carbamic acid tert- butyl ester
Figure imgf000094_0002
(l-Cyano-piperidin-4-yl)-cyclopropyl-carbamic acid tert-butyl ester (Preparation 20, 0.514g, 1.94mmol) was dissolved in DMF (l.OmL) and ammonium chloride (0.207 g, 3.87mmol) was added followed by sodium azide (0.252g, 3.87mmol). The reaction mixture was heated at 100 °C for 18 h before the addition of water (15mL), then brine. The aqueous layer was extracted with EtOAc (3 x) and the combined organic extracts were filtered through a hydrophobic frit and concentrated in vacuo. Purification of the residue by column chromatography (6:94 MeOH:DCM) afforded the title compound: RT = 0.9min; m/z (ES+) = 309.40 [M+ H]+ (LCMS Method - 2). Preparation 219: Cyclopropyl-[l-(2-ethyl-2H-tetrazol-5-yl)-piperidin-4-yl]-carbamic acid tert-butyl ester
Figure imgf000095_0001
Cyclopropyl-[l -(2H-tetrazol-5-yl)-piperidin-4-yl]-carbamic acid tert-butyl ester (Preparation 218, 0.166g, 0.538mmol) was dissolved in acetone (16mL) and potassium carbonate (0.22g, 1.6mmol) added followed by the dropwise addition of iodoethane (0.056mL, 0.70mmol). The reaction mixture was then heated at 45 °C for 5 h before concentrating in vacuo. The residue was partitioned between EtOAc (50mL) and water (25mL). The organic phase was separated and washed with brine, filtered through a hydrophobic frit and concentrated in vacuo. Purification by column chromatography (3 :7 EtOAc :IH) afforded the title compound: RT = 1.22 min; 337.44m/z (ES+) = [M + H]+ (LCMS Method - 2).
Preparation 220: Cyclopropyl-[l-(2-eth l-2H-tetrazol-5-yl)-piperidin-4-yl]-amine
Figure imgf000095_0002
Cyclopropyl-[ 1 -(2-ethyl-2H-tetrazol-5-yl)-piperidin-4-yl]-carbamic acid tert-butyl ester
(Preparation 219, 0.135g, 0.401mmol) was dissolved in DCM (8.0mL) and cooled to 0 °C. TFA (l.OmL, 13mmol) was added drop wise. Once addition was complete, the reaction was stirred at room temperature for 2 h. The reaction mixture was passed through a SCX cartridge and eluted with 10% ammonia in MeOH solution to obtain the title compound: RT = 0.49min; m/z (ES+) = 237.35 [M+ H]+ (LCMS Method - 2). Preparation 221: [(3/?,45)-l-(5-{Cyclopropyl-[l-(2-ethyl-2H-tetrazol-5-yl)-piperidin-4- yl]-carbamoyl}-pyrimidin-2-yl)-4-(2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid teri-butyl ester
Figure imgf000096_0001
The compound was synthesized from 2-[(3R,45')-3-tert-butoxycarbonylamino-4-(2,5- difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid (Preparation 80) and cyclopropyl-[l-(2-ethyl-2H-tetrazol-5-yl)-piperidin-4-yl]-amine (Preparation 220) employing a procedure similar to that in Preparation 95: RT = 1.24min; m/z (ES ) = 639.64 [M + H]+ (LCMS Method - 2).
Preparation 222: Cyclopropyl-{l-[3-(tetrahydro-pyran-4-yl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl}-amine
Figure imgf000096_0002
The title compound was synthesized from (l-cyano-piperidin-4-yl)-cyclopropyl- carbamic acid tert-butyl ester (Preparation 20) and N-hydroxy-tetrahydro-pyran-4- carboxamidine employing a procedure similar to that outlined in Preparation 27: RT = 0.50min; m/z (ES+) = 293.41 [M+ H]+ (LCMS Method - 2).
Preparation 223: [(3/?,45)-l-[5-(Cyclopropyl-{l-[3-(tetrahydro-pyran-4-yl)- [l,2,4]oxadiazol-5-yl]-piperidin-4-yl}-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro- phenyl)-pyrrolidin-3-yl]-carbamic acid teri-butyl ester
Figure imgf000097_0001
The title compound was synthesized from 2-[(3R,45')-3-tert-butoxycarbonylamino-4- (2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid (Preparation 80) and cyclopropyl- { 1 -[3-(tetrahydro-pyran-4-yl)-[ 1 ,2,4]oxadiazol-5-yl]-piperidin-4-yl} -amine (Preparation 222) employing a procedure similar to that outlined in Preparation 95: RT = 1.19min; m/z (ES+) = 695.65 [M+ H]+ (LCMS Method - 2).
Preparation 224: 4-Fluoro-benzoic acid 1-carboxy-cyclopropyl ester
Figure imgf000097_0002
To a solution of 1 -hydroxy- 1 -eye lopropanecarboxylic acid (0.500g, 4.90mmol) in pyridine (1.25mL) and chloroform (lmL) at 0°C was added 4-fluoro-benzoyl chloride (0.608mL, 5.14mmol) and the resulting reaction mixture was heated at 70°C for 40 min. Upon cooling, the reaction mixture was poured into water (20mL) and extracted with chloroform (3 x 15mL). The combined organic extracts were washed with 2M HC1 (20mL) and then extracted into saturated aqueous aHC03 solution (2 x 20mL). The combined aHC03 extracts were adjusted to pH 1 with 12M HC1 and then extracted into chloroform (3 x 20mL). The combined organic extracts were passed through a hydrophobic frit and the solvent removed in vacuo. Purification by column chromatography (EtOAcTH; 1 :9 to 3 : 10) afforded the title compound: RT = 0.94 min; mlz (ES~) = 223.18 [M - H]+.
Preparation 225: 4-Fluoro-benzoic acid 1-carbamoyl-cyclopropyl ester
Figure imgf000097_0003
To a solution of 4-fluoro-benzoic acid 1-carboxy-cyclopropyl ester (Preparation 224, 0.423g, 1.89mmol) in THF (13mL) was added EDCI (0.434g, 2.26mmol) and HOBt and the resulting reaction mixture was stirred at r.t. for 10 min. ¾ in 1,4-dioxane (0.5M, 37.74mL, 18.87mmol) was added and the reaction mixture stirred at r.t. for 72 h. The solvent was removed in vacuo and the remainder partitioned between water (lOOmL) and EtOAc (200mL). The organic layer was separated and washed with saturated aqueous NaHC(¾ solution and brine, dried (MgSC^), filtered and concentrated in vacuo: RT = 0.82 min; mlz (ES+) = 224.19 [M + H]+.
Preparation 226: 4-Fluoro-benzoic acid l-(5-trichloromethyl-[l,2,4]oxadiazol-3-yl)- cyclopropyl ester
Figure imgf000098_0001
To a solution of 4-fluoro-benzoic acid 1 -carbamoyl-cyclopropyl ester (Preparation
225, 0.290g, 1.30mmol) in THF (lOmL) at 0°C was added triethylamine (0.54mL, 3.9mmol) followed by trifluoroacetic anhydride (0.28mL, 1.9mmol) and the resulting reaction mixture was stirred for 1 h. The reaction was quenched with water (50mL), extracted into DCM (2 x 70mL) and the combined organic extracts were passed through a hydrophobic frit and concentrated in vacuo. The remainder was dissolved in EtOH (l OmL) and hydro xylamine (50% aqueous solution, 87.6μΕ, 1.43mmol) was added. The resulting reaction mixture was heated at 60°C for 1.5 h, then the solvent removed in vacuo. The remainder was dissolved in toluene (20mL), perchloroacetic anhydride (0.237mL, 1.30mmol) was added and the reaction mixture heated at 90°C for 3 h. The solvent was removed in vacuo and the remainder purified by column chromatography (EtOAcTH; 5:95) to afford the title compound: RT = 1.42 min; mlz (ES+) = 365.07 [M+H]+.
