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OA12362A - Substituted imidazoles as tafia inhibitors. - Google Patents

Substituted imidazoles as tafia inhibitors. Download PDF

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OA12362A
OA12362A OA1200300039A OA1200300039A OA12362A OA 12362 A OA12362 A OA 12362A OA 1200300039 A OA1200300039 A OA 1200300039A OA 1200300039 A OA1200300039 A OA 1200300039A OA 12362 A OA12362 A OA 12362A
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OA1200300039A
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Charlotte Moira Norfo Allerton
Julian Blagg
Mark Edward Bunnage
John Steele
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Pfizer
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Priority claimed from GB0020346A external-priority patent/GB0020346D0/en
Priority claimed from GB0027409A external-priority patent/GB0027409D0/en
Priority claimed from GB0029556A external-priority patent/GB0029556D0/en
Application filed by Pfizer filed Critical Pfizer
Publication of OA12362A publication Critical patent/OA12362A/fr

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Description

012362 1
SUBSTITUTED IMIDAZOLES AS TAFIA INHIBITORS
The présent invention describes a sériés of substituted imidazoles as TAFIa inhibitors, usefui in the treatment of disease.
Thrombin Activatable Fibrinolysis Inhibitor, TAFI, is a 60kDa giycoprotein found in 5 human plasma. It is also known as procarboxypeptidase B, carboxypeptidase B,plasma carboxypeptidase B, carboxypeptidase U and carboxypeptidase R. It playsan intrinsic part in the blood coagulation process, during which it is transformed intoan activated form, TAFIa, whereupon it acts upon the fibrin matrix which comprises adeveloping blood clôt to prevent its dissolution. Imbalances in the blood coagulation 10 process are thought to be the origin of a large and disparate number of diseaseconditions, which are linked by an unwanted build up of fibrin. The scale of fibrinbuild up is determined by the délicate equilibrium between two biochemical cascadesin the human body; the coagulation and fibrinolysis cascades. These cascades arean intégral part of maintaining hemostasis. *5 To maintain hemostasis in the blood, mammals hâve developed mechanisms torepair the body in the event of vascular injury. The injured blood vessel will constrictto reduce the blood fiow to the area. Piatelets will aggregate to reduce the loss ofblood from the area, followed by fibrinogen which will polymerise and form a fibrinclôt. This clôt will cover the area of vascular damage, preventing blood loss. Once 20 the blood vessel has been repaired the clôt will then dissolve. The coagulationcascade is responsible for the forming of a clôt; the fibrinolysis cascade isresponsible for the dissolution of the clôt.
Studies hâve shown that these two processes-are intrinsically linked through thegénération of a-thrombin. α-Thrombin is the final product of the blood coagulation 25 cascade and is responsible for the conversion of soluble plasma fibrinogen to aninsoluble fibrin matrix. Polymerised fibrin provides a haemostatic plug which preventsblood loss from the site of vascular injury and provides a provisional matrix whichenhances the subséquent repair process. In addition to mediating coagulation, a-thrombin also reduces the rate at which blood clots are broken down by the serine 012362 2 protease plasmin. The anti-fibrinoiytic activity of α-thrombin résulte from ïts activationof TAFI. TAFI circulâtes in normal plasma at a concentration of about 75nM in aninactive form. Thrombin converts the inactive zymogen to the active TAFI (TAFIa), areaction that is augmented about 1250-fold by thrombomodulin. Once activated,TAFIa cleaves both C-temninal arginine and lysine residues from the developingfibrin clôt. The removal of di-basic amino acids from the surface of the fibrin matrixatténuâtes clôt lysis by inhibiting the binding of the key mediators of fibrinolysis:tissue plasminogen activator (tPA) and its substrate, plasminogen, which is theprecursor of plasmin. Both tPA and plasminogen contain a structural motif called akringle domain which binds tightly to C-terminal lysine residues. The removal ofthese binding sites prevents the formation of a temary complex between tPA,plasminogen and fibrin and this inhibits the conversion of plasminogen to plasmin,thus protecting the clôt from rapid dégradation.
It can be seen that if the equilibrium between coagulation and fibrinolysis is in favourof coagulation, then there will be a larger amount of fibrin présent than normal. Thismakes it more likeiy that the subject will develop one or more of the conditions inwhich thrombus build up is implicated. By the use of a TAFIa inhibitor, TAFIa will notbe able to act upon a developing fibrin clôt as described above to inhibit fibrinolysisof the clôt. Thus a TAFIa inibitor should serve to enhance the fibrinolysis cascade.
The use of TAFI inhibitors to treat certain conditions is known in the art. Whilst theuse of TAFIa inhibitors to treat such conditions is unknown, certain weak, non-specific TAFIa inhibitors hâve been identified. USA 5993815 teaches the use of a peptide that binds to the TAFI zymogen,inhibiting activation of the TAFI zymogen, to treat those disorders where a C-terminallysine or arginine is cleaved from an intact peptide. Suitable disorders are arthritis,sepsis, thrombosis, strokes, deep vein thrombosis and myocardial infarctions. Thepeptide used is an antibody or a functionally active fragment. The peptide should beused in an amount to promote fibrinolysis in vivo. JÜ I z o 6 2 3
McKay et al, Biochemistry, 1978, 17, 401, discloses the testing of a number ofcompounds as compétitive inhibitors of bovine carboxypeptidase B of pancreaticorigin. Inhibition was measured by the inhibitor’s eflïciency in protecting the activecentre tyrosine and glutamic acid of bovine carboxypeptidase B from irréversiblealkylation by bromoacetyl-D-arginine or bromoacetamidobutylguanidine. It issuggested that such inhibitors could act as bradykinin potentiators.
Bovine enzymes of pancreatic origin are very different to those found in humanplasma, so one would not expect inhibitors of one to inhibit the other. Moreover,such inhibitors are directed towards a very different utility. Accordingly the abovereference contains no teaching of TAFIa inhibitors or their utility.
Redlitz et al, J. Clin. Invest. 1995, 96, 2534, teaches the involvement of plasmacarboxypeptidase B (pCPB, or TAFI) in the formation of dots. The lysis of blood clotswas followed in the absence and presence of pCPB, whereupon it was found that thepresence of pCPB slowed clôt lysis. To confirm that pCPB was responsible twocontrat reactions were run; one where the lysis experiment was repeated in thepresence of pCPB and a carboxypeptidase inhibitor, PCI, a second where the lysisreaction was conducted in the presence of plasma from which pCPB was removed.In both cases lysis proceeded uninhibited.
Boffa et al, J. Biol. Chem. 1998, 273,2127, compares plasma and recombinant TAFIand TAFIa with respect to glycosylation, activation, thermal stability and enzymaticproperties. Inhibition constants for three compétitive inhibitors were determined: ε-aminocaproic acid (ε-ACA), 2-guanidinoethylmercaptosuccinic acid (GEMSA) andpotato carboxypeptidase inhibitor (PCI).
There are large numbers of carboxypeptidases, characterised by cleaving the C-terminal amino acid from a peptide. They may be divided into acidic, neutral or basic,depending on the type of amino acid they cleave. Basic carboxypeptidases cleavearginine, lysine and histidine. TAFIa is a spécifie subset of basic carboxypeptidases.In terms of the présent invention the inhibitors disciosed above by Redlitz et al andBoffa et al, are too weak, non-specific or otherwise unsuitable to be considered as 012346 2 * suitable TAFIa inhibitors for therapeutic application. Further, whilst the rôle of TAFIain clôt lysis is expiained, there is no suggestion that TAFIa inhibitors can be used totreat disease. WO00/66550 discusses a broad class of compounds useful as inhibitors of 5 carboxypeptidase U. Inhibitors of carboxypeptidase U are postulated to faciiitatefibrinolysis and thus the compounds are taught as useful in the treatment ofthrombotic conditions. There is no data to support this assertion, though details of asuitable assay are given. WOOO/66152 discloses formulations containing a carboxypeptidase U inhibitor and a 10 thrombin inhibitor. Suitable carboxypeptidase U inhibitors are those of WO00/66550.
The formulations are taught as primarily useful in treating thrombotic conditions.
The présent invention discloses a ciass of TAFIa inhibitors. There are very greatadvantages in using a TAFIa inhibitor over a TAFI inhibitor. TAFI is activated toTAFIa by reaction with thrombin. A TAFI inhibitor must prevent these two largepeptides coming together to react at the appropriate site. To date only large peptideshâve been described which can interfère with this reaction (USA-5993815). However,it has been discovered that the active site on TAFIa, responsible for reacting with adeveloping clôt, is small, and thus can be blocked by a small molécule, one with amolecular weight of below 1000, preferably below 500. It is a great advantage to 20 hâve a low molecular weight compound as the ‘active’ in a médicament. They areassociated with oral bioavailabiiity and patients usually prefer oral formulations.Further there is the potential for peptide therapeutics to induce an immune response.This is unlikely to be an issue with a small molécule. Small molécules are aisogenerally more stable in plasma and thus hâve a greater duration of action. This is 25 unlikely to be the case with large molécules, particularly peptides. For these reasonsa TAFIa inhibitor is preferred. The invention provides a potent class of TAFIainhibitors.
The présent invention provides as à preferred set of TAFIa inhibitors, compounds of•formula (I) 012362 5 R9
(l)
Where:
X is N or CH n is 0 to 3 R1 is: a) C-i-6 alkyl, straight Chain or branched chain, b) Cvs alkenyl, straight chain or branched chain, c) Ci-6 alkynyl, straight chain or branched chain, d) Heterocycie, e) Aromatic heterocycie, f) Aryl; g) hydrogen; said groups (a), (b) and (c) optionally further substituted by: C3-7 cycloalkyl, aryl,aromatic heterocycie, heterocycie, OR11, NR11R12, S(O)PR11, OC(O)R11, CO2R11,CONR11R12, SO2NR11R12, halo and NHSO2R11, where R1 may be attached at any position on the imidazole ring. R2 and R3 are each independently selected from hydrogen, C-|.s alkyl, optionallyfurther substituted by OR11, halo; or wherein R2 and R3 may be joined to form a link, said link is C2-e alkylene. R4 is hydrogen, Ci.6 alkyl, optionally further substituted by C3-7 cycloalkyl, aryl, OR11,halo and R11; or 012362 6 wherein R4 and R10 may be joined to form a iink, wherein said link is Cm alkylene,optionally further substituted by OR11, halo and R11, R5 and R6 are selected from: hydrogen, aryl, Ci.6 alkyl, said alkyl optionally further substituted by C3.7 cycloalkyl,aromatic heterocycle, heterocycle, aryl, OR11, R11 and halo; orwherein R10 and either of R5 or R6 may be joined to form a link, wherein said link is aC1-3 alkylene, optionally further substituted by OR11, halo, R11 and aryl; orwherein R5 and R6 may be joined to form a link, wherein said link is C2-6 alkylene. R7 and R8 are independently selected from: hydrogen, Cm alkyl, optionally further substituted by OR11, halo, aryl and R11; orwherein R7 and R8 may be joined to form a link, wherein said link is C2-6 alkylene. R9 and R10 are independently selected from:
Hydrogen, C(NR11)NR11R12, Cm alkyl, said alkyl optionally substituted by OR11, halo,aryl and R11; or wherein R9 and R10 may be joined to form a link, wherein said link is C2-6 alkylene. R11 and R12 are each independently selected from hydrogen or Cm alkyl; or whenforming a NR11R12 moiety, R11 and R12 may also be joined to form a link whereinsaid link is C2-6 alkylene. p is 0, 1 or 2
Wherein:
Aryl is defined as a 6-14 membered aromatic carbocycle, optionally furthersubstituted by R11, halo, OR11, NR11R12, NR11CO2R12, CO2R11, NR11SO2R12, CN,haloalkyl, O(haioalkyl), S(O)PR11, OC(O)R11, SO2NR11R12, C(O)NR11R12.
Aromatic heterocycle is defined as a 5 to 7 membered ring, containing from 1 to 3heteroatoms, each independently selected from O, S and N, said heterocycle groupoptionally substituted by OR11, NR11R12, CO2R11, NR11eo2R12, R11, halo, CN,haloalkyl, O(haioalkyl), S(O)PR11, OC(O)R11, NR11SO2R12, SO2NR11R12, C(O)NR11R12. 012362 7
Heterocycie is defined as a 3-8 membered ring containing from 1-3 heteroatoms,each independently selected from O, S and N, said ring being saturated or partiallysaturated, said heterocycie group optionally substituted by OR11, NR11R12, CO2R11,NR11CO2R12, R11, halo, CN, haloalkyl, O(haloalkyl), S(O)PR11, OC(O)R11, 5 NR11SO2R12,SO2NR11R12, C(O)NR11R12.
Compounds of formula (I) includes zwitterions, pharmaceutically acceptable salts,prodrugs, solvatés and polymorphs thereof.
Halo includes fluoro, chloro, bromo and iodo groups.
Alkyl includes straight Chain and branched chain. 10 A 6-14 membered aromatic carbocycle includes phenyl, naphthyl, indenyl, anthryland phenanthryl. A pharmaceutically acceptable sait of a compound of the formula (I) may be readilyprepared by mixing together solutions of a compound of the formula (I) and thedesired acid or base, as appropriate. The sait may precipitate from solution and be *5 collected by filtration or may be recovered by évaporation of the solvent
The pharmaceutically acceptable salts of the compounds of the formula (I) includethe acid addition and the base salts thereof.
Suitable acid addition salts are formed from acids which form non-toxic salts andexamples are the hydrochloride, hydrobromide, hydroiodide, sulfate, bisulfate, 20 nitrate, phosphate, hydrogen phosphate, acetate, maleate, fumarate, lactatê,tartrate, citrate, gluconate, succinate, saccharate, benzoate, methanesulphonate,ethanesulphonate, benzenesulphonate, p-toluenesuiphonate and pamoate salts. 012362 8
Suitable base salts are formed from bases which form non-toxic salts and examplesare the sodium, potassium, aluminium, calcium, magnésium, zinc anddiethanolamine salts.
For a review on suitable salts see Berge et al, J. Pharm. Sci., 1977, 66,1. 5 The pharmaceutically acceptable solvatés of the compounds of the formula (!)include the hydrates thereof.
Also included within the présent scope of the compounds of the formula (I) arepoiymorphs thereof.
