HRP20040659A2 - 3-(imidazolyl)-2-alkoxypropanoic acids as tafia inhibitors - Google Patents

Description

This invention relates to a series of new derivatives of 3-(imidazolyl)-2-(ω-aminoalkyloxy)-propanoic acid that act as TAFIa inhibitors and are useful in the treatment of diseases.

The basis of the invention

Mammals have developed sophisticated mechanisms to repair vascular injuries, with the aim of maintaining homeostasis in the body. The damaged blood vessel constricts to reduce blood flow to the area, platelets aggregate to reduce blood loss from the area, and fibrinogen is cleaved to yield fibrin, which then polymerizes to form a clot. This clot covers the area of vascular damage, thus preventing blood loss. Polymerized fibrin also provides a temporary matrix that facilitates subsequent repair processes. When the blood vessel is restored, the clot dissolves. The process that leads to clot formation is the coagulation cascade, and the process that leads to its dissolution is the fibrinolysis cascade. Imbalances in blood clotting processes are considered the cause of a large and diverse number of disease states, which are associated with unwanted fibrin accumulation. Fibrin accumulation is determined by a delicate balance between two biochemical cascades in the body. Agents that can mediate the balance between coagulation and fibrinolysis are therefore useful in the treatment of such disease states.

Studies have shown that coagulation and fibrinolysis are linked through the formation of α-thrombin. α-thrombin is the final product of the blood coagulation cascade and is responsible for the transition of fibrinogen to fibrin. In addition to mediating coagulation, α-thrombin also slows the breakdown of blood clots via the serine protease plasmin. The protein that mediates this antifibrinolytic effect of α-thrombin is TAFI (Thrombin Activatable Fibrinolysis Inhibitor).

TAFI is a 60 kDa glycoprotein found in human plasma. It is also known as procarboxypeptidase B, carboxypeptidase B, plasmatic carboxypeptidase B, carboxypeptidase U and carboxypeptidase R. After the initiation of the coagulation cascade, it is translated into the active form, TAFIa, which acts on the fibrin matrix of the emerging blood clot to prevent its dissolution. TAFI circulates in normal plasma at a concentration of about 75 nM in an inactive form. Thrombin converts inactive zymogen into active TAFI (TAFIa), and this reaction is increased about 1250 times by thrombomodulin. Upon activation, TAFIa cleaves C-terminal arginine and lysine residues from the nascent fibrin clot. Removal of these dibasic amino acids from the surface of the fibrin matrix stops clot lysis by inhibiting the binding of key mediators of fibrinolysis: tissue plasminogen activator (tPA) and its substrate, plasminogen, which is a precursor of plasmin. Both tPA and plasminogen contain a structural motif, called the "kringle" domain, that tightly binds C-terminal lysine residues. The removal of these binding sites prevents the formation of a ternary complex between tPA, plasminogen and fibrin, which inhibits the transition of plasminogen to plasmin, thus protecting the clot from rapid degradation.

In the presence of a TAFIa inhibitor, TAFIa will not be able to act on the nascent fibrin clot as described above to inhibit fibrinolysis of the clot. Thus, the TAFIa inhibitor would act by promoting fibrinolysis.

It can be observed that, in pathological conditions when the normal balance between coagulation and fibrinolysis is disturbed, in favor of coagulation, a larger amount of fibrin is present than is normal. Therefore, individuals are more likely to develop one or more conditions that are implicated in thrombus formation. Such individuals may benefit from treatment with a profibrinolytic agent. McKay et al. (Biochemistry 1978, 17, 401) reported testing various compounds as competitive inhibitors of bovine pancreatic carboxypeptidase B. Inhibition was measured as the effectiveness of the inhibitor in protecting the tyrosine and glutamic acid active center of bovine carboxypeptidase B from irreversible alkylation by bromoacetyl-D-arginine or bromoacetaminobutylguanidine. It has been suggested that such inhibitors could act as potentiators of bradykinin. Bovine pancreatic enzymes are very different from those found in human plasma, so it is difficult to expect that inhibitors of one enzyme will inhibit the other. Furthermore, such inhibitors are directed towards very different applications. Accordingly, this publication does not provide guidance on TAFI inhibitors or their utility.

Redlitz et al. (J. Clin. Invest. 1995, 96, 2534) describe the involvement of plasma carboxypeptidase B (pCPB or TAFI) in clot formation. Blood clot lysis was monitored in the presence or absence of pCPB, where it was found that the presence of pCPB slowed clot lysis. To confirm the responsibility of pCPB, two control reactions were performed; in one, the lysis experiment was repeated in the presence of pCPB and the potato carboxypeptidase inhibitor, PCI, while the other was a lysis reaction in the presence of plasma from which pCPB was removed. In both cases, lysis proceeded without inhibition.

Boffa et al. (J. Biol. Chem. 1998, 273, 2127) compared plasmatic and recombinant TAFI and TAFIa, with respect to glycosylation, activation, thermal stability and enzymatic properties. Inhibition constants were determined for three competitive inhibitors: ε-aminocaproic acid (ε-ACA), 2-guanidinoethylmercaptosuccinic acid (GEMSA) and potato carboxypeptidase inhibitor (PCI).

There are a large number of carboxypeptidases (ie enzymes that cleave the C-terminal amino acid of peptides). They can be classified as acidic, neutral or basic, depending on the type of amino acid they cleave. Basic carboxypeptidases cleave arginine, lysine and histidine. TAFIa is a member of a specific subgroup of basic carboxypeptidases. In view of this invention, the above-disclosed inhibitors of Redlitz et al., and Boff et al., respectively, are too weak, non-specific, or otherwise unsuitable for consideration as suitable inhibitors of TAFI in therapeutic use. Furthermore, while the role of TAFIa in clot lysis has been elucidated, there is no suggestion that TAFIa inhibitors could be used in the treatment of the disease.

US-A-5993815 describes the use of a peptide that binds the TAFI zymogen, thereby inhibiting its activation, in the treatment of diseases in which the C-terminal lysine or arginine is cleaved from the intact peptide. Corresponding diseases are arthritis, sepsis, thrombosis, strokes, deep vein thrombosis and myocardial infarctions. The peptide used is an antibody or a functionally active fragment. The peptide should be used in an amount such that it promotes fibrinolysis in vivo.

WO 00/66550 and WO 00/66557 disclose a wide range of compounds useful as carboxypeptidase U inhibitors. Carboxypeptidase U inhibitors are believed to facilitate fibrinolysis and thus the compounds are considered useful in the treatment of thrombotic conditions. There is no data to support this claim, although details of a suitable test are provided.

WO 00/66152 discloses formulations containing a carboxypeptidase U inhibitor and a thrombin inhibitor. Suitable carboxypeptidase U inhibitors are those of WO 00/66550. The formulations are primarily considered useful in the treatment of thrombotic conditions.

WO 01/19836 discloses a series of phosphonate esters and their analogues as carboxypeptidase B inhibitors, useful in the treatment or prevention of thrombotic diseases.

WO 02/14285 discloses a series of α-imidazolylmethyl-ω-aminocarboxylic acids and Nα-(ω-aminoalkyl)-histidine derivatives that act as TAFIa inhibitors. The compounds are primarily considered useful in the treatment of various conditions.

This invention discloses another group of TAFIa inhibitors.

Description of the invention

In a first aspect, the present invention provides compounds of general formula (I)

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where:

n is 0, 1, 2 or 3;

R1 is chosen between

optionally substituted straight-chain or branched C1-6 alkyl groups,

optionally substituted straight-chain or branched C2-6 alkenyl groups,

optionally substituted straight-chain or branched C2-6 alkynyl groups,

arilla,

aromatic heterocycle,

heterocycle or

hydrogen;

wherein the possible substituents in the above mentioned groups (a), (b) and (c) are selected from C3-7 cycloalkyl, aryl, aromatic heterocycle, heterocycle, OR10, NR10R11, S(O)pR10, OC(O)R11, CO 2 R 10 , CONR 10 R 11 , SO 2 NR 10 R 11 , halo and NHSO 2 R 10 , and p is 0, 1 or 2;

R 2 , R 3 , R 4 , R 6 , R 7 and R 9 are each independently selected from hydrogen and straight or branched C 1-6 alkyl, optionally substituted with OR 10 or halo;

R 5 and R 8 are each independently selected from hydrogen and straight or branched C 1-6 alkyl, optionally substituted with OR 10 or halo, or together form a C 2-6 alkylene chain;

R 10 and R 11 are each independently selected from hydrogen and straight or branched C 1-6 alkyl;

aryl is a 6-14 membered aromatic monocyclic or fused polycyclic carbocyclic group, optionally substituted with one or more groups selected from R12, halo, OR13, NR13R14, NR13CO2R12, CO2R13, NR13SO2R12, CN, haloalkyl, O(haloalkyl), SR13, S (O)R12, SO2R12, OC(O)R13, SO2NR13R14, C(O)NR13R14, C3-7 cycloalkyl, O(C3-7 cycloalkyl), R15 and OR15, and R12 is straight or branched C1-6 alkyl, while R13 and R14 are independently selected from hydrogen and straight-chain or branched C1-6 alkyl, while R15 is phenyl, optionally substituted with R12, OR13, halo or haloalkyl;

aromatic heterocycle is a 5-7 membered aromatic ring containing from 1 to 3 heteroatoms, independently selected from O, S and N, wherein said ring is optionally substituted by one or more groups selected from OR13, NR13R14, CO2R13, NR13CO2R12, R12, halo, CN, haloalkyl, O(haloalkyl), SR13, S(O)R12, SO2R12, OC(O)R13, NR13SO2R12, SO2NR13R14 and C(O)NR13R14; And

heterocycle is a 3-8-membered ring containing from 1 to 3 heteroatoms, independently selected from O, S and N, wherein said ring is saturated or partially saturated, and is optionally substituted by one or more groups selected from OR13, NR13R14, CO2R13, NR13CO2R12, R12, halo, CN, haloalkyl, O(haloalkyl), SR13, S(O)R12, SO2R12, OC(O)R13, NR13SO2R12, SO2NR13R14 and C(O)NR13R14,

or its tautomers or pharmaceutically acceptable salts or solvates of said compound or tautomer.

As used here:

halo includes fluoro, chloro, bromo and iodo groups.

haloalkyl includes monohaloalkyl, polyhaloalkyl and perhaloalkyl, such as 2-bromoethyl, 2,2,2-trifluoroethyl, chlorodifluoromethyl and trichloromethyl.

unless otherwise indicated, alkyl includes straight and branched chain alkyl.

It is understood that in the compounds according to the general formula (I), the groups R1 and C(R2)(R3)(amino acid) can be attached to any atom of the imidazole ring that can form a covalent bond, so it is not intended that the general formula be interpreted as a limitation R1 groups at C2- and N3-positions, nor C(R2)(R3)(amino acid) groups at C4- and C5-positions. Furthermore, it is understandable that two groups cannot be attached to the same atom of the imidazole ring, and that only one of the nitrogen atoms (designated as N1 by convention) of the imidazole ring can form a covalent bond. Thus, 1,2- substitution patterns are possible; 1,4-; 1.5-; 2.4- and 2.5. When the imidazole is 2,4- or 2,5-substituted, a hydrogen atom is attached to the N1-position.

Some compounds of formula (I) may exist in more than one tautomeric form. If the imidazole of the general formula (I) is substituted at the 2- and 4-position, the 2,4-disubstituted imidazole can tautomerize to form the corresponding 2,5-disubstituted imidazole. Furthermore, when the compound includes an aromatic heterocycle substituted with a hydroxyl group, it may exist as a "keto"-tautomer. The tautomeric relationship between 2-hydroxypyridine and 2-pyridone is a well-known example of this phenomenon. All such tautomers of compounds of formula (I), including mixtures thereof, are included within the scope of the present invention.

Compounds of formula (I) contain one or more asymmetric carbon atoms (chiral centers) and can therefore exist in two or more optical stereoisomeric forms, such as enantiomers, diastereomers and epimers. When the compounds of formula (I) contain a carbon-carbon double bond, cis (Z) / trans (E) stereoisomers can also occur. All such individual stereoisomers of compounds of formula (I) and mixtures thereof, including racemates, are included within the scope of this invention.

The individual stereoisomers can be isolated from the mixture by conventional techniques, such as, for example, fractional crystallization or chromatography of the mixture of compounds or its corresponding salt or derivative. Specifically, individual enantiomers of compounds of formula (I) can be prepared by resolution, such as HPLC of the appropriate racemate, using an appropriate chiral support, or by fractional crystallization of diastereoisomeric salts resulting from reaction of the appropriate racemate with a suitable optically active acid or base. Individual enantiomers can also be obtained from the corresponding pure intermediate, prepared by such a resolution method. These general principles are discussed in more detail in J. Jacques and A. Collet ("Enantiomers, Racemates and Resolutions", Wiley, NY, 1981) and W. Liu ("Handbook of Chiral Chemicals", D. Ager (ed.), M .Dekker, NY, 1999; Chapter 8).

It can be noted that the compounds of formula (I) have both acidic and basic functional groups. therefore, apart from the uncharged form shown by the general formula, they can also exist as internal salts (zwitterions). Furthermore, they can form pharmaceutically acceptable salts with acids and bases. Such zwitterions and salts are included within the scope of the present invention.

A pharmaceutically acceptable salt of a compound of formula (I) may be prepared by mixing a solution of a compound of formula (I) with the desired acid or base, as appropriate. The salt can be precipitated out of solution and collected by filtration, or it can be collected by evaporation of the solvent. Salts can also be prepared by ion exchange, such as by equilibrating a solution of a compound of formula (I) with a suitable ion exchange resin. Ion exchange can also be used to convert one form of a salt of a compound of formula (I), such as a non-pharmaceutically acceptable acid-base salt, into another salt form. These procedures are generally well known in the art. Suitable acid addition salts are formed with acids that form non-toxic salts, examples of which are hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, nitrate, phosphate, hydrogenphosphate, acetate, maleate, fumarate, lactate, tartrate, citrate, gluconate, succinate, saccharate, benzoate , methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate and pamoate. Suitable base salts are formed with bases that form non-toxic salts, examples being sodium, potassium, aluminum, calcium, magnesium, zinc and diethanolamine. For a review of pharmaceutically acceptable salts, see Berge et al. (J. Pharm. Sci., 1977, 66, 1).

The compounds of formula (I) can form pharmaceutically acceptable solvates (including hydrates). These solvates are also included within the scope of this invention.

Compounds of formula (I) may exist in one or more crystalline forms. These polymorphs, including mixtures thereof, are also included within the scope of this invention.

The scope of this invention further includes prodrugs of compounds of formula (I), i.e. pharmaceutically acceptable derivatives of compounds in which one or more of the previously mentioned functional groups are modified so that they are converted in vivo into the compounds from which they originate. Suitable prodrugs are discussed in Drugs of Today 1983, 19, 499-538 and Annual Reports in Medicinal Chemistry 1975, 10, 306-326.

The absolute stereochemistry of the compounds of formula (I) may be as shown in formulas (IA) and (IB) below. By convention, the absolute stereochemistry of the chiral center (IA) is denoted as "S" and that of (IB) as "R". Compounds of formula (IA) are particularly preferred.

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Preferred compounds of formula (I) include those in which the imidazole is substituted at the C2 or C4 positions with a C(R2)(R3)(amino acid) group to give compounds of formula (IC) and (ID), respectively. Especially preferred are the compounds of formula (I) where R1 is attached at the C4 position of the imidazole unit, and the group C(R2)(R3)(amino acid) is attached at the C2 position, so that it gives a 2,4-disubstituted imidazole of the formula (IC1) or where R1 is attached at the N1 position of the imidazole unit, and the group C(R2)(R3)(amino acid) is attached at the C4 position, giving a 1,4-disubstituted imidazole of the formula (ID1). The most preferred are the compounds of formula (I) in which R1 is attached at the N1 position of the imidazole unit, and the group C(R2)(R3)(amino acid) is attached at the C4 position, thus giving a 1,4-disubstituted imidazole of the formula (ID1).

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Preferably, n is 0 or 1. Most preferably, n is 0.

It is preferred that R1 is hydrogen, aryl or a C1-6 alkyl or C2-6 alkenyl group, optionally substituted with one or more groups selected from C3-7 cycloalkyl, aryl, aromatic heterocycle, heterocycle, OR10, NR10R11, S(O)pR10 , OC(O)R11, CO2R10, CONR10R11, SO2NR10R11, halo and NHSO2R10. Even more preferably, R1 is hydrogen, aryl, C2-6 alkenyl or C1-6 alkyl, optionally substituted with one or more groups selected from C3-7 cycloalkyl, aryl, aromatic heterocycle, OR10, CO2R10, halo and NHSO2R10. More preferably, R 1 is hydrogen, aryl or a C 1-6 alkyl group, optionally substituted with a group selected from C 3-7 cycloalkyl, aryl, aromatic heterocycle, OR 10 , CO 2 R 10 and NHSO 2 R 10 . Even better, R 1 is hydrogen, aryl or a C 1-6 alkyl group, optionally substituted with a group selected from cyclohexyl and aryl. Even better, R 1 is hydrogen or C 1-3 alkyl. Preferably, R1 is hydrogen.

Preferably, R 2 and R 3 are each independently selected from hydrogen and C 1-6 alkyl. Preferably, R2 and R3 are hydrogen.

Preferably, R4 is hydrogen or C1-6 alkyl. More preferably, R 4 is hydrogen.

Preferably, R 6 , R 7 and R 9 are each independently selected from hydrogen and C 1-6 alkyl. Even more preferably, R 6 , R 7 and R 9 are each independently selected from hydrogen and C 1-3 alkyl. Even more preferably, R 6 , R 7 and R 9 are each independently selected from hydrogen and methyl. Preferably R6, R7 and R9 are hydrogen.

When R5 and R8 do not form a C2-6 alkylene bond, it is preferable that R5 is hydrogen or C1-6 alkyl, preferably hydrogen or C1-3 alkyl, even better hydrogen or methyl, and preferably methyl, while R8 is preferably hydrogen or C1- 6 alkyl, preferably hydrogen or C1-3 alkyl, even more preferably hydrogen or methyl, and most preferably hydrogen.

When R5 and R8 form a C2-6 alkylene bond, it is preferable that the bond is a C2-3 alkylene bond, and even better a C2 alkylene bond.

Preferably, R 10 and R 11 are each independently selected from hydrogen and C 1-3 alkyl. Even better, R 10 and R 11 are each independently selected from hydrogen and methyl.

Aryl includes optionally substituted phenyl, naphthyl, anthracenyl and phenanthrenyl. Preferably, the aryl is phenyl or naphthyl, optionally substituted with 1-3 groups selected from R12, halo, OR13, NR13R14, NR13CO2R12, CO2R13, NR13SO2R12, CN, haloalkyl, O(haloalkyl), SR13, S(O)R12, SO2R12 , OC(O)R13, SO2NR13R14, C(O)NR13R14, C3-7 cycloalkyl, O(C3-7 cycloalkyl), R15 and OR15. It is more preferred that the aryl is phenyl, optionally substituted with C1-6 alkyl, halo, O(C1-6 alkyl), CF3, C3-7 cycloalkyl, O(C3-7 cycloalkyl), R15 or OR15, and R15 is phenyl, by optionally substituted with C1-6 alkyl, halo, O(C1-6 alkyl) or CF3. More preferably, the aryl is phenyl, optionally substituted with C 1-6 alkyl, Cl, F, O(C 1-6 alkyl) or CF 3 . Preferably the aryl is phenyl.

Preferably, the aromatic heterocycle is a 5- or 6-membered aromatic ring containing 1 or 2 heteroatoms, each independently selected from O, S and N, including optionally substituted furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl , imidazolyl, oxadiazolyl, thiadiazolyl, triazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl and triazinyl, optionally substituted with 1-3 groups selected from OR13, NR13R14, CO2R13, NR13CO2R12, R12, halo, CN, haloalkyl, O(haloalkyl), SR13 , S(O)R12, SO2R12, OC(O)R13, NR13SO2R12, SO2NR13R14 and C(O)NR13R14. Preferably, the aromatic heterocycle is a 5- or 6-membered aromatic ring containing 1 or 2 heteroatoms, each independently selected from O, S and N, optionally substituted with 1-3 groups selected from OR13, NR13R14, CO2R13, NR13CO2R12, R12, halo, CN, haloalkyl, O(haloalkyl), SR13, S(O)R12, SO2R12, OC(O)R13, NR13SO2R12, SO2NR13R14 and C(O)NR13R14. Preferably, the aromatic heterocycle is an unsubstituted 5- or 6-membered aromatic ring containing 1 to 2 heteroatoms, each independently selected from O, S, and N.

Preferably, the heterocycle is a 3- to 8-membered ring containing 1 to 2 heteroatoms, each of which is independently selected from O, S, and N, and said ring is saturated or partially saturated, optionally substituted with 1 to 3 groups, selected between OR13, NR13R14, CO2R13, NR13CO2R12, R12, halo, CN, haloalkyl, O(haloalkyl), SR13, S(O)R12, SO2R12, OC(O)R13, NR13SO2R12, SO2NR13R14 and C(O)NR13R14. It is more preferred that the heterocycle is a 5- or 6-membered ring containing 1 to 2 heteroatoms, each of which is independently selected from O, S and N, and said ring is saturated or partially saturated, optionally substituted with 1 to 3 groups, selected between OR13, NR13R14, CO2R13, NR13CO2R12, R12, halo, CN, haloalkyl, O(haloalkyl), SR13, S(O)R12, SO2R12, OC(O)R13, NR13SO2R12, SO2NR13R14 and C(O)NR13R14. Preferably, the heterocycle is an unsubstituted 5- to 6-membered ring containing 1 to 2 heteroatoms, each independently selected from O, S, and N, said ring being saturated or partially saturated, including oxiranyl, azetidinyl, tetrahydrofuranyl, thiolanyl , pyrrolidinyl, dioxolanyl, dihydropyranyl, tetrahydropyranyl, morpholinyl, piperidinyl and piperazinyl.

Preferred compounds of this invention are:

(2S)-(-)-2-(2-aminoethoxy)-3-(1-phenyl-1H-imidazol-4-yl)-propanoic acid (Example 6);

(2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]-propanoic acid (Example 15);

(2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-[1-phenyl-1H-imidazol-4-yl]-propanoic acid (Example 17);

(2S)-2-{[(2S)-2-aminopropyl]oxy}-3-[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]-propanoic acid (Example 34);

(2S)-2-(2-aminoethoxy)-3-(1H-imidazol-4-yl)-propanoic acid (Example 50);

(2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-[1H-imidazol-4-yl]-propanoic acid (Example 51) and

(2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-[1-(2-pyridinyl)-1H-imidazol-4-yl]-propanoic acid (Example 52).

Particularly preferred is (2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-[1H-imidazol-4-yl]-propanoic acid (Example 51).

Compounds of formula (I) are TAFIa inhibitors. Inhibition of TAFI can be demonstrated using an assay based on the method of Boff et al. (J. Biol. Chem. 1998, 273, 2127), as described in more detail below. The activity of compounds is characterized by the calculated Ki value. In general, the compounds of the present invention have a Ki value of about 10 μM or less. Better compounds have a Ki value of 1 μM or less, or even 100 nM and less. The most potent compounds have a Ki value of 25 nM or less.

The compounds of formula (I) are selective for TAFIa against other carboxypeptidases, especially carboxypeptidase N (CPN). Unwanted inhibition of CPN is considered the most likely cause of unwanted side effects in clinical use. Selectivity can be expressed as the ratio of Ki for TAFIa and Ki for CPN. Generally, the compounds of this invention have a selectivity ratio of at least 5. Better compounds have a selectivity ratio of at least 10. The most selective compounds have a selectivity ratio of at least 50.

Compounds of formula (I) can be prepared according to the general procedures described below and in the Examples and Preparations section. These procedures provide an additional aspect to the present invention. However, one skilled in the art will appreciate that the compounds of this invention can be prepared by methods other than those described herein, by adaptation of the methods described herein, and/or by adaptation of many methods known in the art. It is understandable that the synthetic transformation procedures mentioned here can be carried out in different order, with the aim of efficiently assembling the desired compounds. A skilled chemist will exercise his judgment and skill to arrive at the most efficient sequence of reactions for the synthesis of a given target compound.

It will be clear to one skilled in the art that sensitive functional groups should be protected, or deprotected, during the synthesis of a compound of the present invention. This can be achieved by conventional techniques, for example, those described in T.W. Greene and P.G.M. Wuts ("Protective Groups in Organic Synthesis", 3rd ed., Wiley-Interscience, NY, 1999).

Compounds of formula (I) can be prepared from the corresponding esters of formula (II) (where P 1 is a lower alkyl group, a benzyl group, or any carboxyl protecting group).

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Preferably, P1 is a lower alkyl group, such as methyl or ethyl, in which case suitable conditions for this step include treatment with NaOH in dioxane for 1-3 days.

Compounds of formula (II) can be prepared from the corresponding protected amines of formula (III) (wherein P 2 is a tert-butyloxycarbonyl, benzyloxycarbonyl or fluorenylmethyloxycarbonyl group, or some other protecting group for the amine). When R9 is H, preparation involves only unshielding. When R9 is different from H, an additional step is required to introduce R9, such as a reductive amination reaction.

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Alternatively, compounds of formula (III) can be converted to the corresponding acids (IV) prior to deprotection from the amine to give compounds of formula (I).

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Compounds of formula (III) can be prepared from imidazolacetic acid derivatives of formula (V), where X is a leaving group, such as a chlorine, bromine or iodine atom, or a methanesulfonate or trifluoromethanesulfonate group, by reaction with an alcohol of formula (VI).

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Compounds of formula (V) can be prepared from the corresponding hydroxy acid derivatives of formula (VII) or, when X is Br, by direct halogenation of esters of formula (VIII).

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Compounds of formula (VI), (VII) and (VIII) are known or can be prepared by methods analogous to those used in the preparation of such known compounds.

Compounds of formula (III) may alternatively be prepared from α-hydroxyimidazolacetic acid derivatives of formula (VII) by reaction with a compound of formula (IX), where Y is a leaving group, such as a chlorine, bromine or iodine atom, or a methanesulfonate or trifluoromethanesulfonate group.

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Compounds of formula (II) where neither R8 nor R9 is hydrogen can also be prepared from compounds of formula (X). When R 3 is hydrogen, the transformation can be achieved by hydrogenation in the presence of a suitable catalyst. When R 3 is other than hydrogen, the transformation can be achieved using a reagent such as R 3 -M, where M is a metal, eg lithium or magnesium, in the presence of a copper(I) salt.

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Compounds of formula (X) can be prepared from compounds of formula (XI) by dehydration. The transformation can be achieved using, for example, methanesulfonyl chloride and a tertiary amine.

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Compounds of formula (XI) can be prepared by an aldol-type reaction between an alkoxy ester of formula (XII) and an aldehyde or ketone of formula (XIII) in the presence of a strong base, such as lithium diisopropylamide.

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Compounds of formula (XIII) are generally known, or can be prepared by methods analogous to those disclosed. Compounds of formula (XII) can be prepared by reacting the corresponding amino alcohols R8R9NC(R6)(R7)(CH2)nC(R4)(R5)OH with bromoacetate BrCH2CO2P1 in the presence of a base, such as sodium hydride.

Compounds of formula (I) in which R9 is hydrogen can alternatively be prepared by hydrolysis of lactams of formula (XIV).

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Compounds of formula (XIV) can be prepared from corresponding unsaturated compounds of formula (XV). When R3 is hydrogen, the transformation can be achieved by hydrogenation in the presence of a suitable catalyst. When R 3 is other than hydrogen, the transformation can be achieved using a reagent such as R 3 -M, where M is a metal, eg lithium or magnesium, in the presence of a copper(I) salt.

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Compounds of formula (XV) can be prepared by dehydrating alcohols of formula (XVI).

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Compounds of formula (XVI) can be prepared by reacting an aldehyde or ketone of formula (XIII) with a lactam of formula (XVII) in the presence of a strong base, such as lithium diisopropylamide.

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Compounds of formula (XVII) can be prepared from amino alcohols of formula (XVIII) by reaction with chloroacetyl chloride. Compounds of formula (XVIII) are generally known, or can be prepared by adaptation of generally known procedures.

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When R 8 is H, the preceding procedure can be problematic, especially in the step involving the reaction of compounds of formula (XIII) and (XVII). In general, in this case it is customary to use a protected aminoalcohol of the formula (XVIIIa), where P3 is a nitrogen protecting group. A particularly useful embodiment of P3 is a 4-methoxybenzyl group. It can be removed after treatment of the intermediate (XIVa) by treatment with cerium-ammonium-nitrate.

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When R 1 is H, it may be necessary or customary to protect the imidazole as a trityl derivative. Accordingly, when R 1 is H, compounds of formula (XIX), (XX) or (XXI) may be treated by the foregoing procedures to give compounds of formula (XXII) which, after deprotection, give compounds of formula (III).

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This route can also be useful for the preparation of some compounds of formula (I), when R1 is attached to the N1 position of the imidazole ring. Compounds of formula (III) wherein R 1 is H can be alkylated or arylated to give compounds of formula (III) wherein R 1 is other than H and attached at the N 1 position.

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When R 1 is an alkyl, alkenyl or alkynyl group, it can be introduced in an alkylation reaction. Suitable conditions for this step include treatment with 1.1 eq of cesium carbonate and 1.1 eq of alkylating agent in N,N-dimethylformamide, or sodium hydride and 1.1 eq of alkylating agent in THF. Suitable alkylating agents include R1-Cl, R1-Br, R1-I, R1-OSO2CH3 and R1-OSO2CF3. When R1 is aryl or an aromatic heterocycle, it can be introduced in an arylation reaction. Suitable conditions for this step include treatment with 2 eq of aryl-B(OH)2 or aromatic heterocycle-B(OH)2 in the presence of 1.5 eq of copper acetate, 2 eq of pyridine, air, and a 4 Å molecular sieve.