Preparation 227: 4-Fluoro-benzoic acid l-{5-[4-(teri-butoxycarbonyl-cyclopropyl- amino)-piperidin-l-yl]-[l,2,4 oxadiazol-3-yl}-cyclopropyl ester
Figure imgf000098_0002
Cyclopropyl-piperidin-4-yl-carbamic acid tert-butyl ester (Preparation 14, 0.038g, 0.16mmol) and 4-fluoro-benzoic acid l-(5-trichloromethyl-[l,2,4]oxadiazol-3-yl)- cyclopropyl ester (Preparation 226, 0.034g, 0.093mmol) were combined in dimethyl sulfoxide (0.2mL) and the mixture stirred at room temperature for 16 h. The reaction mixture was diluted with water (15mL) and then extracted with EtOAc (3 x 50mL). The combined organic extracts were washed with brine and passed through a hydrophobic frit then concentrated in vacuo. Purification by preparative HPLC (standard method) afforded the title compound: RT = 1.45min; m/z (ES+) = 487.44 [M+ H]+ (LCMS Method - 2).
Preparation 228: Cyclopropyl-{l-[3-(l-hydroxy-cyclopropyl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl}-carbamic acid tert-but l ester
Figure imgf000099_0001
4-Fluoro-benzoic acid l- {5-[4-(?ert-butoxycarbonyl-cyclopropyl-amino)-piperidin-l- yl]-[l ,2,4]oxadiazol-3-yl}-cyclopropyl ester (Preparation 227, 0.034g, 0.070mmol) was dissolved in THF (2.0 mL), MeOH (1.0 mL), water (1.0 mL), lithium hydroxide monohydrate (0.012g, 0.28mmol) added and the mixture stirred at room temperature for 2 h. The reaction mixture was diluted with water and extracted with EtOAc (2 x). The combined organic extracts were filtered through a hydrophobic frit and concentrated in vacuo to afford the title compound: RT = 1.04 min ; m/z (ES+) = 369.39 [M+ H]+ (LCMS Method - 2).
Preparation 229: Cyclopropyl-{l-[3-(l-methoxy-cyclopropyl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl}-amine
Figure imgf000099_0002
Cyclopropyl- { 1 -[3 -(1 -hydroxy-cyclopropyl)-[ 1 ,2,4]oxadiazol-5-yl]-piperidin-4-yl} - carbamic acid tert-butyl ester (Preparation 228, 0.082g, 0.22mmol) was dissolved in THF (6.0mL) and cooled to 0.°C. NaH (60% dispersion in mineral oil, 0.012g, 0.29mmol) was added and the mixture stirred at 0°C for 20 min followed by stirring at room temperature for 10 min. Methyl iodide (0.018mL, 0.29mmol) was added and stirred at r.t. for 2 h. The reaction mixture was diluted with water (30mL) and extracted with EtOAc (3 x 50mL). The combined organic extracts were washed with brine, filtered through a hydrophobic frit and concentrated in vacuo. The residue was dissolved in DCM (lOmL) and cooled to 0°C. TFA (1.5mL, 19mmol) was added and stirred for 2 h. The mixture was passed through a SCX cartridge eluting with 10% ammonia in MeOH solution to afford the title compound: RT = 0.53 min; m/z (ES+) = 279.49 [M + H]+ (LCMS Method - 2).
Preparation 230: [(3/?,45)-l-[5-(Cyclopropyl-{l-[3-(l-methoxy-cyclopropyl)- [l,2,4]oxadiazol-5-yl]-piperidin-4-yl}-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro- phenyl)-pyrrolidin-3-y -carbamic acid tert-but l ester
Figure imgf000100_0001
The title compound was synthesized from 2-[(3R,45')-3-tert-butoxycarbonylamino-4- (2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid (Preparation 80) and cyclopropyl- { 1 -[3-(l -methoxy-cyclopropyl)-[ 1 ,2,4]oxadiazol-5-yl]-piperidin-4-yl} -amine (Preparation 229) employing a procedure similar to that outlined in Preparation 95: RT = 1.27 min; m/z (ES+) = 681.57 [M + H]+ (LCMS Method - 2).
Preparation 231: Cyclopropyl-{l-[(5)-3-(tetrahydro-furan-2-yl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl}-amine
Figure imgf000100_0002
The title compound was synthesized from (l-cyano-piperidin-4-yl)-cyclopropyl- carbamic acid tert-butyl ester (Preparation 20) and (5)-N-hydroxytetrahydrofuran-2- carboxamidine (Preparation 26) employing a procedure similar to that outlined in Preparation 27: RT = 0.48min; m/z (ES+) = 279.35 [M+ H]+ (LCMS Method - 2).
Preparation 232: [(3Λ,45)-1-[5-(€γο1ορΓοργ1-{1-[(5)-3-(ίβίΓ3ΗγΰΓθ-ίηΓ3η-2-γ1)- [l,2,4]oxadiazol-5-yl]-piperidin-4-yl}-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro- phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester
Figure imgf000101_0001
The title compound was synthesized from 2-[(3R,45')-3-tert-butoxycarbonylamino-4- (2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid (Preparation 80) and cyclopropyl- { l-[(5)-3-(tetrahydro-furan-2-yl)-[l,2,4]oxadiazol-5-yl]-piperidin-4-yl} -amine (Preparation 231) employing a procedure similar to that outlined in Preparation 95: RT = 1.19 min; m/z (ES+) = 681.54 [M + H]+ (LCMS Method - 2).
Preparation 233: (/?)-7V-Hydroxytetrahydrofuran-2-carboxamidine
Figure imgf000101_0002
The title compound was synthesized from (R)-tetrahydrofuran-2-carbonitrile employing a procedure similar to that outlined in Preparation 26: ¾ NMR ¾ (300MHz, CDC13): 4.85 (bs, 2H), 4.44 - 4.32 (m, 1H), 4.00 - 3.74 (m, 2H), 2.20 - 1.80 (m, 4H).
Preparation 234: Cyclopropyl-{l-[(/?)-3-(tetrahydro-furan-2-yl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl}-amine
Figure imgf000101_0003
The title compound was synthesized from (l-cyano-piperidin-4-yl)-cyclopropyl- carbamic acid tert-butyl ester (Preparation 20) and (R)-N-hydroxytetrahydrofuran-2- carboxamidine (Preparation 235) employing a procedure similar to that outlined in Preparation 27: RT = 0.48min; m/z (ES+) = 279.34 [M+ H]+ (LCMS Method - 2).