It will also be appreciated that the compounds of the invention will include prodrugs10 thereof: pharmaceutically acceptable dérivatives of (I) in which the functional groupsexplicitly recited above hâve been derivatised to provide prodrugs which can beconverted to the parent compound in vivo. Such prodrugs are discussed in Drugs of
Today, 1983, 19, 499-538 and Annual Reports in Médicinal Chemistry, 1975, Vol.10, Ch 31, 306-326. Suitable prodrugs will include compounds of formula (11) and 15 OH)·
Wherein R1, R2, R3, R4, R5, R6, R7, R8, X and Z are as described above, R9 and R10are as described above orin addition one or both groups may be a suitable nitrogenprotecting group and R13 is an appropriate oxygen protecting group. Suitablenitrogen protecting groups include carbamates, particularly BOC and benzyl groups. 012362 9
Appropriate oxygen protecting groups are known to those skilled in the art andinclude ailyl, aryl and alkyl groups, said aikyl group optionally substituted by aryl orC3.7 cycloaikyl, or more specifically, such groups as benzyl, pivaloyloxymethyl (POM)and Ci„6 alkyl. Référencé is also made herein to "Protective Groups in Organic 5 Synthesis”, 2nd édition, T. W. Greene and P. G. M. Wutz, Wiiey-lnterscience (1991).
Compounds of formula (!) contain one or more asymmetric carbon atoms andtherefore exists in two or more stereoisomeric forms. Where compounds of formula(I) contain an alkenyl or alkenylene group, cis (E) and trans (Z) isomerism may alsooccur. The présent invention includes the individual stereoisomers of the compounds 10 of formula (I) and, where appropriate, the individual tautomeric forms thereof,together with mixtures thereof.
Preferred compounds of formula (1) include those which possess the stereochemistryshown below.
(IA) (IB)
Those compounds of formula (IA) are particularly preferred. 15 Séparation of diastereoisomers or cis and trans isomers may be achieved byconventional techniques, e.g. by fractional crystallisation, chromatography orH.P.L.C. of a stereoisomeric mixture of a compound of the formula (SA) or (IB) or asuitable sait or dérivative thereof. An individual enantiomer of a compound offormulae (IA) or (IB) may also be prepared from a corresponding optically pure 20 intermediate or by resolution, such as by H.P.L.C. of the corresponding racemateusing a suitable chiral support or by fractional crystallisation of the diastereoisomeric 012362 10
«F salts formed by reaction of the corresponding racemate with a suitable opticallyactive acid or base, as appropriate. Reference is made herein to “Enantiomère,Racemates and Resolutions” J. Jacques and A. Collet, published by Wiley, NY,1981; and “Handbook of Chiral Chemicals” chapter 8, Eds D. Ager and M. Dekker, 5 ISBN:0-8247-1058-4.
Preferred compounds of formula (I) include those where the imidazole is substiutedat any position by R1 and at the C2 or C4 positions by the amino acid fragment.Particularly preferred are those compounds of formula (I) where R1 is attached to N1of the imidazole moiety so as to give the (1,4)-disubstituted imidazole and 10 compounds of formula (I) where R1 is attached to C4 of the imidazole so as to givethe (2,4)-disubstituted imidazole.
Preferabiy R1 is an aryl group, a C3.7 cycloalkyl group, a Cm alkenyl group or a Cmalkyl group, said alkyl or alkenyl groups optionaliy substituted by one or more groupsseiected from: a C3-7 cycloalkyl group, heterocycie, aromatic heterocycle, OR11, 15 CO2R11, NR11SO2R12, NR11R12, C(O)NR11R12, SO2NR11R12, halo, OC(O)R11, aryl or S(O)PR11, where p is 0-2.
More preferabiy R1 is an aryl group, Cm alkenyl group or a Cm alkyl group, whereinsaid alkyl is group is optionaly subsituted by one or more groups seiected fromCO2R11, OR11, aryl, C3.7 cycloalkyl, NHSO2R11, halo, aromatic heterocycle. 20 Yet more preferabiy R1 is a CF3 group or a Cm alkyl group, wherein said alkyl isoptionaliy substituted by a C3.7 cycloalkyl group, aromatic heterocycle, OR11, CO2R11,NR11SO2R12 or aryl.
Even more preferabiy R1 is Cm alkyl, optionaliy substituted by a C3.4 cycloalkyl groupor aryl group. 25 Most preferabiy R1 is Cm alkyl R2 and R3 are preferabiy independently seiected from hydrogen and Cm alkyl.Most preferabiy R2 and R3 are hydrogen. 012362 11 R4 is preferably independently selected from hydrogen and C1-6 alkyl, said alkyloptionally substituted by phenyl; or wherein R4 and R10 may be joined to form a link,said link is C2-3 alkylene.
More preferably R4 is independently selected from hydrogen and C1-3 alkyl; or 5 wherein R4 and R10 may be joined to form a link, said link is C2.3 alkylene.
Yet more preferably R4 is independently selected from hydrogen; or wherein R4 andR10 may be joined to form a link, said link is C2-3 alkylene.
Most preferably R4 is hydrogen. R5 and R6 are preferably independently selected from hydrogen and C1.6 alkyl, saidl° alkyl group optionally substituted by phenyl; or wherein R5 and R10 may be joined to form a link, said link is C1-3 alkylene.
More preferably R5 and R6 are independently selected from hydrogen and Ci_3 alkyl,said alkyl group optionally substituted by phenyl; or wherein R5 and R10 may bejoined to form a link, said link is C2 alkylene. ‘5 Yet more preferably R5 and R6 are independently selected from hydrogen and C1-3alkyl.
Most preferably R5 and R6 are hydrogen.
Preferably R7 and R8 are independently selected from hydrogen and Cm alkyl, saidalkyl optionally substituted by phenyl. 20 More preferably R7 and R8 are independently selected from hydrogen and alkyl.Yet more preferably R7 and R8 are independently selected from hydrogen and C1.3alkyl.
Even more preferably R7 and R8 are independently selected from hydrogen and CH3.Most preferably R7 and R8 are hydrogen. 25 Preferably R9 and R10 are independently selected from hydrogen, C(NH)NH2 and C1-6alkyl; or wherein R10 and R4 may be joined to form a link, said link is C2-3 alkylene.More preferably R9 and R10 are independently selected from hydrogen and C1.3 alkyl;or wherein R10 and R4 may be joined to form a link, said link is C2-3 alkylene.
Yet more preferably R9 and R10 are independently selected from hydrogen and C1-3 3° alkyl. 012362 12
Most preferably R9 and R10 are hydrogen.
Preferably R11 and R12 are independently selected from hydrogen and Ci-3alkyl.
More preferably R11 and R12 are independently selected from hydrogen and CH3. X is preferably CH. 5 n is preferably 0 or 1.n is most preferably 0. aryl is preferably phenyl, optionally substituted by 1-3 groups selected from: R11,halo, OR11, NR11R12, CO2R11, NHSO2R11, CN or haloalkyl.
Most preferably aryl is phenyl. 10 Preferably Aromatic heterocycle is defined as a 5 to 6 membered ring, containingfrom 1 to 3 heteroatoms, each independently selected from O, S and N, saidheterocycle group optionally substituted by 1-3 groups selected from: OR11, NR11R12,CO2R11, NR11CO2R12, R11, halo, CN, haloalkyl, O(haloalkyl), S(O)PR11, OC(O)R11,NR11SO2R12, SO2NR11R12, C(O)NR11R12. 15 More preferably Aromatic heterocycle is defined as a 5 to 6 membered ring,containing from 1 to 2 heteroatoms, each independently selected from O, S and N,said heterocycle group optionally substituted by 1-3 groups selected from: OR11,NR11R12, CO2R11, NR11CO2R12, R11, halo, CN, haloalkyl, O(haioalkyl), S(O)PR11,OC(O)R11, NR11SO2R12, SO2NR11R12, C(O)NR11R12. 20 Most preferably Aromatic heterocycle is defined as a 5 to 6 membered ring,containing from 1 to 2 heteroatoms, each independently selected from O, S and N.
Preferably, Heterocycle is defined as a 3-8 membered ring containing from 1-2heteroatoms, each independently selected from O, S and N, said ring beingsaturated or partially saturated, said heterocycle group optionally substituted by 1-3 012362 13 groupe selected from: OR11, NR11R12, CO2R11, NR11CO2R12, R11, halo, CN, haloalkyl,O(haloalkyl), S(O)PR11, OC(0)R11, NR11SO2R12, SO2NR11R12, C(O)NR11R12.
More preferably, Heterocycle is defined as a 5-6 membered ring containing from 1-2heteroatoms, each independently selected from O, S and N, said ring being 5 saturated or partially saturated, said heterocycle group optionally substituted by 1-3groups selected from: OR11, NR11R12, CO2R11, NR11CO2R12, R11, halo, CN, haloalkyl,O(haloalkyl), S(O)PR11, OC(O)R11,NR11SO2R12, SO2NR11R12, C(O)NR11R12
Most preferably, Heterocycle is defined as a 5-6 membered ring containing from 1-2heteroatoms, each independently selected from O, S and N, said ring being 10 saturated or partially saturated.
Preferred compounds of the présent invention include: (±)-5-Amino-2-[( 1 -n-propyl-1 W-imidazol-4-yl)methyllpentanoic acid(Example 2) (+)-(2S)-5-Amino-2-[(1-o-butyl-1/-/-imidazol-4-yl)rnethyl]pentanoic acidif’ (Exampîe 5) (+)-(2S)-5-Amino-2-[(1 -n-propyl-1 H-imidazol-4-yl)methyl]pentanoic acid(Example 7) (+)-(2S)-5-Amino-2-(1 H-imidazol-4-ylmethyl)pentanoic acid(Example 9) 20 (2S)-2-[(2-Aminoethyl)amino]-3-(1 -n-propyl-1 H-imidazol~4-yl)propanoic acid (Example 25) (2S)-2-[(2-Aminoethyl)amino]-3-(1 -n-butyl-1 W-imidazol-4-yl)propanoic acid (Example26) (2S)-2-[(2-Aminoethyl)amino]-3-(1 -n-isobutyl-1 H-ifnîdazol-4-yi)propanoic acid 25 (Example 29) (2S)-2-[(2-Aminoethyl)aminoJ-3-(1 -n-isopentyl-1 H-imidazol-4-yi)propanoic acid(Example 30) 0,2362 14
Particulariy preferred is (+)-(2S)-5-Amino-2-[(1-n-propyl-1/7-imidazol-4-yl)methyljpentanoic acid (Example 7)
The présent invention also includes compounds of formula (XXIII) and (XXIV)
Where R1, R3, R5, R6, R7, R8 and R10 are as described above, R4 is hydrogen, n is 0,X is CH and R9 is as described above or is an appropriate nitrogen protecting group.Appropriate nitrogen protecting groups include carbamates, particulariy BOC andbenzyl groups. These compounds are particulariy usefu, as intermediates in thesynthesis of compounds of formula (!)
The invention further provides Methods for the préparation of the compounds of the 10 invention, which are described below and in the Examples and Préparations section.The skilled man will appreciate that the compounds of the invention could be made bymethods other than those herein described, by adaptation of the methods hereindescribed and/or adaptation of a plethora of methods known in the art. It is to beunderstood that the synthetic transformation methods specifically mentioned herein 15 may be carried out in various different sequences in order that the desiredsubstances can be efficiently assembled. The skilled chemist will exercise hisjudgement and skill as to the most efficient sequence of reactions for synthesis of agiven target substance.
It will be apparent to those skilled in the art that sensitive functional groups may need20 to be protected and deprotected during synthesis of a substance of the invention. Thismay be achieved by conventional techniques, for example as described in “Protective 012362 15
Groups in Organic Synthesis” by T. W. Greene and P. G. M. Wuts, John Wiley andSons Inc, 1991.
Compounds of formula (i) may be prepared by reacting a compound of formula (II)
Wherein R1, R2, R3, R4, R5, R6, R7, R8 and X are as described above, R9, R10 are as5 described above, additionally one or both may be a suitabie nitrogen protecting group and R13 is an appropriate oxygen protecting group,with a suitabie reagent to remove said oxygen protecting group.
Appropriate oxygen protecting groups include allyl groups and alkyl groups, said alkyigroups optionally substituted by aryl groups. 10 Suitabie reagents and conditions to remove said groups are well known to thoseskilled in the art and may include hydrolysis and hydrogénation.
Where R9 and/or R10 is a nitrogen protecting group, it may be necessary to removesaid nitrogen protecting group after reaction of (II) with a suitabie reagent to removesaid oxygen protecting group. Suitabie nitrogen protecting groups are well known to 15 those skilled in the art, as are suitabie conditions for their removal.
Compounds of formula (II), where R1, R3, R4, R5, R6, R7, R8, R9, R10, R13 and X areas described above and. R2 is hydrogen may be prepared from compounds offormula (V) and (VI) in accordance with the followirig réaction scheme 012362 16
012362 17
Compounds of formula (IV) may be formed by process step (a), a Wadsworth-Emmons réaction between compounds of formula (V) and (VI). This may beconducted under standard conditions, such as described in Org. Synth. Coll. Vol.,1988, 6, 358 and 1993, 8, 265. Suitable conditions include formation of thephosphonate anion with a suitable base such as NaH at 0°C, then reacting with 1 eqof the appropriate aldéhyde at room température for 18 hours. A suitable solventwould be tetrahydrofuran.
Compounds of formula (II) may be formed by process step (b), a hydrogénation. Thismay be carried out by a method such as catalytic hydrogénation, e.g. 10% Pd/C at 4atmosphères, in an alcoholic solvent (methanol or éthanol) at room température to60°C for between 4 and 72 hours; or by activated métal hydride réduction, e.g. 30 eqNaBhU, 1.5 to 2.5 eq CuCI, in methanol, at room température for 2 hours. Theprocess may also be conducted to give an asymmetric hydrogénation of the alkenebond. Such methods are well known to those skilled in the art and are discussed in“Asymmetric Synthetic Methodology” chapter 9, Eds D. Ager and M, East, CRCPress, 1996, ISBN: 0-8493-8492-9.