For compounds of formula (I) in which the imidazole is 2,4- or 2,5-disubstituted, it may also be customary or necessary to use a protecting group at the N1 position.

Compounds of formula (I) are useful as therapeutic agents. The compounds will generally be formulated to be suitable for administration to individuals by the chosen route. In a further aspect, therefore, this invention provides a pharmaceutical composition containing a compound of formula (I) or its stereoisomer, tautomer or a pharmaceutically acceptable salt, solvate or prodrug and a pharmaceutically acceptable excipient, diluent or carrier, chosen with regard to the intended method of administration and standard pharmaceutical practice. For example, compounds of formula (I) can be administered orally, buccally or sublingually in the form of tablets, capsules, ovules, elixirs, solutions or suspensions. These formulations may contain flavoring or coloring agents, and may be adapted for immediate, delayed, sustained, sustained, pulsed, or controlled release administration.

Tablets may contain excipients such as microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, dibasic calcium phosphate and glycine, disintegrants such as starch (preferably corn, potato or tapioca starch), sodium starch glycolate, cross-linked sodium carboxymethylcellulose and some complex silicates , and granulation binders such as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin and acacia. Furthermore, lubricants such as magnesium stearate, stearic acid, glyceryl behenate and talc may be added.

Solid preparations of a similar type can also be used as fillers in gelatin capsules. Preferred excipients in this regard include lactose, starch, cellulose and their derivatives, milk sugar and high molecular weight polyethylene glycols.

For solutions, suspensions and elixirs, the compounds of formula (I) can be combined with various agents for adding flavor or color, with emulsifying and/or suspending agents, and with diluents such as water, ethanol, propylene glycol and glycerin and their combinations.

The compounds of formula (I) can also be administered in the form of a capsule of soft or hard gelatin filled with a solution or suspension. Such capsules are generally made from gelatin, glycerin, water and sorbitol. Hard capsules differ from soft capsules in that they contain less water and thus have a correspondingly tighter shell. Additional excipients suitable for use in such capsules include propylene glycol, ethanol, water, glycerol and edible oils.

Compounds of formula (I) can also be administered parenterally, for example, intravenously, intraarterially, intraperitoneally, intrathecally, intraventricularly, intraurethrally, intracranially, intramuscularly or subcutaneously. Such administration can be as a single bolus injection or as a short-term or long-term infusion. For such parenteral administration, it is preferable to formulate the compounds as a sterile solution in water or other suitable solvent or solvent mixture. The solution may also contain other substances, such as salts, especially sodium chloride, and sugars, especially glucose and mannitol, to make the solution isotonic with blood; buffering agents, such as acetic, citric and phosphoric acids and their sodium salts, so that the pH of the solution is between 3 and 9; and preservatives. Preparation of suitable parenteral formulations under sterile conditions is readily accomplished by standard pharmaceutical techniques well known to those skilled in the art.

Compounds of formula (I) may also be administered intranasally or by inhalation, and are typically administered as a dry powder inhaler or aerosol spray from a pressurized container, pump, spray or nebulizer, with or without the use of a suitable filler such as dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane , a hydrofluoroalkane, such as 1,1,1,2,-tetrafluoroethane (HFA 134ATM) or 1,1,1,2,3,3,3-heptafluoropropane (HFA 227EATM), carbon dioxide or other suitable gas. In the case of pressurized aerosols, the dosage unit can be determined by fitting a valve that will deliver a measurable quantity. The container, pump, spray or atomizer under pressure can contain a solution or suspension of the active ingredient, for example with a mixture of ethanol and filler as a solvent, with an additional lubricant, for example sorbitan-trioleate. Capsules and containers (made, for example, of gelatin) for use in an inhaler or insufflator may be formulated to contain a powder mixture of a compound of formula (I) and a suitable powder base, such as lactose or starch.

Alternatively, the compounds of formula (I) can be administered vaginally or rectally in the form of suppositories or pessaries. Compounds of formula (I) can also be administered dermally or transdermally, for example, using skin patches.

Alternatively, the compounds of formula (I) can be applied topically in the form of a gel, hydrogel, lotion, solution, cream, ointment or powder. Suitable ointments may contain the active ingredient suspended or dissolved in, for example, a mixture with one or more of the following: mineral oil, liquid petroleum ether, white petroleum ether, propylene glycol, polyoxyethylene-polyoxypropylene compound, emulsifying wax and water. Suitable lotions and creams may contain the active ingredient suspended or dissolved in, for example, a mixture with one or more of the following: mineral oil, sorbitan monostearate, polyethylene glycol, liquid paraffin, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2- octyldodecanol, benzyl alcohol and water.

Alternatively, the compounds of formula (I) can be administered by the ocular route. For ophthalmic use, the compounds may be formulated as micronized suspensions in isotonic, sterile, pH-adjusted saline, or, preferably, as solutions in isotonic, sterile, pH-adjusted saline, optionally in combination with a preservative such as benzylalkonium chloride. Alternatively, they can be formulated as an ointment, such as petroleum ether.

The compounds of formula (I) can also be used in combination with cyclodextrin. Cyclodextrins are known to form inclusion and non-inclusion complexes with drug molecules. The formation of drug-cyclodextrin complexes can modify the solubility, dissolution rate, bioavailability and/or stability properties of the drug molecule. Drug-cyclodextrin complexes are generally useful for most dosage forms and routes of administration. As an alternative to direct complexation with the drug, cyclodextrin can be used as an auxiliary additive, eg as a carrier, diluent or solubilizer. Alpha-, beta- and gamma-cyclodextrins are most commonly used, suitable examples being described in WO 91/11172, WO 94/02518 and WO 98/55148.

Since the compounds of formula (I) are inhibitors of TAFIa, they are useful as therapeutic agents in pathological conditions in which inhibition of TAFIa is desirable. In a further aspect, therefore, the present invention provides the use of a compound of formula (I) or a stereoisomer, tautomer, solvate, pharmaceutically acceptable salt or prodrug thereof for medicinal purposes. In particular, this invention provides the use of a compound of formula (I) or a stereoisomer, tautomer, solvate, pharmaceutically acceptable salt or prodrug thereof in the preparation of a medicament for the treatment or prevention of a condition selected from thrombotic conditions, atherosclerosis, adhesions, skin scars, cancer, fibrotic conditions, inflammatory diseases and conditions that improve by maintaining or increasing bradykinin levels in the body. The usefulness of TAFIa inhibitors in the treatment of thrombotic conditions stems from their ability to promote fibrinolysis without interfering with coagulation. In most clinically relevant conditions, thrombus formation is subacute, i.e. the thrombus forms slowly. Conventional antithrombotic agents block the coagulation pathway and thereby prevent thrombus growth, but an undesired consequence is blocking the clotting response to vascular damage, resulting in increased bleeding. By promoting fibrinolysis, TAFIa inhibitors accelerate the dissolution of developing thrombus without interfering with the coagulation response. Accordingly, one preferred embodiment of the present invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt, solvate or prodrug thereof in the preparation of a medicament for the treatment of a thrombotic condition selected from myocardial infarction, deep vein thrombosis, stroke, juvenile stroke, cerebral infarction, cerebral thrombosis, cerebral embolism, peripheral vascular disease, angina and other forms of acute coronary syndromes, disseminated intravascular coagulation, sepsis, pulmonary embolism, embolic events secondary to cardiac arrhythmias and prevention of cardiovascular events after surgical revascularization or intervention, or to improve transplant outcomes organs by reducing blood clotting and thereby preserving organ function. Cardiovascular events after interventional surgery include conditions such as restenosis or reocclusion after an intervention such as percutaneous transluminal coronary angioplasty, grafting, stenting, coronary bypass grafting, or other forms of surgical revascularization or intervention. Dispersed intravascular coagulation includes all conditions resulting from intravascular activation of the coagulation process. This can occur acutely through the release of procoagulant substances (e.g. obstetric emergencies, snakebite, blunt trauma), abnormal blood contact (e.g. infections, burns, extracorporeal circulation, transplantation) or the formation of procoagulants in the blood (transfusion reactions, leukemia). ; or chronic (eg toxemia, malignant hypertension, severe liver cirrhosis). Deep vein thrombosis also includes a condition known as "economy class syndrome," where clots occur in people who have to endure cramping conditions for a period of time, such as sitting in economy class seats on an airplane.

The role of thrombus formation in the pathophysiology of atherosclerosis has recently been clarified by several independent groups. Non-occlusive rhombi not only restrict blood flow, thus leading to myocardial ischemia and angina pectoris, but due to incomplete endogenous lysis, they can become embedded in the arterial wall as solid plaque material, thereby accelerating the atherosclerosis process. Long-term use of TAFIa inhibitors promotes the lysis of developing thrombus and thus provides a safe and effective treatment that relieves the symptoms of angina pectoris, while at the same time slowing down the progression of the underlying disease. The usual treatment of myocardial ischemia in clinically stable coronary artery disease is basically aimed at reducing cardiac work and increasing blood flow. Such approaches reduce myocardial ischemia and thereby improve the quality of life. However, these strategies have little effect on the pathogenesis of coronary atherosclerosis, which is a chronic process of continuous remodeling of the vascular tree in response to varying degrees of vascular damage. Accordingly, another preferred embodiment of the present invention provides the use of compounds of formula (I) or pharmaceutically acceptable salts, solvates and prodrugs thereof in the preparation of a medicament for the treatment or prevention of atherosclerosis, including atherosclerosis as a consequence of peripheral vascular disease, insulin resistance and syndrome X, and further including myocardial ischemia and angina pectoris resulting from atherosclerosis. Atherosclerosis includes both primary and secondary coronary artery disease, in which atherosclerosis restricts blood flow to the heart. Primary prevention of coronary artery disease involves preventing the onset of ischemic complications, such as myocardial infarction, in patients who do not have a history of coronary artery disease but have one or more risk factors. Secondary prevention of coronary artery disease involves preventing ischemic complications in patients with established coronary artery disease, such as patients who have already had a myocardial infarction. Syndrome X is a term often used to group several interrelated diseases. The first stage of syndrome X consists of insulin resistance, abnormal cholesterol and triglyceride levels, obesity and hypertension. Any of these conditions can be used to diagnose the onset of syndrome X. The disease may progress so that one condition precedes the development of the others in the group. For example, insulin resistance is associated with high lipid levels, hypertension and obesity. The disease then develops in a cascade, with the development of each additional condition increasing the risk of developing more serious diseases. This can lead to the development of diabetes, kidney or heart disease. These diseases can lead to stroke, myocardial infarction and organ failure. Atherosclerosis is common in patients with syndrome X.

TAFIa inhibitors are also effective in preventing the formation of adhesions in the body. Most surgical procedures and physical trauma result in bleeding into body cavities between tissues. The blood that collects in these places coagulates, forming fibrin-rich clots. These clots bridge the cracks between adjacent tissues and act as focal points for the accumulation of inflammatory cells and fibroblasts. Infiltrating fibroblasts coat a collagen-rich extracellular matrix, which reinforces tissue adhesion and creates tight junctions that can then limit movement. Adhesions are characterized by location and can occur after any surgery, eg abdominal, orthopedic, neurological, cardiovascular and ocular. Inappropriate tissue adhesion after surgery or trauma is the main reason that leads to different outcomes, such as "aches", "stabs", local inflammations, limitation of mobility, pain, intestinal obstruction and, sometimes, in the most severe cases, death. Infertility can occur during gynecological operations. Additional fibrin-rich clots are involved in skin scarring and restenosis. Without being bound by any particular theory, it is believed that adhesion formation may be enhanced when insufficient fibrinolysis leads to increased and sustained clot formation. Treatment with a TAFIa inhibitor before and/or after surgical intervention can promote fibrinolysis of fibrin-rich clots and thus inhibit the formation of thrombus, adhesions and their stabilization, which slows down the development of adhesions. A TAFIa inhibitor, applied either locally as a topical application or systemically, has been observed to improve the outcome of surgical procedures. Furthermore, the use of TAFIa inhibitors may be useful in the treatment of adhesions resulting from other forms of non-surgical physical trauma that have caused internal bleeding. Examples of such traumas may include sports injuries or any other injuries that result in lacerations, cuts, contusions or stiffening of tissues. Accordingly, another advantageous embodiment of this invention is the use of compounds of formula (I) and their pharmaceutically acceptable salts, solvates and prodrugs in the preparation of a medicament for the treatment or prevention of adhesions or skin scars.

TAFIa inhibitors are also effective in inhibiting tumor maturation, progression and metastasis. Without being bound to any particular theory, the homeostatic system is thought to be involved in cancer pathology at multiple levels, including neovascularization, detachment of cells from the primary tumor, invasion into the bloodstream, adhesion to vessel walls, and growth at the metastatic site. The efficacy of TAFIa inhibitors is thought to derive from their ability to reduce fibrin deposition around solid tumors, thus inhibiting the above processes. Accordingly, another advantageous embodiment of this invention is the use of compounds of formula (I) and their pharmaceutically acceptable salts, solvates and prodrugs in the preparation of a drug for the treatment or prevention of cancer.

TAFIa inhibitors are effective in treating any condition that contributes to fibrosis. Relevant fibrotic conditions include cystic fibrosis, pulmonary fibrotic diseases such as chronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome (ARDS), fibromuscular dysplasia and fibrotic lung disease, as well as fibrin deposition in the eye during eye surgery. Accordingly, another advantageous embodiment of this invention is the use of compounds of formula (I) and their pharmaceutically acceptable salts, solvates and prodrugs in the preparation of a drug for the treatment or prevention of fibrotic diseases, in particular for the treatment or prevention of fibrotic conditions selected from cystic fibrosis, pulmonary fibrotic diseases, chronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome (ARDS), fibromuscular dysplasia, fibrotic lung diseases and fibrin deposition in the eye during eye surgery.

TAFIa inhibitors are effective in the treatment of inflammation, inflammatory diseases such as asthma, arthritis, endometriosis, inflammatory bowel disease, psoriasis and atopic dermatitis, as well as neurodegenerative diseases such as Alzheimer's and Parkinson's disease. Accordingly, another advantageous embodiment of this invention is the use of compounds of formula (I) and their pharmaceutically acceptable salts, solvates and prodrugs in the preparation of a drug for the treatment or prevention of inflammation, inflammatory diseases such as asthma, arthritis, endometriosis, inflammatory bowel diseases , psoriasis and atopic dermatitis, as well as neurodegenerative diseases, such as Alzheimer's and Parkinson's disease.

TAFIa binds to and cleaves bradykinin (Tan et al., Biochemistry 1995, 34, 5811). There are several conditions that are known to improve with elevated bradykinin levels, such as hypertension, angina, heart failure, pulmonary hypertension, kidney and other organ failure. Accordingly, another advantageous embodiment of this invention is the use of compounds of formula (I) and their pharmaceutically acceptable salts, solvates and prodrugs in the preparation of a drug for the treatment or prevention of conditions that improve when bradykinin levels are maintained or increased.

In yet another aspect, the present invention provides a method of treating or preventing thrombotic conditions, atherosclerosis, adhesions, skin scars, cancer, fibrotic conditions, inflammatory diseases, and conditions that improve when bradykinin levels are maintained or increased, comprising administering a therapeutically effective amount of a compound of formula ( I) or its stereoisomer, tautomer or pharmaceutically acceptable salt, solvate or prodrug in a patient in need of such treatment.

One preferred embodiment of this invention provides a method of treating or preventing thrombosis, especially myocardial infarction, deep vein thrombosis, stroke, juvenile stroke, cerebral infarction, cerebral thrombosis, cerebral embolism, peripheral vascular disease, angina and other forms of acute coronary syndromes, diffuse intravascular coagulation, sepsis, pulmonary embolism, embolic events secondary to cardiac arrhythmias and prevention of cardiovascular events after surgical revascularization or intervention, which includes the administration of a therapeutically effective amount of a compound of formula (I) or its stereoisomer, tautomer or pharmaceutically acceptable salt, solvate or prodrug to a patient in whom such treatment is necessary. Individuals with thrombotic conditions amenable to treatment in accordance with the present invention include patients with conditions associated with hypercoagulation, such as factor V mutation, antithrombin III deficiency, heparin cofactor II deficiency, protein C, protein S, and polycythemia vera, as well as those exhibiting homocystinemia or homocystinuria.

Another preferred embodiment of this invention provides a method of treating or preventing atherosclerosis, which comprises administering a therapeutically effective amount of a compound of formula (I) or its stereoisomer, tautomer or pharmaceutically acceptable salt, solvate or prodrug to a patient in need of such treatment.

Another preferred embodiment of this invention provides a method of treating or preventing adhesions or skin scars, which comprises administering a therapeutically effective amount of a compound of formula (I) or its stereoisomer, tautomer or pharmaceutically acceptable salt, solvate or prodrug to a patient in need of such treatment.

Another preferred embodiment of this invention provides a method of treating or preventing cancer, which comprises administering a therapeutically effective amount of a compound of formula (I) or a stereoisomer, tautomer or pharmaceutically acceptable salt, solvate or prodrug thereof to a patient in need of such treatment.

Another preferred embodiment of the present invention provides a method of treating or preventing fibrotic conditions, such as chronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome (ARDS), fibromuscular dysplasia and fibrotic lung disease, as well as fibrin deposition in the eye during eye surgery, which comprises administering a therapeutically effective amount of a compound of formula (I) or a stereoisomer, tautomer or pharmaceutically acceptable salt, solvate or prodrug thereof to a patient in need of such treatment.

Another preferred embodiment of this invention provides a method of treating or preventing inflammatory diseases, such as asthma, arthritis, endometriosis, inflammatory bowel diseases, psoriasis and atopic dermatitis, as well as neurodegenerative diseases, such as Alzheimer's and Parkinson's disease, which includes the use of a therapeutically effective amount of a compound of formula (I) or its stereoisomer, tautomer or pharmaceutically acceptable salt, solvate or prodrug in a patient in need of such treatment.

Another preferred embodiment of the present invention provides a method of treating or preventing conditions that improve upon maintenance or elevation of bradykinin levels, comprising administering a therapeutically effective amount of a compound of formula (I) or a stereoisomer, tautomer or pharmaceutically acceptable salt, solvate or prodrug thereof to a patient in whom such treatment is necessary.

It may be noted that all references to treatment in this text include curative, palliative and prophylactic treatment. The amount of compound administered and the frequency of administration will be determined by the physician, taking into account the characteristics of the patient, such as age, weight and health, as well as the degree of TAFI inhibition desired. The total daily dose for a typical 70 kg adult will generally be between 1 mg and 5 g, preferably between 10 mg and 1 g, and more preferably between 50 and 750 mg. The total dose can be given as a single or divided dose.

The compounds of this invention may be used alone or in combination with other therapeutic agents. When used in combination with another therapeutic agent, the application of the two agents can be simultaneous or consecutive. Concomitant administration includes administration of a single dosage form containing both agents and administration of two agents in separate dosage forms at the same time. Consecutive application involves the application of two perm agents in different schedules, with the condition that there is an overlap in the periods during the treatment. Suitable agents with which the compounds of formula (I) may be administered include antithrombotic agents, including antiplatelet agents, anticoagulants and profibrinolytics. Suitable antithrombotics include: aspirin, PlavixTM, ticlopidine, warfarin (CoumadinTM), unfractionated heparin, hirudin (LepirudinTM), streptokinase, urokinase, recombinant tissue plasminogen activator (tPA), dipyridamole, ReoproTM, AggrastatTM and IntegrilinTM. The compounds of formula (I) can also be administered together with antihypertensive agents and with agents for the treatment of dyslipidemia, such as statins, eg Lipitor™. Other suitable classes of drugs for concomitant use include factor X inhibitors and antiarrhythmics, such as amiodarone or digoxin. Accordingly, in another aspect, this invention provides the use of a compound of formula (I) or its stereoisomer, tautomer or pharmaceutically acceptable salt, solvate or prodrug in combination with an antithrombotic agent for the preparation of a medicament for the treatment of thrombosis. In a preferred embodiment, the antithrombotic is a profibrinolytic. In an even more preferred embodiment, the antithrombotic is recombinant tissue plasminogen activator (tPA).

In yet another aspect, the present invention provides a method of treating or preventing thrombosis, which comprises administering a therapeutically effective amount of a compound of formula (I) or a stereoisomer, tautomer thereof, or a pharmaceutically acceptable salt, solvate, or prodrug in combination with an antithrombotic agent to a patient in need of such treatment. In a preferred embodiment, the antithrombotic is a profibrinolytic. In an even more preferred embodiment, the antithrombotic is recombinant tissue plasminogen activator (tPA).

In yet another embodiment, the present invention provides a kit comprising:

a composition comprising a compound of formula (I) or a stereoisomer, tautomer thereof or a pharmaceutically acceptable salt, solvate or prodrug as disclosed herein, and a pharmaceutically acceptable diluent or carrier;

a composition comprising an antithrombotic agent and a pharmaceutically acceptable diluent or carrier;

tank.

The components of this kit can be applied separately, simultaneously or sequentially.

This invention also provides the use of a compound of formula (I) or a stereoisomer, tautomer or a pharmaceutically acceptable salt, solvate or prodrug thereof as a coating on intravascular devices, such as indwelling catheters for dialysis, replacement heart valves or arterial "stents"; and as a coating on extracorporeal devices for blood circulation, such as heart, lung and kidney dialysis machines, to prevent thrombosis, especially myocardial infarction, deep vein thrombosis, stroke, juvenile stroke, cerebral infarction, cerebral thrombosis, cerebral embolism, peripheral vascular disease, angina and other forms of acute coronary syndromes, diffuse intravascular coagulation, sepsis, pulmonary embolism, embolic events secondary to cardiac arrhythmias and prevention of cardiovascular events, such as restenosis after surgical intervention, e.g. percutaneous transluminal coronary angioplasty, grafting, stenting , installation of a coronary bypass or any other form of surgical revascularization or intervention.

The present invention provides intravascular agents, the intravascular portion of which is covered with a compound of formula (I) or a stereoisomer, tautomer or pharmaceutically acceptable salt, solvate or prodrug thereof; and extracorporeal devices for blood circulation, such as heart, lung and kidney dialysis machines, where the part that comes into contact with the patient's blood is coated with a compound of formula (I) or a stereoisomer, tautomer or a pharmaceutically acceptable salt, solvate or prodrug thereof.

The compounds of the present invention are TAFIa inhibitors, the utility of which is based on preventing the reaction between a developing clot and TAFIa. It has been found that the compounds of the present invention can also bind the inactive TAFI molecule at a site involved in the reaction between TAFI and the developing clot. The use of TAFIa inhibitors, as previously described, as field and utility, includes such TAFIa inhibitors that bind TAFI.

The invention is illustrated below by non-limiting examples. Melting points were determined with a Gallenkamp apparatus for determining the melting point, using a glass capillary tube and are uncorrected. Unless otherwise indicated, all reactions were carried out under a nitrogen atmosphere, using commercially available anhydrous solvents. "0.88 ammonia" refers to commercially available aqueous solutions of ammonia with a specific gravity of about 0.88. Thin-layer chromatography was performed on glass pre-coated Merck silica gel (60 F254) plates, and silica gel column chromatography was performed with 40-63 μm silica gel (Merck silica gel 60). Ion-exchange chromatography was performed with a specific ion-exchange resin, previously washed with deionized water. Proton NMR spectra were measured on a Varian Inova 300, Varian Inova 400 or Varian Mercury 400 spectrometer in the indicated solvents. In the NMR spectra, only the unaltered protons that were found to be different from the solvent peaks are listed. Low-resolution mass spectra were recorded either on a Fisons Trio 1000, with thermospray positive ionization, or on a Finnigan Navigator, with electrospray positive or negative ionization. High-resolution mass spectra were recorded on a Bruker Apex II FT-MS, with electrospray positive ionization. Combustion analyzes were carried out at Exeter Analytical UK Ltd., Uxbridge, Middlesex. Optical rotations were determined at 25°C using a Perkin Elmer 341 polarimeter, using the indicated solvents and concentrations. The compounds from the examples were assigned (+) or (-) optical isomer labels based on the sign of the optical rotation when determined in a suitable solvent.

Abbreviations and definitions

ArbocelTM filter media, J. Rettenmaier & Sohne, Germany

Amberlyst® 15 ion exchange resin, available from Aldrich Chemical Company

atm pressure in atmospheres (1 atm = 760 Torr = 101.3 kPa)

BiotageTM chromatography performed using Flash 75 silica gel, from Biotage, UK

BOC tert-butyloxycarbonyl group

No. broad

c concentration used for optical rotation measurements in g per 100 mL (1 mg/mL is c 0.10)

cat catalytic

d doublet

dd double doublet

Degussa® 101 10% by weight palladium on activated carbon, Degussa type E101, available from Aldrich Chemical Company

Dowex® ion exchange resin, from Aldrich Chemical Company

ee enantiomeric excess

High-resolution HRMS mass spectroscopy (electrospray positive ionization)

Hyflo™ Hyflo supercel®, from Aldrich Chemical Company

liq liquid

Low resolution LRMS mass spectrometry (electrospray or thermospray positive ionization)

LRMS (ES-) low resolution mass spectrometry (electrospray negative ionization)

m multiplet

m/z mass spectrum peak

MCITM gel highly porous polymer, CHP20P 75-150 μm, from Mitsubishi Chemical Corporation

psi pounds per square inch (pounds per square inch, 1 psi = 6.9 kPa)

q quartet

Rf retention factor on TLC

with a singlet

Sep-Pak® silica gel C18 reverse phase, Waters Corporation

t triplet

TLC thin layer chromatography

δ chemical shift

Example 1

(2S)-(-)-2-(2-aminoethoxy)-3-(1-propyl-1H-imidazol-4-yl)propanoic acid

[image]

A solution of the compound from Preparation 88 (437 mg, 1.96 mmol) in 6M hydrochloric acid (35 mL) was heated at reflux for 72 hours, then allowed to cool and concentrated under reduced pressure. The residue was dissolved in water (2 mL), and the solution was purified by chromatography on a column of Dowex® 50WX8-200 ion-exchange resin, with an elution gradient of water and 0.88 ammonia (100:0 to 98:2). Fractions containing the product were combined and evaporated under reduced pressure, and the product was freeze-dried to give the title compound, 456 mg. 1H-NMR (CDCl3, 400 MHz) δ: 0.82 /t, 3H), 1.70 (m, 2H), 2.80 (m, 1H), 3.01 (m, 3H), 3.44 (m, 1H), 3.78 (m, 3H), 3.89 (dd, 1H), 6.70 (s, 1H), 7.35 (s, 1H). LRMS: m/z (ES+) 264 [MNa+]. Microanalysis showed: C, 49.04; H, 8.17; N, 15.51. C11H19N3O3⋅1.6H2O requires C 49.04; H, 8.28; N, 15.60%. [α]D = -33.43 (c = 0.193, methanol).

Examples 2 to 4

The following compounds of the general formula

[image]

were prepared from the corresponding morpholino compounds following the procedure from Example 1.

[image]

Example 5

(2S)-(-)-2-(2-aminoethoxy)-3-[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]propanoic acid

[image]

A solution of the compound from Preparation 133 (72 mg, 0.25 mmol) in concentrated hydrochloric acid (5 mL) was heated at 110°C for 18 hours, then allowed to cool and concentrated under reduced pressure. The residue was dissolved in water, and the solution was purified by chromatography on a column of Dowex® 50WX8-200 ion-exchange resin, with an elution gradient of water and 0.88 ammonia (95:5:0 to 90:5:5). The product was dissolved in water (5 mL) and freeze-dried to give the title compound as a sticky mass, 45 mg. 1H-NMR (CD3OD, 400 MHz) δ: 0.99 (m, 2H), 1.21 (m, 4H), 1.61-1.78 (m, 7H), 2.86 (dd, 1H) , 3.02 (m, 3H), 3.58-3.70 (m, 2H), 3.98 (m, 3H), 6.93 (s, 1H), 7.50 (s, 1H). LRMS: m/z (ES+) 310 [MH+]. Microanalysis showed: C, 59.56; H, 8.76; N, 12.91. C16H27N3O3⋅0.75H2O requires C 59.51; H, 8.90; N, 13.01%. [α]D = -23.34 (c = 0.102, methanol).

Example 6

(2S)-(-)-2-(2-aminoethoxy)-3-(1-phenyl-1H-imidazol-4-yl)propanoic acid

[image]

The title compound was obtained as a tan solid in 87% yield from the morpholino of Preparation 104, following the procedure of Example 5. 1H-NMR (D2O, 400 MHz) δ: 2.88 (dd, 1H), 3.00 (dd , 1H), 3.10 (t, 2H), 3.62 (t, 2H), 4.02 (m, 1H), 7.30 (s, 1H), 7.39 (m, 1H), 7 .45 (m, 4H), 7.98 (s, 1H). LRMS: m/z (ES+) 298 [MNa+]. Microanalysis showed: C, 59.15; H, 6.39; N, 14.71. C14H17N3O3⋅0.5H2O requires C 59.14; H, 6.38; N, 14.78%. [α]D = -16.8 (c = 0.10, methanol).

Example 7

(2S)-2-(2-aminoethoxy)-3-{1-[3,5-bis(trifluoromethyl)phenyl]-1H-imidazol-4-yl}-propanoic acid

[image]

The title compound was obtained as a white solid in 45% yield from morpholinone from Preparation 106, following the procedure of Example 5. 1H-NMR (CD3OD, 400 MHz) δ: 3.00-3.18 (m, 3H), 3 .65 (m, 1H), 3.75 (m, 1H), 4.04 (m, 2H), 7.58 (s, 1H, 7.98 (s, 1H), 8.22 (s, 2H ), 8.30 (s, 1H). LRMS: m/z (ES-) 410 [M-H-]. Microanalysis showed: C, 43.95; H, 3.79; N, 9.99. C16H15F6N3O3⋅ 1,25H2O finds C 44.30, H, 4.07, N, 9.69%.