Preparation 235: [(3Λ,45)-1-[5-(€γο1ορΓοργ1-{1-[(Λ)-3-(ίβίΓ3ΗγΰΓθ-1ηΓ3η-2-γ1)- [l,2,4]oxadiazol-5-yl]-piperidin-4-yl}-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro- phenyl)-pyrrolidin-3-yl]-carbamic acid teri-butyl ester
Figure imgf000102_0001
The title compound was synthesized from 2-[(3R,45')-3-tert-butoxycarbonylamino-4- (2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5-carboxylic acid (Preparation 80) and cyclopropyl- { 1 -[(R)-3 -(tetrahydro-furan-2-yl)-[ 1 ,2,4]oxadiazol-5-yl]-piperidin-4-yl} -amine (Preparation 234) employing a procedure similar to that outlined in Preparation 95: RT = 1.19 min; m/z (ES+) = 681.54 [M + H]+ (LCMS Method - 2).
Preparation 236: 5-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2,5-difluoro-phenyl)- pyrrolidin-l-yl]-pyrazine-2-carbox lic acid methyl ester
Figure imgf000102_0002
To a solution of 2-chloro-pyrizine-5-carboxylic acid methyl ester (2.55g, 14.75mmol) and triethylamine (4.2ml, 26.82mmol) in anhydrous THF (240mL) was added [(3R,4S)-4- (2,5-difluorophenyl)pyrrolidin-3-yl]carbamic acid tert-butyl ester (Preparation 193, 6.6g, 22.12mmol) and the resulting reaction mixture was stirred at 50°C for 4 h. The reaction mixture was concentrated in vacuo and the remainder was purified by chromatography on Si02 gel using EtOAc first then DCM/MeOH/NH3 as eluents to afford the title compound: RT = 0.82 min; mlz (ES+) = 435.12 [M + H]+.
Preparation 237: 5-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2,5-difluoro-phenyl)- pyrrolidin-l-yl]-pyrazine-2-carbox lic acid
Figure imgf000102_0003
The title compound was synthesized from 5-[(3R,45')-3-tert-butoxycarbonylamino-4- (2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrazine-2-carboxylic acid methyl ester (Preparation 236) employing a procedure similar to that outlined in Preparation 80: RT = 1.54 min; mlz (ES+) = 421.14 [M + H]+.
Preparation 238: [(3Λ,45)-1-[5-({1-[3-(1,1-ϋίηηοΓθ-β^1)-[1,2,4]οχ3(1ί3ζο1-5^1]- piperidin-4-yl}-methyl-carbamoyl)-pyrazin-2-yl]-4-(2,5-difluoro-phenyl)-pyrrolidin-3- yl]-carbamic acid teri-but l ester
Figure imgf000103_0001
The title compound was synthesized from 5-[(3R,45')-3-iert-butoxycarbonylamino-4- (2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrazine-2-carboxylic acid (Preparation 237) and { 1- [3-(l, l-difluoro-ethyl)-[l,2,4]oxadiazol-5-yl]-piperidin-4-yl} -methyl-amine (Preparation 34) employing a procedure similar to that outlined in Preparation 95: RT = 1.27min; m/z (ES+) = 649.65 [M + H]+ (LCMS Method - 2).
Preparation 239: 6-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2,5-difluoro-phenyl)- pyrrolidin-l-yl] -nicotinic acid meth l ester
Figure imgf000103_0002
To a solution of 2-chloro-pyridine-5-carboxylic acid methyl ester (0.5g, 2.91mmol) and [(3R,45)-4-(2,5-difluorophenyl)pyrrolidin-3-yl]carbamic acid tert-butyl ester (Preparation 193, 0.96g, 3.21mmol) in DMF (lOmL) was added potassium carbonate (0.81g, 5.83mmol) and the resulting reaction mixture was stirred at 50°C for 16 h. The reaction mixture was diluted with DCM (50mL), washed with brine (20mL), dried (MgS04), filtered and concentrated in vacuo. The remainder was purified by chromatography on S1O2 gel using EtOAc/hexane as eluents; the resulting product was triturated with Et20 to afford the title compound: *Η NMR DH (300MHz, CDC13): 8.81 (m, 1H), 8.02 (d, 1H), 7.10-6.90 (m, 3H), 6.35 (d, 1H), 4.46 (m, 1H), 4.10 (m, 2H), 3.85 (s, 3H), 3.60 (m , 2H), 3.35 (t, 1H), 1.40 (s, 9H).
Preparation 240: 6-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2,5-difluoro-phenyl)- pyrrolidin-l-yl] -nicotinic acid
Figure imgf000104_0001
The title compound was synthesized from 6-[(3R,45')-3-iert-butoxycarbonylamino-4- (2,5-difluoro-phenyl)-pyrrolidin-l-yl]-nicotinic acid methyl ester (Preparation 239) employing a procedure similar to that outlined in Preparation 80: RT = 0.60 min; mlz (ES ) = 420.48 [M+ H]+.
Preparation 241 : [(3Λ,45)-1-[5-({1-[3-(1,1-ϋίηηοΓθ-β^1)-[1,2,4]οχ3ΰί3ζο1-5^1]- piperidin-4-yl}-methyl-carbamoyl)-pyridin-2-yl]-4-(2,5-difluoro-phenyl)-pyrrolidin-3- yl]-carbamic acid teri-but l ester
Figure imgf000104_0002
The title compound was synthesized from 6-[(3R,45')-3-iert-butoxycarbonylamino-4- (2,5-difluoro-phenyl)-pyrrolidin-l-yl]-nicotinic acid (Preparation 240) and {l-[3-(l,l- difluoro-ethyl)-[l,2,4]oxadiazol-5-yl]-piperidin-4-yl}-methyl-amine (Preparation 34) employing a procedure similar to that outlined in Preparation 95: RT = 1.11 min; m/z (ES ) = 648.62 [M+ H]+ (LCMS Method - 2).
Preparation 242: 6-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2,5-difluoro-phenyl)- pyrrolidin-l-yl]-5-cyano-nicotinic acid methyl ester
Figure imgf000105_0001
To a solution of 2-hydroxy-3-cyano-pyridine-5-carboxylic acid methyl ester (0.5g, 2.81mmol) and [(3R,45')-4-(2,5-difluorophenyl)pyrrolidin-3-yl]carbamic acid tert-butyl ester (Preparation 193, l .Og, 3.37mmol) in MeCN (lOmL) was added BOP (1.86g„ 4.21mmol) and DBU (0.63ml, 4.21mmol); the resulting reaction mixture was stirred at r.t. for 16 h. The reaction mixture was diluted with EtOAc (50mL) and water (20mL); aqueous layer was back-extracted with more EtOAc, then the organics were combined, washed with brine (20mL), dried (MgS04), filtered and concentrated in vacuo. The remainder was purified by chromatography on S1O2 gel using EtOAc/Hexane as eluents to afford the title compound: ¾ NMR δΗ (300MHz, CDC13): 8.85 (s, 1H), 8.34 (s, 1H), 7.10-6.90 (m, 3H), 4.44 (m, 1H), 3.90 (s, 3H), 3.75 (t, 1H), 3.65 (m, 1H), 1.40 (s, 9H).
Preparation 243: 6-[(3/?,45)-3-teri-Butoxycarbonylamino-4-(2,5-difluoro-phenyl)- pyrrolidin-1 -yl] -5-cyano-nicotini acid
Figure imgf000105_0002
The title compound was synthesized from 6-[(3R,45')-3-tert-butoxycarbonylamino-4- (2,5-difluoro-phenyl)-pyrrolidin-l-yl]-5-cyano-nicotinic acid methyl ester (Preparation 242) employing a procedure similar to that outlined in Preparation 80: RT = 0.63 min; mlz (ES+) = 445.20 [M + H]+.