Compounds of formula (V) are commercially available or may be prepared by anumber of literature methods well known to a man skilled in the art. Reference ismade to the préparations described herein and to G. Shapiro et al, Heterocycles,1995, 41, 215; L. A. Reiter, J. Org. Chem., 1987, 52, 2714; B. H. Lipshutz et al,Tetrahedron Lett. 1986, 27, 4095; F. Aldebbagh et al, Tetrahedron Lett., 1997, 387937; and S. M. Abdelaal, J. Het. Chem. 1995, 32, 903.
Compounds of formula (VI) where R4, R5, R6, R7, R8, R9, R10 and R13 are asdescribed above and X is CH, may be prepared in accordance with the followingscheme. 012362 18 R10 R9 R8 R7
EtO„
EtO- γΎ OR13 R4 (VII) (c)
2/n (CH2),
Re
Rs (VIII) (VI)
Compounds of formula (VI) may be prepared from the compounds of formula (VII)and (VIII) where Y is halo, under the conditions of process step (c), an alkylationreaction. This may be carried out under standard conditions, typically 1 eq of (VII) istreated with 1.1 eq of NaH, before reaction with (VIII), 18-crown-6 (cat) at reflux for 5 18hours.
Compounds of formula (VI) where Rs, R6, R7, R8, R9, R10 and R13 are as describedabove, R4 is a suitable nitrogen protecting group and X is N, may be prepared usingthe reaction scheme described above.
Compounds of formula (I) may aiso be prepared by treating a compound of formula0 (US) under the conditions of a lactam hydrolysis reaction. Suitable conditions includethose of process step (d), a lactam hydrolysis. This may be conducted under 012362 19 standard conditions, typically basic conditions, e.g. aqueous LiOH in tetrahydrofuranat room température for 4-18 hours.
Compounds of formula (III) where R1, R3, R4, R5, R6, R7 *, R®, R9, R13, X and Z are asdescribed above and R2 is hydrogen, may be prepared by the following process
(a)
5 Compounds of formula (IX) may be prepared by reacting compounds of formula (V) and (X) under the conditions of process step (a) described above. Compounds of formula (111) may be prepared by reacting compounds of formula (IX) under the conditions of process step (b) described above. 012362 20
Compounds of formula (X) where R4, R5, R6, R7, R8, R9, R10 and R13 are as describedabove, with the proviso R9 and R10 may not be linked and X is CH may be preparedfrom a compound of formula (XI) where Y is halo, in accordance with the followingréaction scheme.
(c)
5 Compounds of formula (X) may be prepared from compounds of formula (XI) underthe conditions of process step (c) described above.
Compounds of formula (II) where R1, R2, R3, R4, R6, R7, R8, R10 and R13 are asdescribed above, R9 is as above or is a suitable nitrogen protecting group, X is N andR6 is hydrogen may be prepared from a compounds of formula (XII) and (XIII), inaccordance with the following reaction scheme. 10 012362 21
Compounds of formula (11) may be prepared by reacting compounds of formula (Xll)and (Xlll) under the conditions of process step (e), a reductive alkylation reaction,performed under standard conditions known to those skilled in the art. Suitableconditions would include reacting (Xll) and (XIII) in the presence of sodium acetate 5 and sodium cyanoborohydride.
Compounds of formula (II) wherein R9 is H may be obtained from compounds offormulae (II) where R9 is a suitable nitrogen protecting group by optional processstep (k), removal of a nitrogen protecting group; appropriate conditions for theremoval of nitrogen protecting groups P1 are described in "Protective Groups inl0 Organic Synthesis”, 2nd édition, T. W. Greene and P. G. M. Wutz, Wiley-lnterscience (1991). Appropriate conditions include: BOC deprotection: 6N aqueous hydrochloric acid at room température to reflux temp,for between 1 and 3 hours; 012362 22
Benzyl deprotection: dissolving métal réduction, e.g. Na, liq NH3, -78°C.
Compounds of formula (XIII) are commercially availabie or may be prepared bymethods well known to a man skilled in the art.
Compounds of formula (XII) above are commercially availabie. Alternatively where5 R1, R3, R4 and R13 are as described above, and R2 is hydrogen, they may be made by the route disclosed in Helv. Chim. Acta., 1994, 77,1395 or as disclosed below. 012362 23
Compounds of formula (XII) may be prepared by reacting compounds of formula (V)and (XIV) under the conditions of process step (a), described above. Compounds offormula (Xlla) may be prepared by reacting compounds of formula (XIII) under theconditions of process step (b) described above. If a compound of formula (XII) isrequired where R4 is not hydrogen, then compounds of formula (XII) may be 5 012362 24 prepared by reacting compounds of formula (Xlla) under the conditions of process step (e), described above.
Compounds of formula (Xlla) where R1, R2 and R3, are as described above, with theproviso R2 and R3 are not linked and R13 is methyl, may also be asymmetrically 5 prepared from a compound of formula (XVI), where Y is halo, in accordance with thefollowing reaction scheme.
Compounds of formula (XV) may be prepared· by reacting compounds of formula(XVII) and (XVI) under the conditions of process step (f) a Schollkopf asymmetricalkylation reaction, comprising reaction of a halide with a suitable deprotonatedSchollkopf chiral auxiliary (Angew. Chem. Int. Ed. Engl., 1981, 20, 798). Suitableconditions are treating the Schollkopf auxiliary in tetrahydrofuran at -78°C with BuLi, 10 012362 25
A followed by addition of (XVI) then 24 hours at room température. Compounds of» formula (XIla) may be prepared by reacting compounds of formula (XV) under the conditions of process step (g), a hydrolysis reaction, described in Angew. Chem. Int.
Ed. Engl., 1981, 20, 798. Suitable conditions are 5eq of 0.25N aqueous hydrochloric5 acid at room température for 2 hours.
Compounds of formula (XII) may be obtained by methods well known to thoseskilled in the art or as exemplified in the examples. It should be noted thatcompounds of formula (XII) and intermediates thereto wherein R1 is not H maybe produced by coupling compounds of formula (XII) and intermediates thereto 10 where R1 is H, with an appropriate reagent containing R1, where R1 is asdisclosed above.
Compounds of formula (II) where R1, R2, R3, R4, R5, R6, R7, R8, Rs, R10 and R13 areas described above and X is nitrogen may also be prepared from compounds offormula (XIX) and (XVIII) where Y is halo by the method described in the followingreaction scheme. 012362 26
Compounds of formula (II) may be prepared by reacting compounds of formula(XVIII) and (XIX) under the conditions of process step (h) an alkylation reaction,reacting an excess of the amine with the haiide. Suitable conditions are 6eq of (XIX)and 1eq of (XVIII) in acetonitrile at room température for 2 hours followed by 18 5 hours at reflux.
Compounds of formula (XIX) may be prepared by a number of literature routes, wellknown to a man skilled in the art, as well as being commercially available.
Compounds of formula (XX) where R1, R2, R3 and R13 are as described above, withthe proviso R2 and R3 are not linked, may be prepared by the method described in •° the followirig reaction scheme. 012362
Compounds of formula (XX) may be prepared by reacting compounds of formula(Xlla) under the conditions of process step (i) a diazotisation/halogenation reaction,comprising conversion of the amine group to a diazo group, followed by reaction witha suitabie halide, typically in situ. Suitable conditions are treating 1 eq of amine with 3.3 eq of NaNO2 in concentrated hydrochloric acid:water (30:5) at -5°C, then 17hours at room température.
Compounds of formulae (IA) and (IB) where R1, R3, R5, R6, R7 and R8 are asdescribed above, R2, R4 and R10 are hydrogen, R9 is as described above or is asuitable nitrogen protecting group, n is 0 and X is CH may be prepared fromcompounds of formula (XXIII), both the E and Z isomers in accordance with thefollowing scheme. 012362 28
(XXI)
072362 29
Compounds of formula (XXII) may be prepared from compounds of formula (XXIII),under the conditions of process step (b), as described above. Appropriate nitrogenprotecting groups include carbamates, particularly BOC and benzyl groups. Processstep (b) may also be conducted asymmetrically, using techniques known to thoseskilled in the art.
Compounds of formula (XXI) may be prepared from compounds of formula (XXII)under the conditions of process step (d), a lactam hydrolysis reaction which may beconducted under acidic or basic conditions as appropriate.
Compounds of formulae (IA) and (IB) may be prepared from compounds of formula(XXI) under the conditions of process step (j), resolution of the enantiomers, followedby optional process step (k), removal of the nitrogen protecting group when R9 is anitrogen protecting group.
In process step (j), individual enantiomers of a compound of the formulae (IA) or (IB)may be prepared by resolution, such as by H.P.L.C. of the corresponding racemateusing a suitable chiral support or by fractional crystallisation of the diastereoisomericsalts formed by reaction of the corresponding racemate with a suitable opticallyactive acid or base, as appropriate. Reference is made herein to “Enantiomers,Racemates and Resolutions” J. Jacques and A. Collet, published by Wiley, NY,1981; and “Handbook of Chiral Chemicals” chapter 8, Eds D. Ager and M. Dekker,lSBN:0-8247-1058-4.
Compounds of formulae (IA) or (IB) wherein R9 is H may be obtained fromcompounds of formulae (IA) or (IB) where R9 is a suitable nitrogen protecting groupby optional process step (k), removal of a nitrogen protecting group; appropriateconditions for the removal of nitrogen protecting groups R9 are described in"Protective Groups in Organic Synthesis", 2nd édition, T. W. Greene and P. G. M.Wutz, Wiley-lnterscience (1991). Appropriate conditions include: BOC deprotection: 6N aqueous hydrochloric acid at room température to reflux temp,forbetween 1 and 3 hours;
Benzyl deprotection: dissolving métal réduction, e.g. Na, liq NH3, -78°C. 012362 30
Compounds of formulae (IA) and (IB) where R1, R3, R5, R6, R7, R8 and X are as described above and R2, R4, and R10 are hydrogen and R9 is as described above or is an appropriate nitrogen protecting group may also be prepared asymmetrically from compounds of formula (XXIII), where (XXIII) is either the E or Z isomer, in 5 accordance with the reaction scheme shown below. R9
Compounds of formula (XXIV) maÿ be prepared from compounds of formula (XXIII)under the conditions of process step (d), as described above. 2362 31
Compounds of formula (IA) or (IB) may be prepared from compounds of formula(XXIV) under the conditions of process steps (b), a hydrogénation, (j), resolution ofenantiomers and optionally, (k), removal of the nitrogen protecting group P1 when R9is a nitrogen protecting group. Process steps (b), (j) and (k) are described above. 5 |n an alternative embodiment, compounds of formula (IA) where R1, R3, R5, R6, R7,Rb, R10 and X are as described above, R2 and R4 are hydrogen and R9 is asdescribed above or may be an appropriate nitrogen protecting group, may also beprepared asymmetrically from compounds of formula (XXIV). where (XXIV) is eitherthe E or Z isomer, in accordance with the reaction scheme shown below.
Compounds of formulae (IA) or (IB) may be prepared from compounds of formula(XXIV) under the conditions of process steps (I), an asymmetric hydrogénation, (j),resolution of the enantiomers and optionally (k), removal of the nitrogen protectinggroup when R9 is a nitrogen protecting group. Process step (j) is optional and is 012362 32 dépendent upon the degree of enantiomeric selectivity obtained in step (I). Processstep (j) may also be conducted in situ during process step (I). Process steps (j) and(k) are described above and are further exemplified in the examples.
The methods used to conduct process step (I) are weii known to those skilied in the5 art and are discussed in “Asymmetric Synthetic Methodology” chapter 9, Eds D. Agerand M. East, CRC Press, 1996, ISBN: 0-8493-8492-9, as well as being further exemplified in the examples. 10
Compounds of formula (XXIII) where R1, R3, R5, R6, R7, R8 and X are as describedabove and R9 is as described above or a suitable nitrogen protecting group may beprepared from compounds of formula (V) and (XXVI) in accordance with the reactionscheme below.
[3 012362 33
Compounds of formula (XXV) may be prepared from compounds of formula (V) and(XXVI) under the conditions of process step (m), an Aldol type reaction. Suitableconditions for such a reaction are well known to a man skilled in the art. Reference isalso made herein to “Advanced Organic Chemistry” (4th Edition) by Jerry March, 5 John Wiley and Sons Inc.
Compounds of formula (XXIII) may be prepared from compounds of formula (XXV)under the conditions of process step (n), an élimination reaction. (XXV) may betreated such that the hydroxy group is removed directly in a déhydration reaction, orit may be eliminated having first being transformed into a good leaving group such as 10 a tosylate or mesylate group.
Compounds of formula (XXII) where R1, R2, R3 R4 R5, R6, R7, R8 and X are asdisclosed above, R9 is as disclosed above or a nitrogen protecting group and n is 0may also be prepared from compounds of formula (XXX) and (XXVI) in accordancewith the scheme below. (XXX)
_R6 R5 (XXVI) (m) 012362 34
(XXXI) (XXXII) (b) (k)
(XXIIa) (r)
(XXil)
Compounds of formula (XXXI) may be prepared from compounds of formula (XXVI)and formula (XXX), wherein R3 is as described above and P2 is a suitable nitrogenprotecting group, under the conditions of process step (m) as described above. 012362 35
Compounds of formula (XXXII) may be prepared from compounds of formula (XXXI)under the conditions of process step (n) as described above.
Compounds of formula (XXlla) where R2, R3 R4 R5, R6, R7, R8 and X are as disclosedabove, R9 is as disclosed above or a nitrogen protecting group, n is 0 and R1 is 5 hydrogen may be prepared from compounds of formula (XXXII) under the conditionsof process step (b), followed by process step (k), both as described above.
Compounds of formula (XXII) where R1 is not hydrogen may be obtained fromcompounds of formula (XXlla) under the conditions of process step (r), a couplingreaction. Suitable conditions include those described in process steps (h) or (p) t0 regarding alkylation reactions as well as arylation reactions well known to the skilled man. Suitable alkylation conditions may include: 1.5eq of base (eg CS2CO3) and 1.25eq of alkyiating agent, (eg R1 Br), in DMF at 70°Cfor 3 hours.