Example 8

(2RS)-2-[(2-methylamino)ethoxy]-3-(1-propyl-1H-imidazol-4-yl)-propanoic acid

[image]

The title compound was obtained from morpholinone from Preparation 51, following the procedure of Example 1. 1H-NMR (D2O, 400 MHz) δ: 0.70 (t, 3H), 1.62 (m, 2H), 2.56 ( s, 3H), 2.76 (dd, 1H), 2.84 (dd, 1H), 3.05 (m, 2H), 3.53-3.63 (m, 2H), 3.80 (t , 2H), 3.94 (dd, 1H), 6.84 (s, 1H), 7.50 (s, 1H). LRMS: m/z (TSP+) 256.2 [MH+].

Example 9

(2RS)-2-[(2-dimethylamino)ethoxy]-3-(1-propyl-1H-imidazol-4-yl)-propanoic acid

[image]

A solution of the protected acid from Preparation 142 (200 mg, 0.62 mmol) in trifluoroacetic acid (5 mL) and dichloromethane (5 mL) was stirred at room temperature for 18 hours, after which it was concentrated under reduced pressure. The residue was dissolved in water, and the solution was purified by chromatography on a column of Dowex® 50WX8-200 ion-exchange resin, with water and 0.88 ammonia (96:4) as eluent. Fractions containing the product were concentrated under reduced pressure and the residue was dissolved in water and freeze-dried to give the title compound as a sticky mass, 100 mg. 1H-NMR (D2O, 400 MHz) δ: 0.66 (t, 3H), 1.60 (m, 2H), 2.60-2.78 (m, 7H), 2.82 (dd, 1H) , 3.14 (m, 2H), 3.57 (m, 1H), 3.61 (m, 1H), 3.78 (t, 2H), 3.92 (m, 1H), 6.82 ( s, 1H), 7.48 (s, 1H). LRMS: m/z (TSP+) 270.2 [MH+]. Microanalysis showed: C, 54.41; H, 8.72; N, 14.58. C13H23N3O3⋅H2O requires C, 54.34; H, 8.77; N, 14.62%.

Example 10

(2RS)-3-(1-propyl-1H-imidazol-4-yl)-2-[(3R)-pyrrolidin-3-yl-oxy]propanoic acid

[image]

Concentrated hydrochloric acid (3 mL) was added to a solution of the protected amino acid from Preparation 143 (232 mg, 0.55 mmol) in dioxane (2 mL) and the mixture was stirred at room temperature for 18 h, after which it was concentrated under reduced pressure. The residue was dissolved in water (1 mL), and the solution was purified by chromatography on a column of Dowex® 50WX8-200 ion-exchange resin, with an elution gradient of water and 0.88 ammonia (100:0 to 98:2). The product was freeze-dried to give the title compound as a white solid, 67 mg. 1H-NMR (D2O, 400 MHz) (mixture of diastereomers) δ: 0.72 (m, 3H), 1.63 (m, 2H), 1.80, 1.98, 2.08 (3×m, 2H ), 2.65, 2.84 (2×m, 3H), 3.03-3.38 (m, 3H), 3.81 (m, 2H), 3.94 (m, 1H), 4, 14 (m, 1H9, 6.89 (m, 1H), 7.53 (m, 1H). LRMS: m/z (ES + ) 290 [MNa + ].

Example 11

(2RS)-2-(2-aminoethoxy)-3-{1-[2-(4'-ethyl[1,1'-biphenyl]-4-yl)ethyl]-1H-imidazol-4-yl}propanoic acid

[image]

A mixture of the compound from Preparation 121 (170 mg, 0.43 mmol) and concentrated hydrochloric acid (2 mL) was heated at 110 °C for 18 h, then allowed to cool and concentrated under reduced pressure. The residue was azeotroped with ethanol, methanol and dichloromethane and then purified by column chromatography on Dowex® 50WX8-200 ion exchange resin, eluting with a gradient of water and 0.88 ammonia (100:0 to 98:2) to give the title compound , 15 mg. 1H-NMR (CD3OD, 400 MHz) δ: 1.22 (t, 3H), 2.62 (q, 2H), 2.82 (m, 1H), 2.98 (m, 3H), 3.02 (t, 2H), 3.50-3.62 (m, 2H), 3.90 (m, 1H), 4.18 (t, 2H), 6.86 (s, 1H), 7.14 ( d, 2H), 7.20 (d, 2H), 7.30 (s, 1H), 7.43 (m, 4H). Microanalysis showed: C, 64.05; H, 6.99; N, 9.35. C25H29N3O3⋅2.7H2O requires C, 64.28; H, 7.21; N, 8.99%.

Examples 12 to 14

The following compounds of the general formula

[image]

were prepared from the corresponding morpholinones following the procedure of Example 11.

[image]

Example 15

(2RS)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]-propanoic acid

[image]

The title compound was obtained in 45% yield from morpholinone of Preparation 92, following the procedure of Example 11. 1H-NMR (CD3OD, 400 MHz) δ: 0.94 (m, 4H), 1.18 (m, 4H) , 1.59-1.76 (m, 7H), 2.72 (m, 2H), 2.96 (m, 1H), 3.03 (m, 1H), 3.26 (m, 1H), 3.55 (m, 1H), 3.98 (m, 3H), 6.88 (s, 1H), 7.48 (s, 1H). LRMS: m/z (ES+) 324 [MNa+]. Microanalysis showed: C, 58.97; H, 9.01; N, 11.85. C17H29N3O3⋅1.2H2O requires C, 59.18; H, 9.17; N, 12.18%.

Example 16

(2RS)-2-(2-aminoethoxy)-3-(1-phenyl-1H-imidazol-4-yl)-propanoic acid

[image]

A solution of the compound from Preparation 99 (70 mg, 0.27 mmol) in concentrated hydrochloric acid (2 mL) was heated at 110 °C for 18 h, then allowed to cool and concentrated under reduced pressure. The residue was azeotroped with water and then purified by chromatography on a column of Dowex® 50WX8-200 ion exchange resin, with water and 0.88 ammonia as eluent (95:5), to give the title compound as a pale yellow solid, 50 mg. 1H-NMR (CD3OD, 400 MHz) δ: 2.93-3.14 (m, 3H), 3.24 (m, 1H), 3.59-3.73 (m, 2H), 4.00 ( m, 1H), 7.35 (m, 2H), 7.45 (m, 4H), 7.98 (s, 1H). LRMS: m/z (ES+) 298 [MNa+].

Example 17

(2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-(1-phenyl-1H-imidazol-4-yl)-propanoic acid

[image]

The title compound was obtained in 75% yield from morpholinone of Preparation 109, following the procedure of Example 16, except that an elution gradient of methanol:water:0.88 ammonia (20:80:5 to 30: 65:5). 1H-NMR (D2O, 400 MHz) δ: 0.90 (d, 3H), 2.80 (m, 2H), 3.00 (m, 2H), 3.59 (m, 1H), 4.14 (dd, 1H), 7.26 (s, 1H), 7.37 (m, 1H), 7.42 (m, 4H), 7.96 (s, 1H). LRMS: m/z (ES-) 288 [M-H-]. Microanalysis showed: C, 58.76; H, 6.72; N, 13.37. C15H19N3O3⋅H2O requires C 58.63; H, 6.89; N, 13.67%. [α]D = -83.0 (c = 0.1, methanol).

Example 18

(2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-[1-(3',4'-dichloro[1,1'-biphenyl]-3-yl)- 1H-imidazol-4-yl]-propanoic acid

[image]

The title compound was obtained as a white foam from the morpholino of Preparation 115, following the procedure of Example 17, except that an elution gradient of water:methanol:0.88 ammonia (75:20:5 to 15:80: 5). 1H-NMR (CD3OD, 400 MHz) δ: 1.00 (d, 3H), 2.77 (dd, 1H), 2.92 (m, 2H), 3.15 (dd, 1H), 3.59 (m, 1H), 4.10 (m, 1H), 7.42 (s, 1H), 7.50-7.627 (m, 5H), 7.78 (s, 1H), 7.86 (s, 1H), 8.10 (s, 1H). LRMS: m/z (ES+) 434, 436 [MH+].

Example 19

(2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-[1-(4-phenoxyphenyl)-1H-imidazol-4-yl]-propanoic acid

[image]

A solution of morpholinone from Preparation 111 (130 mg, 0.36 mmol) in concentrated hydrochloric acid (10 mL) was heated at 110°C for 18 h, then allowed to cool and concentrated under reduced pressure. The residue was purified by chromatography on a column of Dowex® 50WX8-200 ion-exchange resin, with water:methanol:0.88 ammonia (70:30:5) as eluent. The product was dissolved in water and freeze-dried to give the title compound as an off-white powder, 70 mg. 1H-NMR (CD3OD, 400 MHz) δ: 1.00 (d, 3H), 2.78 (dd, 1H), 2.86-3.00 (m, 2H), 3.17 (dd, 1H) , 3.60 (m, 1H), 4.15 (dd, 1H), 7.02 (d, 2H), 7.08 (d, 2H), 7.17 (m, 1H), 7.30 ( s, 1H), 7.39 (m, 2H), 7.50 (d, 2H), 7.95 (s, 1H). LRMS: m/z (ES+) 404 [MNa+]. Microanalysis showed: C, 62.70; H, 6.22; N, 10.30. C21H23N3O4⋅1.2H2O requires C 62.58; H, 6.35; N, 10.43%. [α]D = -50.9 (c = 0.117, methanol).

Examples 20 to 31

The following compounds of the general formula

[image]

were prepared by heating a solution of the corresponding morpholino (0.2 to 0.4 mmol) in concentrated hydrochloric acid (2 mL) at reflux for 18 hours, concentrating the cooled solution and azeotroping the residue with ethanol, ethyl acetate and dichloromethane.

[image] [image] [image]

Example 32

(2RS)-2-(2-aminoethoxy)-3-[1-(3-[1,1'-biphenyl]-3-yl-propyl)-1H-imidazol-4-yl]-propanoic acid dihydrochloride

[image]

The title compound was prepared as an off-white foam from the compound of Preparation 145, following the procedure of Example 20. 1H-NMR (CD3OD, 400 MHz) δ: 2.25 (m, 2H), 2.75 (m, 2H), 3.14 (m, 2H), 3.27 (m, 1H), 3.74 (m, 1H), 3.78 (m, 2H), 4.22 (m, 3H), 7.18 (m , 1H), 7.25-7.42 (m, 6H), 7.45 (s, 1H), 7.58 (d, 2H), 8.80 (s, 1H). LRMS: m/z (ES+) 394 [MH+]. Microanalysis showed: C, 54.03; H, 6.73; N, 8.09. C23H27N3O3⋅2HCl⋅2.4H2O requires C 54.21; H, 6.68; N, 8.24%.

Example 33

(2RS)-2-(2-aminoethoxy)-3-[1-(3-[1,1'-biphenyl]-2-yl-propyl)-1H-imidazol-4-yl]-propanoic acid dihydrochloride

[image]

The title compound was prepared as an off-white foam in 30% yield from the compound of Preparation 146, following the procedure of Example 20. 1H-NMR (CD3OD, 400 MHz) δ: 1.96 (m, 2H), 2.60 ( t, 2H), 3.05-3.23 (m, 4H), 3.79 (m, 2H), 4.02 (t, 2H), 4.48 (m, 1H), 7.14 (d , 1H), 7.19-7.30 (m, 6H), 7.36 (m, 3H), 8.62 (s, 1H). Microanalysis showed: C, 54.89; H, 6.24; N, 8.35. C23H27N3O3⋅2HCl⋅2H2O requires C 54.98; H, 6.62; N, 8.36%.

Example 34

(2S)-2-{[(2S)-aminopropyl]oxy}-3-[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]-propanoic acid dihydrochloride

[image]

The title compound was obtained in 64% yield from morpholinone from Preparation 95, following the procedure of Example 20. 1H-NMR (CD3OD, 400 MHz) δ: 0.80-1.32 (m, 9H), 1.60- 1.80 (m, 7H), 3.20 (m, 1H), 3.57 (m, 3H), 3.80 (m, 1H), 4.20 (m, 2H), 4.35 (m , 1H), 7.48 (s, 1H), 8.86 (s, 1H). HRMS: m/z (ESI) 324.2280 [MH + ].

Example 35

(2R)-2-{[(2R)-aminopropyl]oxy}-3-[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]-propanoic acid dihydrochloride

[image]

The title compound was obtained from morpholinone from Preparation 96, following the procedure of Example 20. 1H-NMR (CD3OD, 400 MHz) δ: 0.80-1.32 (m, 9H), 1.60-1.80 (m , 7H), 3.20 (m, 1H), 3.57 (m, 3H), 3.80 (m, 1H), 4.20 (m, 2H), 4.35 (m, 1H), 7 .48 (s, 1H), 8.86 (s, 1H). HRMS: m/z (ESI) 324.2275 [MH + ].

Example 36

(2RS)-2-(2-aminoethoxy)-3-[1-(2-methylphenyl)-1H-imidazol-4-yl]-propanoic acid dihydrochloride

[image]

A solution of morpholinone from Preparation 100 (100 mg, 0.37 mmol) in concentrated hydrochloric acid (2 mL) was heated at 110 °C for 36 hours, after which it was cooled and concentrated under reduced pressure. The residue was azeotroped with methanol and dichloromethane to give the title compound. 1H-NMR (CD3OD, 400 MHz) δ: 2.10 (s, 3H), 3.19 (m, 2H), 3.26 (m, 1H), 3.39 (m, 1H), 3.82 (t, 2H), 4.42 (m, 1H), 7.40 (m, 2H), 7.46 (m, 2H), 7.62 (s, 1H), 9.14 (s, 1H) . HRMS: m/z 290.1502 [MH+].

Example 37

(2R)-2-{[(1S)-amino-1-methylethyl]oxy}-3-[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]-propanoic acid dihydrochloride

[image]

The title compound was obtained from morpholinone from Preparation 91, following the procedure of Example 20. 1H-NMR (CD3OD, 400 MHz) δ: 1.01 (m, 5H), 1.21 (m, 4H), 1.74 ( m, 7H), 2.93-3.14 (m, 3H), 3.25 (m, 1H), 3.79 (m, 1H), 4.21 (t, 2H), 4.38 (m , 1H), 7.55 (s, 1H), 8.90 (s, 1H). LRMS: m/z (ES-) 322 [M-H-]. Microanalysis showed: C, 47.73; H, 7.85; N, 9.66. C17H29N3O3⋅2HCl⋅1.6H2O requires C 48.02; H, 8.11; N, 9.88%.

Example 38

(2S)-2-{[(1R)-amino-1-methylethyl]oxy}-3-{1-[2-(4,4-dimethylcyclohexyl)ethyl]-1H-imidazol-4-yl}-propanoic acid dihydrochloride

[image]

The title compound was obtained in 83% yield from morpholinone from Preparation 93, following the procedure of Example 20. 1H-NMR (CD3OD, 400 MHz) δ: 0.88 (s, 6H), 1.04 (d, 3H) , 1.17-1.34 (m, 5H), 1.40 (m, 2H), 1.60 (m, 2H), 2.97-3.18 (m, 3H), 3.30 (m , 1H), 3.81 (m, 1H), 4.24 (m, 2H), 4.40 (m, 1H9, 7.58 (s, 1H), 8.94 (s, 1H). HRMS: m/z (ES+) 374.2484 [MNa+].

Example 39

(2S)-2-{[(1R)-amino-1-methylethyl]oxy}-3-{1-(3-cyclohexyl-3-methylbutyl)-1H-imidazol-4-yl]-propanoic acid dihydrochloride

[image]

The title compound was obtained from morpholinone from Preparation 94, following the procedure of Example 20. 1H-NMR (CD3OD, 400 MHz) δ: 0.94 (s, 6H), 0.98-1.30 (m, 10H), 1.62 (m, 1H), 1.78 (m, 6H), 2.90-3.15 (m, 3H), 3.78 (m, 1H), 4.20 (m, 2H), 4 .38 (m, 1H), 7.52 (s, 1H), 8.90 (s, 1H). LRMS: m/z (ES+) 366 [MH+].

Example 40

2-(2-aminoethoxy)-3-[1-(3-phenoxyphenyl)-1H-imidazol-4-yl]propanoic acid dihydrochloride

[image]

A solution of morpholinone from Preparation 102 (25 mg, 0.07 mmol) in concentrated hydrochloric acid (5 mL) was heated at 110 °C for 18 hours, after which it was cooled and concentrated under reduced pressure. The residue was dissolved in water and freeze-dried to give the title compound as a tan solid, 35 mg. 1H-NMR (D2O, 400 MHz) δ: 3.04 (m, 3H), 3.18 (dd, 1H), 3.63 (t, 2H), 4.19 (m, 1H), 6.98 (d, 2H), 7.06 (m, 3H), 7.19 (d, 1H), 7.46 (dd, 2H), 7.50 (dd, 1H), 7.54 (s, 1H) , 8.87 (s, 1H). LRMS: m/z (ES+) 368 [MH+].

Examples 41 to 44

The following compounds of the general formula

[image]

were prepared from the corresponding morpholinones following the procedure of Example 40.

[image]

Example 45

(2S)-(-)-2-(2-aminoethoxy)-3-[1-(4-tert-butylphenyl)-1H-imidazol-4-yl]-propanoic acid dihydrochloride

[image]

The title compound was obtained as a tan solid from morpholinone from Preparation 105, following the procedure of Example 40. 1H-NMR (D2O, 400 MHz) δ: 1.20 (s, 9H), 3.04-3.35 (m, 4H), 3.78 (m, 2H), 4.38 (m, 1H), 7.41 (m, 2H), 7.58 (m, 2H), 7.61 (s, 1H), 8, 98 (s, 1H). LRMS: m/z (ES+) 332 [MH+]. Microanalysis showed: C, 49.58; H, 6.75; N, 9.84. C18H25N3O3⋅2HCl⋅1.75H2O requires C 49.58; H, 7.06; N, 9.64%. [α]D = -2.00° (c = 0.30, methanol).

Examples 46 to 48

The following compounds of the general formula

[image]

were prepared from the corresponding morpholinones following the procedure of Example 40.

[image]

Example 49

(2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-{1-[4-(cyclohexyloxy)phenyl]-1H-imidazol-4-yl]-propanoic acid

[image]

A solution of the protected amino acid from Preparation 120 in a mixture of 2N hydrochloric acid (1 mL), water (1 mL) and dioxane (1 mL) was stirred at room temperature for 2 hours, after which it was concentrated under reduced pressure. The residue was dissolved in water and purified by chromatography on a column of Dowex® 50WX8-200 ion-exchange resin, with water:methanol:0.88 ammonia (48:48:4) as eluent. The product was dissolved in water and freeze-dried to give the title compound, 25 mg. 1H-NMR (D2O, 400 MHz) δ: 0.94 (d, 3H), 1.35 (m, 4H), 1.55 (m, 3H), 1.72-2.01 (m, 3H) , 2.78-2.97 (m, 2H), 3.10 (m, 2H), 3.60 (m, 1H), 4.01 (m, 1H), 4.21 (m, 1H), 6.92 (d, 2H), 7.01 (s, 1H), 7.20 (d, 2H), 7.62 (s, 1H). LRMS: m/z (ES+) 388 [MH+]. Microanalysis showed: C, 59.33; H, 7.65; N, 9.80. C21H29N3O4⋅2H2O requires C 59.56; H, 7.85; N, 9.92%.

Example 50

(2S)-2-(2-aminoethoxy)-3-(1H-imidazol-4-yl)-propanoic acid dihydrochloride

[image]

A suspension of morpholinone from Preparation 118 (90 mg, 0.24 mmol) and lithium hydroxide (32 mg, 1.33 mmol) in a mixture of water (2 mL) and tetrahydrofuran (1 mL) was stirred at room temperature for 18 hours, after which is concentrated under reduced pressure. The residue was suspended in concentrated hydrochloric acid (3 mL) and the mixture was heated at 110°C for 18 hours, then cooled and concentrated under reduced pressure. The residue was triturated in acetone and the resulting solid was collected and dried, then recrystallized from methanol/acetone to give the title compound as a white solid, 12 mg. 1H-NMR (CD3OD, 400 MHz) δ: 3.20 (m, 2H), 3.30 (m, 2H), 3.83 (m, 2H), 4.37 (m, 0.5H), 4 .41 (m, 0.5H), 7.41 (s, 1H), 8.81 (s, 1H). LRMS: m/z (ES-) 198 [M-H-].

Example 51

(2S)-2-{[(1R)-amino-1-methylethyl]oxy}-3-(1H-imidazol-4-yl)-propanoic acid

[image]

A solution of morpholinone from Preparation 105 (320 mg, 1.64 mmol) in concentrated hydrochloric acid (5 mL) was heated at 110°C for 18 h, then allowed to cool and diluted with water (80 mL). The resulting solution was purified by chromatography on a column of Dowex® 50WX8-200 ion-exchange resin, with an elution gradient of water:0.88 ammonia (100:0 to 95:5). The product was dissolved in water and freeze-dried to give the title compound as a colorless foam, 290 mg. 1H-NMR (D2O, 400 MHz) δ: 0.90 (d, 3H), 2.80 (dd, 2H), 2.98 (dd, 2H), 3.57 (m, 1H), 4.04 (m, 1H), 6.84 (s, 1H), 7.60 (s, 1H). LRMS: m/z (ES+) 236 [MNa+]. Microanalysis showed: C, 46.00; H, 7.41; N, 17.81. C9H15N3O4⋅1.25H2O requires C 45.85; H, 7.48; N, 17.82%. [α]D = -94.62° (c = 1.72, water).

Alternative synthesis

A mixture of the protected lactone from Preparation 55a (1.58 g, 4.42 mmol) and methanesulfonic acid (6.5 mL) was heated at 70°C for 2.5 hours. The cooled solution was diluted with ether (40 mL), the mixture was stirred, and the ether was decanted. This procedure was repeated twice. Water was added, and the mixture was vigorously stirred and filtered. The filtrate was left to stand at room temperature for 24 hours, and then it was purified by chromatography on a column of Dowex® 50WX8-200 ion-exchange resin, with water:0.88 ammonia (95:5) as eluent, and the resulting fractions were evaporated under reduced pressure (with bath water temperature below 33°C). The obtained product was dissolved in concentrated hydrochloric acid (5 mL), and the solution was heated at 100°C for 18 hours, after which it was cooled and diluted with water (30 mL). This solution was purified by chromatography on a column of Dowex® 50WX8-200 ion-exchange resin, with water:0.88 ammonia (95:5) as eluent. The product was stirred in acetonitrile (10 mL) for 1 hour, filtered and dried in vacuum for 18 hours. Microanalysis showed: C, 46.58; H, 7.50; N, 17.98. C9H15N3⋅1H2O requires C 45.75; H, 7.41; N, 18.17%.

Example 52

(2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-[1-(2-pyridinyl)-1H-imidazol-4-yl]-propanoic acid dihydrochloride

[image]

A solution of morpholinone from Preparation 152 (72 mg, 0.26 mmol) in concentrated hydrochloric acid (3 mL) was heated at 100°C for 18 hours, then allowed to cool, diluted with water and concentrated under reduced pressure. The residue was purified by chromatography on a column of Dowex® 50WX8-200 ion-exchange resin, eluting with a gradient of water:0.88 ammonia (100:0 to 95:5). The product was dissolved in 2N hydrochloric acid, and the resulting solution was concentrated under reduced pressure. The residue was dissolved in water (10 mL) and freeze-dried to give the title compound as a solid, 58 mg. 1H-NMR (D2O, 400 MHz) (mixture of imidazole regioisomers) δ: 0.79, 0.99 (2×d, 3H), 2.56, 2.92 (2×m, 1H), 3.02- 3.44 (m, 3H), 3.58, 3.78 (2×m, 1H), 7.55 (m, 1H), 7.71 (m, 1H), 7.96-8.19 ( m, 2H), 8.52, 8.61 (2×m, 1H), 9.03, 9.38 (2×s, 1H). LRMS: m/z (ES+) 291 [MH+].

Example 53

(2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-(1-propyl-1H-imidazol-4-yl)-propanoic acid

[image]

A solution of the compound from Preparation 157 (560 mg, 2.36 mmol) in concentrated hydrochloric acid (10 mL) was stirred at 110°C for 18 hours. The cooled solution was concentrated under reduced pressure and the residue was purified by column chromatography on Dowex® 50WX8-200 ion exchange resin, eluting with a gradient of water:0.88 ammonia (100:0 to 95:5) to give the title compound as foam, 350 mg. 1H-NMR (D2O, 400 MHz) δ: 0.66 (t, 3H), 0.81 (d, 3H), 1.60 (m, 2H), 2.60-2.78 (m, 2H) , 2.80-2.98 (m, 2H), 3.45 (m, 1H), 3.78 (t, 2H), 3.98 (dd, 1H), 6.82 (s, 1H), 7.43 (s, 1H). LRMS: m/z (ES+) 278 [MNa+]. Microanalysis showed: C, 53.44; H, 8.13; N, 15.40. C12H21N3O3⋅0.8H2O requires C 53.44; H, 8.45; N, 15.58%. [α]D = -86.93° (c = 0.11, methanol).

Example 54

(2R)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-(1H-imidazol-4-yl)-propanoic acid

[image]

A mixture of lactam from Preparation 158 (80 mg, 0.50 mmol) and 2N hydrochloric acid (2 mL) was heated at 110°C for 16 hours, after which it was allowed to cool and diluted with water (10 mL). This solution was purified by chromatography on a column of Dowex® 50WX8-200 ion-exchange resin, with water:0.88 ammonia (95:5) as eluent. The product was triturated in acetone and the resulting solid was dried in vacuo to give the title compound as a brown solid, 68 mg. 1H-NMR (D2O, 400 MHz) δ: 1.00 (d, 3H), 2.76 (dd, 2H), 2.83 (m, 2H), 3.60 (m, 1H), 3.86 (m, 1H), 6.79 (s, 1H), 7.57 (s, 1H).

Preparation 1

1-n-propyl-1H-imidazole-4-carboxaldehyde

[image]

Imidazole-4-carboxaldehyde (30 g, 0.31 mol) was added portionwise to a solution of sodium hydride (13.9 g, 60% dispersion in mineral oil, 0.348 mol) in tetrahydrofuran (450 mL), and the solution mixed for 45 minutes. Then n-propyl bromide (31.2 mL, 0.344 mol) was added portionwise, followed by 18-crown-6 (150 mg) and the mixture was heated at reflux for 18 hours. Aqueous ammonium chloride solution was added to the cooled solution and the mixture was extracted with ethyl acetate (2×) and dichloromethane (2×). The combined organic extracts were dried (MgSO4), filtered and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography, eluting with ethyl acetate:pentane (40:60), to give the title compound, 20.2 g, 47% yield. 1H-NMR (DMSO-d6, 400 MHz) δ: 0.80 (t, 3H), 1.76 (m, 2H), 3.98 (t, 2H), 7.84 (s, 1H), 8 .04 (s, 1H), 9.70 (s, 1H). LRMS: m/z (TSP+) 277.3 [2M+H+].

Preparation 2

1-n-butyl-1H-imidazole-4-carboxaldehyde

[image]

The title compound was obtained in 28% yield from imidazole-4-carboxaldehyde and n-butyl bromide, following a procedure similar to that described in Preparation 1. 1H-NMR (CDCl3, 300 MHz) δ: 0.97 (t, 3H ), 1.37 (m, 2H), 1.80 (m, 2H), 4.00 (t, 2H), 7.55 (s, 1H), 7.62 (s, 1H), 9.88 (s, 1H). LRMS: m/z (TSP+) 153.3 [MH+].

Preparation 3

1-(2-cyclohexylethyl)-1H-imidazole-4-carboxaldehyde

[image]

Imidazole-4-carboxaldehyde (4.8 g, 50 mmol) was added portionwise to a suspension of sodium hydride (2.20 g, 60% dispersion in mineral oil, 55 mmol) in tetrahydrofuran (150 mL) and the mixture was stirred at room temperature for 1 hour. 2-Cyclohexylethyl bromide (8.6 mL, 55 mmol) was added and the mixture was heated at reflux for 18 h. The cooled mixture was evaporated under reduced pressure and the residue was partitioned between water (500 mL) and dichloromethane (500 mL). The layers were separated, and the organic phase was dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of toluene:ethyl acetate (100:0 to 96:4) to give the title compound, 1.78 g. 1H-NMR (CDCl3, 400 MHz) δ: 0, 98 (m, 2H), 1.20 (m, 4H), 1.68 (m, 7H), 4.00 (t, 2H), 7.4 (s, 1H), 7.60 (s, 1H ), 9.80 (s, 1H). LRMS: m/z (TSP+) 207.2 [MH+].

Preparation 4

1-(2-Phenylethyl)-1H-imidazole-4-carboxaldehyde

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Imidazole-4-carboxaldehyde (6.73 g, 70 mmol) was added portionwise to a suspension of sodium hydride (1.68 g, 60% dispersion in mineral oil, 70 mmol) in tetrahydrofuran (280 mL), and the solution stirred for 30 minutes at room temperature. (2-Bromoethyl)benzene (9.56 mL, 70 mmol) was added and the mixture was stirred at room temperature for 72 hours. The mixture was evaporated under reduced pressure, and the residue was partitioned between water (300 mL) and dichloromethane (500 mL), and the layers were separated. The organic phase was dried (MgSO4) and evaporated under reduced pressure. The crude product was pre-adsorbed on silica gel and purified by silica gel column chromatography, eluting with a gradient of ethyl acetate:pentane (50:50 to 100:0), to give the title compound, 1.44 g. 1H-NMR (CDCl3 , 400 MHz) δ: 3.16 (t, 2H), 4.23 (t, 2H), 7.02 (d, 2H), 7.28 (m, 3H), 7.36 (s, 1H) , 7.50 (s, 1H), 9.83 (s, 1H). LRMS: m/z (ES+) 223 [MNa+].