Preparation 244: [(S^^^-l-^-C ano-S-Cil- -Cl^-difluoro-eth ^-Il^^loxadiazol-S- yl]-piperidin-4-yl}-methyl-carbamoyl)-pyridin-2-yl]-4-(2,5-difluoro-phenyl)-pyrrolidin- 3-yl]-carbamic acid teri-butyl ester
Figure imgf000106_0001
The title compound was synthesized from 6-[(3R,45')-3-tert-butoxycarbonylamino-4- (2,5-difluoro-phenyl)-pyrrolidin-l-yl]-5-cyano-nicotinic acid (Preparation 243) and {l -[3- (1,1 -difluoro-ethyl)-[ 1 ,2,4]oxadiazol-5 -yl]-piperidin-4-yl} -methyl-amine (Preparation 34) employing a procedure similar to that outlined in Preparation 95: RT = 1.32 min; m/z (ES+) = 673.56 [M + H]+ (LCMS Method - 2).
Example 1: 2-[(3/?,45)-3-Amino-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine- 5-carboxylic acid cyclopropyl-[l-(3-isopropyl-[l,2,4]oxadiazol-5-yl)-piperidin-4-yl]- amide hydrochloride
To a solution of [(3R,45)-l - {5-[(l-cyano-piperidin-4-yl)-cyclopropyl-carbamoyl]- pyrimidin-2-yl}-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 93, 61mg, 100μmol) and N-hydroxy-isobutyramidine (13mg, 120μmol) in EtOH (4mL) was added dropwise zinc dichloride in Et20 (1M, 120μΕ, 120mmol) and the resulting reaction mixture was stirred at r.t. for 80 min. HC1 in 1,4-dioxane (4M, 130μΕ, 521 μιηο1) was added and the reaction mixture was heated at 60 °C for 16 h. Upon cooling, the reaction mixture was diluted with MeOH and loaded onto an SCX cartridge eluting with MeOH followed by ¾ in MeOH (7M). The basic fraction was concentrated in vacuo and purified by preparative HPLC. The product containing fractions were concentrated in vacuo and the remainder dissolved in MeOH, added to an SCX cartridge and eluted with MeOH followed by ¾ in MeOH (7M). The basic fraction was concentrated in vacuo, dissolved in MeCN:MeOH (1 :2) and HC1 in 1 ,4-dioxane (4M, 19μΕ) was added. The resulting mixture was stirred at r.t. for 10 min and the solvent removed in vacuo to afford the title compound: RT = 0.81 min; mlz (ES+) = 571.52 [M + H]+ (LCMS Method - 2).
Example 2 : 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid [l-(3-iso ropyl-[l,2,4]oxadiazol-5-yl)-piperidin-4-yl]-methyl-amide
Figure imgf000107_0001
To a solution of [(3R,45')-4-(2,5-difluoro-phenyl)-l -(5- {[l-(3-isopropyl- [ 1 ,2,4]oxadiazol-5-yl)-piperidin-4-yl]-methyl-carbamoyl} -pyrimidin-2-yl)-pyrrolidin-3 -yl]- carbamic ester tert-butyl ester (Preparation 94, 143mg, 228μmol) in DCM (2.9mL) was added TFA (352μΙ^, 4.56mmol) and the resulting reaction mixture was stirred at r.t. for 1 h. Further TFA (320μί, 4.15mmol) was added and the reaction mixture stirred at r.t. for 1.5 h. The reaction mixture was added to an SCX cartridge and eluted with MeOH followed by H3 in MeOH (7M). The basic fraction was concentrated in vacuo to afford the title compound: RT = 2.50 min; mlz (ES+) = 527.29 [M+ H]+.
Example 3 : 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid ethyl-[l-(3-isopropyl-[l,2,4]oxadiazol-5-yl)-piperidin-4-yl]-amide hydrochloride
Figure imgf000107_0002
To a solution of [(3R,45)-4-(2,5-difluoro-phenyl)-l -(5- {ethyl-[l-(3-isopropyl- [ 1 ,2,4]oxadiazol-5-yl)-piperidin-4-yl]-carbamoyl} -pyrimidin-2-yl)-pyrrolidin-3 -yl]-carbamic acid tert-butyl ester (Preparation 96, 95mg, 150μιηο1) in DCM (2.3mL) was added TFA (485μί, 6.30mmol) and the resulting reaction mixture stirred at r.t. for 1.5 h. The reaction mixture was added to an SCX cartridge and eluted with MeOH followed by H3 in MeOH (7M). The basic fraction was concentrated in vacuo, dissolved in MeOH (5mL) and HC1 in Et20 (2M, 380μΚ) was added. The mixture was stirred at r.t. for 15 min, then concentrated in vacuo to afford the title compound: RT = 0.78 min; mlz (ES+) = 541.52 [M + H]+ (LCMS Method - 2).
Example 4: 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid {l-[3-(l-fluoro-l-methyl-ethyl)-[l,2,4]oxadiazol-5-yl]-piperidin-4-yl}- methyl-amide hydrochloride
Figure imgf000108_0001
To a solution of {(3R,45')-4-(2,5-difluoro-phenyl)-l -[5-({ l-[3-(l -fluoro-l-methyl- ethyl)-[ 1 ,2,4]oxadiazol-5-yl]-piperidin-4-yl} -methyl-carbamoyl)-pyrimidin-2-yl]-pyrrolidin- 3-yl}carbamic acid tert-butyl ester (Preparation 95, 85mg, 130μηιο1) in MeOH (3mL) 5 under argon was added HCl in Et20 (2M, 660μΙ.) at 0°C. The reaction mixture was allowed to warm to r.t. and stirred for 18 h. The mixture was concentrated in vacuo to afford the title compound: RT = 0.77 min; mlz (ES+) = 545.42 [M+ H]+ (LCMS Method - 2).