Suitable arylation conditions may include: 15 2eq of R1-B(OH)2,1.5eq of Cu(ll)acetate catalyst, 2 eq of pyridine in DCM, for 2days, under a stream of compressed air. (P.Y.S. Lam et al, Tetrahedron Lett. 39;2941; 1998)
Compounds of formula (I), where R1, R2, R3, R5, R6, R7, R8, R9 and R10 are asdescribed above, R4 is hydrogen and X is nitrogen, with the proviso one of R9 and 20 R10 is not hydrogen and R1 is attached to an imidazole N atom, may be prepared from compounds of formula (XXIX) in accordance with the reaction scheme below. 012362 36 R9
(P)
V R8 R7 R1
>2
[3 O (XXVII) 012362 37 R1 R8 R7
Compounds of formula (XXVIII) may be prepared from compounds of formula(XXIX), where R4 is hydrogen and one of R9 or R10 is not hydrogen, by process step(o), a carbonylation reaction. The reaction may be performed under standardconditions, such as described in Tetrahedron 1996, 52, 5363. Appropriate conditions 5 include reacting 1eq of (XXIX) with 1eq of carbonyldiimidazole in /V,/V-dimethylformamide at 60°C for 17 hours.
Compounds of formula (XXVii) may be prepared from compounds of formula(XXVill) by process step (p), an alkylation reaction. This may be conducted understandard conditions, e.g. reacting (XXVIII) with an alkylating agent such as an alkyl 10 halide, optionaily in the presence of a catalyst, in a suitable solvent. Suitableconditions include treating 1eq of (XXVIII) with 2eq of R1-Cl in acetonitrile at reflux for18 hours.
Compounds of formula (!) may be prepared from compounds of formula (XXVII)under the conditions of process step (q), a hydrolytic deprotection reaction. The '5 starting material is treated with an aqueous acid, preferably hydrochloric or sulfuricacid.
Compounds of formula (XXIX) may be prepared by the routes disclosed in thisdocument, wherein R1 is instead hydrogen.
All of the above reactions and the préparations of novel starting matériels used in the 20 preceding methods are conventions! and appropriate reagents and reaction 012362 38 conditions for their performance or préparation as well as procedures for isolating thedesired products will be well-known to those skilled in the art with référencé toliterature precedents and the Examples and Préparations hereto.
The présent invention provides for the compounds of formula (I) and 5 pharmaceutically acceptable salts, solvatés and prodrugs thereof for use as amédicament.
The invention further provides for the use of a TAFIa inhibitor in the préparation of amédicament for the treatment or prévention of a condition selected from thrombosis,atherosclerosis, adhesions, dermal scarring, cancer, fibrotic conditions, inflammatory 10 diseases and those conditions which benefit from maintaining or enhancingbradykinin leveis in the body.
Preferably the TAFIa inhibitor is a compound of formula (I) as described herein.Accordingly the présent invention provides for the use of a compound of formula (I)or a pharmaceutically acceptable sait, solvaté or prodrug thereof in the préparation 15 of a médicament for the treatment or prévention of a condition selected from thrombosis, atherosclerosis, adhesions, dermal scarring, cancer, fibrotic conditions,inflammatory diseases and those conditions which benefit from maintaining orenhancing bradykinin leveis in the body.
Additionally the invention provides a method of treating or preventing thrombosis, 20 atherosclerosis, adhesions, dermal scarring, cancer, fibrotic conditions, inflammatorydiseases and those conditions which benefit from maintaining or enhancingbradykinin leveis in the body which comprises administering a therapeuticallyeffective amount of a TAFIa inhiitor and pharmaceutically acceptable salts, solvatésand prodrugs thereof to a patient in need of such treatment. 25 Preferably the TAFIa inhibitor is a compound of formula (I) as described herein.Accordingly the présent invention provides a method of treating or preventingthrombosis, atherosclerosis, adhesions, dermal scarring, cancer, fibrotic conditions, 012362 39 inflammatory diseases and those conditions which benefit from maintaining orenhancing bradykinin levels in the body which comprises administering atherapeutically effective amount of a compound of formula (I) and pharmaceuticallyacceptable salts, solvatés and prodrugs thereof to a patient in need of such 5 treatment.
Thrombotic conditions are amongst the most common cause of death in thedeveloped worid. There are large numbers of anti-thrombotic agents available totreat these conditions. Most agents work by reducing thrombus formation. AU theseagents are associated with varying degrees of adverse hémorrhagie side effects. 10 Accordingly, patients being treated in this manner will require regular monitoring inorder to avoid adverse bleeding events.
There is a need for an antithrombotic that is efficacious but does not cause bleeding.However this would seem impossible given the inhérent contradiction betweenstopping clôt formation to prevent thrombotic disease, and allowing clôt formation so -15 as to prevent the patient hemorrhaging.
Surprisingly this has been solved by the compounds of the présent invention whichare a class of TAFla inhibitors. Most conventional thérapies act to inhibit coagulationor platelet activation. TAFla inhibitors work by enhancing fibrinolysis and thereforethe rate at which the clôt is dissolved. This has the effect of shifting the equilibrium 20 between coagulation and fibrinolysis, in favour of fibrinolysis. Most clinically relevantthrombus are sub acute, that is they form slowly over time. The effect of shifting theequilibrium in favour of fibrinolysis is that these clots are dissolved before theybecome clinically significant.
In the case of vascular injury, the equilibrium moves back in favour of coagulation. 25 The body’s first responses of vasoconstriction and platelet agglutination remainunimpaired by the use of TAFla inhibitors. The body then rapidly activâtes thecoagulation cascade. The effect of this is to temporarily shîffc the equilibrium towardscoagulation and allow formation of a hemostatic plug using fibrin. Once the vascularinjury is sealed the body will revert to its pre-injury equilibrium. 0 T 236 2 40
> A
The présent invention also provides for the use of TAFIa inhibitors in the préparationof a médicament for the treatment or prévention of thrombosis, particulariymyocardial infarction, deep vein thrombosis, stroke, young stroke, peripheraivascular disease, angina and other forms of acute coronary syndromes,disseminated intravascular coagulation, sepsis, pulmonary embolism, embolie eventssecondary to cardiac arrhythmias and the prévention of cardiovascular eventsfollowing intervention surgery. Preferably said TAFIa inhibitor should hâve a Ki ofless than 20μΜ, using the assay described below. Preferably said TAFIa inhibitorshould hâve a selectivity for TAFIa over carboxypeptidase N of >50:1, preferably>1000:1, using the assay described below. Preferably said TAFIa inhibitors are non-peptidic.
Preferably the TAFIa inhibitor is a compound of formula (I) as disclosed herein.Accordingly the présent invention provides for the use of a compound of formula (I)in the préparation of a médicament for the treatment of a thrombotic conditionselected from myocardial infarction, deep vein thrombosis, stroke, young stroke,cérébral infarction, cérébral thrombosis, cérébral embolism, peripherai vasculardisease, angina and other forms of acute coronary syndromes, disseminatedintravascular coagulation, sepsis, pulmonary embolism, embolie events secondary tocardiac arrhythmias and the prévention of cardiovascular events following surgicalrevascularisation or intervention.
The invention also provides for a method of treating or preventing thrombosis,particulariy myocardial infarction, deep vein thrombosis, stroke, young stroke,cérébral infarction, cérébral thrombosis, cérébral embolism, peripherai vasculardisease, angina and other forms of acute coronary syndromes, disseminatedintravascular coagulation, sepsis, pulmonary embolism, embolie events secondary tocardiac arrhythmias and the prévention of cardiovascular events followingintervention surgery which comprises administering a therapeuticaily effectiveamount of a compound of formula (I) and pharmaceutically acceptable salts, solvatésand prodrugs thereof to a patient in need of such treatmënt. 012362 41
Subjects with thrombotic conditions which are suitable for treatment by the présent invention include those having conditions associated with hypercoaguiability. These would include though not limited to: factor V mutation, antithrombin lll deficiency, protein C and protein S deficiencies, polycythemia vera, heparin cofactor 11 and subjects exhibiting hyperhomocysteinaemia or homocysteinuria.
The présent invention also includes as a thrombotic indication the improvement oforgan function seen after transplantation, by reducing blood clotting and thuspreserving function.
Cardiovascular events following intervention surgery include conditions such asrestenosis or reocclusion following interventions such as percutaneous transluminalcoronary angioplasty, grafting, stent in-placement, coronary bypass surgery or anyother forms of surgical revascularisation or intervention
In the présent invention, disseminated intravascuiar coagulation includes ailconditions resulting from intravascuiar activation of the coagulation process. Thismight occur acutely through the release of procoagulant substances (eg. obstetricemergencies, snakebite, crush injury maiignancy), by abnormal contact of the blood(eg.infections, bums, extracorporeal circulation, grafts) or though génération ofprocoagulants in the blood (transfusion reactions, leukemia)·, or chronically, (eg.toxemia, maiignant hypertension, severe liver cirrhosis).
Deep vein thrombosis also encompasses what is known as ‘economy classsyndrome’, where dots form in subjects forced to endure cramped conditions for aperiod of time, such as those sitting in cramped economy class seats on a plane.
The présent invention also provides for the use of TAFIa inhibitors and/or TAF1inhibitors as a coating on intravascuiar devices such as indwelling cathéters fordialysis, replacement heart valves or arterial. stents; and as a coating on extracorporéal blood circulation devices such as heart, lung and kidney dialysis machines, 012362 42 to prevent thrombosis, particularly myocardial infarction, deep vein thrombosis,stroke, young stroke, cérébral infarction, cérébral thrombosis, cérébral embolism,peripheral vascular disease, angina and other forms of acute coronary syndromes,disseminated intravascular coagulation, sepsis, pulmonary embolism, embolie eventssecondary to cardiac arrhythmias and the prévention of cardiovascular events suchas restenosis following intervention surgery such as percutaneous transluminalcoronary angioplasty, grafting, stent in-placement, coronary bypass surgery or anyother forms of surgica, revascularisation or intervention. Particularly preferred as acoating are compounds of formula (I) and pharmaceutically acceptable salts,solvatés and prodrugs thereof.
Accordingly the présent invention provides for the use of TAFIa inhibitors and/orTAFI inhibitors as a coating on intravascular devices.
Further the présent invention provides for the use of a compound of formula (I) as acoating on intravascular devices.
The invention includes intravascular devices, of which the intravascular portion iscoated with a TAFIa inhibitor and/or a TAFI inhibitor; and extra corporéal bloodcirculation devices such as heart, lung and kidney dialysis machines, where theportion coming into contact with the subjects blood are coated with a TAFIa inhibitorand/or a TAFI inhibitor. Particularly preferred are those intravascular or extracorporéal blood circulation devices coated with compounds of formula (I) andpharmaceutically acceptable salts, solvatés and prodrugs thereof. Preferably saidTAFIa inhibitor should hâve a Ki of less than 20μΜ, using the assay described below.Preferably said TAFIa inhibitor should hâve a selectivity for TAFIa overcarboxypeptidase N of >50:1, preferably >1000:1, using the assay described below.Preferably said TAFIa inhibitors are non-peptidic.
Accordingly the présent invention provides an intravascular device coated with aTAFIa inhibitor. 012362 43
Further, the présent invention provides an intravascular device coated with acompound of formula (1).
The compounds of the présent invention were tested in a model of coronary arteryreperfusion using a method similar to that described by W. E. Rote et al, J. 5 Cardiovasc. Pharmacol., 1994, 23, 203, and were found to be efficacious. TAFIa inhibitors are also useful in the treatment of atherosclerosis. Atherosclerosis isa common condition in subjects suffering from peripheral vascular disease, insulinrésistance and the group of conditions commonly referred to as ‘Syndrome X’.Syndrome X is a term often used to group together a number of interrelated 10 diseases. The first stage of syndrome X consists of insulin résistance, abnormalcholestérol and triglycéride levels, obesity and hypertension. Any one of theseconditions may be used to diagnose the start of Syndrome X. The disease maythen progress with one condition leading to the development of another in the group.For example insulin résistance is associated with high lipid levels, hypertension andobesity. The disease then cascades, with the development of each additionalcondition increasing the risk of developing more serious diseases. This can progressto the development of diabètes, kidney disease and heart disease. These diseasesmay lead to stroke, myocardial infarction and organ failure.
Conventional treatment of myocardial ischaemia in clinicaliy stable coronary artery 20 disease is predominately designed to reduce cardiac workload and enhance bloodflow. Such approaches clearly reduce myocardial ischaemia thus increasing qualityof life. However, these strategies hâve little effect on the pathogenesis of coronaryatherosclerosis which is a chronic process of continuous remodeling of the vasculartrée in response to varying degrees of vascular injury. 25 A rôle for thrombus formation in the pathophysiology of stable angina pectoris hasrecently been highlighted by several independent groups. The formation of non-occlusive thrombi not only restrict blood flow, but due to incomplète endogenouslysis may be incorporated-by the arterial wall as solidified plaque matériel enhancingthe atherosclerotic process. Long term administration of a TAFIa inhibitor prevents 44 012362 the formation of thrombi and therefore provides a safe and efficacious treatment which alieviates the symptoms of angina pectoris. Without thrombi présent, they cannot be incorporated into the arterial wall and thus a TAFla inhibitor impairs the progression of the disease.
The présent invention also provides for the use of compounds of formula (I) andpharmaceutically acceptable salts, solvatés and prodrugs thereof in the préparationof a médicament for the treatment or prévention of atherosclerosis.
The invention also provides for a method of treating or preventing atherosclerosiswhich comprises administering a therapeutically effective amount of a compound offormula (I) and pharmaceutically acceptable salts and prodrugs thereof to a patient inneed of treatment.
Further the invention also provides for the use of a TAFla inhibitor in the préparationof a medicàment for the treatment or prévention of atherosclerosis. Preferably saidTAFla inhibitor should hâve a Ki of less than 20μΜ, using the assay described below.Preferably said TAFla inhibitor should hâve a selectivity for TAFla overcarboxypeptidase N of >50:1, preferably >1000:1, using the assay described below.Preferably said TAFla inhibitors are non-peptidic.