Preparation 5

1-Trityl-1H-imidazole-4-carboxaldehyde

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A solution of trityl chloride (9.5 g, 34.3 mmol) in N,N-dimethylformamide (50 mL) was added dropwise to an ice-cold solution of imidazole-4-carboxaldehyde (3 g, 31.2 mmol) and triethylamine ( 17 mL, 125 mmol) in N,N-dimethylformamide (30 mL) and the solution was stirred for 2 hours. The reaction mixture was allowed to warm to room temperature and was stirred for another 18 hours. Water (200 mL) was added, and the resulting pink solid was collected and dried, after which it was dissolved in dichloromethane (200 mL). The resulting solution was washed with water (2×100 mL), dried (MgSO4) and evaporated under reduced pressure. The product was recrystallized from ethanol to give the title compound as a solid, 7.8 g. 1H-NMR (CDCl 3 , 400 MHz) δ: 7.06 (m, 6H), 7.32 (m, 9H), 7, 48 (s, 1H), 7.58 (s, 1H), 9.82 (s, 1H). Microanalysis showed: C, 81.54; H, 5.37 N, 8.24. C23H18N2O requires C 81.63; H, 5.36; N, 8.28%.

Preparation 6

2-[(4-methoxybenzyl)amino]ethanol

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Acetic acid (ca. 150 mL) was added to a solution of p-anisaldehyde (58.2 g, 0.42 mol) and ethanolamine (152 mL, 2.52 mol) in methanol (1 L) to achieve pH 6. Portion per sodium triacetoxyborohydride (100 g, 0.47 mol) was added in one portion, and after the addition was complete, the mixture was stirred at room temperature for 72 hours. The mixture was concentrated under reduced pressure, basified with 1N sodium hydroxide solution and extracted with dichloromethane (10 x 300 mL). The combined extracts were evaporated and the crude product was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol (98:2 to 90:10) to give the title compound, 42 g. 1H-NMR (CDCl3, 400 MHz) δ: 2.78 (t, 2H), 3.62 (t, 2H), 3.75 (m, 5H), 4.24 (s, 2H), 6.81 (d, 2H), 7.22 (d , 2H). LRMS: m/z (ES+) 182 [MH+].

Preparation 7

(2R)-1-[(4-Methoxybenzyl)amino]-2-propanol

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A solution of (R)-(-)-1-amino-2-propanol (9.00 g, 0.12 mol) in tetrahydrofuran (40 mL) and acetic acid (5 mL) was added dropwise to a solution of p-anisaldehyde ( 5.45 g, 0.04 mol) in tetrahydrofuran (40 mL), and after the addition was complete, the solution was stirred at room temperature for 2 hours. The solution was cooled in a water bath and sodium triacetoxyborohydride (9.50 g, 0.045 mol) was added portion by portion and the mixture was stirred at room temperature for 18 hours. The reaction mixture was concentrated under reduced pressure and the residue was partitioned between dichloromethane (150 mL) and sodium hydroxide solution (150 mL, 0.5 N). The layers were separated and the aqueous phase was saturated with sodium chloride, after which it was extracted with additional dichloromethane (3 × 30 mL). The combined organic solutions were dried (MgSO4) and evaporated under reduced pressure. The residue was purified by silica gel chromatography (elution gradient 98:2:0.2 to 97:3:0.3 dichloromethane:methanol:0.88 NH 3 ) to give an orange oil (4.9 g). 1H-NMR (CDCl3, 400 MHz) δ: 1.12 (d, 3H), 2.39 (dd, 1H), 2.70 (dd, 1H), 3.62-3.79 (m, 6H) , 6.82 (d, 2H), 7.19 (d, 2H). LRMS: m/z (ES+) 196 [MH+].

Preparation 8

(2S)-1-[(4-Methoxybenzyl)amino]-2-propanol

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A mixture of (S)-(+)-1-amino-2-propanol (9 g, 0.12 mol), p-anisaldehyde (5.45 g, 0.04 mol), acetic acid (5 mL), and sodium triacetoxyborohydride (9.5 g, 0.045 mol) in methanol (80 mL) was stirred at room temperature for 72 hours. The reaction mixture was concentrated under reduced pressure and the residue was partitioned between dichloromethane (150 mL) and sodium hydroxide solution (100 mL, 0.5 N). The layers were separated and the aqueous phase was extracted with additional dichloromethane (4 × 30 mL). The combined organic solutions were dried (MgSO4) and concentrated under reduced pressure. The remaining yellow oil was purified by silica gel column chromatography, eluting with a gradient of dichloromethane:methanol:0.88 ammonia (98:2:0.2 to 95:5:0.5) to give the title compound, 6.2 g. 1H-NMR (CDCl3, 400 MHz) δ: 1.10 (d, 3H), 2.24-2.40 (m, 2H), 2.65 (dd, 1H), 3.62-3.80 ( m, 6H), 6.82 (d, 2H), 7.19 (d, 2H). LRMS: m/z (ES+) 218 [MNa+].

Preparation 9

(2R)-2-[(4-Methoxybenzyl)amino]-1-propanol

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(R)-(-)-2-amino-1-propanol (10.36 mL, 133 mmol) was added dropwise to a solution of p-anisaldehyde (5.85 g, 42.9 mmol) in methanol (90 mL). and the solution was cooled in an ice bath. Acetic acid (2.5 mL) and sodium triacetoxyborohydride (10.0 g, 47.2 mol) were added and the reaction mixture was allowed to warm to room temperature over one hour. The solution was heated to 40°C and stirred for another 48 hours, after which it was concentrated under reduced pressure. The residue was partitioned between saturated sodium bicarbonate solution (50 mL) and dichloromethane (100 mL) and the layers were separated. The aqueous phase was extracted with additional dichloromethane (10 × 50 mL), and the combined organic solutions were dried (MgSO4) and evaporated under reduced pressure. The residue was purified twice by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (97:3:0.3 to 90:10:1) to give the title compound, 6.0 g. 1H-NMR ( CDCl3, 400 MHz) δ: 1.04 (d, 3H), 2.80 (m, 1H), 3.22 (dd, 1H), 3.58 (dd, 1H), 3.62 (d, 1H ), 3.78 (m, 4H), 6.82 (d, 2H), 7.20 (d, 2H). LRMS: m/z (ES+) 196 [MH+]. [α]D = -34.85° (c = 0.137, methanol).

Preparation 10

(2S)-2-[(4-Methoxybenzyl)amino]-1-propanol

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The title compound was obtained in 67% yield from (S)-(+)-2-amino-1-propanol and p-anisaldehyde following the procedure described in Preparation 9. 1H-NMR (CDCl3, 400 MHz) δ:1, 08 (d, 3H), 2.83 (m, 1H), 3.30 (dd, 1H), 3.59 (dd, 1H), 3.66 (d, 1H), 3.77 (s, 3H ), 3.82 (d, 1H), 4.08 (bs, 2H), 6.82 (d, 2H), 7.22 (d, 2H). [α]D = +39.19° (c = 0.146, methanol).

Preparation 11

4-(4-Methoxybenzyl)-3-morpholinone

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A solution of sodium hydroxide (7.08 g, 0.177 mol) in water (150 mL) was added to a solution of the amino alcohol from Preparation 6 (32 g, 0.177 mol) in dichloromethane (250 mL) and the mixture was cooled to 0°C. A solution of chloroacetyl chloride (14.3 mL, 0.177 mol) in dichloromethane (50 mL) was added dropwise over 30 minutes and the mixture was stirred at room temperature for 18 hours. The phases were separated and the organic layer was washed with sodium hydroxide (2N, 150 mL), 2N hydrochloric acid (150 mL) and brine (50 mL), dried (MgSO4) and concentrated under reduced pressure. The remaining oil was dissolved in ethanol (200 mL), a solution of potassium hydroxide (9.93 g, 0.177 mol) in ethanol (200 mL) was added and the mixture was stirred at room temperature for 18 h. The mixture was filtered, the filtrate was evaporated under reduced pressure and the residue was triturated in diethyl ether/pentane to give the title compound as a white powder, 26 g. 1H-NMR (CDCl 3 , 400 MHz) δ: 3.20 (t, 2H), 3.78 (m, 5H), 4.19 (s, 2H), 4.54 (s, 2H), 6.82 (d, 2H), 7.18 (d, 2H). LRMS: m/z (ES+) 244 [MNa+].

Preparation 12

(6R)-4-(4-Methoxybenzyl)-6-methyl-3-morpholinone

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The title compound was obtained in a yield of 76% from the alcohol from Preparation 7 and chloroacetyl chloride, following a procedure similar to that described in Preparation 11, except that the compound was further purified by chromatography on a silica gel column, with dichloromethane:methanol:0.88 ammonia ( 98:2:0.2) as eluent. 1H-NMR (CDCl3, 400 MHz) δ: 1.18 (d, 3H), 2.99-3.14 (m, 2H), 3.79 (m, 4H), 4.17 (d, 1H) , 4.25 (d, 1H), 4.38 (d, 1H), 4.61 (d, 1H), 6.82 (d, 2H), 7.17 (d, 2H). LRMS: m/z (ES+) 258 [MNa+].

Preparation 13

(6S)-4-(4-Methoxybenzyl)-6-methyl-3-morpholinone

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The title compound was obtained as a yellow oil in 61% yield from the amino alcohol of Preparation 8, following the procedure described in Preparation 11. 1H-NMR (CDCl3, 400 MHz) δ: 1.18 (d, 3H), 2, 98-3.10 (m, 2H), 3.78 (m, 4H), 4.18 (d, 1H), 4.25 (d, 1H), 4.38 (d, 1H), 4.60 (d, 1H), 6.61 (d, 2H), 7.17 (d, 2H). LRMS: m/z (ES+) 258 [MNa+].

Preparation 14

(5S)-4-(4-Methoxybenzyl)-5-methyl-3-morpholinone

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A solution of chloroacetyl chloride (2.34 mL, 29 mmol) in dichloromethane (25 mL) was added dropwise over 10 minutes to an ice-cold mixture of the amino alcohol from Preparation 10 (5.6 g, 28.7 mmol) in solution of sodium hydroxide (1.16 g, 29 mmol in water (20 mL)) and dichloromethane (50 mL) with stirring. The mixture was stirred at room temperature for 18 hours and the layers were then separated. The organic phase was washed with 1N sodium hydroxide solution (25 mL), 2M hydrochloric acid (20 mL) and brine (20 mL), then dried (MgSO4) and evaporated under reduced pressure. The residue was dissolved in ethanol (40 mL), the solution was cooled in an ice bath and a solution of potassium hydroxide (1.63 g, 29 mmol) in ethanol (40 mL) was added dropwise over 5 minutes. The mixture was then allowed to warm to room temperature and stirred for an additional 18 hours. The resulting precipitate was filtered off, the filtrate was concentrated under reduced pressure and the residue was dissolved in dichloromethane (150 mL). This solution was dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (97:3:0.3 to 95:5:0.5) to give the title compound, 4 g. 1H-NMR ( CDCl3, 400 MHz) δ: 1.26 (d, 3H), 3.37 (m, 1H), 3.62 (dd, 1H), 3.70 (dd, 1H), 3.80 (s, 3H ), 3.90 (d, 1H), 4.20 (d, 1H), 4.24 (d, 1H), 5.33 (d, 1H), 6.83 (d, 2H), 7.19 (d, 2H). [α]D = -109.66° (c = 0.139, methanol).

Preparation 15

(5R)-4-(4-Methoxybenzyl)-5-methyl-3-morpholinone

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The title compound was obtained in 49% yield from the alcohol of Preparation 9, following a procedure similar to that described in Preparation 14. 1H-NMR (CDCl3, 400 MHz) δ: 1.24 (d, 3H), 3.30 (m , 1H), 3.60 (dd, 1H), 3.70 (dd, 1H), 3.78 (s, 3H), 3.85 (d, 1H), 4.18 (d, 1H), 4 .22 (d, 1H), 5.28 (d, 1H), 6.81 (d, 2H), 7.18 (d, 2H).

Preparation 16

4-methyl-3-morpholino

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A solution of chloroacetyl chloride (3.81 L, 50 mmol) in dichloromethane (100 mL) was added dropwise over 30 min to a suspension of 2-(methylamino)ethanol (4 mL, 50 mmol) and sodium hydroxide (2 g, 50 mmol). ) in dichloromethane (50 mL) and water (50 mL) and the mixture was stirred at room temperature for 72 hours, after which it was evaporated under reduced pressure. The residue was dissolved in ethanol (100 mL), potassium hydroxide (2.8 g, 50 mmol) was added and the mixture was stirred at 40 °C for 18 h and then filtered. The filtrate was concentrated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of pentane:ethyl acetate (100:0 to 50:50 to 0:100) to give the title compound, 3.42 g. 1H-NMR (CDCl3, 400 MHz ) δ: 2.98 (s, 3H), 3.34 (t, 2H), 3.84 (t, 2H), 4.14 (s, 2H). Found: C, 51.55; H, 8.02 N, 12.01. C5H9NO2 requires C 51.36; H, 7.93; N, 11.98%.

Preparation 17

tert-butyl-[2-(dimethylamino)ethoxy]acetate

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N,N-dimethylethanolamine (5.02 mL, 50 mmol) was added dropwise over 5 min to an ice-cold suspension of sodium hydride (2.2 g, 60% dispersion in mineral oil, 55 mmol) in tetrahydrofuran (100 mL) and the solution was stirred for 30 minutes. Tert-butyl bromoacetate (7.38 mL, 50 mmol) was added dropwise over 5 min and the mixture was allowed to warm to room temperature and stirred for a further 18 h. The mixture was preadsorbed on silica gel and purified by silica gel column chromatography eluting with a gradient of pentane:ethyl acetate:methanol (50:50:0 to 0:100:0 to 0:80:20) to give the title compound as a yellow oil. , 1.46 g. 1H-NMR (CDCl 3 , 400 MHz) δ: 1.50 (s, 9H), 2.30 (s, 6H), 2.59 (t, 2H), 3.64 (t, 2H), 4.00 (s, 2H). LRMS: m/z (ES+) 204 [MH+].

Preparation 18

tert-butyl-(3R)-3-(2-tert-butoxy-2-oxoethoxy)pyrrolidine-1-carboxylate

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Sodium hydride (704 mg, 60% in mineral oil, 17.6 mmol) was added to an ice-cold solution of tert-butyl-(3R)-3-hydroxypyrrolidine-1-carboxylate (J. Med. Chem. 1998, 41(25 ), 4983) (5 g, 26.7 mmol) in tetrahydrofuran (10 mL), and the mixture was allowed to warm to room temperature and stirred for 20 minutes. Tert-butyl bromoacetate (5.2 g, 26.7 mmol) was added and the mixture was heated at reflux for 18 h, then cooled and concentrated under reduced pressure. The residue was partitioned between ethyl acetate and water, and the phases were separated. The organic layer was dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of ethyl acetate:pentane (0:100 to 20:80) to give the title compound, 1.95 g. 1H-NMR (CDCl3, 400 MHz) δ: 1, 42 (s, 9H), 1.44 (s, 9H), 1.85-2.05 (m, 2H), 3.40 (m, 4H), 3.94 (m, 2H), 4.10 (m, 1H). LRMS: m/z (ES+) 324 [MNa+].

Preparation 19

Ethyl 3-cyclohexyl-3-methylbutanoate

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Trimethylsilyl chloride (1.3 mL, 10.2 mmol), copper(I) chloride (30 mg, 0.3 mmol) and cyclohexylmagnesium chloride (4.6 mL, 2N in diethyl ether, 9.2 mmol) were slowly added to an ice-cold solution of ethyl 3,3-dimethylacrylate (1 g, 8.5 mmol) in tetrahydrofuran (10 mL). The solution was stirred for 10 minutes, then allowed to warm to room temperature and stirred for another hour. Saturated aqueous ammonium chloride (10 mL) was added and the mixture was partitioned between water (10 mL) and diethyl ether (20 mL). The layers were separated and the aqueous phase was extracted with diethyl ether (2 x 10 mL). The combined organic extracts were dried (MgSO4) and concentrated under reduced pressure. The crude product was purified by silica gel column chromatography with ethyl acetate:pentane (5:95) as eluent to give the title compound, 1.2 g. 1H-NMR (CDCl3, 400 MHz) δ: 0.75 (m , 8H), 1.05-1.28 (m, 7H), 1.62 (m, 1H), 1.78 (m, 4H), 2.20 (s, 2H), 4.10 (q, 2H).

Preparation 20

3-cyclohexyl-3-methyl-1-butanol

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Lithium borohydride (1.23 g, 56.6 mmol) was added to a solution of the ester from Preparation 19 (4 g, 18.9 mmol) in tetrahydrofuran (30 mL) and the mixture was stirred at 50 °C for 18 h. Aqueous ammonium chloride solution (15 mL) was carefully added to the cooled solution and the mixture was extracted with ethyl acetate (3 x 30 mL). The combined organic extracts were washed with brine, dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography with ethyl acetate:pentane (20:80) as eluent to give the title compound, 1 g. 1H-NMR (CDCl3, 400 MHz) δ: 0.80 (s, 6H) , 0.88-1.18 (m, 6H), 1.50 (t, 2H), 1.60 (m, 1H), 1.70 (m, 4H), 3.64 (m, 2H).

Preparation 21

(3-bromo-1,1-dimethylpropyl)cyclohexane

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Triphenylphosphine (1.8 g, 7.1 mmol) was added portionwise to an ice-cold solution of alcohol from Preparation 20 (1 g, 5.9 mmol) and carbon tetrabromide (2.9 g, 8.8 mmol) in dichloromethane ( 15 mL). When the addition was complete, the mixture was stirred at room temperature for 72 hours. The solution was concentrated under reduced pressure and the residue was suspended in a mixture of pentane:ethyl acetate (5:1, by volume). The resulting precipitate was filtered through a pad of silica gel and washed with pentane:ethyl acetate (5:1, by volume, 300 mL). The combined filtrates were concentrated under reduced pressure and the product was purified by silica gel column chromatography, eluting with pentane, to give the title compound, 1.1 g. 1H-NMR (CDCl3, 400 MHz) δ: 0.80 (2× s, 6H), 0.90-1.20 (m, 6H), 1.62 (m, 3H), 1.75 (m, 2H), 1.81 (m, 2H), 3.36 (m , 2H).

Preparation 22

4-(2-bromoethyl)-1,1-dimethylcyclohexane

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A mixture of 2-(4,4-dimethylcyclohexyl)ethanol (WO 99/59971) (2 g, 12.8 mmol), concentrated sulfuric acid (750 μL) and 48% hydrobromic acid (3 mL) was stirred at 90°C. 7 o'clock. The cooled mixture was then carefully quenched with water (25 mL) and the mixture was extracted with dichloromethane (3 × 30 mL). The combined organic extracts were washed with 2M sodium carbonate solution and brine (30 mL), dried (MgSO4) and concentrated under reduced pressure. The remaining black gum was purified by distillation to give the title compound, 330 mg 1H-NMR (CDCl 3 , 400 MHz) δ: 0.82 (2×s, 6H), 1.02-1.20 (m, 4H) , 1.35 (m, 3H), 1.50 (m, 2H), 1.78 (m, 2H), 3.40 (t, 2H).

Preparation 23

2-[hydroxy-(1-propyl-1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-3-morpholinone

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A solution of the compound from Preparation 11 (13 g, 58.7 mmol) in tetrahydrofuran (100 mL) was added dropwise to a solution of lithium diisopropylamide (35.3 mL, 2M in tetrahydrofuran/heptane/ethylbenzene, 70.5 mmol) at -78 °C and the solution was stirred at -78°C for 20 minutes. The aldehyde from Preparation 1 (9.75 g, 70.5 mmol) was added dropwise and the mixture was allowed to warm to room temperature, after which it was stirred for 1.5 h. Ammonium chloride solution (100 mL) was added and the mixture was diluted with water (100 mL) and tetrahydrofuran (300 mL). The layers were separated, the aqueous phase was extracted with tetrahydrofuran (250 mL), and the combined organic solutions were dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of ethyl acetate:dichloromethane:methanol:0.88 ammonia (100:0:0:0 to 0:95:5:0.5) to give the title compound, 14 g. 1H-NMR (CDCl 3 , 400 MHz) (mixture of diastereomers) δ: 0.90 (t, 3H), 1.76 (m, 2H), 3.00 (m, 1H), 3.32-3, 43 (m, 1H), 3.61-3.81 (m, 6H), 3.98 (m, 1H), 4.42-4.58 (m, 3H), 4.75 (m, 0, 5H), 5.03 (m, 0.5H), 6.81 (m, 3H), 7.15 (m, 2H), 7.38 (s, 1H). LRMS: m/z (ES+) 360.0 [MH+].

Preparation 24

2-[hydroxy-(1-propyl-1H-imidazol-4-yl)methyl]-4-methyl-3-morpholinone

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Lithium diisopropylamide (17.4 mL, 2M in heptane/tetrahydrofuran/ethylbenzene, 34.8 mmol) was added over 10 min to a cooled (-78°C) solution of the compound from Preparation 10 (3.42 g, 29 mmol) in tetrahydrofuran ( 100 mL) and the resulting solution was stirred for 20 minutes. The aldehyde from Preparation 1 (4.81 g, 34.8 mmol) was added and the mixture was allowed to slowly warm to room temperature, after which it was stirred for 18 hours. Aqueous ammonium chloride solution (20 mL) was added and the mixture was evaporated under reduced pressure. The residue was purified by silica gel column chromatography eluting with a gradient of pentane:ethyl acetate:methanol:diethylamine (50:50:0:0 to 0:100:0:0 to 0:90:5:5) to give the title compound. as a yellow gum, 5.47 g. 1H-NMR (CDCl 3 , 400 MHz) (mixture of diastereomers) δ: 0.90 (t, 3H), 1.78 (m, 2H), 2.98 (2×s, 3H), 3.10 (m, 1H), 3.57 (m, 2H), 3.80 (m, 3H), 3.98-4.08 (m, 1H), 4.39, 4.49 , 4.86, 4.98, 5.22 (5×m, 2H), 6.88 (s, 1H), 7.38 (s, 1H). LRMS: m/z (TSP+) 254.2 [MH+].

Preparation 25

2-[(1-butyl-1H-imidazol-4-yl)(hydroxy)methyl]-4-(4-methoxybenzyl)-3-morpholinone

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A solution of the compound from Preparation 11 (3.63 g, 16.4 mmol) in tetrahydrofuran (40 mL) was added dropwise over 5 min to a solution of lithium diisopropylamide (9.8 mL, 2M in tetrahydrofuran/heptane/ethylbenzene, 19.7 mmol) at -78°C and the solution was stirred at -78°C for 30 minutes. The aldehyde from Preparation 2 (3.0 g, 19.7 mmol) was added dropwise and the mixture was allowed to warm to room temperature, after which it was stirred for 18 hours. The mixture was partitioned between ammonium chloride solution and ethyl acetate (300 mL). The layers were separated, and the organic solution was dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of ethyl acetate:methanol (100:0 to 90:10) to give the title compound, 4.35 g. 1H-NMR (CDCl 3 , 400 MHz) (mixture of diastereomers). δ: 0.96 (t, 3H), 1.35 (m, 2H), 1.78 (m, 2H), 3.03 (m, 1H), 3.42 (m, 1H), 3.66 -3.80 (m, 5H), 3.87 (m, 2H), 4.00 (m, 1H), 4.50 (m, 0.5H), 4.57 (m, 2.5H), 4.83 (m, 0.5H), 5.06 (m, 0.5H), 6.90 (m, 3H), 7.20 (m, 2H), 7.41 (s, 1H). LRMS: m/z (TSP+) 374.0 [MH+].

Preparation 26

2-[[1-(2-cyclohexylethyl)-1H-imidazol-4-yl](hydroxy)methyl]-4-(4-methoxybenzyl)-3-morpholino

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The title compound was obtained as a sticky mass in 66% yield from the compound of Preparation 11 and the aldehyde of Preparation 3, following a procedure similar to that described in Preparation 25. 1H-NMR (CDCl3, 400 MHz) (mixture of diastereomers) δ: 0, 98 (m, 2H), 1.22 (m, 4H), 1.58-1.78 (m, 7H), 3.03 (m, 1H), 3.38-3.50 (m, 1H) , 3.70-3.82 (m, 4H), 3.90 (m, 2H), 4.01 (m, 1H), 4.50 (d, 0.5H), 4.58 (m, 2 ,5H), 4.87 (m, 0.5H), 5.08 (m, 0.5H), 6.82-6.95 (m, 3H), 7.19 (m, 2H), 7, 41 (s, 1H). LRMS: m/z (TSP+) 428.1 [MH+].

Preparation 27

2-{hydroxy[1-(2-phenylethyl)-1H-imidazol-4-yl]methyl}-4-(4-methoxybenzyl)-3-morpholinone

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The title compound was obtained in 54% yield from the compound from Preparation 11 and the aldehyde from Preparation 4, following a procedure similar to that described in Preparation 25, except that the elution gradient was pentane:ethyl-acetate:methanol (50:50:0 to 0:100:0 to 0:87:13). 1H-NMR (CDCl3, 400 MHz) (mixture of diastereomers) δ: 3.02 (m, 3H), 3.42 (m, 1H), 3.74 (m, 4H), 3.95-4.05 ( m, 1H), 4.12 (m, 2H), 4.45 (m, 0.5H), 4.58 (m, 2.5H), 4.81 (m, 0.5H), 5.06 (d, 0.5H), 6.84 (m, 3H), 7.07 (d, 2H), 7.19 (m, 2H), 7.27 (m, 4H). LRMS: m/z (ES+) 422 [MH+].

Preparation 28

2-[hydroxy(1-trityl-1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-3-morpholinone

[image]

A solution of lithium diisopropylamide (88.5 mL, 1.5M in cyclohexane, 133 mmol) and tetrahydrofuran (100 mL) was cooled to -78°C. A solution of the compound from Preparation 11 (26 g, 118 mmol) in tetrahydrofuran (100 mL) was added dropwise and the solution was stirred for 30 minutes. Then, a solution of imidazole from Preparation 5 (39.9 g, 118 mmol) in tetrahydrofuran (350 mL) was added drop by drop over the course of 1 hour, and upon completion of the addition, the reaction was allowed to slowly warm to room temperature with stirring for 3 an hour. Saturated ammonium chloride solution (200 mL) and water (100 mL) were added, the phases were separated and the aqueous layer was extracted with ethyl acetate (100 mL). The combined organic solutions were dried (MgSO4) and evaporated under reduced pressure. The remaining orange oil was dissolved in ethyl acetate/methanol, the solution was sonicated and the resulting white precipitate was filtered off, washed with diethyl ether and dried to give the title compound, 21 g. The filtrate was evaporated under reduced pressure and the residue was purified by chromatography on silica gel column with ethyl acetate:methanol (96:4) as eluent to give more product, 28 g. 1H-NMR (CDCl3, 400 MHz) (mixture of diastereoisomers) δ: 2.98 (m, 1H), 3, 35 (m, 1H), 3.66 (m, 1H), 3.73 (s, 3H), 3.96 (m, 1H), 4.40 (d, 1H), 4.54 (s, 1H ), 4.60 (d, 1H), 5.21 (m, 1H), 6.79 (m, 3H), 7.08 (m, 6H), 7.15 (m, 2H), 7.26 (m, 9H), 7.37 (s, 1H). LRMS: m/z (TSP+) 560.2 [MH+].

Preparation 29

(6R)-2-[hydroxy(1-trityl-1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-6-methyl-3-morpholinone

[image]

The title compound was obtained in 62% yield from the morpholinone of Preparation 12 and the imidazole of Preparation 5, following a procedure similar to that described in Preparation 28. 1H-NMR (CDCl3, 400 MHz) (mixture of diastereomers) δ: 1.00, 1 .06 (2×m, 3H), 2.87-3.04 (m, 2H), 3.76 (m, 3H), 3.80-4.74 (m, 4H), 4.98-5 .21 (m, 1H), 6.78 (m, 3H), 7.05-7.20 (m, 8H), 7.23-7.40 (m, 10H). LRMS: m/z (ES+) 574 [MH+].

Preparation 30

(6S)-2-[hydroxy(1-trityl-1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-6-methyl-3-morpholinone

[image]

The title compound was obtained as a yellow foam in 53% yield from the morpholinone of Preparation 13 and the imidazole of Preparation 5, following a procedure similar to that described in Preparation 28. 1H-NMR (CDCl3, 400 MHz) (mixture of diastereomers) δ: 1, 00, 1.05 (2×m, 3H), 2.85-3.04 (m, 2H), 3.76 (m, 3H), 3.80-3.96 (m, 1H), 4, 39-4.70 (m, 3H), 4.98-5.20 (m, 1H), 6.78 (m, 3H), 7.05-7.20 (m, 8H), 7.25 ( m, 9H), 7.39 (m, 1H). LRMS: m/z (ES+) 574 [MH+].

Preparation 31

(5S)-2-[hydroxy(1-trityl-1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-5-methyl-3-morpholinone

[image]

The title compound was obtained in 57% yield from morpholinone from Preparation 14 and imidazole from Preparation 5, following a procedure similar to that described in Preparation 28. 1H-NMR (CDCl3, 400 MHz) (mixture of diastereomers) δ: 1.03-1 .19 (m, 3H), 2.99-4.83 (m, 8H), 5.02-5.38 (m, 2H), 6.78-6.88 (m, 3H), 7.10 -7.21 (m, 8H), 7.22-7.42 (m, 10H).