The following compounds were prepared from the appropriate tert-butoxy carbonyl 0 protected amino intermediate employing a procedure similar to that outlined in Example 2:
Figure imgf000108_0002
The following compounds were prepared from the appropriate tert-butoxy carbonyl protected amino intermediate employing a procedure similar to that outlined in Example 3:
Figure imgf000109_0001
hydrochloride
Figure imgf000110_0001
Figure imgf000111_0001
Figure imgf000112_0001
hydrochloride
Ill h d hl id
Figure imgf000113_0001
Figure imgf000115_0001
Figure imgf000116_0001
Figure imgf000117_0001
The following compounds were prepared from the appropriate tert-butoxy carbonyl protected amino intermediate employing a procedure similar to that outlined in Example 4:
Figure imgf000117_0002
Figure imgf000118_0001
Figure imgf000119_0001
Figure imgf000120_0001
Figure imgf000121_0001
hydrochloride
Figure imgf000122_0001
piperidin-4-yl]-amide
Figure imgf000123_0001
hydrochloride
Figure imgf000124_0001
Example 84 : 2- [(3/?,45)-3-Amino-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-l -yl] - pyrimidine-5-carboxylic acid [l-(3-ethyl-[l,2,4]oxadiazol-5-yl)-piperidin-4-yl]-methyl- amide hydrochloride
Figure imgf000124_0002
To a solution of [(3R,45")-l-(5- {[l-(3-ethyl-[l,2,4]oxadiazol-5-yl)-piperidin-4-yl]- methyl-carbamoyl} ^yrimidin-2-yl)-4-(2,4,5-trifluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 118, 115mg, 182μmol) in 1 ,4-dioaxne (5mL) was added HCl in 1 ,4-dioxane (4M, 182μί, 729μιηο1) and the resulting reaction mixture was stirred at r.t. for 75 min. Further HCl in 1,4-dioxane (4M, 1.5mL, 6mmol) was added and stirring at r.t. continued for 16 h. The solvent was removed in vacuo and the remainder purified by preparative HPLC. The product containing fractions were loaded onto an SCX cartridge and eluted with MeOH followed by NH3 in MeOH (7M). The basic fraction was concentrated in vacuo, then the remainder was dissolved in 1,4-dioxane and HC1 in 1,4-dioxane (4M, 0.2mL) was added. The resulting mixture was stirred at r.t. for 1 h and concentrated in vacuo to afford the title compound: RT = 0.71 min; mlz (ES ) = 531.5 [M+ H]+ (LCMS Method - 2).
Example 85: 2-[(3/?,4S)-3-Amino-4-(2-fluoro-5-methyl-phenyl)-pyrrolidin-l-yl]- pyrimidine-5-carboxylic acid [l-(3-ethyl-[l,2,4]oxadiazol-5-yl)-piperidin-4-yl]-methyl- amide hydrochloride
Figure imgf000125_0001
To a solution of [(3R,45")-l-(5- {[l-(3-ethyl-[l,2,4]oxadiazol-5-yl)-piperidin-4-yl]- methyl-carbamoyl} -pyrimidin-2-yl)-4-(2-fluoro-5-methyl-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 141, 109mg, 179μmol) in Et20 (5mL) at 0°C was added HC1 in 1 ,4-dioxane (4M, 1 mL, 4mmol) and the resulting reaction mixture was stirred at r.t. for 4 h. The solvent was removed in vacuo and the remainder was dissolved in the minimum amount of hot EtOH and allowed to cool to r.t. Et20 was added dropwise until a white precipitate formed, then the yellow solution was decanted off and the solid dried in vacuo to afford the title compound: RT = 2.48 min; mlz (ES ) = 509.18 [M + H]+.
Example 86: 2-[(3/?,4S)-3-Amino-4-(2-fluoro-5-methyl-phenyl)-pyrrolidin-l-yl]- pyrimidine-5-carboxylic acid [l-(3-isopropyl-[l,2,4]oxadiazol-5-yl)-piperidin-4-yl]- methyl-amide hydrochloride
Figure imgf000125_0002
The title compound was synthesized from [(3R,45)-4-(2-fluoro-5-methyl-phenyl)-l- (5- {[1 -(3-isopropyl-[l ,2,4]oxadiazol-5-yl)-piperidin-4-yl]-methyl-carbamoyl} -pyrimidin-2- yl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 139, 180mg, 290μιηο1) employing a procedure similar to that outlined in Example 85: RT = 2.70 min; mlz (ES+) = 523.19 [M+ H]+.
Example 87: 2-[(3/?,4S)-3-Amino-4-(2-fluoro-5-methyl-phenyl)-pyrrolidin-l-yl]- pyrimidine-5-carboxylic acid ethyl-[l-(3-isopropyl-[l,2,4]oxadiazol-5-yl)-piperidin-4- yl]-amide hydrochloride
Figure imgf000126_0001
To a solution of [(3R,45')-l-(5- {ethyl-[l -(3-isopropyl-[l,2,4]oxadiazol-5-yl)- piperidin-4-yl]-carbamoyl} -pyrimidin-2-yl)-^^
carbamic acid tert-butyl ester (Preparation 140, 210mg, 330μmol) in Et20 (5mL) and DCM (2.5mL) at 0°C was added HCl in 1 ,4-dioxane (4M, 2.0mL, 8.0mmol) and the resulting reaction mixture was stirred at r.t. for 16 h. The solvent was removed in vacuo, then the remainder dissolved in EtOAc, washed with aqueous NaOH solution (2M), dried (MgSC^), filtered and concentrated in vacuo. The remainder was purified by preparative HPLC (basic method) and the product dissolved in MeOH prior to the addition of HCl in Et20 (2M, 2.0mL). The solvents were removed in vacuo to afford the title compound: RT = 2.72 min; m/z (ES+) = 537.25 [M+ H]+. Example 88: 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid {l-[5-(l,l-difluoro-ethyl)-[l,2,4]oxadiazol-3-yl]-piperidin-4-yl}-methyl- amide hydrochloride
Figure imgf000126_0002
The title compound was synthesized from [(3R,45)-l -[5-({ l-[5-(l, l-difluoro-ethyl)- [l,2,4]oxadiazol-3-yl]-piperidin-4-yl} -methyl-carbamoyl)-pyrimidin-2-yl]-4-(2,5-difluoro- phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 202, 357mg, 0.55mmol) employing a procedure similar to that outlined in Example 4: RT = 0.81 min; mlz (ES+) = 549.51 [M+ H]+.
Example 89: 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl-{l-[5-(l-methoxy-l-methyl-ethyl)-[l,2,4]oxadiazol-3-yl]- piperidin-4-yl}-amide hydrochloride
Figure imgf000127_0001
The title compound was synthesized from [(3R,4S)-l-[5-(cyclopropyl- { l-[5-(l- methoxy- 1 -methyl-ethyl)- [ 1 ,2,4]oxadiazol-3 -yl]-piperidin-4-yl} -carbamoyl)-pyrimidin-2- yl]-4-(2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 205, 91mg, 0.13mmol) employing a procedure similar to that outlined in Example 3: RT = 0.81 min; mlz (ES+) = 583.6 [M + H]+.
Example 90: 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl-{l-[5-(l,l-difluoro-ethyl)-[l,2,4]oxadiazol-3-yl]-piperidin- 4-yl}-amide hydrochloride
Figure imgf000127_0002
The title compound was synthesized from [(SR^^-l-fS-icyclopropyl- l l -fS-il,!- difluoro-ethyl)-[l,2,4]oxadiazol-3-yl]-piperidin-4-yl}-carbamoyl)-pyrimidin-2-yl]-4-(2,5- difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 209, 31mg, 0.046mmol) employing a procedure similar to that outlined in Example 4: RT = 0.81 min; m/z (ES+) = 583.6 [M + H]+.
Example 91: 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl-{l-[(/?)-5-(tetrahydro-furan-3-yl)-[l,2,4]oxadiazol-3-yl]- piperidin-4-yl}-amide h drochloride
Figure imgf000127_0003
The title compound was synthesized from [(3R,45')-l-[5-(cyclopropyl- { l -[(R)-5- (tetrahydro-furan-3 -yl)-[ 1 ,2,4]oxadiazol-3 -yl]-piperidin-4-yl} -carbamoyl)-pyrimidin-2-yl]-4- (2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid iert-butyl ester (Preparation 210, 36mg, 0.053mmol) employing a procedure similar to that outlined in Example 4: RT = 0.72 min; mlz (ES+) = 581.6 [M + H]+ (LCMS Method - 2).