Atherosclerosis is taken to include both primary and secondary coronary arterydisease, in which atherosclerosis restricts the blood supply to the heart. Primaryprévention of coronary artery disease means preventing the onset of ischémiecomplications such as myocardial infarction in patients with no history of coronaryartery disease but who hâve one or more risk factors. Secondary prévention ofcoronary artery disease means preventing ischémie complications in patients withestabiished coronary artery disease, such as patients who hâve had a previousmyocardial infarction. TAFla inhibitors are also effective in inhibiting tumour maturation and progression.Metastasis is a complex and multifactorial process which is not yet fully understood.Accordingly, whilst not wishing to be bound by any theorÿ, it is believed that the 012362 45 haemostatic System is involved at several levels of cancer pathology, including neovascularisation, shedding of cells from the primary tumour, invasion of the blood supply, adhérence to the vessel wall and growth at the metastatic site. It is thought that the efficacy of TAFIa inhibitors stems from an ability to reduce fibrin déposition 5 around soiid tumours and thereby inhibit the above processes.
The présent invention also provides for the use of compounds of formula (I) andpharmaceutically acceptable salts, solvatés and prodrugs thereof in the préparationof a médicament for the treatment or prévention of cancer.
The invention also provides for a method of treating or preverrting cancer which W comprises administering a therapeutically effective amount of a compound of formula(I) and pharmaceutically acceptable salts, solvatés and prodrugs thereof to a patientin need of such treatment.
Further the invention also provides for the use of a TAFIa inhibitor in the préparationof a médicament for the treatment or prévention of cancer. Preferably said TAFIainhibitor should hâve a Ki of less than 20μΜ, using the assay described below.Preferably said TAFIa inhibitor should hâve a selectivity for TAFIa overcarboxypeptidase N of >50:1, preferably >1000:1, using the assay described below.Preferably said TAFIa inhibitors are non-peptidic. TAFIa inhibitors are also effective in preventing the formation of adhesions in the 20 body. Most surgica, procedures and physical trauma resuit in bleeding into the cavitybetween tissues. The blood which collects at these sites then clots forming fibrin richthrombi. These thrombi bridge the gaps between adjacent tissues and act as a focifor the accumulation of inflammatory cells and fibroblasts. Invading lïbroblasts laydown a collagen rich extracellular matrix which strengthens the adhesion of the 25 tissues producing a firm bond which may then restrict movement. Adhesions hâvebeen characterised according to their location and may resuit following any surgerye.g. abdominal, orthopaedic, neurological, cardiovascular and ocular. This,inappropriate, adhesion of tissues post surgery or trauma is a major issue which can -2362 46 lead to various outcomes e.g. "aches and pains", "twinges", local inflammation,restriction in mobility, pain, intestinal obstruction and sometimes in the most severecases death. In the case of gynaecological surgery, infertility may resuit. Additionallyclots forming fibrin rich thrombi are implicated in derma, scarring and restenosis.
Without being bound by any theory, it is believed that adhesion formation may beenhanced due to a deficiency in fibrinolysis resulting in enhanced and maintainedclôt formation. Treatment with a TAFIa inhibitor péri- and/or post-surgical interventionmay enhance fibrinolysis of the fibrin rich thrombi and hence inhibit thrombiformation, accretion, stabilisation and therefore inhibit adhesion formation. A TAFIainhibitor given either systemically, or locally as a topical application, may be seen tobe of benefit in a range of surgical procedures. In addition, administration of a TAFIainhibitor may be seen to treat adhesions resulting from other forms of non surgicalphysical trauma where this has caused internai bleeding. Examples of such traumamight include sporting injuries, or anything else resulting in a tear, eut, bruise orinduaration.of the body.
The présent inventio* also provides for the use of compounds of formula (I) andpharmaceuticaily acceptable salts, solvatés and prodrugs thereof in the préparationof a médicament for the treatment or prévention of adhesions or dermal scarring.
The invention also provides for a method of treating or preventing adhesions ordermal scarring which comprises administering a therapeuticaliy effective amount ofa compound of formulae (I) and pharmaceuticaily acceptable salts, solvatés andprodrugs thereof to a patient in need of such treatment.
Further the invention also provides for the use of. a TAFIa inhibitor in the préparationof a médicament for the treatment or prévention of adhesions or dermal scarring.Preferably said TAFIa inhibitor should hâve a Ki of less than 20μΜ, using the assaydescribéd below. Preferably said TAFIa inhibitor should hâve a selectivity for TAFIaover carboxypeptidase N of >50:1, preferably >1000:1, using the assay describedbelow. Preferably said TAFIa inhibitors are non-peptidic.. 012362 47 TAFIa binds to and breaks down bradykinin (Tan et al. Biochemistry 1995, 34, 5811).There are many conditions which are known to benefit from maintaining orenhancing leveis of bradykinin. Accordingly the présent invention also provides forthe use of compounds of formula (I) and pharmaceuticafly acceptable salts, solvatésand prodrugs thereof in the préparation of a médicament for the treatment orprévention of conditions which benefit from maintaining or enhancing leveis ofbradykinin.
The invention also provides for a method of treating or preventing conditions whichbenefit from maintaining or enhancing leveis of bradykinin which comprisesadministering a therapeutically effective amount of a compound of formula (I) andpharmaceutically acceptable salts, solvatés and prodrugs thereof to a patient in needof such treatment.
Conditions known to benefit from maintaining or enhancing bradykinin leveis include:diseases such as hypertension, angina, heart failure, pulmonary hypertension, rénalfailure and organ failure. TAFIa inhibitors are efficacious in treatment of any condition in which fibrosis is acontributing factor. Accordingly the présent invention also provides for the use ofTAFIa inhibitors in the préparation of a médicament for the treatment or préventionof fibrotic disease. Preferably said TAFIa inhibitor should hâve a Ki of less than20μΜ, using the assay described below. Preferably said TAFIa inhibitor should hâvea selectivity for TAFIa over carboxypeptidase N of >50:1, preferably >1000:1, usingthe assay described below. Preferably said TAFIa inhibitors are non-peptidic.Particularly preferred are compounds of formula (I) and pharmaceutically acceptablesalts, solvatés and prodrugs thereof.
Suitable fibrotic conditions include cystic fibrosis, pulmonary fibrotic diseases egchronic obstructive pulmonary disease (COPD), adult respiratory distress syndrome(ARDS), fibromuscular dysplasia, fibrotic lung disease and fibrin deposits in the eyeduring opthalmic surgery. 012362 48
Accordingly the présent invention provides for the use of a compound of formula (I)as disclosed herein in the préparation of a médicament for the treatment orprévention of a fibrotic condition seiected from cystic fibrosis, pulmonary fibroticdiseases, chronic obstructive pulmonary disease (COPD), adult respiratory distresssyndrome (ARDS), fibromuscular dysplasia, fibrotic lung disease and fibrin depositsin the eye during opthalmic surgery.
The invention also provides for a method of treating or preventing a fibrotic conditionseiected from cystic fibrosis, pulmonary fibrotic diseases, chronic obstructivepulmonary disease (COPD), adult respiratory distress syndrome (ARDS),fibromuscular dysplasia, fibrotic lung disease and fibrin deposits in the eye duringopthalmic surgery which comprises administering a therapeutically effective amountof a compound of formula (l) and pharmaceutically acceptable salts and prodrugsthereof to a patient in need of treatment. TAFIa inhibitors are efficacious in treatment of inflammation. Accordingly the présentinvention also provides for the use of TAFIa inhibitors in the préparation of amédicament for the treatment or prévention of inflammation. Preferably said TAFIainhibitor should hâve a Ki of less than 20μΜ, using the assay described beiow.Preferably said TAFIa inhibitor should hâve a selectivity for TAFIa overcarboxypeptidase N of >50:1, preferably >1000:1, using the assay described below.Preferably said TAFIa inhibitors are non-peptidic. Particularly preferred arecompounds of formula (i) and pharmaceutically acceptable salts, solvatés andprodrugs thereof.
In particular the invention may be used for the treatment or prévention ofinflammatory diseases such as asthma, arthritis, endometriosis, inflammatory boweldiseases, psoriasis and atopie dermatitis and for neurodegenerative diseases suchas Alzheimers and Parkinsons.
Accordingly the présent invention provides for the use of a compound of formula (l)and pharmaceutically acceptable salts, solvatés and prodrugs thereof in thepréparation of a médicament for the treatment of an inflammatory disease seiected 012362 49 from asthma, arthritis, endometriosis, inflammatory bowel diseases, psoriasis andatopie dermatitis and neurodegenerative diseases, Alzbeimers and Parkinsons.
The invention also provides for a method of treating or preventing an inflammatorydisease selected from asthma, arthritis, endometriosis, inflammatory bowel diseases,psoriasis and atopie dermatitis and neurodegenerative diseases, Alzheimers andParkinsons which comprises administering a therapeutically effective amount of acompound of formula (I) and pharmaceutically acceptable salts and prodrugs thereofto a patient in need of treatment.
It is to be appreciated -that ail references herein to treatment include curative,palliative and prophylactic treatment.
The compounds of the présent invention hâve been tested using the following assay.To détermine the degree of TAFla inhibition, compounds were incubated withactivated TAFI, and the amount of inhibition expressed in terms of Ki. This assay isbased on that disclosed in Boffa et al, J. Biol. Chem., 1998, 273, 2127.
Assay for TAFla inhibition. i) TAFI activation
Human TAFI (recombinant or purified) was activated by incubating 20μΙ of stocksolution (360pg/m!) with 10μΙ of human thrombin (10NIH units/ml), 10μΙ of rabbitthrombomodulin (30pg/ml), 6μΙ calcium chloride (50mM) in 50μΙ_ of 20mM HEPES(N-[2-hydroxyethyl]piperazine-N-[2-ethanesulfonic acid]) buffer containing 150mMsodium chloride and 0.01% TWEEN 80 (polyoxyethylene-sorbitan monooleate) pH7.6 for 20 minutes at 22°C. At the end of the incubation period, thrombin wasneutralised by the addition of 10μΙ_ of PPACK (D-Phe-Pro-Arg chloromethylketone)(100nM). TAFla solution was stored on ice for 5 minutes and finally dilutedwith 175μΙ of HEPES buffer. ii) Ki Détermination (TAFla)
Calcuiated Ki 012362 50 A number of different dilutions of the test compound in water were made up. Το 20μΙ of each dilution was added 150μΙ of HEPES buffer and 10μΙ of TAFIa, which was then pre-incubated for 15 minutes at 24’C. To each dilution was then added 20μΙ furylacryloyl-alanyl-lysine (FAAL) at a standard concentration. Substrate tum over 5 was measured by reading the absorbance of the reaction mixture at 330nm every 15seconds for 30 minutes. Reaction was performed at 24°C and sampies were mixedfor 3 seconds prior to each absorbance reading. A graph of % inhibition against test compound concentration was then plotted; fromwhich was calculated the IC5o value. The Ki value may then be caiculated using the 10 Cheng-Prusoff équation.
Two Controls, positive and négative, were used to check the accuracy of the resultsin each case. For the first contrai, the assay was performed as above, but with 20μΙof water rather than a dilution of the test compound. This showed minimal inhibition.For the second contrai, the assay was performed as. above, but with an effective 15 amount of a non spécifie carboxypeptidase inhibitor rather than a dilution of the testcompound. This showed maximal inhibition.
Should the two contrais not demonstrate minimal and maximal inhibition respectiveiythen the results were discounted and the test compound reanalysed.
Using the above assay the compounds of the présent invention were found to be 20 potent and sélective inhibitors of TAFIa. Ail compounds had a Ki value less than20μΜ. The spécifie Ki value of certain compounds are detailed below:
(±)-6-Amino-2-[(1 -n-propyl-1 B-imidazo!-4-y!)methyl]hexanoic acid(Example 3) Ki = 310 nM (+)-(2S)-5-Amino-2-[(1 -n-propyl-1 W-imidazoI-4-yl)methyf]pentanoic acid
25 (Example 7) Ki = 13 nM
(2S)-2-[(2-Aminoethyl)amino3-3-(1H-imidazol-4-yl)propanoic acid(Example 11) Ki = 344nM 012362 51
(2S)-2-[(2-Aminoethyl)amino]-3-[1 -(1,3-thiazol-5-ylmethyl)-1 H-imidazol-4-yi]propanoicacid (Example 45) Ki = 197 nM
The selectivity of the compounds of the présent invention for TAFIa overcarboxypeptidase N has aiso been determined. This was done by calculating the Ki 5 of the compounds of the présent invention for carboxypeptidase N, then comparing itto the Ki for TAFIa. The Ki was calculated using the assay for the calculation ofTAFIa Ki, but substituting 10μΙ of human carboxypeptidase N for 10μΙ of TAFIa.
The compounds of the présent invention exhibit a strong selectivity for TAFIa overcarboxypeptidase N of the order of >50:1. 1° The compounds of the présent invention are TAFIa inhibitors, whose utility is basedupon preventing the reaction between a developing thrombus and TAFIa.
It has been found that the compounds of the présent invention are also capable ofbinding to a TAFI molécule, at the site implicated in the reaction between TAFIa andthe developing clôt. The use of TAFIa inhibitors as described above in terms of 15 scope and utility, includes such TAFIa inhibitors which bind to TAFI.
The compounds of the formula (I) can also be administered together with otherantithrombotics, including antiplateiet, anticoagulants and profibrinolytics. Suitableantithrombotics include: aspirin, Plavix™, ticlopidine, warfarin (coumarin™),unfractionated heparin, hirudin (Lepirudin™), streptokinase, urokinase, recombinant 20 tissue plasminogen activator (tPA), dipyridamole, Reopro™, Aggrastat™, andIntegrilin™. The compounds of the formula (l) can also be administered together withantihypertensives and with agents to treat dyslipidaemia such as statins eg Lipitor™.Further suitable drug classes for coadministration include Factor X inhibitors andantiarrhythmics such as amiodarone or digoxin. 25 The présent invention provides for the use of a TAFIa inhibitor in the préparation of amédicament in combination with an antithrombotic for the treatment of thrombosis. 012362 52
The présent invention provides for the use of a compound of formula (l) as described above in the préparation of a médicament in combination with an antithrombotic for the treatment of thrombosis.
In a preferred embodiment the antithrombotic is an profibrinoiytic.
In a more preferred embodiment the antithrombotic is recombinant tissueplasminogen activator (tPA).
The présent invention provides a method of treating or preventing thrombosis, whichcomprises administering a therapeutically effective amount of a TAFla inhibitor incombination with an antifibrinolytic to a patient in need of such treatment.