Preparation 32

(5R)-2-[hydroxy(1-trityl-1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-5-methyl-3-morpholinone

[image]

The title compound was obtained as a yellow foam in 80% yield from the morpholinone of Preparation 15 and the imidazole of Preparation 5, following a procedure similar to that described in Preparation 28. 1H-NMR (CDCl3, 400 MHz) (mixture of diastereomers) δ: 1, 00-1.16 (m, 3H), 3.16-4.94 (m, 8H), 5.00-5.37 (m, 2H), 6.72-6.83 (m, 3H), 7.04-7.19 (m, 8H), 7.21-7.40 (m, 10H).

Preparation 33

tert-butyl-2-[2-(dimethylamino)ethoxy]-3-hydroxy-3-(1-propyl-1H-imidazol-4-yl)propanoate

[image]

Lithium diisopropylamide (4.3 mL, 2M in heptane/tetrahydrofuran/ethylbenzene, 8.6 mmol) was added dropwise over 5 min to a solution of the amine from Preparation 17 (1.46 g, 7.2 mmol) in tetrahydrofuran ( 20 mL) and the solution was stirred at -78°C for 20 minutes. The aldehyde from Preparation 1 (1.18 g, 8.6 mmol) was added and the mixture was stirred for 3 hours, after which it was allowed to warm to -20°C. Water was added and the mixture was preadsorbed on silica gel. The product was purified by silica gel column chromatography eluting with a gradient of ethyl acetate:methanol:diethylamine (100:0:0 to 96:2:2) to give the title compound, 1.36 g. 1H-NMR (CDCl3, 400 MHz ) (mixture of diastereomers) δ: 0.88 (m, 3H), 1.35 (2×s, 9H), 1.75 (m, 2H), 2.22 (s, 6H), 2.42 (m , 1H), 2.58 (m, 1H), 3.55 (m, 1H), 3.80 (m, 2H), 3.90 (m, 1H), 4.17 (m, 1H), 4 .82, 5.00 (m, 1H), 6.90 (2×s, 1H), 7.35 (2×s, 1H). LRMS: m/z (TSP+) 342.2 [MH+].

Preparation 34

tert-butyl-(3S)-3-{1-tert-butoxycarbonyl-2-hydroxy-2-(1-propyl-1H-imidazol-4-yl)-ethoxy}pyrrolidine-1-carboxylate

[image]

A solution of the compound from Preparation 18 (5.67 g, 18.8 mmol) in tetrahydrofuran (20 mL) was added dropwise to a solution of lithium diisopropylamide (11.3 mL, 2M in heptane/tetrahydrofuran/ethylbenzene, 22.6 mmol) at -78°C and the solution was stirred at -78°C for 20 minutes. Aldehyde from Preparation 1 (3.12 g, 22.6 mmol) was added portion by portion and the mixture was allowed to warm to room temperature, after which it was stirred for 18 hours. Ammonium chloride solution (50 mL) was carefully added and the mixture was extracted with tetrahydrofuran (2 x 200 mL). The combined organic solutions were dried (MgSO4) and evaporated under reduced pressure. The remaining orange oil was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (100:0:0 to 90:10:1) to give the title compound, 3.7 g. 1H-NMR (CDCl3 , 400 MHz) (mixture of diastereomers) δ: 0.92 (t, 3H), 1.42 (s, 18H), 1.79 (m, 2H), 1.94-2.14 (m, 1H), 2.75-3.50 (m, 5H), 3.84 (m, 2H), 4.03-4.35 (m, 2H), 4.81-5.08 (m, 2H), 6, 88 (m, 1H), 7.39 (s, 1H). LRMS: m/z (ES+) 440 [MH+].

Preparation 35

(2EZ)-4-(4-methoxybenzyl)-2-[(1-propyl-1H-imidazol-4-yl)methylidene]-3-morpholinone

[image]

Triethylamine (9.19 mL, 65.9 mmol) was added to a solution of the alcohol from Preparation 23 (15.8 g, 44.0 mmol) in dichloromethane (300 mL). The solution was cooled on ice, methanesulfonyl chloride (5.1 mL, 65.9 mmol) was added and the solution was stirred for 2 hours at room temperature. More triethylamine (3.06 mL, 22 mmol) was added and the mixture was stirred at 40°C for 18 hours, after which it was cooled. The mixture was diluted with dichloromethane (1000 mL) and washed with sodium bicarbonate solution (200 mL). The aqueous wash was extracted with dichloromethane (400 mL), and the combined organic solutions were dried (MgSO4) and evaporated under reduced pressure. The residue was purified by silica gel column chromatography with an elution gradient of ethyl acetate:dichloromethane:methanol:0.88 ammonia (100:0:0:0 to 0:95:5:0.5 to 0:90:10:1) to would give the title compound, 8.3 g. 1H-NMR (CDCl 3 , 400 MHz) (mixture of diastereomers) δ: 0.90 (t, 3H), 1.78 (m, 2H), 3.39 (t, 2H), 3.77 (s, 3H), 3.83 (t, 2H), 4.15 (t, 2H), 4.61 (s, 2H), 6.81 (d, 2H), 7, 00 (s, 1H, 7.19 (d, 2H), 7.28 (s, 1H), 7.41 (s, 1H). LRMS: m/z (ES + ) 342 [MH + ].

Preparation 36

(2EZ)-4-(4-methoxybenzyl)-2-[(1-trityl-1H-imidazol-4-yl)methylidene]-3-morpholinone

[image]

The title compound was obtained in 77% yield as a yellow foam from the alcohol of Preparation 28, following the procedure described in Preparation 35. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 3.34 (t, 2H) , 3.78 (s, 3H), 4.00 (t, 2H), 4.59 (s, 2H), 6.80 (d, 2H), 6.98 (s, 1H), 7.10 ( m, 6H), 7.17 (d, 2H), 7.28 (m, 10H), 7.39 (s, 1H). LRMS: m/z (ES+) 542 [MH+].

Preparation 37

(2EZ,6R)-4-(4-methoxybenzyl)-6-methyl-2-[(1-trityl-1H-imidazol-4-yl)methylidene]-3-morpholinone

[image]

The title compound was obtained in 57% yield as a pale yellow foam from the alcohol of Preparation 29, following the procedure described in Preparation 35. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 1.02 (d, 3H ), 3.08 (dd, 1H), 3.20 (dd, 1H), 3.77 (s, 3H), 4.10 (m, 1H), 4.50 (d, 1H), 4.60 (d, 1H), 6.80 (d, 2H), 6.99 (s, 1H), 7.14 (m, 8H), 7.28 (m, 10H), 7.38 (s, 1H) . LRMS: m/z (ES+) 556 [MH+].

Preparation 38

(2EZ,6S)-4-(4-methoxybenzyl)-6-methyl-2-[(1-trityl-1H-imidazol-4-yl)methylidene]-3-morpholinone

[image]

Methanesulfonyl chloride (911 μL, 11.78 mmol) was added dropwise to an ice-cold solution of the alcohol from Preparation 30 (4.5 g, 7.85 mmol) in dichloromethane (40 mL) and triethylamine (1.64 mL and 11 .78 mmol) and the solution was stirred at room temperature for 1 hour. More triethylamine (546 μL, 3.93 mmol) was added and the mixture was stirred at 40°C for 18 hours. The cooled mixture was partitioned between dichloromethane (50 mL) and water (50 mL) and the layers were separated. The organic phase was dried (MgSO4) and concentrated under reduced pressure. The remaining orange oil was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (99:1:0.1 to 98:2:0.2) to give the title compound as a yellow foam, 2.5 g. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 1.02 (d, 3H), 3.05 (m, 1H), 3.20 (m, 1H), 3.78 (s , 3H), 4.06 (m, 1H), 4.47-4.63 (m, 2H), 6.80 (d, 2H), 6.98 (s, 1H), 7.12 (m, 8H), 7.27 (m, 10H), 7.38 (s, 1H). LRMS: m/z (ES+) 556 [MH+].

Preparation 39

(2EZ,5S)-4-(4-methoxybenzyl)-5-methyl-2-[(1-trityl-1H-imidazol-4-yl)methylidene]-3-morpholinone

[image]

The title compound was obtained in 28% yield from the alcohol of Preparation 31, following a procedure similar to that described in Preparation 38. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 1.22 (d, 3H), 3.38 (m, 1H), 3.78 (s, 3H), 3.81 (d, 1H), 3.95 (m, 2H), 5.28 (d, 1H), 6.80 (d , 2H), 6.95 (s, 1H), 7.10-7.19 (m, 9H), 7.28 (m, 9H), 7.40 (s, 1H).

Preparation 40

(2EZ,5R)-4-(4-methoxybenzyl)-5-methyl-2-[(1-trityl-1H-imidazol-4-yl)methylidene]-3-morpholinone

[image]

The title compound was obtained in 27% yield from the alcohol of Preparation 32, following a procedure similar to that described in Preparation 38. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 1.25 (d, 3H), 3.41 (m, 1H), 3.78 (s, 3H), 3.83 (dd, 1H), 3.98 (m, 2H), 5.30 (d, 1H), 6.82 (d , 2H), 6.98 (s, 1H), 7.18 (m, 9H), 7.32 (m, 9H), 7.41 (s, 1H).

Preparation 41

(2EZ)-2-[(1-butyl-1H-imidazol-4-yl)methylidene]-4-(4-methoxybenzyl)-3-morpholinone

[image]

Triethylamine (1.78 mL, 12.8 mmol) was added to a solution of alcohol from Preparation 25 (4.34 g, 11.6 mmol) in dichloromethane (50 mL). The solution was cooled in ice and methanesulfonyl chloride (990 μL, 12.8 mmol) was added. The solution was stirred for 30 minutes, more triethylamine (1.78 mL, 12.8 mmol) was added and the mixture was stirred at room temperature for 18 hours. The mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with a gradient of dichloromethane:ethyl acetate:methanol (100:0:0 to 0:90:10) to give the title compound as a sticky mass, 1, 12 g 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 0.95 (t, 3H), 1.34 (m, 2H), 1.78 (m, 2H), 3.42 ( t, 2H), 3.80 (s, 3H), 3.94 (t, 2H), 4.19 (t, 2H), 4.63 (s, 2H), 6.84 (d, 2H), 7.02 (s, 1H), 7.22 (d, 2H), 7.32 (s, 1H), 7.44 (s, 1H). LRMS: m/z (TSP+) 356.2 [MH+].

Preparation 42

(2EZ)-2-{[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]methylidene}-4-(4-methoxybenzyl)-3-morpholinone

[image]

The title compound was obtained as a sticky mass in 57% yield from the alcohol of Preparation 26, following the procedure described in Preparation 41. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 0.94 (m, 2H) , 1.18 (m, 4H), 1.64 (m, 7H), 3.40 (t, 2H), 3.78 (s, 3H), 3.90 (t, 2H), 4.17 ( t, 2H), 4.61 (s, 2H), 6.81 (d, 2H), 6.99 (s, 1H), 7.20 (d, 2H), 7.25 (s, 1H), 7.40 (s, 1H). LRMS: m/z (TSP+) 410.1 [MH+].

Preparation 43

(2EZ)-4-(4-methoxybenzyl)-2-{[1-(2-phenylethyl)-1H-imidazol-4-yl]methylidene}-3-morpholinone

[image]

Triethylamine (0.98 mL, 7.03 mmol) was added to a solution of the alcohol from Preparation 27 (1.41 g, 3.35 mmol) in dichloromethane (15 mL). The solution was cooled in ice, methanesulfonyl chloride (311 μL, 4.02 mmol) was added and the mixture was heated to 40°C and stirred for 18 hours, after which it was concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with a gradient of pentane:ethyl acetate:methanol (75:25:0 to 0:100:0 to 0:95:5) to give the title compound as an orange oil, 449 mg. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 3.03 (t, 2H), 3.42 (t, 2H), 3.80 (s, 3H), 4.18 (m, 4H ), 4.63 (s, 2H), 6.85 (d, 2H), 7.02 (s, 1H), 7.06 (d, 2H), 7.25 (m, 7H). LRMS: m/z (ES+) 404 [MH+].

Preparation 44

tert-butyl-(2EZ)-2-[2-(dimethylamino)ethoxy]-3-(1-propyl-1H-imidazol-4-yl)-2-propenoate

[image]

Methanesulfonyl chloride (340 μL, 4 .4 mol). The solution was stirred at room temperature for 1 hour, more triethylamine (616 μL, 4.4 mmol) was added and the solution was stirred at room temperature for 18 hours. TLC analysis showed retention of the starting material, so the solution was heated to reflux and stirred for another 3 hours. The cooled mixture was preadsorbed onto silica gel and purified by silica gel column chromatography eluting with a gradient of ethyl acetate:diethylamine:methanol (100:0:0 to 96:2:2) to give the title compound, 650 mg. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 0.98 (t, 3H), 1.56 (s, 9H), 1.84 (m, 2H), 2.34 (s, 6H ), 2.65 (t, 2H), 3.94 (t, 2H), 4.04 (t, 2H), 7.08 (s, 1H), 7.44 (s, 1H), 7.98 (s, 1H). LRMS: m/z (TSP+) 324.2 [MH+].

Preparation 45

tert-butyl-(3S)-3-{[(EZ)-1-(tert-butoxycarbonyl)-2-(1-propyl-1H-imidazol-4-yl)ethenyl]oxy}pyrrolidine-1-carboxylate

[image]

The title compound was obtained as an orange oil in 36% yield from the alcohol of Preparation 34, following the procedure described in Preparation 44. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 0.93 (t, 3H) , 1.35-1.56 (m, 18H), 1.80 (m, 2H), 1.98 (m, 1H), 2.17 (m, 1H), 3.26-3.66 (m , 4H), 3.86 (m, 2H), 5.15 (m, 1H), 7.06, 7.15 (2×s, 1H), 7.35, 7.39 (2×s, 1H ), 7.41 (s, 1H). LRMS: m/z (ES+) 422 [MH+].

Preparation 46

(2EZ)-4-methyl-2-[(1-propyl-1H-imidazol-4-yl)methylidene]-3-morpholinone

[image]

To a solution of alcohol from Preparation 24 (5.47 g, 21.6 mmol) in dichloromethane (80 mL) was added triethylamine (3.32 mL, 23.8 mmol) and the solution was cooled on ice. A solution of methanesulfonyl chloride (1.84 mL, 23.8 mmol) in dichloromethane (3 mL) was added over 5 minutes and the solution was stirred at room temperature for 1 hour. The mixture was evaporated under reduced pressure, the residue was dissolved in N,N-dimethylformamide (15 mL), triethylamine (3.32 mL, 23.8 mmol) was added and the solution was heated at reflux and stirred for 20 min. The cooled solution was concentrated under reduced pressure and the residue was dissolved in methanol (100 mL), then adsorbed on silica gel and purified by chromatography on silica gel with an elution gradient of ethyl acetate:methanol:diethylamine (100:0:0 to 95:5:0 ,5) to give the title compound, 2.5 g. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 0.98 (t, 3H), 1.82 (m, 2H), 3 .14 (s, 3H), 3.59 (t, 2H), 3.90 (t, 2H), 4.26 (t, 2H), 7.00 (s, 1H), 7.35 (s, 1H), 7.45 (s, 1H). LRMS: m/z (TSP+) 236.2 [MH+].

Preparation 47

(-)-(2S)-4-(4-Methoxybenzyl)-2-[(1-propyl-1H-imidazol-4-yl)methyl]-3-morpholinone

[image]

A mixture of the alkene from Preparation 35 (8.3 g, 24.3 mmol) and 10% Pd/C (Degussa® 101) (800 mg) in ethanol (240 mL) was hydrogenated at 100 psi (90 kPa) and 50°C. for 18 hours, then cooled and filtered through Arbocel®. The filtrate was evaporated under reduced pressure to give an orange oil. This product was purified by chromatography on a Chiralcel® OJ column, with hexane:isopropyl-alcohol:diethylamine (70:30:0.5) as eluent, to give enantiomer 1, 1.65 g, and then enantiomer 2, the compound from title, 1.54 g. 1H-NMR (CDCl3, 400 MHz) δ: 0.92 (t, 3H), 1.78 (m, 2H), 3.07 (m, 2H), 3.38 (m , 2H), 3.74 (m, 1H), 3.79 (m, 5H), 3.98 (m, 1H), 4.52 (m, 3H), 6.72 (s, 1H), 6 .82 (d, 2H), 7.18 (d, 2H), 7.38 (s, 1H). LRMS: m/z (ES+) 344 [MH+]. [α]D = -66.10° (c = 0.368, methanol).

Preparation 48

(2RS)-2-[(1-butyl-1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-3-morpholinone

[image]

A mixture of the alkene from Preparation 41 (2.5 g, 6.96 mmol) and 10% Pd/C (Degussa® 101) (250 mg) in ethanol (100 mL) was hydrogenated at 50°C and 60 psi (410 kPa) during 18 hours. The cooled mixture was filtered through Arbocel® and the filtrate was evaporated under reduced pressure to give the title compound as an oil, 2.44 g. 1H-NMR (CDCl3, 400 MHz) δ: 0.95 (t, 3H), 1 .32 (m, 2H), 1.73 (m, 2H), 3.04 (m, 2H), 3.34-3.42 (m, 2H), 3.73 (m, 1H), 3, 78 (s, 3H), 3.82 (t, 2H), 3.98 (m, 1H), 4.45-4.60 m, 3H), 6.74 (s, 1H), 6.82 ( d, 2H), 7.18 (d, 2H), 7.38 (s, 1H). LRMS: m/z (TSP+) 358.2 [MH+].

Preparations 49 to 50

Compounds of the following general formula:

[image]

were prepared from the corresponding alkenes (Preparations 42 and 43), following a procedure similar to that described in Preparation 48.

[image]

1 = purified by chromatography on a silica gel column with an elution gradient of ethyl acetate:methanol (100:0 to 90:10).

Preparation 51

(2RS)-4-methyl-2-[(1-propyl-1H-imidazol-4-yl)methyl]-3-morpholinone

[image]

A mixture of the alkene from Preparation 46 (2.5 g, 1.06 mmol) and 10% Pd/C (Degussa® 101) (250 mg) in ethanol (50 mL) was hydrogenated at 50°C and 60 psi (410 kPa) during 18 hours. The cooled mixture was filtered through Arbocel®, and the filtrate was evaporated under reduced pressure. The residue was purified by silica gel column chromatography eluting with an ethyl acetate:methanol:diethylamine gradient (100:0:0 to 96.5:1.75:1.75) to give the title compound as a colorless oil, 2.08 g. 1H-NMR (CDCl 3 , 400 MHz) δ: 0.95 (t, 3H), 1.80 (m, 2H), 3.00 (m, 4H), 3.18 (m, 1H), 3 .37 (m, 1H), 3.57 (m, 1H), 3.82 (m, 3H), 4.03 (m, 1H), 4.46 (dd, 1H), 6.78 (s, 1H), 7.39 (s, 1H). LRMS: m/z (ES+) 238 [MH+].

Preparation 52

(2RS)-2-[(1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-3-morpholinone

[image]

A mixture of the protected imidazole from Preparation 36 (25 g, 46 mmol) and Pd/C (Degussa® 101) catalyst (2.5 g) in ethanol (500 mL) was hydrogenated at 50°C and 60 psi (410 kPa) for 18 hours. TLC analysis showed residual starting material, so the mixture was filtered through Arbocel®, and the filtrate was hydrogenated with fresh catalyst (2.5 g) in ethanol (500 mL) at 50°C and 60 psi (410 kPa) for a further 18 hours. The mixture was filtered through Arbocel®, and the filtrate was evaporated under reduced pressure. The product was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (98:2:0.2 to 95:5:0.5) to give the title compound as a white foam, 9 g. 1H- NMR (CDCl3, 400 MHz) δ: 3.01 (m, 1H), 3.18 (m, 2H), 3.38 (m, 1H), 3.70 (dd, 1H), 3.78 (s , 3H), 3.97 (dd, 1H), 4.32 (m, 1H), 4.50 (dd, 2H), 6.81 (d, 2H), 6.86 (s, 1H), 7 .02 (d, 2H), 7.46 (s, 1H). LRMS: m/z (ES+) 302 [MH+].

Preparation 53

(-)-(2S)-2-[(1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-3-morpholinone

and

Preparation 54

(+)-(2S)-2-[(1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-3-morpholinone

The racemic compound from Preparation 52 was further purified by HPLC with a Chiralpak® OD column and hexane:isopropyl alcohol (80:20) as eluent to give the title compound from Preparation 53, >99% ee. 1H-NMR (CDCl3, 400 MHz) δ: 3.01 (m, 1H), 3.18 (m, 2H), 3.38 (m, 1H), 3.70 (dd, 1H), 3.78 (s, 3H), 3.97 (dd, 1H), 4.32 (m, 1H), 4.50 (dd, 2H), 6.81 (d, 2H), 6.86 (s, 1H) , 7.02 (d, 2H), 7.46 (s, 1H). LRMS: m/z (ES+) 324 [MNa+]. [α]D = -529.3° (c = 0.05, methanol).

Additional elution gave the title compound Preparation 54, >99% ee. 1H-NMR (CDCl3, 400 MHz) δ: 3.01 (m, 1H), 3.18 (m, 2H), 3.38 (m, 1H), 3.66-3.79 (m, 4H) , 3.97 (dd, 1H), 4.32 (t, 1H), 4.50 (dd, 2H), 6.81 (d, 2H), 6.86 (s, 1H), 7.02 ( d, 2H), 7.46 (s, 1H). LRMS: m/z (ES+) 324 [MNa+].

Preparation 55a

(2S,6R)-2-[(1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-6-methyl-3-morpholinone

and

Preparation 55b

(2R,6R)-2-[(1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-6-methyl-3-morpholinone

[image]

A mixture of the alkene from Preparation 154 (41 g, 131 mmol) and 10% palladium on carbon (Degussa® type 101) (8 g) in ethanol (500 mL) was hydrogenated at 50 psi (345 kPa) and 60°C for 24 hours. The mixture was filtered through Arbocel®, the filtrate was concentrated under reduced pressure and the residue was azeotroped with dichloromethane. The crude product was purified by chromatography on a silica gel column with a Biotage column, eluting with a solvent gradient of dichloromethane:methanol:0.88 ammonia (97.5:2.5:0.1 to 90:10:1). The runoff, the major product, was further purified by silica gel column chromatography with dichloromethane:methanol:0.88 ammonia (95:5:0.25) to give the title compound of Preparation 55a as a white foam. 1H-NMR (CDCl3, 400 MHz) δ: 1.19 (d, 3H), 2.97-3.22 (m, 4H), 3.78 (s, 3H), 3.84 (m, 1H) , 4.32 (t, 1H), 4.40 (d, 1H), 4.50 (d, 1H), 6.80 (d, 2H), 6.86 (s, 1H), 7.00 ( d, 2H), 7.46 (s, 1H). LRMS: m/z (ES-) 314 [M-H]-.

The slower flow byproduct was further purified by silica gel column chromatography with ether:methanol:0.88 ammonia (90:10:1) as eluent and the product was azeotroped with dichloromethane to give the title compound of Preparation 55b as a white foam, 220 mg. 1H-NMR (CDCl3, 400 MHz) δ: 1.19 (d, 3H), 2.97-3.22 (m, 4H), 3.78 (s, 3H), 3.84 (m, 1H) , 4.32 (t, 1H), 4.40 (d, 1H), 4.50 (d, 1H), 6.80 (d, 2H), 6.86 (s, 1H), 7.00 ( d, 2H), 7.46 (s, 1H).

Preparation 56

(2R,6S)-2-[(1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-6-methyl-3-morpholinone

[image]

The title compound was obtained in 20% yield from the protected alkene of Preparation 38, following the procedure described in Preparation 52. 1H-NMR (CDCl3, 400 MHz) δ: 1.19 (d, 3H), 2.99 (m, 2H), 3.17 (m, 2H), 3.78 (s, 3H), 3.83 (m, 1H), 4.30 (m, 1H), 4.44 (dd, 2H), 6, 80 (d, 2H), 6.85 (s, 1H), 7.00 (d, 2H), 7.45 (s, 1H). LRMS: m/z (ES+) 316 [MH+].

Preparation 57

(2S,5S)-2-[(1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-5-methyl-3-morpholinone

[image]

The title compound was obtained in 47% yield from the protected alkene of Preparation 39, following a procedure similar to that described in Preparation 52. 1H-NMR (CDCl3, 400 MHz) (~3:1 mixture of C-2 stereoisomer shown) δ: 1 .15 (d, 3H), 3.22 (m, 3H), 3.48, 3.70-4.00 (2×m, 6H), 4.38 (m, 1H), 5.18-5 .37 (m, 1H), 6.83 (d, 2H), 6.98 (m, 1H), 7.18 (d, 2H), 7.62 (s, 1H). LRMS: m/z (ES-) 314 [M-H-].

Preparation 58

(2R,5R)-2-[(1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-5-methyl-3-morpholinone

[image]

The title compound was obtained in 67% yield from the protected imidazole of Preparation 40, following a procedure similar to that described in Preparation 52. 1H-NMR (CDCl3, 400 MHz) (~3:1 mixture of the C-2 stereoisomer shown) δ: 1 .16 (d, 3H), 3.22 (m, 3H), 3.46, 3.70-3.98 (2×m, 6H), 4.35, 4.39 (2×m, 1H) , 5.20, 5.30 (2×d, 1H), 6.82, 6.94 (2×m, 3H), 7.18 (d, 2H), 7.53 (2×s, 1H) . LRMS: m/z (ES-) 314 [M-H-].

Preparation 59

(2RS)-({1-[2-(4-bromophenyl)ethyl]-1H-imidazol-4-yl}methyl)-4-(4-methoxybenzyl)-3-morpholinone

[image]

Sodium hydride (836 mg, 60% dispersion in mineral oil, 20.9 mmol) was added portionwise to an ice-cold solution of imidazole from Preparation 52 (5 g, 19.9 mmol) in tetrahydrofuran (100 mL) and the solution was stirred for 15 minutes. 4-Bromophenylethyl methanesulfonate (Bioorg. Med. Chem. 1996; 4(5); 645) (6.1 g, 21.9 mmol) was then added and the mixture was stirred at room temperature for 3 days. The reaction was quenched with water (50 mL), and the mixture was extracted with ethyl acetate (2×100 mL). The combined organic extracts were dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (99:1:0.1 to 95:5:0.5) to give the title compound, 4.4 g. 1H- NMR (CDCl 3 , 400 MHz) δ: 2.97 (t, 2H), 3.04 (m, 2H), 3.36 (m, 2H), 3.70 (m, 1H), 3.78 (s , 3H), 3.97 (m, 1H), 4.03 (t, 2H), 4.45 (m, 1H), 4.54 (dd, 2H), 6.62 (s, 1H), 6 .82 (d, 2H), 6.90 (d, 2H), 7.18 (d, 2H), 7.20 (s, 1H), 7.39 (d, 2H).

Preparation 60

(-)-(2S)-2-{[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]methyl}-4-(4-methoxybenzyl)-3-morpholinone

[image]

A mixture of the compound from Preparation 53 (400 mg, 1.32 mmol), cesium carbonate (472 mg, 1.45 mmol) and 2-cyclohexylethyl bromide (227 μL, 1.45 mmol) in N,N-dimethylformamide (4 mL ) was mixed at 80°C for 18 hours. The cooled mixture was partitioned between ethyl acetate (250 mL) and water (100 mL) and the layers were separated. The organic phase was washed with water (3 x 100 mL), dried (MgSO4) and evaporated under reduced pressure. The crude product was purified twice by chromatography. on a silica gel column, using first an elution gradient dichloromethane:methanol:0.88 ammonia (100:0:0 to 95:5:0.5), and then an elution gradient ethyl acetate:diethylamine (100:0 to 95: 5) to give the title compound as a colorless gum, 191 mg 1H-NMR (CDCl 3 , 400 MHz) δ: 0.90 (m, 2H), 1.04-1.24 (m, 4H), 1, 60 (m, 7H), 3.02 (m, 2H), 3.26-3.40 (m, 2H), 3.70 (m, 1H), 3.78 (s, 3H), 3.81 (t, 2H), 3.97 (m, 1H), 4.42-4.58 (m, 3H), 6.68 (s, 1H), 6.80 (d, 2H), 7.14 ( d, 2H), 7.35 (s, 1H). LRMS: m/z (ES+) 412 [MH+]. [α]D = -50.37° (c = 0.112, methanol).

Preparation 61

(2R,6S)-2-{[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]methyl}-4-(4-methoxybenzyl)-6-methyl-3-morpholinone

[image]

A mixture of the compound from Preparation 56 (200 mg, 0.635 mmol), cesium carbonate (248 mg, 0.762 mmol) and 2-cyclohexylethyl bromide (109 μL, 0.70 mmol) in N,N-dimethylformamide (8 mL) was stirred at 70°C for 18 hours. The cooled mixture was partitioned between ethyl acetate (10 mL) and water (10 mL) and the layers were separated. The organic phase was washed with water (2 x 20 mL) and brine (20 mL), then dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (99:1:0.1 to 98:2:0.2) to give the title compound as a colorless gum, 120 mg. 1H-NMR (CDCl3, 400 MHz) δ: 0.90 (m, 2H), 1.18 (m, 7H), 1.62 (m, 7H), 2.94-3.08 (m, 3H) , 3.32 (dd, 1H), 3.78 (s, 3H), 3.80 (m, 3H), 4.43 (m, 3H), 6.66 (s, 1H), 6.80 ( d, 2H), 7.14 (d, 2H), 7.32 (s, 1H). LRMS: m/z (ES+) 426 [MH+].