Example 92: 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl-{l-[(5)-3-(tetrahydro-furan-3-yl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl}-amide
Figure imgf000128_0001
The title compound was synthesized from [(SR^^-l-fS-icyclopropyl- l l-fi^-S- (tetrahydro-mran-3-yl)-[l,2,4]oxadiazol-5-yl]^iperidin-4-yl}-carbamoyl)-pyrimidin-2-yl]-4- (2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamicacid tert-butyl ester (Preparation 215) employing a procedure similar to that outlined in Example 4: RT = 0.74 min; mlz (ES+) = 581.46 [M+ H]+ (LCMS Method - 2).
Example 93: 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid (l-cyclopropanesulfonyl-piperidin-4-yl)-cyclopropyl-amide hydrochloride
Figure imgf000128_0002
The title compound was synthesized from [(3R,45)-l - {5-[(l-cyclopropanesulfonyl- piperidin-4-yl)-cyclopropyl-carbamoyl]^yrimidin-2-yl} -4-(2,5-difluoro-phenyl)-pyrrolidin- 3-yl]-carbamic acid tert-butyl ester (Preparation 217) employing a procedure similar to that outlined in Example 3: RT = 0.73 min; m/z (ES+) = 547.51 [M + H]+ (LCMS Method - 2).
Example 94: 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl-[l-(2-ethyl-2H-tetrazol-5-yl)-piperidin-4-yl]-amide hydrochloride
Figure imgf000128_0003
The title compound was synthesized from [(3R,45)-l-(5- {cyclopropyl-[l -(2-ethyl-2H- tetrazol-5-yl)^iperidin-4-yl]-carbamoyl} ^yrimidin-2-yl)-4-(2,5-difluoro-phenyl)- pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 221) employing a procedure similar to that outlined in Example 4: RT = 0.75min; m/z (ES ) = 539.57 [M + H]+ (LCMS Method - 2).
Example 95: 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl-{l-[3-(tetrahydro-pyran-4-yl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl}-amide
Figure imgf000129_0001
The title compound was synthesized from [(3R,45)-l -[5-(cyclopropyl- { l -[3- (tetrahydro-pyran-4-yl)-[ 1 ,2,4]oxadiazol-5-yl]-piperidin-4-yl} -carbamoyl)-pyrimidin-2-yl]- 4-(2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 223) employing a procedure similar to that outlined in Example 3: RT = 0.73min; m/z (ES+) = 595.62 [M+ H]+ (LCMS Method - 2).
Example 96: 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl-{l-[3-(l-methoxy-cyclopropyl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl}-amide
Figure imgf000129_0002
The title compound was synthesized from [(3R,4,S)-l-[5-(cyclopropyi- { l-[3-(l- methoxy-cyclopropyl)-[ 1 ,2,4]oxadiazol-5-yl]-piperidin-4-yl} -carbamoyl)-pyrimidin-2-yl]-4- (2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 230) employing a procedure similar to that outlined in Example 3: RT = 0.76min; m/z (ES+) = 581.57 [M+ H]+ (LCMS Method - 2).
Example 97: 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl-{l-[(5)-3-(tetrahydro-furan-2-yl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl}-amide hydrochloride
Figure imgf000130_0001
The title compound was synthesized from [(SR^^-l-fS-icyclopropyl-l l-fi^-S- (tetrahydro-mran-2-yl)-[l,2,4]oxadiazol-5-yl]^iperidin-4-yl}-carbamoyl)-pyrimidin-2-yl]-4- (2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 232) employing a procedure similar to that outlined in Example 3: RT = 0.75min; m/z (ES ) = 581.51 [M+ H]+ (LCMS Method - 2).
Example 98: 2-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrimidine-5- carboxylic acid cyclopropyl-{l-[(/?)-3-(tetrahydro-furan-2-yl)-[l,2,4]oxadiazol-5-yl]- piperidin-4-yl}-amide h drochloride
Figure imgf000130_0002
The compound was synthesized from [(3R,45)-l-[5-(cyclopropyl-{l-[(R)-3- (tetrahydro-furan-2-yl)-[l,2,4]oxadiazol-5-yl]^iperidin-4-yl}-carbamoyl)^yrimidin-2-yl]-4- (2,5-difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 235) employing a procedure similar to that outlined in Example 3: RT = 0.75 min; m/z (ES ) = 581.46 [M+ H]+ (LCMS Method - 2).
Example 99: 5-[(3/?,4S)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-pyrazine-2- carboxylic acid{l-[3-(l,l-difluoro-ethyl)-[l,2,4]oxadiazol-5-yl]-piperidin-4-yl}-methyl- amide hydrochloride
Figure imgf000130_0003
The title compound was synthesized from [(3R,45)-l-[5-({l-[3-(l,l-difluoro-ethyl)- [l,2,4]oxadiazol-5-yl]-piperidin-4-yl}-methyl-carbamoyl)-pyrazin-2-yl]-4-(2,5-difluoro- phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 238) employing a procedure similar to that outlined in Example 4: RT = 0.79min; m/z (ES ) = 549.55 [M + H]+ (LCMS Method - 2).
Example 100: 6-[(3JR,45)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-7V-{l-[3-(l,l- difluoro-ethyl)-[l,2,4]oxadiazol-5-yl]-piperidin-4-yl}-7V-methyl-nicotinamide hydrochloride
Figure imgf000131_0001
The title compound was synthesized from [(SR^^-l -fS-d l-tS-ilj l-difluoro-ethyl)- [l,2,4]oxadiazol-5-yl]-piperidin-4-yl} -methyl-carbamoyl)-pyridin-2-yl]-4-(2,5-difluoro- phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 241) employing a procedure similar to that outlined in Example 3: RT = 0.74min; m/z (ES ) = 548.60 [M + H]+ (LCMS Method - 2).
Example 101 : 6-[(3/?,45)-3-Amino-4-(2,5-difluoro-phenyl)-pyrrolidin-l-yl]-5-cyano-iV-
{l-[3-(l,l-difluoro-ethyl)-[l,2,4]oxadiazol-5-yl]-piperidin-4-yl}-7V-methyl-nicotinamide hydrochloride
Figure imgf000131_0002
The title compound was synthesized from [(SR^^-l-fS-cyano-S-d l-fS-ilj l-difluoro- ethyl)-[l,2,4]oxadiazol-5-yl]-piperidin-4-yl} -methyl-carbamoyl)-pyridin-2-yl]-4-(2,5- difluoro-phenyl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (Preparation 244) employing a procedure similar to that outlined in Example 3: RT = 0.79min; m/z (ES+) = 573.54 [M+ H]+ (LCMS Method - 2).
The biological activity of the compounds of the invention may be tested in the following assay systems:
GPR119 cAMP Assay A stable cell line expressing recombinant human GPR1 19 was established and this cell line was used to investigate the effect of compounds of the invention on intracellular levels of cyclic AMP (cAMP). The cell monolayers were washed with phosphate buffered saline and stimulated at 37°C for 30 min with various concentrations of compound in stimulation buffer plus 1% DMSO. Cells were then lysed and cAMP content determined using the Perkin Elmer AlphaScreen™ (Amplified Luminescent Proximity Homogeneous Assay) cAMP kit. Buffers and assay conditions were as described in the manufacturer's protocol.