The présent invention also provides for a method of treating or preventingthrombosis, which comprises administering a therapeutically effective amount of acompound of formula (I) and pharmaceutically acceptable salts, solvatés andprodrugs thereof in combination with an profibrinoiytic to a patient in need of suchtreatment.
In a preferred embodiment the antithrombotic is an profibrinoiytic.
In a more preferred embodiment the antithrombotic is recombinant tissueplasminogen activator (tPA).
The présent invention provides for a kit comprising: a) a composition comprising a compound of formula (I) as disclosed hereinand a pharmaceutically acceptable diluent or carrier, b) a composition comprising an antithrombotic and a pharmaceuticallyacceptable diluent or carrier; and c) a container
The components of this kit may be administered separately, simultaneously orsequentially. 012362 53
The ability of a TAFla inhibitor used in conjunction with an antithrombotic to lyse thrombi was investigated using surgical procedures similar to those outiined in J.
Cardiovasc. Pharmacol. 1994 Feb; 23(2)194-202 and 203-211.
The study was designed with 4 groupe (8 dogs/group): 5 (i) aspirin pre-treatment/vehicle infusion; (ii) no pre-treatment/vehicle infusion; (iii) no pre-treatment/ TAFla inhibitor; and (iv) aspirin pre-treatment/ TAFla inhibitor.
Method 1° Aspirin pre-treatment was 325mg daily for 3 days. TAFla inhibitor (compound of Ex7) was given as a loading dose followed by a continuous infusion with the aim ofachieving a steady State free plasma concentration of 4000nM (220x IC50 for TAFla,in vitro). Thirty minutes after initiating vehicle or compound infusion a continuouselectrical current was delivered to the lumen of the left circumflex (LCX) coronaryartery to cause endothélial damage and stimulate the production of a thrombus.Thrombi were allowed to mature for 1 hour prior to attempting to lyse the thrombusand cause vessel reperfusion with t-PA. A total of 4 bolus injections of t-PA (each0.45 mg/kg i.v.) were given sequentially at 15 minute intervals. Blood flow throughthe coronary artery was then monitored for a further 2 hours so as to assess vessel 20 patency. Time to vessel occlusion, and reperfusion were measured and frie quantityand quality of blood flow analysed post-vesse, re-perfusion. In addition, the effect oftreatment on surgical bleeding, activated clotting time, cutaneous bleeding andplatelet aggregation was aiso assessed.
Results 25 Data is described in Fig 1. From Fig 1 it can be seen that: 012362 54 1) TP A alone is superior to the combination of tPA and aspirin. 2) The combination of a TAFIa inhibitor and tPA is far superior to tPA alone. 3) The improvement in coronary blood flow caused by TAFIa inhibitor wasmaintained for the whole of the reperfusion period (165 minutes) with significantlygreater flow compared to respective Controls. Notably, TAFIa inhibitorsignificantly increased the proportion of animais in which flow was >75% ofbaseline at the end of the protocol. At the end of the experiment only 2/8 dogs inthe no pre-treatment/vehide group and 1/8 dogs in the aspirin pre-treatment/vehicle group were patent. In contrast, the injured vessels were patentin 8/8 dogs in the TAFIa inhibitor treatment group. 4) There was no effect of any of the treatments on surgical bleeding, cutaneousbleeding time, activated clotting time or ADP induced platelet aggregation eitherpre- or post t-PA treatment.
Combination (iv) is not considered here.
The présent invention provides for a composition comprising a compound of formula(i) and a pharmaceuticaliy acceptable excipient, diluent or carrier.
The compounds of formula (I) can be administered alone but will generally beadministered in admixture with a suitable pharmaceutical excipient, diluent or carrierselected with regard to the intended route of administration and standardpharmaceutical practice.
For example, the compounds of formula (I) can be administered orally, buccally orsublingually in the form of tablets, capsules, ovules, élixirs, solutions or suspensions,which may contain flavouring or colouring agents, for immédiate-, delayed-, modified-, sustained-, pulsed- or controiled-release applications.
Such tablets may contain excipients such as microcrystalline cellulose, lactose,sodium citrate, calcium· carbonate, dibasiG calcium phosphate and glycine,disintegrants such as starch (preferably corn, potato or tapioca starch), sodium 012362 55 starch glycollate, croscarmellose sodium and certain complex silicates, and granulation binders such as polyvinylpyrrolidone, hydroxypropylmethylcellulose (HPMC), hydroxypropylcellulose (HPC), sucrose, gelatin and acacia. Additionally, lubricating agents such as magnésium stéarate, stearic acid, glyceryl behenate and talc may be included.
Solid compositions of a similar type may also be employed as fillers in gelatincapsules. Prèferred excipients in this regard include lactose, starch, a cellulose, milksugar or high molecular weight polyethylene glycols. For aqueous suspensionsand/or élixirs, the compounds of the formula (I) may be combined with varioussweetening or flavouring agents, colouring matter or dyes, with emulsifying and/orsuspending agents and with diluents such as water, éthanol, propylene glycol andglycerin, and combinations thereof.
The compounds of formula (I) may also be administered in the form of a iiquid orsuspension filled soft or hard gelatin capsule. Such capsules are generally made ofgelatin, glycerin, water and sorbitol. Hard capsules are distinguished from softcapsules by containing less water and thus having a correspondingly stronger shell.Additional excipients suitable for use in such capsules include propylene glycol,éthanol, water, glycerol and edible oils.
The compounds of formula (I) can also be administered parenterally, for example,intravenously, intra-arterially, intraperitoneally, intrathecally, intraventricularly,intraurethrally, intrasternally, intracranially, intramusculariy or subcutaneousiy, orthey may be administered by infusion techniques. For such parentéral administrationthey are best used in the form of a stérile aqueous solution which may contain othersubstances, for example, enough salts or glucose to make the solution isotonie withbiood. The aqueous solutions should be suitably buffered (preferably to a pH of from3 to 9), if necessary. The préparation of suitable parentéral formulations understérile conditions is readily accomplished by standard pharmaceutical techniqueswell known to those skilled in the art. 012362 56 • *
The compounds of formula (I) can also be administered intranasally or by inhalationand are conveniently delivered in the form of a dry powder inhaler or an aérosol ’spray présentation from a pressurised container, pump, spray, atomiser or nebuliser,with or without the use of a suitable propellant, e.g. dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, a hydrofluoroalkane such as 1,1,1,2-tetrafluoroethane (HFA 134A™) or 1,1,1,2,3,3,3-heptafluoropropane (HFA227EA™), carbon dioxide or other suitable gas. In the case of a pressurisedaérosol, the dosage unit may be determined by providing a valve to deliver ametered amount. The pressurised container, pump, spray, atomiser or nebulisermay contain a solution or suspension of the active compound, e.g. using a mixture oféthanol and the propellant as the solvent, which may additionally contain a lubricant,e.g. sorbitan trioleate. Capsules and cartridges (made, for example, from gelatin) foruse in an inhaler or insufflator may be formulated to contain a powder mix of acompound of the formula (I) and a suitable powder base such as lactose or starch.
Altematively, the compounds of the formula (I) can be administered in the form of asuppository or pessary, or they may be applied topically in the form of a gel,hydrogel, lotion, solution, cream, ointment or dusting powder. The compounds of theformula (I) may also be dermally or transdermally administered, for example, by theuse of a skin patch. They may also be administered by the pulmonary or rectalroutes.
They may also be administered by the ocular route. For ophthalmic use, thecompounds can be formulated as micronised suspensions in isotonie, pH adjusted,stérile saline, or, preferably, as solutions in isotonie, pH adjusted, stérile saline,optionally in combination with a preservative such as a benzylalkonium chloride.Altematively, they may be formulated in an ointment such as petrolatum.
For topical application, the compounds of the formula (I) can be formulated as asuitable ointment containing the active compound suspended or dissolved in, forexample, a mixture with one or more of the following: minerai oil, liquid petrolatum,white petrolatum, propylene glycol, polyoxyethyiene pôlyoxypropylene compound,emulsifÿing wax and water. Altematively, they can be formulated as a suitable lotion 012362 57 or cream, suspended or dissolved in, for example, a mixture of one or more of the following: minerai oil, sorbitan monostearate, a polyethylene glycol, liquid paraffin, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.
The compounds of the formula (I) may also be used in combination with acyclodextrin. Cyclodextrins are known to form inclusion and non-inclusion complexeswith drug molécules. Formation of a drug-cyclodextrin complex may modify thesolubility, dissolution rate, bioavailabiiity and/or stability property of a drug molécule.Drug-cyclodextrin complexes are generally useful for most dosage forms andadministration routes. As an alternative to direct complexation with the drug thecyclodextrin may be used as an auxiliary additive, e.g. as a carrier, diluent orsolubiliser. Alpha-, beta- and gamma-cyclodextrins are most commonly used andsuitable examples are described in WO-A-91/11172, WO-A-94/02518 and WO-A-98/55148.
The invention is further illustrated by the following, non-limiting exemples.
Melting points were determined on a Gallenkamp melting point apparatus usingglass capillary tubes and are uncorrected. Unless otherwise indicated ail reactionswere carried out under a nitrogen atmosphère, using commercially availableanhydrous solvents. Ό.88 Ammonia’ refers to commercially-available aqueousammonia solution of about 0.88 spécifie gravity. Thin-layer chromatography wasperformed on glass-backed pre-coated Merck silica gel (60 F254) plates, and silicagel column chromatography was carried out using 40-63/m silica gel (Merck silicagel 60). Ion exchange chromatography was performed using with the specified ionexchange resin which had been pre-washed with deionised water. Proton NMRspectra were measured on a Varian Inova 300, Varian Inova 400, or Varian Mercury400 spectrometer in the solvents specified. In the NMR spectra, only exchangeableprotons which appeared distinct from the solvent peaks are reported. Low resolutionmass spectra were recorded on either a Fisons Trio 1000, using thermospraypositive ionisation, or a Finnigan Navigator, using electrospray positive or négative
ionisation. High resolution mass spectra were recorded on a Bruker Apex II FT-MS 072362 58 using electrospray positive ionisation. Combustion analyses were conducted by
Exeter Analytical UK. Ltd., Uxbridge, Middlesex. Optical rotations were determined at 25°C using a Perkin Elmer 341 polarimeter using the solvents and concentrations specified. Example compounds designated as (+) or (-) optical isomers are assigned based on the sign of optical rotation when determined in deionised water.
Abbfeviations and Définitions Arboce,™ Filtration agent, from J. Rettenmaier & Sohne, Germany Amberlyst® 15 Ion exchange resin, available from Aldrich ChemicalCompany atm Pressure in atmosphères (1 atm = 760 Torr) Biotage™ Chromatography performed using Flash 75 silica gelcartridge, from Biotage, UK BOC fe/f-Butyloxycarbonyl group br 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 Doublet of doublets
Degussa ® 10 wt% palladium on activated carbon, Degussa type E101101 Company available from Aldrich Chemical CompanyDOWEX® Ion exchange resin, from Aldrich Chemical Company ee Enantiomeric excess HRMS High Resolution Mass Spectrocopy (electrospray ionisationpositive scan) Hyflo™ liq LRMS Hyflo super cel®, from Aldrich Chemical Companyliquid Low Resolution Mass Spectroscopy (electrospray orthermospray ionisation positive scan) LRMS (ES") Low Resolution Mass Spectroscopy (electrospray ionisation 012362 59 négative scan) m Multiplet m/z Mass spectrum peak MCI™ gel High porous polymer, CHP20P 75-1 δΟμιτι, from Mitsubishi 5 Chemical Corporation q Quartet
Rf Rétention factor on TLC s Singlet
Sep-Pak® Reverse phase C18 silica gel cartridge, Waters Corporation 1° t Triplet TLC Thin Layer Chromatography δ Chemical shift
Example 1 (±)-5-Amino-2-( 1 H-irnidazol-4-vlmethvQpentanoic acid
15 A mixture of the ester from Préparation 1 (150mg, 0.25mmol) in dioxane (2ml) andaqueous sodium hydroxide (2ml, 2N) was stirred at room température for 1.5 hours.Aqueous hydrochioric acid (6ml, 6N) was carefuily added, and the réaction heatedunder reflux for 24 hours. The cooled mixture was purified by ion exchange columnchromatography (DOWEX® 50WX8-200), using an elution gradient of deionised 20 water : 0.88 ammonia (100:0 to 97:3). The product was triturated with methanol togive the title compound as a white solid, 28mg, 57% yield. 1H-NMR (CD3OD, 300MHz) δ: 1.44-1.75 (m, 4H), 2.48 (m, 1H), 2.62 (dd, 1H), 2.90(m, 3H), 6.81 (s, 1H), 7.55 (s, 1H). 012362 60 HRMS : m/z 198.1242 (MH+), calcd 198.1237.
Example 2 (±)-5-Amino-2-i( 1 -n-propyl-1 H-imidazoi-4-vl)methvnpentanoic acid
A mixture of the ester from Préparation 2 (85mg, 0.17mmol) in dioxane (1m!) and 5 aqueous sodium hydroxide (1ml, 2N) was stirred at room température for 72 hours.TLC analysis showed starting matériel remaining, so the reaction was heated at70°C for 3 hours. Aqueous hydrochloric acid (2ml, 6N) was added to the cooledsolution and the reaction stirred at room température for 18 hours. TLC analysisshowed starting matériel remaining, so the reaction was stirred at 70°C for a further 2 10 hours. The cooled mixture was extracted with hexane, and the remaining aqueoussolution was purified by ion exchange column chromatography (DOWEX® 50WX8-200) eluting with a solvent gradient of deionised water : 0.88 ammonia (100:0 to97:3). The product was dissolved in a minimum volume of deionised water, andfreeze-dried to give the title compound as a gum, 18mg, 43% yield. 15 1H-NMR (CD3OD, 300MHz) δ: 0.92 (t, 3H), 1.45-1.70 (m, 4H), 1.79 (m, 2H), 2.43- 2.60 (m, 2H), 2.76-2.95 (m, 3H), 3.90 (t, 2H), 6.86 (s, 1H), 7.45 (s, 1H). HRMS : m/z 240.1713 (MH+), calcd 240.1706.