Preparation 62

(2S,6R)-2-{[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]methyl}-4-(4-methoxybenzyl)-6-methyl-3-morpholinone

[image]

The title compound was obtained as a colorless oil in 38% yield from imidazole from Preparation 55 and 2-cyclohexylethyl bromide, following the procedure described in Preparation 61. 1H-NMR (CDCl3, 400 MHz) δ: 0.90 (m, 2H ), 1.17 (m, 7H), 1.61 (m, 7H), 2.94-3.08 (m, 3H), 3.32 (dd, 1H), 3.78 (s, 3H) , 3.81 (m, 3H), 4.43 (m, 3H), 6.66 (s, 1H), 6.80 (d, 2H), 7.12 (d, 2H), 7.32 ( with, 1H). LRMS: m/z (ES+) 448 [MNa+].

Preparation 63

(2S,5S)-2-{[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]methyl}-4-(4-methoxybenzyl)-5-methyl-3-morpholinone

[image]

The title compound was obtained in 21% yield from the imidazole of Preparation 57, following a procedure similar to that described in Preparation 61, except that ethyl acetate:methanol:diethylamine (98:1:1) was used as the eluent from the column. 1H-NMR (CDCl3, 400 MHz) (mixture of C-2 diastereoisomers) δ: 0.98 (m, 2H), 1.20 (m, 7H), 1.68 (m, 7H), 3.05-3 .98 (m, 11H), 4.50 (m, 1H), 5.22-5.40 (m, 1H), 6.76, 6.84 (s and m, 3H), 7.14, 7.20 (2×d, 2H), 7.39 (m, 1H). LRMS: m/z (ES+) 426 [MH+].

Preparation 64

(2R,5R)-2-{[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]methyl}-4-(4-methoxybenzyl)-5-methyl-3-morpholinone

[image]

The title compound was obtained in 40% yield from the imidazole of Preparation 58, following a procedure similar to that described in Preparation 61. 1H-NMR (CDCl3, 400 MHz) (mixture of C-2 diastereomers) δ: 0.90 (m, 2H ), 1.18 (m, 7H), 1.63 (m, 7H), 3.01-3.33 (m, 3H), 3.61-3.90 (m, 8H), 4.42 ( m, 1H), 5.22 (m, 1H), 6.72 (s, 1H), 6.80 (d, 2H), 7.16 (d, 2H), 7.35 (s, 1H). LRMS: m/z (ES+) 426 [MH+].

Preparation 65

(2S,6R)-2-{[1-(3-cyclohexyl-3-methylbutyl)-1H-imidazol-4-yl]methyl}-4-(4-methoxybenzyl)-6-methyl-3-morpholinone

[image]

A mixture of bromide from Preparation 21 (420 mg, 1.8 mmol), imidazole from Preparation 55 (470 mg, 1.5 mmol) and cesium carbonate (586 mg, 1.8 mmol) in N,N-dimethylformamide (5 mL) it was stirred at 70°C for 18 hours. The cooled mixture was concentrated under reduced pressure and partitioned between ethyl acetate (20 mL) and water (20 mL), and the layers were separated. The aqueous phase was extracted with ethyl acetate (3 × 15 mL), and the combined organic solutions were washed with brine (15 mL), dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by chromatography on a Biotage® silica column, with toluene:diethylamine (99:1 to 85:15) as eluent to give the title compound, 60 mg. 1H-NMR (CDCl3, 400 MHz) δ: 0.82 (s, 6H), 0.90-1.10 (m, 8H), 1.63 (m, 3H), 1.78 (m, 5H) , 2.94-3.06 (m, 3H), 3.30 (dd, 1H), 3.79 (m, 6H), 4.40-4.55 (m, 3H), 6.68 (s , 1H), 6.80 (d, 2H), 7.10 (d, 2H), 7.32 (s, 1H). LRMS: m/z (ES+) 468 [MH+].

Preparation 66

(2S,6R)-2-({1-[-2-(4,4-dimethylcyclohexyl)ethyl]-1H-imidazol-4-yl}methyl)-4-(4-methoxybenzyl)-6-methyl-3 -morpholino

[image]

The title compound was obtained in 10% yield from the bromide of Preparation 22 and the imidazole of Preparation 55, following a procedure similar to that described in Preparation 65. 1H-NMR (CDCl3, 400 MHz) δ: 0.84 (2×s, 6H ), 1.16 (m, 8H), 1.37 (m, 2H), 1.50 (m, 2H), 1.63 (m, 2H), 2.97-3.10 (m, 3H) , 3.35 (dd, 1H), 3.79 (s, 3H), 3.85 (m, 3H), 4.48 (m, 3H), 6.70 (s, 1H), 6.82 ( d, 2H), 7.16 (d, 2H), 7.35 (s, 1H). LRMS: m/z (ES+) 454 [MH+].

Preparation 67

(2RS)-2-({1-[(2EZ)-3-bromo-2-propenyl]-1H-imidazol-4-yl}methyl)-4-(4-methoxybenzyl)-3-morpholinone

[image]

Sodium hydride (133 mg, 60% dispersion in mineral oil, 3.32 mmol) was added to an ice-cold solution of morpholinone from Preparation 52 (1 g, 3.32 mmol) in tetrahydrofuran (10 mL). 1,3-dibromo-1-propene (mixture of cis and trans isomers) (332 μL, 3.32 mmol) was added dropwise over 5 min and the mixture was allowed to warm to room temperature and stir for 2 h. The mixture was partitioned between water (50 mL) and ethyl acetate (50 mL) and the phases were separated. The aqueous layer was extracted with ethyl acetate (10 mL), and the combined organic solutions were dried (Na2SO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography with dichloromethane:methanol:0.88 ammonia (98:2:0.2) as eluent to give the title compound as a colorless oil, 1.1 g. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 3.02 (m, 2H), 3.26-3.40 (m, 2H), 3.68 (m, 1H), 3.78 (s, 3H), 3 .96 (m, 1H), 4.40 (d, 1H), 4.42-4.55 (m, 3H), 4.60 (m, 1H), 6.22 (m, 1H), 6, 70 (2×s, 1H), 6.80 (m, 3H), 7.15 (m, 2H), 7.38 (2×s, 1H). LRMS: m/z (ES+) 420, 422 [MH+].

Preparation 68

(2RS)-4-(4-methoxybenzyl)-2-[(1-phenyl-1H-imidazol-4-yl)methyl]-3-morpholinone

[image]

To a solution of imidazole from Preparation 52 (500 mg, 1.66 mmol) and pyridine (269 μL, 3.32 mmol) in dichloromethane (20 mL) was added copper (II)-acetate (452 mg, 2.49 mmol), molecular 4 Å sieves and benzeneboronic acid (405 mg, 3.32 mmol) and the mixture was stirred at room temperature for 18 hours. Compressed air was then blown through the solution for 10 hours. The solvent was retained using a "cold finger" condenser. The solution was then stirred under a nitrogen atmosphere for the next 18 hours. A solution of ethylenediaminetetraacetic acid (800 mg) in saturated sodium bicarbonate solution (35 mL) was added and the mixture was stirred for 20 minutes, after which it was extracted with dichloromethane (70 mL). The aqueous phase was further extracted with dichloromethane (20 mL), and the combined organic solutions were washed with water (10 mL), dried (MgSO4) and concentrated under reduced pressure. The remaining black solid was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (98:2:0.2 to 97:3:0.3) to give the title compound as a pale yellow oil, 175 mg . 1H-NMR (CDCl3, 400 MHz) δ: 3.02 (m, 1H), 3.18 (m, 1H), 3.38 (m, 2H), 3.70-3.79 (m, 4H) , 3.99 (m, 1H), 4.42 (d, 1H), 4.50 (m, 1H), 4.60 (d, 1H), 6.77 (m, 2H), 7.10 ( m, 3H), 7.32 (m, 3H), 7.41 (m, 2H), 7.78 (bs, 1H). LRMS: m/z (ES+) 400 [MNa+].

Preparation 69

(2RS)-2-{[1-(2-methylphenyl)-1H-imidazol-4-yl]methyl}-4-(4-methoxybenzyl)-3-morpholinone

[image]

The title compound was obtained in 16% yield from imidazole from Preparation 52 and 2-methylbenzeneboronic acid, following the procedure described in Preparation 68. 1H-NMR (CDCl3, 400 MHz) δ: 2.15 (s, 3H), 3, 02 (m, 1H), 3.18 (m, 1H), 3.38 (m, 2H), 3.77 (m, 4H), 3.98 (m, 1H), 4.54 (m, 3H ), 6.78 (d, 2H), 6.85 (bs, 1H), 7.15 (m, 3H), 7.21 (s, 1H), 7.25 (m, 2H), 7.45 (bs, 1H). LRMS: m/z (TSP+) 470.2 [MH+].

Preparation 70

(2RS)-2-{[1-(3-phenoxyphenyl)-1H-imidazol-4-yl]methyl}-4-(4-methoxybenzyl)-3-morpholinone

[image]

To a solution of imidazole from Preparation 52 (240 mg, 0.8 mmol) and pyridine (130 μL, 1.6 mmol) in dichloromethane (2 mL) was added copper (II)-acetate (174 mg, 0.96 mmol), molecular 4 Å sieves (50 mg) and 3-phenoxyphenylboronic acid (J. Chem. Soc. 1970; 488) (350 mg, 1.6 mmol. Compressed air was then blown through the solution, maintaining 20-25°C in an aqueous bath, for 7 hours. The solvent was retained using a "cold finger" condenser. The solution was then stirred under a nitrogen atmosphere for the next 18 hours. The mixture was partitioned between dichloromethane (80 mL) and a solution of the tetrasodium salt of ethylenediaminetetraacetic acid (1 g) in saturated sodium chloride of bicarbonate (30 mL) and the phases were separated. The organic layer was dried (MgSO4) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography with an elution gradient of ethyl acetate:diethylamine (100:0 to 95:5). The product was additionally purified by chromatography on a Biotage® silica gel column, with an elution gradient of toluene:diethylamine (1 00:0 to 88:12) to give the title compound as a pale yellow gum, 120 mg. 1H-NMR (CDCl3, 400 MHz) δ: 3.01 (m, 1H), 3.15 (dd, 1H), 3.38 (m, 2H), 3.72 (m, 4H), 3.97 (m, 1H), 4.45 (d, 1H), 4.50 (m, 1H), 4.58 (d, 1H), 6.77 (d, 2H), 6.90 (m, 1H) , 6.97 (m, 1H), 7.00-7.18 (m, 7H), 7.37 (m, 3H), 7.75 (s, 1H). LRMS: m/z (TSP+) 470.2 [MH+].

Preparations 71 to 73

The following compounds have a general structure

[image]

were prepared from the morpholinones of Preparation 52 and the corresponding boronic acids, following a procedure similar to that described in Preparation 70.

[image]

Preparations 74 to 76

The following compounds have a general structure

[image]

were prepared from the morpholinones of Preparation 53 and the corresponding boronic acids, following a procedure similar to that described in Preparation 70.

[image]

Preparations 77 to 80

The following compounds have a general structure

[image]

were prepared from the morpholinones of Preparation 55 and the corresponding boronic acids, following a procedure similar to that described in Preparation 70.

[image]

1 = 3-phenoxyphenylboronic acid was used (J. Chem. Soc. 1970; 488).

Preparation 81

(2S,6R)-2-{[1-(4-cyclohexylphenyl)-1H-imidazol-4-yl]methyl}-4-(4-methoxybenzyl)-6-methyl-3-morpholinone

[image]

A mixture of copper(II)-acetate (398 mg, 2.2 mmol), 4-cyclohexylbenzeneboronic acid (980 mg, 4.8 mmol), imidazole from Preparation 55 (790 mg, 2.4 mmol) and pyridine (390 μL, 4.8 mmol) in dichloromethane (20 mL) was stirred at room temperature, while compressed air was blown through the solution for 8 hours, and the solvent was retained using a "cold finger" condenser. The solution was then stirred under a nitrogen atmosphere for a further 18 hours. The mixture was partitioned between dichloromethane (200 mL) and water (200 mL) containing the tetrasodium salt of ethylenediaminetetraacetic acid (1 g) and aqueous sodium bicarbonate (35 mL) and the layers were separated. The organic phase was dried (MgSO4) and concentrated under reduced pressure. The residue was purified by chromatography on a Biotage® silica gel column, eluting with a gradient of toluene:diethylamine (95:5 to 92:8), to give the title compound, 140 mg. 1H-NMR (CDCl3, 400 MHz) δ: 1.15-1.47 (m, 9H), 1.72-1.94 (m, 4H), 2.55 (m, 1H), 2.99 ( dd, 1H), 3.03-3.20 (m, 2H), 3.39 (dd, 1H), 3.73 (s, 3H), 3.90 (m, 1H), 4.42 (d , 1H), 4.56 (m, 2H), 6.78 (d, 2H), 7.02-7.19 (m, 5H), 7.20 (m, 2H), 7.74 (s, 1H). LRMS: m/z (ES+) 474 [MH+].

Preparation 82

4-(cyclohexyloxy)phenylboronic acid

[image]

A solution of 4-(cyclohexyloxy)phenylbromide (J. Am. Chem. Soc. 1978; 3559) in tetrahydrofuran (40 mL) was deaerated and then cooled to -78°C. n-Butyllithium (13.47 mL, 1.45M in hexanes, 19.5 mmol) was added dropwise and the mixture was stirred for 30 minutes. Triisopropyl borate (5.01 mL, 26.6 mmol) was added dropwise over 10 minutes and the mixture was allowed to slowly warm to room temperature with stirring, after which it was poured into sodium hydroxide solution (0.25 M , 300 mL). This mixture was stirred for 15 min and then extracted with diethyl ether (2 x 150 mL). The aqueous layer was acidified to pH 1 with concentrated hydrochloric acid and extracted with dichloromethane (2 x 150 mL). These combined organic extracts were dried (MgSO 4 ) and evaporated under reduced pressure to give the title compound as a white solid, 3.1 g. 1H-NMR (CDCl 3 , 400 MHz) δ: 0.88-1.62 (m, 7H), 1.80 (m, 2H), 2.00 (m, 2H), 4.39 (m, 1H), 6.98 (d, 2H), 8.17 (d, 2H).

Preparation 83

(2S,6R)-2-({1-(4-cyclohexyloxy)phenyl]-1H-imidazol-4-yl}methyl)-4-(4-methoxybenzyl)-6-methyl-3-morpholinone

[image]

The title compound was obtained in 41% yield from imidazole from Preparation 55 and boronic acid from Preparation 82 following a procedure similar to that described in Preparation 81. 1H-NMR (CDCl3, 400 MHz) δ: 1.19 (d, 3H), 1.38 (m, 3H), 1.57 (m, 3H), 1.80 (m, 2H), 1.99 (m, 2H), 2.99 (dd, 1H), 3.04-3 .20 (m, 2H), 3.39 (dd, 1H), 3.77 (s, 3H), 3.88 (m, 1H); 4.22 (m, 1H), 4.42 (d, 1H), 4.56 (m, 1H), 4.60 (d, 1H), 6.79 (d, 2H), 6.96 (d , 2H), 7.02 (s, 1H), 7.10-7.28 (m, 4H), 7.65 (s, 1H). LRMS: m/z (ES+) 490 [MH+].

Preparation 84

(2RS)-2-{[1-(4'-chloro[1,1'-biphenyl]-3-yl)-1H-imidazol-4-yl]methyl}-4-(4-methoxybenzyl)-3- morpholino

[image]

A mixture of bromide from Preparation 73 (200 mg, 0.44 mmol), 4-chlorophenyl-boronic acid (207 mg, 1.32 mmol), lithium chloride (56 mg, 1.32 mmol), cesium carbonate (433 mg, 1 .32 mmol) and tetrakis(triphenylphosphine)palladium(0) (51 mg, 0.044 mmol) in water (2 mL) and tetrahydrofuran (5 mL) was stirred at 75°C for 18 hours. The cooled mixture was partitioned between dichloromethane and a 2M sodium carbonate solution containing 6% v/v 0.88 ammonia. The organic phase was separated, dried (MgSO4) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with a gradient of toluene:diethylamine (95:5 to 93:7) to give the title compound as a pale yellow oil, 150 mg. 1H-NMR (CDCl3, 400 MHz) δ: 3.05 (m, 1H), 3.20 (dd, 1H), 3.42 (m, 2H), 3.72 (s, 3H), 3.78 (m, 1H), 4.00 (m, 1H), 4.48 (d, 1H), 4.58 (dd, 1H), 4.62 (d, 1H), 6.78 (d, 2H) , 7.14 (d, 2H), 7.20 (s, 1H), 7.35-7.59 (m, 8H), 7.82 (s, 1H).

Preparation 85

(2S,6R)-2-{[1-(3'-chloro[1,1'-biphenyl]-3-yl)-1H-imidazol-4-yl]methyl}-4-(4-methoxybenzyl)- 6-methyl-3-morpholinone

[image]

The title compound was obtained as a white solid in 63% yield from the bromide of Preparation 77 and 3-chlorobenzeneboronic acid, following a procedure similar to that described in Preparation 84. 1H-NMR (CDCl3, 400 MHz) δ: 1.20 (d, 3H), 3.00 (dd, 1H); 3.07-3.21 (m, 2H), 3.41 (dd, 1H), 3.72 (s, 3H), 3.90 (m, 1H), 4.44 (d, 1H), 4 .58 (m, 2H), 6.78 (d, 2H), 7.13 (d, 2H), 7.20-7.58 (m, 9H), 7.82 (s, 1H). LRMS: m/z (ES+) 502, 504 [MH+].

Preparation 86

(2RS)-2-{[1-(3',4'-dichloro[1,1'-biphenyl]-3-yl)-1H-imidazol-4-yl]methyl}-4-(4-methoxybenzyl) -3-morpholino

[image]

A mixture of bromide from Preparation 73 (200 mg, 0.44 mmol), 3,4-dichlorobenzeneboronic acid (102 mg, 0.53 mmol), lithium chloride (56 mg, 1.32 mmol), cesium carbonate (433 mg, 1 .32 mmol) and tetrakis(triphenylphosphine)palladium(0) (26 mg, 0.022 mmol) in water (2 mL) and tetrahydrofuran (5 mL) was stirred at 75°C for 2.5 hours. TLC analysis showed a retention of the starting material, so more 3,4-dichlorobenzeneboronic acid (204 mg, 1.06 mmol) and tetrakis(triphenylphosphine)palladium(0) (26 mg, 0.022 mmol) were added and the mixture was heated for another 18 hours at reflux. The cooled mixture was partitioned between dichloromethane and 2M sodium carbonate solution with 6% v/v 0.88 ammonia. The organic phase was separated, dried (MgSO4) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with a gradient of toluene:diethylamine (95:5 to 93:7) to give the title compound as a crystalline solid, 165 mg. 1H-NMR (CDCl3, 400 MHz) δ: 3.06 (m, 1H), 3.20 (dd, 1H), 3.41 (m, 2H), 3.75 (m, 4H), 4.00 (m, 1H), 4.48 (d, 1H), 4.58 (m, 1H), 4.62 (d, 1H), 6.58 (2×s, 2H), 7.18 (m, 3H), 7.40 (m, 2H), 7.52 (m, 4H), 7.65 (s, 1H), 7.82 (s, 1H). LRMS: m/z (ES+) 522, 524 [MH+].

Preparation 87

(2S,6R)-2-{[1-(3',4'-chloro[1,1'-biphenyl]-3-yl)-1H-imidazol-4-yl]methyl}-4-(4- methoxybenzyl)-6-methyl-3-morpholinone

[image]

A mixture of bromide from Preparation 77 (200 mg, 0.425 mmol), 3,4-dichlorobenzeneboronic acid (243 mg, 1.27 mmol), lithium chloride (54 mg, 1.27 mmol), cesium carbonate (416 mg, 1.32 mmol) and tetrakis(triphenylphosphine)palladium(0) (23 mg, 0.02 mmol) in water (2 mL) and tetrahydrofuran (5 mL) was stirred at 75°C for 3 hours. The cooled mixture was partitioned between ethyl acetate (100 mL) and water (50 mL). The organic phase was separated, dried (MgSO4) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with a gradient of pentane:ethyl acetate:methanol (50:50:0 to 0:100:0 to 0:95:5) to give a white foam. It was further purified by column chromatography on Biotage® silica gel eluting with a gradient of ethyl acetate:methanol (100:0 to 93:7) to give the title compound as a white foam, 140 mg. 1H-NMR (CDCl3, 400 MHz) δ: 1.20 (d, 3H), 3.00-3.21 (m, 3H), 3.40 (m, 1H), 3.75 (s, 3H) , 3.90 (m, 1H), 4.42 (m, 1H), 4.59 (m, 2H), 6.78 (d, 2H), 7.13 (d, 2H), 7.19 ( s, 1H), 7.38 (m, 1H), 7.40 (m, 1H), 7.50 (m, 4H), 7.65 (s, 1H), 7.81 (s, 1H). LRMS: m/z (ES+) 536, 538 [MH+].

Preparation 88

(2S)-2-[(1-propyl-1H-imidazol-4-yl)methyl]-3-morpholinone

[image]

Ammonium cerium (IV) nitrate (4.55 g, 8.30 mmol) was added to a solution of the compound from Preparation 47 (1.43 g, 4.15 mmol) in acetonitrile (9 mL) and water (9 mL) and the mixture was stirred at room temperature for 18 hours, after which it was concentrated under reduced pressure, and the residue was dissolved in methanol. This solution was purified by silica gel column chromatography, eluting with a gradient of dichloromethane:methanol:0.88 ammonia (95:5:0.5 to 90:10:1) to give an orange oil. It was further purified by column chromatography on Dowex® 50WX8-200 ion exchange resin, eluting with a gradient of water:0.88 ammonia (100:0 to 98:2) to give the title compound, 522 mg. 1H-NMR (CDCl3, 400 MHz) δ: 0.88 (t, 3H), 1.75 (m, 2H), 3.00 (dd, 1H), 3.23 (m, 2H), 3.50 (m, 1H), 3.74 (m, 1H), 3.79 (t, 2H), 4.00 (m, 1H), 4.43 (dd, 1H), 5.94 (bs, 1H) , 6.73 (s, 1H), 7.36 (s, 1H). LRMS: m/z (ES+) 224 [MH+].

Preparation 89

(2RS)-2-[(1-butyl-1H-imidazol-4-yl)methyl]-3-morpholinone

[image]

Ammonium cerium (IV) nitrate (5.7 g, 10.4 mmol) was added to a solution of the compound from Preparation 48 (1.87 g, 5.2 mmol) in acetonitrile (50 mL) and water (50 mL) and the mixture was stirred at room temperature for 18 hours. The solvents were evaporated under reduced pressure, and the residue was purified on a Dowex® 50WX8-200 ion-exchange resin column with 5% 0.88 ammonia as eluent. This product was further purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (100:0:0 to 90:10:1) to give the title compound, 300 mg. 1H-NMR (CDCl3, 400 MHz) δ: 0.96 (t, 3H), 1.34 (m, 2H), 1.75 (m, 2H), 3.02 (dd, 1H), 3.28 (m, 2H), 3.56 (m, 1H), 3.78 (m, 1H), 3.82 (t, 2H), 4.02 (m, 1H), 4.48 (dd, 1H) , 5.98 (bs, 1H), 6.77 (s, 1H), 7.38 (s, 1H). LRMS: m/z (ES+) 238 [MH+].

Preparation 90

(2RS)-2-{[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]methyl}-3-morpholinone

[image]

Ammonium cerium (IV) nitrate (1.1 g, 2.0 mmol) was added to a solution of the compound from Preparation 49 (411 mg, 1.0 mmol) in acetonitrile (5 mL) and water (5 mL) and the mixture was stirred at room temperature for 18 hours, after which it was concentrated under reduced pressure. The residue was preadsorbed on silica gel and purified by chromatography on a silica gel column with an elution gradient of ethyl acetate: dichloromethane: methanol: 0.88 ammonia (100:0:0:0 to 75:0:25:0 to 0:90:10:1 ). The product was further purified by reverse phase chromatography on polystyrene gel, eluting with a gradient of water:methanol (100:0 to 0:100) to give the title compound, 522 mg. 1H-NMR (CDCl 3 , 400 MHz) δ: 1.01 (m, 2H), 1.24 (m, 4H), 1.76 (m, 7H), 3.22 (m, 3H), 3.40 (m, 1H), 3.80 (m, 1H), 4.02 (m, 1H), 4.22 (m, 2H), 4.38 (m, 1H), 7.43 (s, 1H) , 8.85 (s, 1H). LRMS: m/z (TSP+) 292.2 [MH+].

Preparation 91

(2R,6S)-2-{[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]methyl}-6-methyl-3-morpholinone

[image]

Ammonium cerium (IV) nitrate (387 mg, 0.758 mmol) was added to a solution of the compound from Preparation 61 (120 mg, 0.283 mmol) in acetonitrile (8 mL) and water (5 mL) and the mixture was stirred at 40°C 18 hours, after which it is concentrated under reduced pressure. The residue was preadsorbed on silica gel and purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (98:2:0.2 to 95:5:0.5) to give the title compound as a colorless oil, 66 mg. 1H-NMR (CDCl3, 400 MHz) δ: 0.92 (m, 2H), 1.18 (m, 7H), 1.62 (m, 7H), 2.98 (dd, 1H), 3.18 (m, 2H), 3.23 (dd, 1H), 3.83 (m, 3H), 4.41 (m, 1H), 6.19 (bs, 1H), 6.70 (s, 1H) , 7.30 (s, 1H). LRMS: m/z (ES+) 328 [MH+].

Preparation 92

(2S,6R)-2-{[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]methyl}-6-methyl-3-morpholinone

[image]

The title compound was obtained as a pale yellow solid in 63% yield from the protected morpholinone of Preparation 62, following the procedure described in Preparation 91. 1H-NMR (CDCl3, 400 MHz) δ: 0.92 (m, 2H), 1, 18 (m, 7H), 1.62 (m, 7H), 2.98 (dd, 1H), 3.19 (m, 2H), 3.23 (dd, 1H), 3.83 (m, 3H ), 4.41 (m, 1H), 6.03 (bs, 1H), 6.70 (s, 1H), 7.34 (s, 1H). LRMS: m/z (ES+) 328 [MH+].

Preparation 93

(2S,6R)-2-({1-[2-(4,4-dimethylcyclohexyl)ethyl]-1H-imidazol-4-yl}methyl}-6-methyl-3-morpholinone

[image]

The title compound was obtained in a 53% yield from the protected morpholino from Preparation 66, following a procedure similar to that described in Preparation 91, except that ethyl acetate:methanol:diethylamine (99:0.5:0, 5 to 95:2.5:2.5). 1H-NMR (CDCl3, 400 MHz) δ: 0.82 (2×s, 6H), 1.10 (d, 3H), 1.18-1.38 (m, 8H), 1.48 (m, 1H), 1.63 (m, 2H), 2.98 (m, 1H), 3.19 (m, 2H), 3.23 (dd, 1H), 3.84 (m, 3H), 4, 41 (m, 1H), 6.97 (bs, 1H), 7.21 (s, 1H), 7.34 (s, 1H). LRMS: m/z (ES+) 334 [MH+].

Preparation 94

(2S,6R)-2-{[1-(3-cyclohexyl-3-methylbutyl)-1H-imidazol-4-yl]methyl}-6-methyl-3-morpholinone

[image]

The title compound was obtained in 52% yield from the protected morpholinone of Preparation 65, following a procedure similar to that described in Preparation 91, except that dichloromethane:methanol:0.88 ammonia (99:1:0.1 to 93:7:0.7). 1H-NMR (CD3OD, 400 MHz) δ: 0.86 (s, 6H), 1.00 (m, 2H), 1.03-1.28 (m, 7H), 1.70 (m, 7H) , 2.90 (m, 1H), 3.03 (m, 1H), 3.18 (m, 2H), 3.82 (m, 1H), 3.95 (m, 2H), 4.36 ( m, 1H), 5.42 (s, 1H, 6.86 (s, 1H), 7.50 (s, 1H). LRMS: m/z (ES + ) 348 [MH + ].

Preparation 95

(2S,5S)-2-{[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]methyl}-5-methyl-3-morpholinone

[image]

A mixture of the protected morpholino from Preparation 63 (170 mg, 0.4 mmol) and ammonium cerium (IV) nitrate (550 mg, 1.0 mmol) in water (1 mL) and acetonitrile (1 mL) was stirred at 40° C for 18 hours, after which it was concentrated under reduced pressure. The residue was purified by column chromatography on Biotage® silica gel, eluting with a gradient of ethyl acetate:diethylamine (95:5 to 80:20) to give the title compound, 18 mg. 1H-NMR (CDCl3, 400 MHz) δ: 0.97 (m, 2H), 1.20 (m, 7H), 1.66 (m, 7H), 3.06 (dd, 1H), 3.25 (dd, 1H), 3.58 (m, 1H), 3.65 (dd, 1H), 3.83 (m, 3H), 4.43 (dd, 1H), 6.15 (bs, 1H) , 6.78 (s, 1H), 7.38 (s, 1H).

Preparation 96

(2R,5R)-2-{[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]methyl}-5-methyl-3-morpholinone

[image]

The title compound was obtained in 34% yield from the protected morpholinone of Preparation 64, following the procedure described in Preparation 95. 1H-NMR (CDCl3, 400 MHz) δ: 0.78-0.98 (m, 2H), 1, 03-1.22 (m, 7H), 1.62 (m, 7H), 3.02 (m, 1H), 3.18 (dd, 1H), 3.55 (m, 1H), 3.61 (dd, 1H), 3.77 (dd, 1H), 3.82 (t, 2H), 4.38 (m, 1H), 6.24 (bs, 1H), 6.73 (s, 1H) , 7.37 (s, 1H).