A number of representative examples of compounds of the invention were tested and found to produce a concentration-dependent increase in intracellular cAMP levels and generally had an EC50 of <10 μΜ; some of the examples showed an EC50 of less than 1 μΜ in the cAMP assay.
DPP-IV Assay Method
DPP-IV activity was measured by monitoring the cleavage of the fluorogenic peptide substrate, H-Gly-Pro-7-amino-4-methylcoumarin (GP-AMC) whereby the product 7-amino-
4-methylcoumarin is quantified by fluorescence at excitation 380 nm and emission 460 nm.
Assays were carried out in 96-well plates (Black OptiPlate-96F) in a total volume of 100 per well consisting of 50 mM Tris pH 7.6, 100 μΜ GP-AMC, 10-25 μΙΙ recombinant human
DPP-IV and a range of inhibitor dilutions in a final concentration of 1% DMSO. Plates were read in a fluorimeter after 30 min incubation at 37°C. Recombinant human DPP-IV residues
Asn29-Pro766 was purchased from BioMol.
All of Examples 1 to 101 above were tested and found to show activity in this assay having an IC50 of <20 μΜ. Some of the examples were found to have a DPP-IV EC50 value of less than Ι μΜ; others an EC50 value of less than 20μΜ.
Activities of a representative group of compounds that were tested in the cAMP assay and the DPP-IV assay are shown in Table 1 below:
Table 1.
Figure imgf000132_0001
7 + +
10 + +
51 + +
65 + +
71 + +
Anti-diabetic effects of compounds of the invention in an in-vitro model of pancreatic beta cells (HIT-T15)
Cell Culture
HIT-T15 cells (passage 60) were obtained from ATCC, and were cultured in
RPMI1640 medium supplemented with 10% fetal calf serum and 30 nM sodium selenite. All experiments were done with cells at less than passage 70, in accordance with the literature, which describes altered properties of this cell line at passage numbers above 81 (Zhang HJ, Walseth TF, Robertson RP. Insulin secretion and cAMP metabolism in HIT cells. Reciprocal and serial passage-dependent relationships. Diabetes. 1989 Jan;38(l):44-8). cAMP assay
HIT-T15 cells were plated in standard culture medium in 96-well plates at 100,000 cells/ 0.1 mL/ well and cultured for 24 h and the medium was then discarded. Cells were incubated for 15min at room temperature with Ι ΟΟμΙ stimulation buffer (Hanks buffered salt solution, 5mM HEPES, 0.5mM IBMX, 0.1% BSA, pH 7.4). This was discarded and replaced with compound dilutions over the range 0.001, 0.003, 0.01, 0.03, 0.1, 0.3, 1, 3, 10, 30 μΜ in stimulation buffer in the presence of 0.5% DMSO. Cells were incubated at room temperature for 30 min. Then 75 uL lysis buffer (5mM HEPES, 0.3% Tween-20, 0.1% BSA, pH 7.4) was added per well and the plate was shaken at 900 rpm for 20 min. Particulate matter was removed by centrifugation at 3000rpm for 5 min, then the samples were transferred in duplicate to 384-well plates, and processed following the Perkin Elmer AlphaScreen cAMP assay kit instructions. Briefly 25 μΕ reactions were set up containing 8 μΕ sample, 5 μΕ acceptor bead mix and 12 μί detection mix, such that the concentration of the final reaction components is the same as stated in the kit instructions. Reactions were incubated at room temperature for 150 min, and the plate was read using a Packard Fusion instrument. Measurements for cAMP were compared to a standard curve of known cAMP amounts (0.01, 0.03, 0.1, 0.3, 1, 3, 10, 30, 100, 300, 1000 nM) to convert the readings to absolute cAMP amounts. Data was analysed using XLfit 3 software. Representative compounds of the invention were found to increase cAMP at an EC50 of less than 10 μΜ. Compounds showing an EC50 of less than 1 μΜ in the cAMP assay may be preferred.
Insulin secretion assay
HIT-T15 cells are plated in standard culture medium in 12-well plates at 106 cells/ 1 ml/ well and cultured for 3 days and the medium then discarded. Cells are washed x 2 with supplemented Krebs-Ringer buffer (KRB) containing 1 19 mM NaCl, 4.74 mM KC1, 2.54 mM CaCl2, 1.19 mM MgS04, 1.19 mM KH2P04, 25 mM NaHC03, 10 mM HEPES at pH 7.4 and 0.1% bovine serum albumin. Cells are incubated with 1ml KRB at 37°C for 30 min which is then discarded. This is followed by a second incubation with KRB for 30 min, which is collected and used to measure basal insulin secretion levels for each well. Compound dilutions (0, 0.1, 0.3, 1, 3, 10 μΜ) are then added to duplicate wells in 1ml KRB, supplemented with 5.6 mM glucose. After 30 min incubation at 37°C samples are removed for determination of insulin levels. Measurement of insulin was done using the Mercodia Rat insulin ELISA kit, following the manufacturers' instructions, with a standard curve of known insulin concentrations. For each well, insulin levels are corrected by subtraction of the basal secretion level from the pre-incubation in the absence of glucose. Data is analysed using XLfit 3 software.
Compounds of the invention preferably increase insulin secretion at an EC50 of less than 10 μΜ.
Oral Glucose Tolerance Tests
The effects of compounds of the invention on oral glucose (Glc) tolerance may be evaluated in male Sprague-Dawley rats. Food is withdrawn 16 h before administration of Glc and remains withdrawn throughout the study. Rats have free access to water during the study. A cut is made to the animals' tails, then blood (1 drop) is removed for measurement of basal Glc levels 60 min before administration of the Glc load. Then, the rats are weighed and dosed orally with test compound or vehicle (20% aqueous hydroxypropyl-yS- cyclodextrin) 45 min before the removal of an additional blood sample and treatment with the Glc load (2 g kg 1 p.o.). Blood samples are taken from the cut tip of the tail 5, 15, 30, 60, 120, and 180 min after Glc administration. Blood glucose levels are measured just after collection using a commercially available glucose-meter (OneTouch® UltraTM from Lifescan). Compounds of the invention preferably statistically reduce the Glc excursion at doses <100 mg kg-1. The effects of compounds of the invention on oral glucose (Glc) tolerance were evaluated in male C57B1/6 or male ob/ob mice. Food was withdrawn 5 h before
administration of Glc and remained withdrawn throughout the study. Mice had free access to water during the study. A cut was made to the animals' tails, then blood (20 μΐ,) was removed for measurement of basal Glc levels 45 min before administration of the Glc load. Then, the mice were weighed and dosed orally with test compound or vehicle (20% aqueous hydroxypropyl-yS-cyclodextrin or 25% aqueous Gelucire 44/14) 30 min before the removal of an additional blood sample (20 μί) and treatment with the Glc load (2-5 g kg 1 p.o.). Blood samples (20 μΚ) were then taken 25, 50, 80, 120, and 180 min after Glc administration. The 20 blood samples for measurement of Glc levels were taken from the cut tip of the tail into disposable micro-pipettes (Dade Diagnostics Inc., Puerto Rico) and the sample added to 480 μΐ, of haemolysis reagent. Duplicate 20 μΐ, aliquots of the diluted haemolysed blood were then added to 180 μΐ, of Trinders glucose reagent (Sigma enzymatic (Trinder) colorimetric method) in a 96-well assay plate. After mixing, the samples were left at room temperature for 30 min before being read against Glc standards (Sigma glucose/urea nitrogen combined standard set). Compounds of the invention statistically reduced the Glc excursion at doses <100 mg kg-1, for example at a dose of 30 mg kg 1 the compound of Example 6 showed a > 40% reduction in the Glc excursion.