Exampie 3 (±)-6-Amino-2-f( 1 -n-propyl-1 H-imidazoi-4-vl)methvnhexanoic acid
O h3c
%zN
OH ο I23S2 61 A mixture of the protected amine from Préparation 3 (17mg, O.OSmmol) in aqueous hydrochloric acid (2ml, 6N) was stirred at room température for 3 hours. The solution was purified directly by ion exchange chromatography (DOWEX® 5QWX8-200), eluting with a solvent gradient of deionised water : 0.88 ammonia (100:0 to 97:3), to 5 give the title compound, 7mg, 55% yield. 1H-NMR (CD3OD, 300MHz) δ: 0.88 (t, 3H), 1.42 (m, 3H), 1.62 (m, 3H), 1.78 (m, 2H),2.54 (m, 2H), 2.89 (m, 3H), 3.90 (t, 2H), 6.85 (s, 1H), 7.46 (s, 1H). HRMS : m/z 254.1870 (MH+), calcd 254.1863.
Example 4 10 (-)-(2ff)-5-Amino-2-f (1 -n-butvl-1 H-imidazol-4-vftmethvnpentanoic acid
A mixture of the ester from Préparation 6 (185mg, 0.35mmol) in dioxane (6ml) andaqueous sodium hydroxide (6ml, 2N) was stirred at 50°C for 3 hours. Aqueoushydrochloric acid (12ml, 6N) was carefully added, and the reaction stirred at 70°C fora further 18 hours. The cooled mixture was washed with ether, and the aqueoussolution purified by ion exchange chromatography (DOWEX® 50WX8-200) elutingwith a solvent gradient of deionised water : 0.88 ammonia (100:0 to 95:5). Theproduct was azeotroped well with ether and dried in vacuo to give the title compoundas an off-white solid, 45mg, 51% yield. °’23S2 62 1H-NMR (CD3OD, 300MHz) δ: 0.97 (t, 3H), 1.33 (m, 2H), 1.48-1.79 (m, 6H), 2.45- 2.61 (m, 2H), 2.79-2.95 (m, 3H), 3.95 (t, 2H), 6.88 (s, 1H), 7.45 (s, 1H). HRMS : m/z 254.1873 (MH+), calcd 254.1863.
Example 5 5 (+)-(2S)-5-Amino-2-f(1-n-butvl-1H-imidazol-4-vnmethvnpentanoic acid
The title compound was obtained in 35% yield, from the ester from Préparation 7,following a similar procedure to that described in Example 4. 1H-NMR (CD3OD, 300MHz) δ: 0.97 (t, 3H), 1.33 (m, 2H), 1.48-1.79 (m, 6H), 2.45- 2.61 (m, 2H), 2.79-2.95 (m, 3H), 3.95 (t, 2H), 6.88 (s, 1H), 7.45 (s, 1H). 10 HRMS : m/z 254.1874 (M+), calcd 254.1863.
[<xfo = +3.7 (c 0.14, deionised water) (a]o = -5.2 (c 0.15, methanol)
Example 6 (-)-(2R)-5-Amino-2-i( 1 -n-propyl-1 rt-imidazol-4-vl)methvllpentanoic acid
15 A solution of the protected amine from Préparation 9 (1.01g, 2.97mmof) in aqueoushydrochloric acid (15ml, 6N) was stirred at room température for 18 hours. Thesolution was purified directly bÿ ion exchange chromatography (DOWEX® 50WX8- 012362 63 200), eluting with a solvent gradient of deionised water : 0.88 ammonia (100:0 to 97:3), to give the title compound, 680mg, 94% yield. 1H-NMR (CD3OD, 400MHz) δ: 0.84 (t, 3H), 1.48 (m, 1H), 1.55-1.68 (m, 3H), 1.76 (m, 2H), 2.42-2.57 (m, 2H), 2.86 (m, 3H), 3.83 (t, 2H), 6.82 (s, 1H), 7.42 (s, 1H). 5 HRMS : m/z 262.1533 (MNa+), calcd 262.1526.
Anal. Found: C, 58.04; H, 8.93; N, 16.92. C12H2iN3O2-0.5H2O requires C, 58.04; H,8.93; N, 16.92%.
[oJd = -2.53 (c 0.15, deionised water)
Example 7 10 (+)-(2S)-5-Arnino-2-f(1-/?-propyl-1 H-imidazol-4-vt)methvnpentanoic acid
Lithium hydroxide monohydrate (1.1g, 28mmol) and water (28ml) were added to asolution of the lactam from Préparation 11 (3g, 9.33mmol) in tetrahydrofuran (45ml),and the reaction stirred at room température for 18 hours. The solution wasneutralised using aqueous hydrochloric acid (6N), then further acid (15ml, 6N) was 15 added, and the solution stirred at room température for 4 hours. The mixture waspurified directly by ion exchange chromatography (DOWEX® 50WX8-2Û0), elutingwith a solvent gradient of deionised water : 0.88 ammonia (100:0 to 97:3), to give thetitle compound as a solid, 2.1g, 94% yield. This was triturated well with acetone, thesupematant removed, and the residual solid -dried in vacuo, to give the title 20 compound as a white solid. 1H-NMR (D2O, 400MHz) δ: 0.60 (t, 3H), 1.30 (m, 2H), 1.40 (m, 2H), 1.55 (m, 2H),2.26-2.40 (m, 2H), 2.57 (dd, 1H), 2.76 (m, 2H), 3.68 (t, 2H), 6.66 (s, 1H), 7.36 (s,1H). HRMS : m/z 240.1699 (MH+), calcd 240.1706., 0)2362
Anal. Found: C, 58.90; H, 8.90; N, 17.17. C12H2iN3O2*0.3H2O requires C, 58.88; H, 8.92; N, 16.99%.
[α]ο = +2.80 (c 0.14, deionised water) [a]D - -4·θ (o 0.16, methanol) [a]D --5.0(c 0.10, éthanol)
Alternative method for Example 7 A slurry of the quinidine sait from préparation 110 (19g, 28.6mmol) in water (95ml)was adjusted to pH 10 using 5N sodium hydroxide solution, and the mixtureextracted with dichloromethane (1x40ml, 2x20ml). The remaining aqueoussuspension was acidified using 5N hydrochloric acid to pH 0.5, and the solutionstirred at room température for 18 hours. The solution was purified on a Dowex®HCR-S ion-exchange resin column (40g), using an elution gradient of water:0.88ammonia (100:0 to 97:3). The resulting foam was slurried with acetone (20ml), thesolid filtered and dried in vacuo at 40°C to afford the title compound as a white solid,4.6g, 68% yield. 1H-NMR (CD3OD, 400MHz) δ: 0.87 (t, 3H), 1.50 (m, 1H), 1.58-1.72 (m, 3H), 1.78 (m,2H), 2.44-2.59 (m, 2H), 2.90 (m, 3H), 3.88 (t, 2H), 6.84 (s, 1H), 7.46 (s, 1H). LRMS : m/z 240 (MH*) HRMS : m/z 240.1705 (MH*), calcd 240.1706.
Anal. Found: C, 49.10; H, 9.34; N, 14.31. Ci2H21N3O2»3H2O requires C, 49.13; H,9.28; N, 14.32%.
Exampie 8 (-)-(2/?)-5-Amino-2-(1 H-imidazol-4-vlmethvi)pentanoic acid
A mixture of the protected amine from Préparation 12 (85mg, 0.14mmol) in aqueoussodium hydroxide (1ml, 2N) and dioxane (1ml) was stirred at room température for 3 012362 65 days. TLC analysis showed starting matériel remaining, so additional aqueous sodium hydroxide (1ml, 2N) was added, and the reaction stirred at 50°C for 18 hours.
The mixture was cooled and treated with aqueous hydrochloric acid (5ml, 6N). The solution was then stirred at 80°C for 18 hours, cooled to room température, hexane 5 added and the mixture stirred for an hour. The layers were separated, and theaqueous phase purified directly by ion exchange chromatography (DOWEX®50WX8-200), eluting with a solvent gradient of deionised water : 0.88 ammonia(100:0 to 97:3), to give the title compound, 20mg, 73% yield. 1H-NMR (CD3OD, 300MHz) Ô: 1.40-1.68 (m, 4H), 2.45 (m, 1H), 2.62 (dd, 1H), 2.78 10 (m, 2H), 2.90 (m, 1H), 6.78 (s, 1H), 7.50 (s, 1H). HRMS : m/z 198.1243 (MH+), calcd 198.1237.
[α]ο = -6.0 (c 0.1 mg/ml, deionised water)
Example 9 (+)-(2S)-5-Amino-2-( 1 H-imidazol-4-vimethvl)pentanoic acid
15 The title compound was obtained in 96% yield from the protected amine fromPréparation 13, following the procedure described in Example 8. 1H-NMR (CD3OD, 300MHz) δ: 1.45 (m, 1H), 1.59 (m, 3H), 2.47 (m, 1H), 2.62 (dd,1H), 2.78 (m, 2H), 2.90 (dd, 1H), 6.80 (s, 1H), 7.50 (s, 1H). HRMS : m/z 220.1064 (MNa+), calcd 220.1056.
Example 10 (±)-5-Amino-2-f(4-n-propvl-1H-imidazol-2-vl)methvnpentanoic acid 012362
A mixture of the protected amine from Préparation 14 (108mg, 0.23mmol) inaqueous hydrochloric acid (1.5ml, 6N) was stirred under reflux for 1.5 hours. Thecooled solution was purified directly by ion exchange chromatography (DOWEX®50WX8-200), eluting with a solvent gradient of deionised water : 0.88 ammonia 5 (100:0 to 96:4), to give the title compound as a white soiid, 30mg, 55% yield. 1H-NMR (CD3OD, 400MHz) δ: 0.95 (t, 3H), 1.45 (m, 1H), 1.62 (m, 5H), 2.48 (t, 2H), 2.58 (m, 1H), 2.76 (dd, 1H), 2.86 (m, 2H), 2.98 (dd, 1H), 6.60 (s, 1H). HRMS : m/z 240.1718 (MH+), calcd 240.1707.
Anal. Found: C, 54.04; H, 8.97; N, 15.68. C12H2iN3O2»1.5H2O requires C, 54.12; H, 1° 9.08; N, 15.78%.
Example 11 (2S)-2-i(2-Aminoethvl)amino1-3-( 1 rt-imidazol-4-vl)propanoic acid h2n
15
Trifluoroacetic acid (17ml) was added dropwise to a stirred solution of the productfrom Préparation 16 (2.58g, 8.2mmol) in methanol : water (27ml : 14ml). Thereaction was slightly exothermic with évolution of carbon dioxide gas. The mixturewas stirred at room température for 4 hours and the solvent was removed byévaporation under reduced pressure to give a colourless oi, which was dried in vacuoovemight. The résultant oil was treated with aqueous sodium hydroxide solution(1N) until solution was at pH=8. A further portion of aqueous sodium hydroxidesolution (1N, 30ml) was added and the solution was stirred at room température for 20 012362 67 72 hours. The solution was concentrated under reduced pressure to 10ml andpurified by ion exchange chromatography (DOWEX® 50WX8-200) eluting with asolvent gradient of deionised water : 0.88 ammonia solution (100:0 to 97:3). Thesolvent was removed by évaporation under reduced pressure to afford a yellow oil 5 which was dissolved in deionised water (15ml) and freeze-dried overnight to afford afoam. This material was dissolved in deionised water : methanol (95:5) and furtherpurified using MCI™ gel (55g) chromatography, eluting with a solvent gradient ofdeionised water : methanol (95:5) to afford the trüe compound, 1.13g, 69% yield.1H-NMR (D2O, 300 MHz) δ: 2.61-2.87 (m, 4H), 2.92 (m, 2H), 3.25 (t, 1H), 6.81 (s, 0 1H),7.59(s, 1H). LRMS : m/z 199.2 (MH+)
Anal. Found: C, 43.36; H, 7.51; N, 25.12. Ο8Ηι4Ν4Ο2·1 .3H2O requires C, 43.35; H,7.54; N, 25.28%.
[gc]d = +1.74 (c 0.12, deionised water) 15 Example 12 (2ff)-2-F(2-Aminoethyl)amino1-3-( 1 /7-imidazol-4-yl)propanoic acid
The title compound was prepared from the product of Préparation 17 using theprocedure described for Example 11. 1H-NMR (D2O, 300 MHz) δ: 2.57-2.82 (m, 4H), 2.89 (m, 2H), 3.22 (t, 1H), 6.77 (s,1H), 7.55 (s, 1 H).
[«]□ - -1.0 (c 0.10, deionised water) 012362 68
Example 13 (t)-2-f (2-Aminoethyl)amino1-3-( 1 H-imidazol-2-vl)propanoic acid
o
Trifluoroacetic acid (0.5ml) was added dropwise to a stirred solution the product fromPréparation 18 (105mg, 0.34mmol) in methanol : water (2ml : 1ml) and the mixture 5 was stirred at room température for 4 hours. The solvent was then removed byévaporation under reduced pressure and the residue was treated with aqueoussodium hydroxide solution (1N) until solution was at pH=7. A further portion ofaqueous sodium hydroxide solution (1N, 5ml) was added and the solution was stirredat room température for 72 hours. The reaction solution was then submitted to ionexchange chromatography (DOWEX® 50WX8-200) eluting with deionised water :0.88 ammonia (97:3). The solvent was removed by évaporation under reducedpressure to afford a white solid residue. This material was dissolved in deionisedwater : methanol (95:5) and was further purified using MCI™ gel chromatography,eluting with deionised water:methanol (95:5) to afford the title compound, 4mg, 6% 15 yield. 1H-NMR (CD3OD, 300 MHz) δ: 2.74-2.98 (m, 4H), 3.13 (m, 1H), 3.35 (m, 2H), 6.95(s, 2H).
Example 14 (2SV-2-f(2-Aminoethvl)amino1-3-( 1 W-imidazol-2-vl)propanoic acid o
The product from Préparation 19 (200mg, 0.45mmoi) was treated with aqueoushydrochloric acid (6N, 4ml) and heated at reflux for 3 hours. The solvent was then 012362 69 removed by évaporation under reduced pressure and the residue was purified by ion exchange chromatography (DOWEX® 50WX8-200) eluting with an elution gradient of deionised water : 0.88 ammonia (100:0 to 97:3). The isolated material was then freeze-dried to afford the title compound as a foam, 62mg, 69% yield. 1H-NMR (CD3OD, 300 MHz) δ: 2.71-2.98 (m, 4H), 3.13 (m, 1H), 3.34 (m, 2H), 6.92(s, 2H). HRMS: m/z 199.1184 (MH+), calcd 199.1190.