Preparation 97

(2RS)-2-{[1-(2-phenylethyl)-1H-imidazol-4-yl]methyl}-3-morpholinone

[image]

Ammonium cerium (IV) nitrate (883 mg, 1.6 mmol) was added to a solution of the compound from Preparation 50 (326 mg, 0.81 mmol) in acetonitrile (2.4 mL) and water (2.4 mL) and the mixture was stirred at room temperature for 5 days, after which it was concentrated under reduced pressure. The residue was dissolved in methanol and adsorbed onto silica gel, then purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (100:0:0 to 90:10:1) to give the title compound as orange oil, 97 mg. 1H-NMR (CDCl3, 400 MHz) δ: 3.02 (m, 3H), 3.26 (m, 2H), 3.52 (m, 1H), 3.77 (m, 1H), 4.00 (m, 1H), 4.10 (t, 2H), 4.43 (dd, 1H), 5.99 (bs, 1H), 6.70 (s, 1H), 7.05 (d, 2H) , 7.24 (m, 4H). LRMS: m/z (ES+) 286 [MH+].

Preparation 98

(2RS)-2-({1-[2-(-4-bromophenyl)ethyl]-1H-imidazol-4-yl}methyl}-3-morpholinone

[image]

A solution of ammonium cerium (IV) nitrate (1.35 g, 2.48 mmol) in water (5 mL) was added to a solution of bromide from Preparation 59 (600 mg, 1.24 mmol) in acetonitrile (10 mL) and the mixture was stirred at 40°C for 18 hours. TLC analysis showed that the starting material remained, so more ammonium-cerium (IV)-nitrate (308 mg, 0.56 mmol) was added and the mixture was stirred at 40°C for another 2 hours. The cooled mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (98:2:0.2 to 95:5:0.5) to give the title compound. , 250 mg. 1H-NMR (CDCl 3 , 400 MHz) δ: 2.98 (t, 2H), 3.03 (m, 1H), 3.25 (m, 2H), 3.52 (m, 1H), 3.77 (m, 1H), 4.00 (m, 1H), 4.05 (t, 2H), 4.42 (m, 1H), 5.99 (bs, 1H), 6.65 (s, 1H) , 6.93 (d, 2H), 7.22 (s, 1H), 7.40 (d, 2H).

Preparations 99 to 101

The following compounds have a general structure:

[image]

were prepared from the corresponding protected morpholinones following a procedure similar to that described in Preparation 98.

[image]

1 = ethyl acetate:methanol:diethylamine (100:0:0 to 90:5:5) was used as eluent from the column

Preparation 102

(2RS)-2-{[1-(3-phenoxyphenyl)-1H-imidazol-4-yl]methyl}-3-morpholinone

[image]

A solution of ammonium cerium (IV) nitrate (350 mg, 0.63 mmol) in water (2 mL) was added to a solution of the protected morpholine from Preparation xx (120 mg, 0.256 mmol) in acetonitrile (2 mL) and the mixture was stirred at 40°C for 18 hours. TLC analysis showed that the starting material remained, so more ammonium-cerium (IV)-nitrate (500 mg, 0.91 mmol) was added and the mixture was stirred at 40°C for another 8 hours. The mixture was evaporated under reduced pressure and the residue was partitioned between dichloromethane (50 mL) and a solution of ethylenediaminetetraacetic acid (1 g) in saturated sodium bicarbonate solution (50 mL). The phases were separated, and the organic layer was dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of ethyl acetate:methanol:diethylamine (100:0:0 to 96:2:2) to give the title compound, 25 mg. 1H-NMR (CDCl3, 400 MHz) δ: 3.05 (dd, 1H), 3.21-3.18 (m, 2H), 3.57 (m, 1H), 3.77 (m, 1H) , 4.00 (m, 1H), 4.48 (dd, 1H), 6.15 (bs, 1H), 6.90 (d, 1H), 6.98 (s, 1H), 7.02 ( m, 3H), 7.14 (m, 2H), 7.37 (m, 3H), 7.74 (s, 1H). LRMS: m/z (TSP+) 350.0 [MH+].

Preparation 103

(2RS)-2-{[1-(2-naphthyl)-1H-imidazol-4-yl]methyl}-3-morpholinone

[image]

The title compound was obtained as a yellow gum in 56% yield from the protected morpholinone of Preparation 72, following a procedure similar to that described in Preparation 102, except that dichloromethane:methanol:0.88 ammonia (100:0: 0 to 90:10:1). 1H-NMR (CDCl3, 400 MHz) δ: 3.15 (dd, 1H), 3.23 (m, 1H), 3.38 (dd, 1H), 3.58 (m, 1H), 3.78 (m, 1H), 4.02 (m, 1H), 4.53 (dd, 1H), 6.01 (bs, 1H), 7.22 (d, 2H), 7.48 (m, 3H) , 7.75-7.94 (m, 4H). LRMS: m/z (ES+) 308 [MH+].

Preparation 104

(-)-(2S)-2-{[1-phenyl-1H-imidazol-4-yl]methyl}-3-morpholinone

[image]

Ammonium cerium (IV) nitrate (1.43 g, 2.61 mmol) was added to a solution of the protected morpholino from Preparation 74 (330 mg, 0.87 mmol) in water (2 mL) and acetonitrile (2 mL) and the mixture was stirred at 40°C for 4 hours. TLC analysis showed that the starting material remained, so more ammonium-cerium (IV)-nitrate (1.43 g, 2.61 mmol) was added and the mixture was stirred at 40°C for another 2 hours. The mixture was partitioned between dichloromethane (200 mL) and a solution of ethylenediaminetetraacetic acid (1 g) in saturated sodium bicarbonate solution (50 mL). The phases were separated, and the organic layer was dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by chromatography on a silica gel column with an elution gradient of ethyl acetate:methanol:diethylamine (100:0:0 to 96:2:2). The product was azeotroped with toluene and dichloromethane to give the title compound as an oil, 173 mg. 1H-NMR (CDCl3, 400 MHz) δ: 3.14 (dd, 1H), 3.28 (m, 1H), 3.38 (dd, 1H), 3.58 (m, 1H), 3.80 (m, 1H), 4.05 (m, 1H), 4.56 (dd, 1H), 5.98 (bs, 1H), 7.17 (s, 1H), 7.37 (m, 3H) , 7.45 (m, 2H), 7.79 (s, 1H). LRMS: m/z (TSP+) 258.1 [MH+]. [α]D = -70.59° (c = 0.104, methanol).

Preparation 105

(2S)-2-{[1-(4-tert-butylphenyl)-1H-imidazol-4-yl]methyl}-3-morpholinone

[image]

Ammonium cerium (IV) nitrate (297 mg, 0.55 mmol) was added to a solution of the protected morpholino from Preparation 75 (94 mg, 0.22 mmol) in water (2 mL) and acetonitrile (2 mL) and the mixture was stirred at 40°C for 15 hours. Ethylenediaminetetraacetic acid (0.5 g) in saturated sodium bicarbonate solution (5 mL) was added and the mixture was extracted with dichloromethane (2 x 50 mL). The combined organic extracts were dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography with an elution gradient of ethyl acetate:methanol:diethylamine (98:1:1 to 94:3:3) to give the title compound as an oil, 22 mg. 1H-NMR (CDCl3, 400 MHz) δ: 1.37 (s, 9H), 3.12 (dd, 1H), 3.26 (m, 1H), 3.38 (dd, 1H), 3.58 (m, 1H), 3.79 (m, 1H), 4.04 (m, 1H), 4.55 (m, 1H), 6.22 (bs, 1H), 7.10 (s, 1H) , 7.25 (d, 2H), 7.42 (d, 2H), 7.77 (s, 1H). LRMS: m/z (TSP+) 314.1 [MH+].

Preparation 106

(-)-(2S)-2-{[1-[3,5-bis(trifluoromethyl)phenyl]-1H-imidazol-4-yl}methyl)-3-morpholinone

[image]

The title compound was obtained as a solid in 81% yield from the protected morpholinone of Preparation 76, following the procedure described in Preparation 105. 1H-NMR (CDCl3, 400 MHz) δ: 3.18 (dd, 1H), 3.30 ( m, 1H), 3.39 (dd, 1H), 3.60 (m, 1H), 3.80 (m, 1H), 4.06 (m, 1H), 4.55 (m, 1H), 5.88 (bs, 1H), 7.20 (s, 1H), 7.81 (s, 2H), 7.84 (s, 1H), 7.87 (s, 1H). LRMS: m/z (TSP+) 394.0 [MH+]. [α]D = -40.35° (c = 0.116, methanol).

Preparation 107

(2RS)-2-{[1-(4'-chloro[1,1'-biphenyl]-3-yl)-1H-imidazol-4-yl]methyl}-3-morpholinone

[image]

The title compound was obtained as a solid in 91% yield from the protected morpholinone of Preparation 84, following the procedure described in Preparation 105. 1H-NMR (CDCl3, 400 MHz) δ: 3.10 (dd, 1H), 3.25 ( m, 1H), 3.38 (dd, 1H), 3.57 (m, 1H), 3.78 (m, 1H), 4.02 (m, 1H), 4.52 (dd, 1H), 5.96 (bs, 1H), 7.17 (s, 1H), 7.37 (m, 3H), 7.42 (m, 1H), 7.50 (m, 3H), 7.57 (s , 1H), 7.80 (s, 1H). LRMS: m/z (ES-) 366 [M-H-].

Preparation 108

(2RS)-2-{[1-(3',4'-dichloro[1,1'-biphenyl]-3-yl)-1H-imidazol-4-yl]methyl}-3-morpholinone

[image]

The title compound was obtained in 49% yield from the protected morpholino from Preparation 86, following the procedure described in Preparation 105. 1H-NMR (CDCl3, 400 MHz) δ: 3.10 (dd, 1H), 3.25 (m, 1H), 3.38 (dd, 1H), 3.57 (m, 1H), 3.78 (m, 1H), 4.02 (m, 1H), 4.52 (dd, 1H), 5, 86 (bs, 1H), 7.18 (s, 1H), 7.37 (m, 2H), 7.50 (m, 4H), 7.62 (s, 1H), 7.80 (s, 1H ). LRMS: m/z (ES+) 402, 404 [MH+].

Preparations 109 to 114

The following compounds have a general structure

[image]

were prepared from the corresponding protected morpholinones, following a procedure similar to that described in Preparation 105.

[image] [image]

1 = acetonitrile:water (3:1, by volume) was used as the reaction solvent

2 = acetonitrile:water (2:1, by volume) was used as the reaction solvent

Preparation 115

(2S,6R)-2-{[1-(3',4'-dichloro[1,1'-biphenyl]-3-yl)-1H-imidazol-4-yl]methyl}-6-methyl-3 -morpholino

[image]

The title compound was obtained as a white foam in 63% yield from the protected morpholinone of Preparation 87, following a procedure similar to that described in Preparation 105. 1H-NMR (CDCl3, 400 MHz) δ: 1.24 (d, 3H), 3 .14 (dd, 1H), 3.23 (m, 2H), 3.40 (dd, 1H), 3.97 (m, 1H), 4.56 (dd, 1H), 5.80 (bs, 1H), 7.10 (s, 1H), 7.40 (m, 2H), 7.55 (m, 4H), 7.68 (s, 1H), 7.81 (s, 1H). LRMS: m/z (TSP+) 416.1, 420.1 [MH+].

Preparation 116

(2S)-2-(1H-imidazol-4-yl-methyl)-3-morpholinone

[image]

A mixture of the protected morpholinone from Preparation 53 (500 mg, 1.66 mmol) and ammonium cerium (IV) nitrate (2.5 g, 4.5 mmol) in water (6 mL) and acetonitrile (6 mL) was stirred at 40°C for 18 hours. Potassium carbonate (1.5 g) was added and the mixture was stirred for 10 minutes and then adsorbed onto silica gel. The product was isolated by silica gel column chromatography with ethyl acetate:methanol:diethylamine (96:2:2 to 80:10:10) and further purified by silica gel column chromatography with dichloromethane:methanol (90:10 to 85:15) to would give the title compound, 240 mg. 1H-NMR (CD3OD, 400 MHz) δ: 3.02-3.43 (m, 4H), 3.78 (m, 1H), 4.00 (m, 1H), 4.38 (m, 1H) , 6.75 (s, 1H), 7.78 (s, 1H). HRMS: m/z (ES+) 182.0924 [MH+].

Preparation 117

(-)-(2S,6R)-2-(1H-imidazol-4-yl-methyl)-6-methyl-3-morpholinone

[image]

A mixture of the protected morpholinone from Preparation 55 (1 g, 3.2 mmol) and ammonium cerium (IV) nitrate (5.2 g, 9.6 mmol) in water (20 mL) and acetonitrile (30 mL) was stirred at 40°C for 18 hours. The solvent was evaporated under reduced pressure. The residue was suspended in a mixture of dichloromethane:methanol:0.88 ammonia (99:1:0.1, by volume) and purified twice by chromatography on a silica gel column with an elution gradient of dichloromethane:methanol:0.88 ammonia (90:10:1). The resulting oil was azeotroped with diethyl ether to give the title compound as a colorless foam, 380 mg. 1H-NMR (CD3OD, 400 MHz) δ: 1.21 (d, 3H), 3.02 (m, 2H), 3.19 (m, 2H), 3.90 (m, 1H), 4.36 (m, 1H), 6.81 (s, 1H), 7.54 (s, 1H). LRMS: m/z (ES+) 196 [MH+]. [α]D = -104.56° (c = 0.19, methanol).

Preparation 118

tert-butyl-(2S)-2-{[1-(tert-butoxycarbonyl)-1H-imidazol-4-yl]methyl}-3-oxo-4-morpholinecarboxylate

[image]

A solution of morpholinone from Preparation 116 (70 mg, 0.39 mmol), dimethylaminopyridine (3 mg) and di-tert-butyl dicarbonate (354 mg, 1.62 mmol) in acetonitrile (5 mL) was stirred at room temperature for 42 hours . The mixture was concentrated under reduced pressure, and the residue was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (99:1:0.1 to 95:5:0.5) to give the title compound , 96 mg. 1H-NMR (CDCl3, 400 MHz) δ: 1.58 (s, 9H), 1.61 (s, 9H), 3.04 (dd, 1H), 3.35 (dd, 1H), 3.78 (m, 3H), 4.05 (m, 1H), 4.50 (m, 1H), 7.20 (s, 1H), 8.00 (s, 1H). HRMS: m/z (ES+) 382.1972 [MH+].

Preparation 119

tert-butyl-(2S,6R)-2-({1-[4-(cyclohexyloxy)phenyl]-1H-imidazol-4-yl}methyl)-6-methyl-3-oxomorpholine-4-carboxylate

[image]

4-Dimethylaminopyridine (49 mg, 0.4 mmol) and di-tert-butyl dicarbonate (174 mg, 0.8 mmol) were added to a solution of morpholinone from Preparation 114 (135 mg, 0.37 mmol) in acetonitrile (5 mL ) and the mixture was stirred at room temperature for 5 hours. TLC analysis showed retention of the starting material, so more di-tert-butyl dicarbonate (87 mg, 0.4 mmol) was added and the mixture was stirred at room temperature for another 18 hours. The reaction mixture was concentrated under reduced pressure and the residue was purified by silica gel column chromatography with ethyl acetate as eluent to give the title compound, 95 mg. LRMS: m/z (ES+) 470 [MH+].

Preparation 120

Lithium (2S)-2-({(1R)-2-[(tert-butoxycarbonyl)amino]-1-methylethyl}oxy)-3-{1-[4-(cyclohexyloxy)phenyl]-1H-imidazole-4 -yl}propanoate

[image]

A mixture of the protected morpholino from Preparation 119 (87 mg, 0.19 mmol) and lithium hydroxide (24 mg, 0.56 mmol) in tetrahydrofuran (0.5 mL) and water (1 mL) was stirred at room temperature for 18 hours. The reaction mixture was evaporated under reduced pressure to give the title compound. 1H-NMR (D2O, 400 MHz) δ: 0.60 (m, 2H), 1.00-1.38 (m, 16H), 1.50 (m, 2H), 1.70 (m, 2H) , 2.58 (m, 1H), 2.80 (m, 2H), 2.94 (m, 1H), 3.30 (m, 1H), 3.82 (m, 1H), 4.00 ( m, 1H), 6.63 (d, 2H), 6.82 (s, 1H), 7.00 (d, 2H), 7.56 (s, 1H). LRMS: m/z (ES-) 486 [M-H-].

Preparation 121

(2RS)-2-({1-[2-(4'-Ethyl[1,1'-biphenyl]-4-yl)ethyl]-1H-imidazol-4-yl}methyl)-3-morpholinone

[image]

A mixture of the bromo compound from Preparation 98 (250 mg, 0.69 mmol), 4-ethylbenzeneboronic acid (154 mg, 1.03 mmol), tetrakis(triphenylphosphine)-palladium(0) (78 mg, 0.068 mmol) and a solution of sodium carbonate (411 μL, 2M, 0.823 mmol) in water (1 mL) and dioxane (5 mL) was heated at 100°C for 3 hours. The cooled reaction mixture was diluted with water (10 mL), and the mixture was extracted with ethyl acetate (3 × 15 mL). The combined organic extracts were dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (99:1:0.1 to 95:5:0.5) to give the title compound, 170 mg. 1H-NMR (CDCl 3 , 400 MHz) δ: 1.22 (t, 3H), 2.64 (q, 2H), 3.02 (m, 3H), 3.22 (m, 2H), 3.44 (m, 1H), 3.72 (m, 1H), 3.98 (m, 1H), 4.10 (t, 2H), 4.42 (m, 1H), 5.90 (bs, 1H) , 6.70 (s, 1H), 7.08 (d, 2H), 7.22 (m, 3H), 7.44 (m, 4H).

Preparations 122 to 131

[image]

Aryl-boronic acids (R-B(OH)2), (0.74 mmol) were added to a solution of the bromo-compound from Preparation 98 (180 mg, 0.49 mmol), tetrakis(triphenyphosphine)-palladium(0) (180 mg, 0.49 mmol) and a solution of sodium carbonate (295 μL, 2M, 0.593 mmol) in water (1 mL) and dioxane (5 mL). The reaction mixtures were heated at 100°C for 4 hours, after which they were allowed to cool. Water (15 mL) was added and the mixture was extracted with ethyl acetate (3 x 15 mL). The combined organic extracts were dried (MgSO4) and evaporated under reduced pressure. The crude products were purified by silica gel chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (97:3:0.3 to 95:5:0.5) to give the desired products, shown in the following table.

[image] [image]

Preparation 132

(2RS)-2-({1-[(2EZ)-3-bromo-2-propenyl]-1H-imidazol-4-yl}methyl)-3-morpholinone

[image]

A mixture of ammonium cerium(IV) nitrate (2.6 g, 4.75 mmol) and the compound from Preparation 67 (1 g, 2.38 mmol) in acetonitrile (10 mL) and water (5 mL) was stirred at 40 °C 18 hours. TLC analysis showed a lag of the starting material, so more ammonium-cerium(IV)-nitrate (650 mg, 1.19 mmol) was added and the mixture was stirred for another 3 hours at 40°C. The mixture was concentrated under reduced pressure and azeotroped with methanol. The crude product was preadsorbed on silica gel and purified twice by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (99:1:0.1 to 95:5:0.5) to give the title compound, 370 mg . 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 3.00 (m, 1H), 3.23 (m, 2H), 3.50 (m, 1H), 3.74 (m, 1H ), 4.00 (m, 1H), 4.42 (m, 2H), 4.62 (d, 1H), 5.98 (bs, 1H), 6.26 (m, 1.5H), 6 .42 (d, 0.5H), 6.75 (2×s, 1H), 7.40 (2×s, 1H). LRMS: m/z (ES+) 300, 302 [MH+].

Preparation 133

(-)-(2S)-2-{[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]methyl}-3-morpholinone

[image]

Ammonium cerium(IV) nitrate (482 mg, 0.88 mmol) and water (1 mL) were added to a solution of the compound from Preparation 60 (181 mg, 0.44 mmol) in acetonitrile (1 mL) and the mixture was stirred at 40 °C 18 hours. TLC analysis showed the retention of the starting material, so more ammonium-cerium(IV)-nitrate (250 mg, 0.46 mmol) was added and the mixture was stirred at 40°C for another 5 hours. The mixture was partitioned between dichloromethane (75 mL) and a solution of ethylenediaminetetraacetic acid (1 g) in aqueous sodium bicarbonate solution (30 mL) and the phases were separated. The organic layer was dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (100:0:0 to 94:6:0.6) to give the title compound as a sticky gum, 80 mg. 1H-NMR (CDCl3, 400 MHz) δ: 0.97 (m, 2H), 1.20 (m, 4H), 1.63 (m, 7H), 3.02 (dd, 1H), 3.26 (m, 2H), 3.56 (m, 1H), 3.78 (m, 1H), 3.86 (t, 2H), 4.02 (m, 1H), 4.45 (m, 1H) , 5.83 (bs, 1H), 6.76 (s, 1H), 7.38 (s, 1H). LRMS: m/z (TSP+) 292.1 [MH+]. [α]D = -60.01° (c = 0.05, methanol).

Preparation 134

(2RS)-2-({1-[(2EZ)-3-[1,1'-biphenyl]-4-yl-2-propenyl]-1H-imidazol-4-yl}methyl)-3-morpholinone

[image]

A mixture of the bromo compound from Preparation 132 (185 mg, 0.62 mmol), 4-biphenylboronic acid (183 mg, 0.925 mmol), tetrakis(triphenylphosphine)palladium(0) (72 mg, 0.062 mmol) and sodium carbonate (78 mg , 0.74 mmol) in water (3 mL) and dioxane (6 mL) was heated at 100°C for 3 hours, after which it was cooled and partitioned between water (20 mL) and ethyl acetate (20 mL). The layers were separated and the aqueous phase was extracted with ethyl acetate (10 mL). The combined organic extracts were dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (99:1:0.1 to 98:2:0.2) to give the title compound as a white foam, 100 mg. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 3.00 (m, 1H), 3.22 (m, 2H), 3.54 (m, 1H), 3.75 (m, 1H ), 4.00 (m, 1H), 4.44 (m, 1H), 4.62 (m, 1H), 4.78 (m, 1H), 5.80, 6.26 (2×m, 1H), 5.93 (bs, 1H), 6.54, 6.66-6.80 (2×m, 2H), 7.23-7.60 (m, 10H). LRMS: m/z (ES+) 374 [MH+].

Preparations 135 to 137

The following compounds have a general structure

[image]

were prepared from the bromide of Preparation 132 and the corresponding boronic acid, following a procedure similar to that described in Preparation 134.

[image]

1 = isolated without column chromatography

Preparation 138

(2RS)-2-({1-[(2EZ)-3-(4-bromophenyl)-2-propenyl]-1H-imidazol-4-yl}methyl)-3-morpholinone

[image]

The title compound was obtained in 42% yield from the compound from Preparation 67 and 4-bromobenzeneboronic acid following the procedure described in Preparation 134. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 3.00 (m, 1H ), 3.25 (m, 2H), 3.54 (m, 1H), 3.76 (m, 1H), 4.00 (m, 1H), 4.43 (m, 1H), 4.59 -4.79 (m, 2H), 5.80, 6.22 (2×m, 2H), 6.40, 6.62 (2×m, 1H), 6.72, 6.78 (2× s, 1H), 7.06 (d, 1H), 7.19 (d, 1H), 7.38-7.58 (m, 3H). LRMS: m/z (TSP + ) 376.1, 378.1 [MH + ].

Preparation 139

(2RS)-2-({1-[(2EZ)-3-(4'-methyl[1,1'-biphenyl]-4-yl)-2-propenyl]-1H-imidazol-4-yl}methyl )-3-morpholino

[image]

A mixture of the bromo compound from Preparation 138 (132 mg, 0.35 mmol), 4-methylbenzeneboronic acid (72 mg, 0.53 mmol), tetrakis(triphenylphosphine)-palladium(0) (50 mg, 0.04 mmol) and of sodium carbonate (270 μL, 2M, 0.53 mmol) in dioxane (6 mL) was heated at 100°C for 1.5 hours. The cooled reaction mixture was partitioned between water (20 mL) and ethyl acetate (20 mL) and the layers were separated. The aqueous phase was extracted with ethyl acetate (10 mL), and the combined organic extracts were dried (MgSO4) and evaporated under reduced pressure. The remaining yellow oil was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (99:1:0.1 to 98:2:0.2) to give the title compound as a white foam, 77 mg. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 2.38 (2×s, 3H), 3.00 (m, 1H), 3.25 (m, 2H), 3.54 (m , 1H), 3.75 (m, 1H), 4.00 (m, 1H), 4.43 (m, 1H), 4.62 (m, 1H), 4.78 (m, 1H), 5 .78, 6.28 2×m, 2H), 6.55, 6.68-6.80 (2×m, 2H), 7.22 (m, 3H), 7.38-7.63 (m , 6H). LRMS: m/z (ES+) 388 [MH+].

Preparation 140

(2RS)-2-({1-[(2EZ)-3-(4'-chloro[1,1'-biphenyl]-4-yl)-2-propenyl]-1H-imidazol-4-yl}methyl )-3-morpholino

[image]

A mixture of the bromo compound from Preparation 138 (100 mg, 0.27 mmol), 4-chlorobenzeneboronic acid (63 mg, 0.4 mmol), tetrakis(triphenylphosphine)-palladium(0) (31 mg, 0.027 mmol) and a solution of sodium carbonate (400 μL, 2M, 0.79 mmol) in ethanol (1 mL) and toluene (4 mL) was heated at 100°C for 3 hours. TLC analysis showed residual starting material, so dioxane (3 mL), additional 4-chlorobenzeneboronic acid (21 mg, 0.13 mmol) and tetrakis(triphenylphosphine)palladium (0) (15 mg, 0.013 mmol) were added and the mixture was stirred at 100°C for another 6 hours. The cooled reaction mixture was partitioned between water (10 mL) and ethyl acetate (20 mL) and the layers were separated. The aqueous phase was extracted with ethyl acetate (10 mL) and the combined organic extracts were dried (MgSO4) and evaporated under reduced pressure. The crude product was purified by silica gel column chromatography with dichloromethane:methanol:0.88 ammonia (98:2:0.2) as eluent to give the title compound as a white solid, 60 mg. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 3.02 (m, 1H), 3.27 (m, 2H), 3.57 (m, 1H), 3.78 (m, 1H ), 4.02 (m, 1H), 4.47 (m, 1H), 4.65, 4.79 (2×d, 2H), 5.82, 6.32 (m, 1H), 6, 18 (bs, 1H), 6.57, 6.82 (2×m, 2H), 7.30 (d, 1H), 7.39-7.79 (m, 8H). LRMS: m/z (ES+) 430 [MNa+].

Preparation 141

(2RS)-2-({1-[(2EZ)-3-(2',5'-difluoro[1,1'-biphenyl]-4-yl)-2-propenyl]-1H-imidazole-4- yl}methyl)-3-morpholino

[image]

The title compound was obtained from the compound from Preparation 138 and 2,5-difluorobenzeneboronic acid, following the procedure described in Preparation 140. 1H-NMR (CDCl3, 400 MHz) (mixture of geometric isomers) δ: 3.02 (m, 1H), 3.28 (m, 2H), 3.57 (m, 1H), 3.78 (m, 1H), 4.00 (m, 1H), 4.45 (m, 1H), 4.63 (d , 1H), 4.79 (d, 1H), 5.82, 6.36 (m, 1H), 6.14 (bs, 1H), 6.58, 6.79 (2×m, 2H), 7.00 (m, 1H), 7.14 (m, 2H), 7.32 (d, 1H), 7.40-7.70 (m, 4H). LRMS: m/z (ES+) 432 [MNa+].

Preparation 142

tert-butyl-(2RS)-2-[2-(dimethylamino)ethoxy]-3-(1-propyl-1H-imidazol-4-yl)propanoate

[image]

A mixture of the alkene from Preparation 44 (650 mg, 2.01 mmol) and 10% palladium on carbon (Degussa®101) (60 mg) in ethanol (20 mL) was hydrogenated at 50°C and 60 psi (410 kPa). 18 hours. The cooled mixture was filtered through Arbocel® and the filtrate was evaporated under reduced pressure. The residue was purified by silica gel column chromatography, eluting with a gradient of ethyl acetate:diethylamine:methanol (100:0:0 to 97:1.5:1.5) to give the title compound, 502 mg. 1H-NMR (CDCl3, 400 MHz) δ: 0.92 (t, 3H), 1.42 (s, 9H), 1.77 (m, 2H), 2.21 (s, 6H), 2.48 (t, 2H), 2.90-3.03 (m, 2H), 3.42 (m, 1H), 3.70 (m, 1H), 3.80 (t, 2H), 4.08 ( m, 1H), 6.79 (s, 1H), 7.37 (s, 1H). LRMS: m/z (TSP+) 326.2 [MH+].

Preparation 143

tert-Butyl-(3S)-3-{(1RS)-2-tert-butoxy-2-oxo-1-[(1-propyl-1H-imidazol-4-yl)methyl]ethoxy}-1-pyrrolidinecarboxylate

[image]

A mixture of alkene from Preparation 45 (1.19 g, 2.83 mmol) and Degussa®101 catalyst (120 mg) in ethanol (12 mL) was hydrogenated at 50°C and 60 psi (410 kPa) for 18 hours. TLC analysis showed retention of the starting material, so more catalyst (120 mg) was added and the mixture was hydrogenated at 50°C and 60 psi (410 kPa) for another 48 hours. The mixture was filtered through Arbocel®, the catalyst was washed with ethanol, and the combined filtrates were evaporated under reduced pressure. The remaining oil was purified by silica gel column chromatography with ethyl acetate as eluent to give the title compound as a colorless oil, 227 mg. 1H-NMR (CDCl3, 400 MHz) δ: 0.92 (t, 3H), 1.42 (m, 18H), 1.79 (m, 2H), 2.00-2.30 (m, 2H) , 2.80-2.95 (m, 1H), 3.00-3.48 (m, 5H), 3.84 (t, 2H), 4.05-4.20 (m, 2H), 6 .75 (m, 1H), 7.50 (m, 1H). LRMS: m/z (ES+) 424 [MH+].