Claims

1.
Figure imgf000136_0001
wherein A is a para-linked phenyl, pyridinyl, pyrimidinyl, pyrazinyl or triazinyl;
R1 is hydrogen, halo, cyano, C 1.4 alkyl, C\ ^haloalkyl, Ci _4alkoxy or
C 1 _5alkoxyC 1 _4alkyl;
is selected from the group consisting of:
(a) phenyl optionally substituted by one or more groups independently selected from halo, methyl or halomethyl groups,
(b) pyridyl optionally substituted by one or more groups independently selected from halo, methyl or halomethyl groups
and (c) N-pyridonyl optionally substituted by one or more groups independently selected from halo, methyl or halomethyl groups
R3 is C3_5cycloalkyl, 4 to 6 membered saturated heterocyclyl (comprising 1 or 2 ring heteroatoms selected from N, O and S) or Ci .4 alkyl wherein the C3_ cycloalkyl and Ci _4alkyl are optionally substituted by one to three groups independently selected from halo, cyano, hydroxy or Ci .2 alkoxy;
p and q are each 0, 1 or 2, provided that 0 < p+q < 2;
Z is selected from the group consisting of:
(a) C(0)0 R4,
(b) C(O) R4
(c) S(0)2 R4,
(d) 5- or 6-membered N-containing heteroaryl ring optionally
containing 1 to 3 additional heteroatoms selected from N, O and S, or a 8 to 10 membered fused bicyclic system optionally containing 1 to 3 heteroatoms selected from N, O and S, wherein the heteroaryl ring or fused bicyclic system is optionally substituted by one or two groups independently selected from halo, cyano, C1 .4 alkyl, C1 .4 haloalkyl, C1 .4 alkoxy,
Ci _4hydroxyalkyl, Ci _4alkoxyCi _4alkyl, heterocyclyl (a 4 to 6 membered saturated heterocycle comprising 1 or 2 ring heteroatoms selected from N , O and S), heterocyclylCi .4 alkyl (wherein the heterocyclyl is a 4 to 6 membered saturated heterocycle comprising 1 or 2 ring heteroatoms selected from N, O and S) and C3.5 cycloalkyl optionally substituted by C1 .4 alkyl , C1 .4 alkoxy or halo; R^ is selected from the group consisting of:
a) Ci _6alkyl,
b) phenyl,
c) Ci _ alkoxyC2-6alkyl,
d) C3_ cycloalkyl optionally substituted by Ci .4 alkyl,
e) C3 _5cycloalkylC 1 _4alkyl optionally substituted by C 1 .4 alkyl,
f) 4 to 6 membered saturated heterocycle comprising 1 or 2 ring heteroatoms selected from N, O and S, and
g) 5 or 6 membered heteroaryl ring containing 1 to 4 heteroatoms selected from N, O and S optionally substituted by one or two groups selected from halo, cyano, C 1.4 alkyl, Ci _
4haloalkyl, Ci _4alkoxy, C2-4alkoxyCi _4alkyl, a 4 to 6 membered saturated heterocycle comprising 1 or 2 ring heteroatoms selected from N, O and S and C3_ cycloalkyl wherein the cycloalkyl is optionally substituted by Ci .4 alkyl or halo;
each of R5 and R6 is independently hydrogen, halo, Ci _2alkyl, Ci _2 haloalkyl or
Ci _3 alkoxy;
R7 is hydrogen, halo,
Figure imgf000137_0001
and pharmaceutically acceptable salts thereof.
2. A compound as claimed in Claim 1 wherein said compound has the stereochemistry:
Figure imgf000137_0002
wherein R2 is as defined in Claim 1.
3. A compound as claimed in any one of Claims 1 or 2 wherein p and q are both 1, and has the formula;
Figure imgf000138_0001
4. A compound as claimed in Claim 1 to 3 wherein R1 is hydrogen.
5. A compound as claimed in any one of Claims 1 to 4 wherein R3 is C3_ cycloalkyl or Ci _4 alkyl, wherein the C3_ cycloalkyl and Ci .4 alkyl are optionally substituted by one to three groups independently selected from halo, cyano, hydroxy or Ci ^alkoxy.
6. A compound as claimed in any one of Claims 1 to 5 wherein A is pyridine, pyrimidine or pyrazine.
7. A compound as claimed in any one of Claims 1 to 6 wherein Z is a heteroaryl group optionally substituted by one or two groups selected from halo, Ci .4 alkyl, Ci _4halo alkyl, C\_
4alkoxy, Ci _4alkoxyCi _4alkyl, heterocyclyl, heterocyclylCi _4alkyl and C3_ cycloalkyl, wherein the cycloalkyl is optionally substituted by Ci .4 alkyl, Ci .4 alkoxy or halo.
8. A compound as claimed in any one of Claims 1 to 7 wherein the heteroaryl group is oxadiazole, pyrimidine, pyridazine, thiazole, tetrazole, benzothiazole or thiadiazole.
9. A compound as claimed in Claims 1 to 8 wherein Z comprises optionally substituted l,2,4-oxadiazol-3-yl, l ,2,4-oxadiazol-5-yl or pyrimidin-2-yl.
10. A compound as claimed in Claims 1 to 6 wherein Z is -C(0)OR^.
1 1. A compound as claimed in Claims 1 to 6 wherein R^ is alkyl, C\ _galkoxyC2-6alkyl, C3_ gcycloalkyl or C3_5cycloalkylCi _4alkyl, wherein the C3_5cycloalkylC 1 .4 alkyl is optionally substituted by Ci _4alkyl.
12. A compound as claimed in Claim 10 wherein R^ is propyl, preferably isopropyl.
13. A compound as claimed in any preceding claim wherein is substituted phenyl or pyridyl.
14. A compound as claimed in any preceding claim, wherein is phenyl substituted by one, two or three halo or methyl groups.
15. A compound as claimed in any preceding claim, wherein is phenyl substituted by two or three halo groups.
16. A compound according to any of Examples 1 to 101 or a pharmaceutically acceptable salt thereof.
17. A compound as claimed in any preceding claim for use in a method of treatment of a metabolic disorder, including type II diabetes.
18. A pharmaceutical formulation or composition comprising a compound as claimed in any of claims 1 to 17 and a pharmaceutically acceptable carrier therefor.
19. A method for the treatment or prevention of diabetes, including type II diabetes, or a method for the treatment of obesity, metabolic syndrome (syndrome X), impaired glucose tolerance, hyperlipidemia, hypertriglyceridemia, hypercholesterolemia, low HDL levels or hypertension comprising a step of administering to a human or non-human animal in need thereof an effective amount of a compound according to any one of claims 1 to 17 or a pharmaceutically acceptable salt thereof, or a pharmaceutical formulation or composition as claimed in claim 18.
PCT/EP2012/057964 2011-08-22 2012-05-01 1,4 disubstituted pyrrolidine - 3 - yl -amine derivatives and their use for the treatment of metabolic disorders WO2013026587A1 (en)

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