Preparation 144

(2RS)-2-{[1-(3-[1,1'-biphenyl]-4-yl-propyl)-1H-imidazol-4-yl]methyl}-3-morpholinone

[image]

A mixture of alkene from Preparation 134 (100 mg, 0.268 mmol) and Degussa®101 catalyst (15 mg) in ethanol (12 mL) was hydrogenated at 50°C and 60 psi (410 kPa) for 6 hours. TLC analysis showed a lag of the starting material, so more Degussa®101 catalyst (20 mg) was added and the mixture was hydrogenated for another 18 hours. The mixture was filtered through Arbocel®, the catalyst was washed with ethanol, and the combined filtrates were evaporated under reduced pressure. The residue was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (99:1:0.1 to 98:2:0.2) to give the title compound as a colorless oil, 70 mg. 1H-NMR (CDCl3, 400 MHz) δ: 2.10 (m, 2H), 2.61 (t, 2H), 3.02 (dd, 1H), 3.23 (m, 2H), 3.49 (m, 1H), 3.74 (m, 1H), 3.86 (t, 2H), 4.00 (m, 1H), 4.42 (m, 1H), 6.20 (bs, 1H) , 7.20 (d, 2H), 7.29 (m, 1H), 7.39 (m, 4H), 7.50 (d, 2H), 7.54 (d, 2H). LRMS: m/z (ES+) 398 [MNa+].

Preparations 145 to 150

The following compounds have a general structure

[image]

were prepared from the corresponding alkenes, following procedures similar to that described in Preparation 144.

[image] [image]

1 = isolated without column chromatography

Preparation 151

(2S,6R)-4-(4-methoxybenzyl)-6-methyl-2-{[1-(2-pyridinyl)-1H-imidazol-4-yl]methyl}-3-morpholinone

[image]

A mixture of imidazole from Preparation 55 (566 mg, 1.8 mmol), copper (I) oxide (20 mg, 0.14 mmol) and potassium carbonate (372 mg, 2.7 mmol) in 2-bromopyridine (1 mL) it was heated at 100°C for 18 hours. The cooled mixture was purified by column chromatography on Biotage® silica gel, eluting with a gradient of toluene:diethylamine (93:7 to 86:14) to give the title compound as a foam, 482 mg. 1H-NMR (CDCl3, 400 MHz) (5:1 mixture of regioisomers) δ: 0.94, 1.20 (2×d, 3H), 2.83-3.23 (m, 4H), 3.63, 3.78 (2×s, 3H), 3.97 (m, 1H), 4.16, 4.22 (2×d, 1H), 4.50-4.82 (m, 2H), 6, 64, 6.82 (2×d, 2H), 7.00, 7.18 (2×d, 2H), 7.35, 7.44 (2×m, 1H), 7.59 (m, 2H ), 7.96 (m, 1H), 8.40-8.57 (m, 2H). HRMS: m/z (ES+) 393.1926 [MH+].

Preparation 152

(2S,6R)-6-methyl-2-{[1-(2-pyridinyl)-1H-imidazol-4-yl]methyl}-3-morpholinone

[image]

A mixture of the protected morpholino from Preparation 151 (454 mg, 1.16 mmol) and ammonium cerium(IV) nitrate (1.585 g, 2.9 mmol) in water (8 mL) and acetonitrile (16 mL) was heated at 40° C 6 hours. The cooled mixture was diluted with methanol (100 mL) and the solution was adsorbed onto silica gel. The product was isolated by chromatography on a silica gel column with dichloromethane:methanol:0.88 ammonia (95:5:1) as eluent and further purified by chromatography on a Biotage® silica gel column with toluene:diethylamine (92:8) and dichloromethane:methanol: 0.88 ammonia (95:5:1) as eluent to give the title compound, 240 mg. 1H-NMR (CD3OD, 400 MHz) (7:1 mixture of regioisomers) δ: 1.01, 1.21 (2×d, 3H), 2.92-3.36 (m, 4H), 3.78, 3.93 (2×m, 1H), 4.27, 4.46 (2×m, 1H), 7.37, 7.45 (2×m, 1H), 7.58-7.70 (m , 2H), 7.96, 8.00 (2×m, 1H), 8.40-8.18 (m, 2H).

Preparation 153

(6R)-2-[hydroxy(1-propyl-1H-imidazol-4-yl)methyl]-4-(4-methoxybenzyl)-6-methyl-3-morpholinone

[image]

A solution of the compound from Preparation 12 (6.81 g, 29.0 mmol) in tetrahydrofuran was added dropwise to a solution of lithium diisopropylamide (23.2 mL, 1.5 M in cyclohexanes, 34.8 mmol) at -78°C and the solution was stirred for another 20 minutes at -78°C. Then, a solution of the aldehyde from Preparation 1 (4 g, 29.0 mmol) in tetrahydrofuran (total volume 80 mL) was added dropwise and the mixture was allowed to slowly warm to room temperature. Saturated ammonium chloride solution (50 mL) was added, followed by water (100 mL) and the mixture was extracted with ethyl acetate. The combined organic extracts were dried (MgSO4) and concentrated under reduced pressure. The remaining orange oil was purified by silica gel column chromatography eluting with a gradient of ethyl acetate:methanol (98:2 to 95:5) to give the title compound as an orange oil, 5.71 g. 1H-NMR (CDCl 3 , 400 MHz) (mixture of diastereomers) δ: 0.92 (m, 3H), 1.14 (m, 3H), 1.58 (m, 2H), 2.96-3.18 (m, 2H), 3.78- 4.00 (m, 6H), 4.22-4.76 (m, 3H), 5.06-5.30 (m, 1H), 6.81-6.95 (m, 3H), 7, 18 (m, 2H), 7.42 (d, 1H). LRMS: m/z (ES+) 374 [MH+].

Preparation 154

(2EZ,6R)-4-(4-methoxybenzyl)-6-methyl-2-[(1H-imidazol-4-yl)methylidene]-3-morpholinone

[image]

A mixture of the compound from Preparation 37 (91 g, 164 mmol) and water (90 mL) in glacial acetic acid (900 mL) was heated at 40°C for one hour. The cooled mixture was concentrated under reduced pressure, diluted with water (400 mL), and the resulting precipitate was filtered off. The filtrate was washed with ether (2 x 400 mL), then neutralized with sodium bicarbonate and extracted with ethyl acetate (1000 mL). This organic solution was washed with water, dried (Na2SO4) and evaporated under reduced pressure to give the title compound as a gum, 46.4 g. 1H-NMR (CDCl3, 400 MHz) δ: 1.41 (d, 3H), 3.24 (dd, 1H), 3.38 (dd, 1H), 3.80 (s, 3H), 4.34 (m, 1H), 4.58 (d, 1H), 4.68 (d , 1H), 6.84 (d, 2H), 6.97 (s, 1H), 7.20 (d, 2H), 7.30 (s, 1H).

Preparation 155

(2EZ,6R)-4-(4-methoxybenzyl)-6-methyl-2-[(1-propyl-1H-imidazol-4-yl)methylidene]-3-morpholinone

[image]

Triethylamine (3 mL, 21.75 mmol) was added dropwise to a solution of alcohol from Preparation 153 (5.41 g, 14.50 mmol) in dichloromethane (60 mL) and the solution was cooled to 0°C. Methanesulfonyl chloride (1.68 mL, 21.75 mmol) was added and the mixture was allowed to warm to room temperature, after which it was stirred for an additional 2 hours. More triethylamine (2 mL, 14.50 mmol) was added and the mixture was heated to 35°C, then stirred for 18 hours. The solution was washed with water (100 mL), sodium bicarbonate solution (100 mL) and brine (50 mL), then dried (MgSO4) and concentrated under reduced pressure. The residue was purified by silica gel column chromatography eluting with a gradient of dichloromethane:methanol:0.88 ammonia (99:1:0.1 to 98:2:0.2) to give one isomer of the title compound as an orange oil, 1.8 g, and the second isomer, 260 mg. 1H-NMR (CDCl3, 400 MHz, major isomer) δ: 0.96 (t, 3H), 1.38 (d, 3H), 1.80 (m, 2H), 3.20 (dd, 1H), 3.32 (dd, 1H), 3.78 (s, 3H), 3.86 (t, 2H), 4.25 (m, 1H), 4.57 (d, 1H), 4.65 (d , 1H), 6.84 (d, 2H), 7.02 (s, 1H), 7.20 (d, 2H), 7.35 (s, 1H), 7.46 (s, 1H). LRMS: m/z (ES+) 356 [MH+]. Microanalysis showed: C, 63.99; H, 6.88; N, 11.00. C20H25N3O3⋅H2O requires C 64.32; H, 7.29; N, 11.25%.

Preparation 156

(2S,6R)-4-(4-methoxybenzyl)-6-methyl-2-[(1-propyl-1H-imidazol-4-yl)methyl]-3-morpholino

[image]

A mixture of the alkene from Preparation 155 (1.8 g, 5.07 mmol) and 10% palladium on carbon (Degussa® type 101) (200 mg) in ethanol (50 mL) was hydrogenated at 60 psi (410 kPa) and 50°C for 18 hours. TLC analysis showed lag of the starting material. The mixture was filtered, the filtrate was evaporated under reduced pressure and the residue was redissolved in ethanol (50 mL). 10% palladium on carbon (Degussa® type 101) (200 mg) was added and the mixture was hydrogenated at 60 psi (410 kPa) and 50°C for 18 hours, then filtered. The filtrate was evaporated under reduced pressure and the residue was purified by silica gel column chromatography with dichloromethane:methanol:0.88 ammonia (98:2:0.2) to give the title compound as a colorless oil. 1H-NMR (CDCl3, 400 MHz) δ: 0.92 (t, 3H), 1.19 (d, 3H), 1.78 (m, 2H), 2.98-3.16 (m, 3H) , 3.58 (dd, 1H), 3.82 (m, 6H), 4.50 (m, 3H), 6.75 (s, 1H), 6.82 (d, 2H), 7.18 ( d, 2H), 7.58 (s, 1H). LRMS: m/z (ES+) 358 [MH+]. Microanalysis showed: C, 62.12; H, 7.58; N, 10.89. C20H27N3O3⋅1.5H2O requires C 62.48; H, 7.86; N, 10.93%.

Preparation 157

(2S,6R)-6-methyl-2-[(1-propyl-1H-imidazol-4-yl)methyl]-3-morpholinone

[image]

A solution of the compound from Preparation 156 (1.2 g, 3.36 mmol) in methanesulfonic acid (5 mL) was stirred at 70°C for 2 hours. The cooled mixture was washed with ether (2 × 20 mL) by decantation. Water (20 mL) was added and the mixture was basified with 0.88 ammonia, after which it was washed with ethyl acetate (20 mL). The aqueous phase was evaporated under reduced pressure, the residue was suspended in acetonitrile and the mixture was heated to 50°C. The acetonitrile solution was separated by decantation and evaporated under reduced pressure to give an oil. It was purified by silica gel column chromatography, eluting with a gradient of dichloromethane:methanol:0.88 ammonia (98:2:0.2 to 96:4:0.4) to give the title compound as a colorless oil, 560 mg. 1H-NMR (CDCl3, 400 MHz) δ: 0.94 (t, 3H), 1.22 (d, 3H), 1.59 (m, 2H), 3.04 (dd, 1H), 3.18 -3.37 (m, 3H), 3.85 (m, 3H), 4.42 (m, 1H), 6.50 (s, 1H), 6.79 (s, 1H), 7.68 ( with, 1H). LRMS: m/z (ES+) 238 [MH+].

Preparation 158

(2R,6R)-2-[(1H-imidazol-4-yl)methyl]-6-methyl-3-morpholinone

[image]

Ammonium cerium(IV) nitrate (1.1 g, 2 mmol) was added to a solution of the protected lactam from Preparation 55b (200 mg, 0.63 mmol) in water (4 mL) and acetonitrile (4 mL) and the mixture was stirred at room temperature for 3 hours. The solution was diluted with acetonitrile (5 mL) and 0.88 ammonia (4 mL), and the mixture was filtered through Arbocel®, washing with a solution of acetonitrile:water (50:50, 10 mL). The filtrate was concentrated under reduced pressure, and the aqueous residue was washed with ether, after which it was evaporated under reduced pressure. The crude product was purified by silica gel column chromatography, eluting with a gradient of dichloromethane:methanol:0.88 ammonia (95:5:0.25 to 92:8:0.4) to give the title compound as a foam, 88 mg. 1H-NMR (D2O, 400 MHz) δ: 1.20 (d, 3H), 3.02-3.30 (m, 6H), 4.01 (m, 1H), 4.40 (dd, 1H) , 6.86 (s, 1H), 7.58 (s, 1H).

Compounds of the present invention can be tested using the following assay, which is based on that described in Boffa et al., J. Biol. Chem. 1998, 273, 2127. Compounds are incubated with activated TAFI and a standard substrate for TAFIa, the rate of substrate hydrolysis is determined and compared to the rate of hydrolysis in the absence of compounds, and the amount of inhibition is expressed as Ki.

TAFI inhibition assay

1. Activation of TAFI

Human TAFI (recombinant or purified) is activated by incubating 20 μL stock solution (360 μg/mL) with 10 μL human thrombin (10 NIH units/mL), 10 μL rabbit thrombomodulin (30 μg/mL), and 6 μL calcium chloride (50 mM) in 50 μL of 20 mM HEPES (N-[2-hydroxyethyl]piperazine-N-[2-ethanesulfonic acid]) buffer containing 150 mM sodium chloride and 0.01% TWEEN 80 (polyoxyethylene-sorbitan-monooleate) , pH 7.6 for 20 minutes at 22°C. At the end of the incubation period, thrombin is neutralized by adding 10 μL of PPACK (D-Phe-Pro-Arg-chloromethyl-ketone) (100 nM). The resulting TAFIa solution is stored in ice for 5 minutes and finally diluted with 175 μL of HEPES buffer.

2. Determination of Ki (TAFIa)

Calculation of Ki

A certain number of different dilutions of the tested compound in water were prepared. 150 μL HEPES buffer and 10 μL TAFIa were added to 20 μL of each dilution, which was then preincubated for 15 minutes at 24°C. 20 μL of standard concentration furyl-acryloyl-alanyl-lysine (FAAL) is then added to each dilution. Substrate turnover is measured by reading the absorbance of the reaction mixture at 330 nm every 15 seconds for 30 minutes. The reaction is carried out at 24°C, and the samples are mixed for 3 seconds before each absorbance reading.

Then a graph is drawn of the percentage of inhibition in relation to the concentration of the tested compound, from which the IC50 value is calculated. The Ki value is then calculated from the Cheng-Prusoff equation.

To check the accuracy of the results, two controls, positive and negative, were always used. For the first control, the assay was performed as described above, but with 20 μL of water instead of diluting the test compound. This shows minimal inhibition. For the second control, the assay was performed as described above, but with an effective amount of a non-specific carboxypeptidase inhibitor instead of the test compound dilution. This shows maximum inhibition. When the two controls did not show minimum or maximum inhibition, the results were discarded and the test compound was reanalyzed.

Using the assay described above, the compounds of the Examples were shown to be potent and selective inhibitors of TAFI. All tested compounds had a Ki value of less than 20 μM. The specific Ki values of some compounds are shown below:

Compound from Example Ki (TAFIa)

4 10 nM

5 10 nM

40 14 nM

49 9 nM

50 26 nM

The selectivity of the compounds of the present invention for TAFIa towards CPN was evaluated by determining the Ki of the compounds of the present invention for CPN and comparing this value with the Ki value for TAFIa. Ki was calculated using the Ki calculation assay for TAFIa, but substituting 10 μL of human CPN for 10 μL of TAFIa. The compounds of the present invention showed marked selectivity for TAFIa to CPN of the order of >50:1. Specific Ki values and calculated selectivities of some compounds are detailed below:

Compound from Example Ki (CPN) Selectivity

5 >10 μM >1000

51 >10 μM >380

Claims (30)

1. Compound according to formula (I): [image] indicated that: n is 0, 1, 2 or 3; R1 is chosen between optionally substituted straight-chain or branched C1-6 alkyl groups, optionally substituted straight-chain or branched C2-6 alkenyl groups, optionally substituted straight-chain or branched C2-6 alkynyl groups, arilla, aromatic heterocycle, heterocycle or hydrogen; wherein the possible substituents in the above mentioned groups (a), (b) and (c) are selected from C3-7 cycloalkyl, aryl, aromatic heterocycle, heterocycle, OR10, NR10R11, S(O)pR10, OC(O)R11, CO 2 R 10 , CONR 10 R 11 , SO 2 NR 10 R 11 , halo and NHSO 2 R 10 , and p is 0, 1 or 2; R 2 , R 3 , R 4 , R 6 , R 7 and R 9 are each independently selected from hydrogen and straight or branched C 1-6 alkyl, optionally substituted with OR 10 or halo; R 5 and R 8 are each independently selected from hydrogen and straight or branched C 1-6 alkyl, optionally substituted with OR 10 or halo, or together form a C 2-6 alkylene chain; R 10 and R 11 are each independently selected from hydrogen and straight or branched C 1-6 alkyl; aryl is a 6-14-membered aromatic monocyclic or fused polycyclic carbocyclic group, optionally substituted with one or more groups selected from R12, halo, OR13, NR13R14, NR13CO2R12, CO2R13, NR13SO2R12, CN, haloalkyl, O(haloalkyl), SR13, S (O)R12, SO2R12, OC(O)R13, SO2NR13R14, C(O)NR13R14, C3-7 cycloalkyl, O(C3-7 cycloalkyl), R15 and OR15, and R12 is straight or branched C1-6 alkyl, while R13 and R14 are independently selected from hydrogen and straight-chain or branched C1-6 alkyl, while R15 is phenyl, optionally substituted with R12, OR13, halo or haloalkyl; aromatic heterocycle is a 5-7 membered aromatic ring containing from 1 to 3 heteroatoms, independently selected from O, S and N, wherein said ring is optionally substituted by one or more groups selected from OR13, NR13R14, CO2R13, NR13CO2R12, R12, halo, CN, haloalkyl, O(haloalkyl), SR13, S(O)R12, SO2R12, OC(O)R13, NR13SO2R12, SO2NR13R14 and C(O)NR13R14; And heterocycle is a 3-8-membered ring containing from 1 to 3 heteroatoms, independently selected from O, S and N, wherein said ring is saturated or partially saturated, and is optionally substituted by one or more groups selected from OR13, NR13R14, CO2R13, NR13CO2R12, R12, halo, CN, haloalkyl, O(haloalkyl), SR13, S(O)R12, SO2R12, OC(O)R13, NR13SO2R12, SO2NR13R14 and C(O)NR13R14, or its tautomers or pharmaceutically acceptable salts or solvates of said compound or tautomer. 2. The compound according to claim 1, characterized in that the imidazole substitution pattern is as shown by the formula (ID1): [image] . 3. A compound according to claim 1, characterized in that its stereochemistry is as shown by formula (IA): [image] . 4. A compound according to any of the preceding claims, characterized in that n is 0 or 1. 5. Compound according to claim 4, characterized in that n is 0. 6. Compound according to any of the preceding claims, characterized in that R1 is hydrogen, aryl, C2-6 alkenyl or C1-6 alkyl group, optionally substituted by one or more groups selected from C3-7 cycloalkyl, aryl, aromatic heterocycle, OR10 , CO2R10, halo and NHSO2R10. 7. Compound according to claim 6, characterized in that R1 is hydrogen, aryl or C1-6 alkyl group, optionally substituted by a group selected from cyclohexyl, and aryl R1 is hydrogen, aryl or C1-6 alkyl, optionally substituted by cyclohexyl or aryl. 8. A compound according to claim 7, characterized in that R1 is hydrogen or C1-3 alkyl. 9. Compound according to claim 8, characterized in that R1 is hydrogen. 10. A compound according to any one of the preceding claims, characterized in that R 2 and R 3 are each independently hydrogen or C 1-6 alkyl. 11. Compound according to claim 10, characterized in that both R2 and R3 are hydrogen. 12. A compound according to any of the preceding claims, characterized in that R4 is hydrogen or C1-6 alkyl. 13. Compound according to claim 12, characterized in that R4 is hydrogen. 14. A compound according to any one of the preceding claims, characterized in that R 6 , R 7 and R 9 are each independently hydrogen or C 1-3 alkyl. 15. A compound according to claim 14, characterized in that R 6 , R 7 and R 9 are each independently hydrogen or methyl. 16. Compound according to claim 15, characterized in that both R6 and R7 and R9 are hydrogen. 17. A compound according to any of the preceding claims, characterized in that R5 is hydrogen or C1-3 alkyl. 18. Compound according to claim 17, characterized in that R5 is hydrogen or methyl. 19. Compound according to claim 18, characterized in that R5 is hydrogen. 20. A compound according to any one of claims 17, 18 and 19, characterized in that R8 is hydrogen or methyl. 21. Compound according to claim 20, characterized in that R8 is hydrogen. 22. Compound according to claim 1, characterized in that it is selected from the following compounds: (2S)-(-)-2-(2-aminoethoxy)-3-(1-phenyl-1H-imidazol-4-yl)-propanoic acid; (2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]-propanoic acid; (2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-[1-phenyl-1H-imidazol-4-yl]-propanoic acid; (2S)-2-{[(2S)-2-aminopropyl]oxy}-3-[1-(2-cyclohexylethyl)-1H-imidazol-4-yl]-propanoic acid; (2S)-2-(2-aminoethoxy)-3-(1H-imidazol-4-yl)-propanoic acid; (2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-[1H-imidazol-4-yl]-propanoic acid and (2S)-2-{[(1R)-2-amino-1-methylethyl]oxy}-3-[1-(2-pyridinyl)-1H-imidazol-4-yl]-propanoic acid, and their pharmaceutically acceptable salts. 23. A compound according to any one of claims 1 to 22, characterized in that it is used as a medicine. 24. A compound according to any one of claims 1 to 22, characterized in that it is used as a medicament for the treatment of a condition selected from thrombotic conditions, atherosclerosis, adhesions, skin scars, cancer, fibrotic conditions, inflammatory diseases and conditions that are improved by maintaining or increasing the level of bradykinin in the body. 25. Pharmaceutical preparation, characterized in that it contains a compound according to any one of claims 1 to 22 and a pharmaceutically acceptable carrier. 26. Use of a compound according to any one of claims 1 to 22, characterized in that it is used in the preparation of a medicament for the treatment of a condition selected from thrombotic conditions, atherosclerosis, adhesions, skin scars, cancer, fibrotic conditions, inflammatory diseases and conditions that improve with maintenance or by increasing the level of bradykinin in the body. 27. Use according to claim 26, characterized in that the drug is used for the treatment of a thrombotic condition. 28. A method of treating a condition selected from thrombotic conditions, atherosclerosis, adhesions, skin scars, cancer, fibrotic conditions, inflammatory diseases and conditions that are improved by maintaining or increasing the level of bradykinin in the body, characterized in that it comprises the administration of a compound according to any one of claims 1 to 22 to an individual who needs such a procedure. 29. Procedure for preparing the compound according to formula (I) [image] where: n is 0, 1, 2 or 3; R1 is chosen between optionally substituted straight-chain or branched C1-6 alkyl groups, optionally substituted straight-chain or branched C2-6 alkenyl groups, optionally substituted straight-chain or branched C2-6 alkynyl groups, arilla, aromatic heterocycle, heterocycle or hydrogen; wherein the possible substituents in the above mentioned groups (a), (b) and (c) are selected from C3-7 cycloalkyl, aryl, aromatic heterocycle, heterocycle, OR10, NR10R11, S(O)pR10, OC(O)R11, CO 2 R 10 , CONR 10 R 11 , SO 2 NR 10 R 11 , halo and NHSO 2 R 10 , and p is 0, 1 or 2; R 2 , R 3 , R 4 , R 6 , R 7 and R 9 are each independently selected from hydrogen and straight or branched C 1-6 alkyl, optionally substituted with OR 10 or halo; R 5 and R 8 are each independently selected from hydrogen and straight or branched C 1-6 alkyl, optionally substituted with OR 10 or halo, or together form a C 2-6 alkylene chain; R 10 and R 11 are each independently selected from hydrogen and straight or branched C 1-6 alkyl; aryl is a 6-14-membered aromatic monocyclic or fused polycyclic carbocyclic group, optionally substituted with one or more groups selected from R12, halo, OR13, NR13R14, NR13CO2R12, CO2R13, NR13SO2R12, CN, haloalkyl, O(haloalkyl), SR13, S (O)R12, SO2R12, OC(O)R13, SO2NR13R14, C(O)NR13R14, C3-7 cycloalkyl, O(C3-7 cycloalkyl), R15 and OR15, and R12 is straight or branched C1-6 alkyl, while R13 and R14 are independently selected from hydrogen and straight or branched C1-6 alkyl, while R15 is phenyl, optionally substituted with R12, OR13, halo or haloalkyl; aromatic heterocycle is a 5-7 membered aromatic ring containing from 1 to 3 heteroatoms, independently selected from O, S and N, wherein said ring is optionally substituted by one or more groups selected from OR13, NR13R14, CO2R13, NR13CO2R12, R12, halo, CN, haloalkyl, O(haloalkyl), SR13, S(O)R12, SO2R12, OC(O)R13, NR13SO2R12, SO2NR13R14 and C(O)NR13R14; And heterocycle is a 3-8-membered ring containing from 1 to 3 heteroatoms, independently selected from O, S and N, wherein said ring is saturated or partially saturated, and is optionally substituted by one or more groups selected from OR13, NR13R14, CO2R13, NR13CO2R12, R12, halo, CN, haloalkyl, O(haloalkyl), SR13, S(O)R12, SO2R12, OC(O)R13, NR13SO2R12, SO2NR13R14 and C(O)NR13R14, or its tautomer, characterized in that it comprises the following steps: preparation of the compound according to formula (II) [image] where is: P1 is an optionally substituted C1-6 alkyl group, an optionally substituted C4-7 cycloalkyl group, an optionally substituted benzyl group or a tri(C1-6 alkyl)silyl group; And R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and n are defined as for formula (I); you treating said compound of formula (II) with a reagent or combination of reagents suitable for removing the P1 group. 30. Procedure for preparing the compound according to formula (I): [image] where: n is 0, 1, 2 or 3; R1 is chosen between optionally substituted straight-chain or branched C1-6 alkyl groups, optionally substituted straight-chain or branched C2-6 alkenyl groups, optionally substituted straight-chain or branched C2-6 alkynyl groups, arilla, aromatic heterocycle, heterocycle or hydrogen; wherein the possible substituents in the above mentioned groups (a), (b) and (c) are selected from C3-7 cycloalkyl, aryl, aromatic heterocycle, heterocycle, OR10, NR10R11, S(O)pR10, OC(O)R11, CO 2 R 10 , CONR 10 R 11 , SO 2 NR 10 R 11 , halo and NHSO 2 R 10 , and p is 0, 1 or 2; R 2 , R 3 , R 4 , R 6 , R 7 and R 9 are each independently selected from hydrogen and straight or branched C 1-6 alkyl, optionally substituted with OR 10 or halo; R 5 and R 8 are each independently selected from hydrogen and straight or branched C 1-6 alkyl, optionally substituted with OR 10 or halo, or together form a C 2-6 alkylene chain; R 10 and R 11 are each independently selected from hydrogen and straight or branched C 1-6 alkyl; aryl is a 6-14-membered aromatic monocyclic or fused polycyclic carbocyclic group, optionally substituted with one or more groups selected from R12, halo, OR13, NR13R14, NR13CO2R12, CO2R13, NR13SO2R12, CN, haloalkyl, O(haloalkyl), SR13, S (O)R12, SO2R12, OC(O)R13, SO2NR13R14, C(O)NR13R14, C3-7 cycloalkyl, O(C3-7 cycloalkyl), R15 and OR15, and R12 is straight or branched C1-6 alkyl, while R13 and R14 are independently selected from hydrogen and straight or branched C1-6 alkyl, while R15 is phenyl, optionally substituted with R12, OR13, halo or haloalkyl; aromatic heterocycle is a 5-7 membered aromatic ring containing from 1 to 3 heteroatoms, independently selected from O, S and N, wherein said ring is optionally substituted by one or more groups selected from OR13, NR13R14, CO2R13, NR13CO2R12, R12, halo, CN, haloalkyl, O(haloalkyl), SR13, S(O)R12, SO2R12, OC(O)R13, NR13SO2R12, SO2NR13R14 and C(O)NR13R14; And heterocycle is a 3-8-membered ring containing from 1 to 3 heteroatoms, independently selected from O, S and N, wherein said ring is saturated or partially saturated, and is optionally substituted by one or more groups selected from OR13, NR13R14, CO2R13, NR13CO2R12, R12, halo, CN, haloalkyl, O(haloalkyl), SR13, S(O)R12, SO2R12, OC(O)R13, NR13SO2R12, SO2NR13R14 and C(O)NR13R14, or its tautomer, characterized in that it comprises the following steps: preparation of the compound according to formula (XIV) [image] where are they: R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , R 8 , R 9 and n are defined as for formula (I); you treating said compound of formula (XIV) with a reagent or combination of reagents suitable for pyrolysis of the amide bond of the lactam ring.