Exampte 15 (2S)-2-ff(1R or S)-1-(Aminomethvl)propvnamino)-3-(1H-imidazol-4-vl)propanoic acid
10 Trifluoroacetic acid was added dropwise to a stirred solution of the product fromPréparation 21 (91 mg, 0.26mmol) in dichloromethane (1ml) and the mixture wasstirred at room température for 17 hours under a nitrogen atmosphère. The solventwas then removed by évaporation under reduced pressure and the residue wasazeotroped with toluene. The résultant material was dissolved in aqueous sodium 15 hydroxide solution (5ml, 2N) and stirred at room température for 72 hours. Solutionwas then purified by ion exchange chromatography (DOWEX® 50WX8-200), elutingwith a solvent gradient of deionised water : 0.88 ammonia (100:0 to 95:5), to affordthe title compound, 37.3mg, 62% yield. 1H-NMR (CD3OD, 400 MHz) δ: 0.81 (t, 3H), 1.37 (m, 1H), 1.50 (m, 1H), 2.62 (m, 1H), 20 2.67 (m, 1H), 2.78(m, 1H), 2.90 (dd, 1H), 2.98 (dd, 1H), 3.33 (dd, 1H), 6.87 (s, 1H), 7.57 (s, 1H). HRMS : m/z 227.1511 (MH+), calcd 227.1503. 012362 70
Example 16 (2S)-2-([(7S or R)-1-(Aminomethyl)propvnamino)-3-(1H->midazol-4-vl)propanoic acid ·
Trifluoroacetic acid was added dropwise to a stirred solution the product fromPréparation 22 (167mg, 0.49mmol) in dichloromethane (1ml) and the mixture was 5 stirred at room température for 17 hours under a nitrogen atmosphère. Solvent wasremoved by évaporation under reduced pressure and residue azeotroped w'rthtoluene. The résultant material was dissolved in aqueous sodium hydroxide solution(5ml, 2N) and stirred at room température for 72 hours. Solution was then purified byion exchange chromatography (DOWEX® 50WX8-200) eluting with a solvent 10 gradient of deionised water : 0.88 ammonia (100:0 to 95:5) to afford the titlecompound, 38.7mg, 35% yield. 1H-NMR (CD3OD, 400 MHz) δ: 0.73 (t, 3H), 1.35 (m, 2H), 2.43 (m, 1H), 2.53 (t, 1H), 2.70 (m, 1H), 2.95 (dd, 1H), 3.10 (dd, 1H), 3.40 (dd, 1H), 6.90 (s, 1H), 7.60 (s, 1H).HRMS : m/z 227.1500 (MH+), calcd 227.1502. 15 Example 17 (2S)-2-ff(1 RS)-1 -(Aminomethvl)-2-methvlpropyllamino)-3-( 1 H-imidazol-4-vl)propanoic
o 012362 71
Trifluoroacetic acid (2ml) was added to a stirred solution of the product fromPréparation 23 (100mg, 0.28mmol) in dichloromethane (1ml) and the mixture wasstirred at room température for 17 hours. The solvent was then removed byévaporation under reduced pressure and the residue azeotroped with toluene. Theresidue was then dissolved in aqueous sodium hydroxide solution (2M, 2ml) andstirred at room température for 72 hours. The solution was then purified by ionexchange chromatography (DOWEX® 50WX8-200) eluting with a solvent gradient ofdeionised water : 0.88 ammonia (100:0 to 97:3). The isolated material (35mg) wasfurther purified by chromatography on reverse phase silica gel (C18 Sep-Pak®),eluting with deionised water, and then freeze-dried to afford the title compound(mixture of diastereoisomers), 20mg, 30% yield. 1H-NMR (CD3OD, 300 MHz), mixture of diastereoisomers, δ: 0.67-0.90 (4x d, 6H),2.40-3.40 (m, 7H), 6.85-6.95 (2x s, 1H), 7.72-7.62 (2x s, 1H). HRMS : m/z 241.1661 (MH+), calcd 241.1659. TLC : methanol : ethyl acetate : 0.88 ammonia : acetic acid : water (60 :12:4:4 :8)Rf=0.52 and 0.44.
Example 18 (2S)-2-ff(1 RS)-2-Amino-1-benzvlethvnamino)-3-( 1 H-imtdazol-4-vl)propanoic acid o
Trifluoroacetic acid (2ml) was added to a stirred solution of the product fromPréparation 24 (100mg, 0.25mmol) in dichloromethane (1ml) and stirred at roomtempérature for 17 hours. The solvent was then removed by évaporation underreduced. pressure and the residue azeotroped with toluene. The residue wasdissolved in aqueous sodium hydroxide solution (2N, 2ml) and stirred at roomtempérature for 17 hours. The solution was then purified by ion exchangechromatography (DOWEX® 50WX8-200) eluting with a solvent gradient of deionised 012362 72 water : 0.88 ammonia (100:0 to 97:3) and isolated material was freeze-dried to affordthe title compound, 41 mg, 58% yield. 1H-NMR (CD3OD, 300 MHz) δ: 2.48-2.72 (m, 2H), 2.77-3.10 (m, 3H), 3.25-3.47 (2xm, 1H), 3.31 (d, 2H), 6.80 (2x s, 1H), 6.91 (d, 1H), 7.10-7.30 (m, 4H), 7.55-7.63 (2x 5 s, 1H). HRMS : m/z 289.1662 (MH+), caicd 289.1659.
Example 19 (2S)-3-( 1 H-lmidazol-4-vl)-2-f(3RS)-pyiTOÎidinvlarrtino)1propanoic acid
Aqueous sodium hydroxide solution (1.7ml, 5N) was added dropwise to a stirred 10 solution of the product from Préparation 20 (200mg, 0.8mmol) in deionised water(20ml) and the solution was stirred at room température ovemight. The solution wasthen purified by ion exchange chromatography (DOWEX® 50WX8-200) eluting witha solvent gradient of deionised water : 0.88 ammonia (100:0 to 95:5) to afford thetitle compound as a pink foam, 90mg, 50% yield. 15 1H-NMR (D2O, 300 MHz), mixture of diastereoisomers, δ: 1.67 (m, 1H), 2.05 (m,1H), 2.70 (m, 2H), 2.90 (m, 1 H), 3.05-3.38 (m, 5H), 6.69 (s, 1H), 7.59 (s, 1H). LRMS : m/z 225.3 (MH+) [α]ο = +1.57 (c 0.076, deionised water)
Example 20 20 (2S)-2-ff(1 R.2S)-2-Amino-1-methvlpropvnamino)-3-(1 B-imidazoI-4-vl)propanoic acid 012362 73
Aqueous sodium hydroxide solution (2ml, 2N) was added to a stirred solution of theproduct from Préparation 26 (260mg, 7.64mmol) in dioxane (2ml) and the mixturewas stirred for 2.5 hours at room température. Aqueous hydrochloric acid (50% byvolume, 4ml) was added and the mixture was stirred at room température for 17 5 hours. The solution was then purified by ion exchange chromatography (DOWEX®50WX8-200) eluting with a solvent gradient of deionised water : 0.88 ammonia(100:0 to 97:3) to afford a white solid which was dissolved in deionised water andfurther purified by chromatography on reverse phase silica gel (C18 Sep-Pak®),eluting with deionised water, to afford the title compound, 15mg, 9% yield. 10 1H-NMR (CD3OD, 300 MHz) δ: 0.93 (d, 3H), 1.17 (d, 3H), 2.62-2.80 (m, 2H), 3.08 (m,1H), 3.20 (m, 1H), 3.37 (m, 1H), 6.92 (s, 1H), 7.61 (s, 1H). HRMS : m/z 227.1506 (MH+), calcd 227.1502.
Example 21 (2S)-2-F(2-Aminoethv»(methvl)aminoT-3-f1H-imidazoi-4-vl)propanoic acid
Trifluoroacetic acid (10ml) was added to a stirred solution of the product fromPréparation 27 (9Q0mg, 2.8 mmol) in methanol : deionised water (10ml : 8ml) andthe mixture was stirred for 2 hours. The solvent was removed by évaporation underreduced pressure to afford a light brown oil which was dissolved in excess aqueous 15 012362 74 «!· sodium hydroxide solution (1N) and stirred for 17 hours. The solution wasconcentrated under reduced pressure and purified by ion exchange chromatography *(DOWEX® 50WX8-200) eluting with a solvent gradient of deionised water : 0.88ammonia (100:0 to 96:4) to afford the title compound as a white foam, 381 mg, 60% 5 yield. 1H-NMR (D2O, 300 MHz) δ: 2.25 (s, 3H), 2.50 (m, 1H), 2.60-3.37 (m, 6H), 6.78 (s, 1 H), 7.58 (s, 1 H).
Example 22 (2S)-3-( 1 H-lmidazol-4-vl)-2-( 1 -piperazinvDpropanoic acid
10 Aqueous sodium hydroxide solution (5N, 170μΙ) was added to a stirred solution of theproduct from Préparation 28 (50mg, 0.012mmol) in water (a few drops) and thesolution was stirred at room température for 18 hours. The solution was thensubmitted to ion exchange chromatography (DOWEX® 50WX8-200), eluting with asolvent gradient of deionised water : 0.88 ammonia (100:0 to 95:5), and the solvent 15 then removed by évaporation under reduced pressure. The residue was suspendedin diethyl ether and then re-evaporated to afford the title compound as a white solid,17mg, 73% yield. 1H-NMR (D2O, 300 MHz) δ: 2.62-2.98 (m, 6H), 3.05-3.30 (m, 5H), 6.80 (s, 1H), 7.60(s, 1H). 20 HRMS : m/z 225.1338 (MH+), calcd 225.1346.
[a]D « +14.84 (c 0.062, deionised water) TLC : methanol : ethyl acetate : 0.88 ammonia : acetic acid : water (60:12:4:4:8)Rf = 0.20. 012362 75
Example 23 (2S)-2-(1.4-Diazepan-1-yl)-3-(1B-imidazol-4-yl)propanoic acid
Homopiperazine (1.86g, 18.6mmol) was added to a stirred solution of the productfrom Préparation 61 (350mg, 1.86mmoi) in acetonitrile (40ml) and the solution was 5 stirred for 2 hours at room température then heated at reflux for 18 hours. Thesolvent was removed under reduced pressure and the residue was dissolved indichloromethane and washed with water (3x20ml). The organic phase wasconcentrated under reduced pressure and the résultant oil was dissolved indeionised water and purified by ion exchange chromatography (DOWEX® 50WX8- 10 200) eluting with a solvent gradient of deionised water : 0.88 ammonia (100:0 to 95:5) to afford the title compound as a beige solid, 300mg, 68% yield. ’H-NMR (D2O, 300 MHz) δ: 1.83 (m, 2H), 2.70-3.23 (m, 10H), 3.40 (t, 1K), 6.80 (s,1H), 7.60 (s, 1H). LRMS : m/z 239.2 (MH+) 15 Anal. Found: C, 50.79; H, 7.85; N, 21.31. CnH^N^I .25H2O requires C, 50.66; H,7.92; N, 21.48%.
[a]p = +2.47 (c 0.24, deionised water)
Example 24 (2S)-2-f(2-Aminoethvl)arnino1-3-( 1 -ethvl-1 B-imidazol-4-vl)propanoic acid 012362 76
Concentrated hydrochloric acid (5ml) was added to a stirred solution of the productfrom Préparation 30 (118mg, 0.32mmol) in water (5ml) and the mixture was heatedat reflux for 17 hours. The mixture was allowed to cool to room température and thesolvent was removed by évaporation under reduced pressure. The residue was 5 purified by ion exchange chromatography (DOWEX® 50WX8-200) eluting withdeionised water : 0.88 ammonia (97:3). The isolated material was freeze-dried toafford the title compound, 34mg, 47% yield. 1H-NMR (CD3OD, 300 MHz) δ: 1.40 (t, 3H), 2.75-3.02 (m, 6H), 3.33 (m, 1H), 3.98 (q,2H), 6.95 (s, 1 H), 7.53 (s, 1 H). HRMS : m/z 227.1492 (MH+), calcd 227.1503. 10 012362 77
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Footnotes: 012362 82 1. Concentrated sulfuric acid (4M) used instead of concentrated hydrochloricacid (6M). 2. Sulfuric acid (2M) used instead of concentrated hydrochloric acid (6M). 3. The isolated product was further purified using a δμίτι Hypersil Hypercarb™ 5 column, using an elution gradient of water : trifluoroacetic acid : acetonitrile (100 : 0.1 : 0 to 50: 0.05 : 50), and then re-subjected to ion-exchangechromatography (as in Example 24). 4. The isolated product was further purified as described in note (3) but using anelution gradient of water : trifluoroacetic acid : methanol (100:0.1:0 to 1° 50:0.05:50).
Example 41 (2S)-2-[(2-Aminoethvl)amino1-3-f1-(carboxvmethvl)-lÎÎ-/-imidazol-4-v!1propanoic acid
The product from Préparation 47 (145mg, 0.296mmol) was dissolved in 15 concentrated sulfuric acid (4ml) and the solution heated under reflux for 18hours. The cooled mixture was purified directly by ion exchange chromatography(DOWEX® 50WX8-200), eluting with 0.88 ammonia : water (3:97). The resultingoil was triturated with methanol, to give a solid which was freeze-dried to affordthe title compound as a white foam, 61 mg, 77% yield. 2θ 1H-NMR (D2O, 400MHz) δ: 2.80 (m, 2H), 2.88 (m, 2H), 2.98 (m, 2H), 3.40 (m,1 H), 4.52 (s, 2H), 6.92 (s, 1 H), 7.81 (s, 1 H). HRMS : m/z 257.1255 (MH+), calcd 257.1245.
Anal. Found: C, 42.66; H, 6.63; N, 20.29. 0·,οΗι6Ν4θ4·1.3Η20 requires C, 42.95;H, 6.70; N, 20.03%. 012362
Example 42 (2SV3-K1 -n-propyl-1 H-imidazoi-4-vl)methyïl-2-piperidinone
The compound from Préparation 11 (500mg, 1.6mmol) in dichloromethane(15ml) was treated with trifluoroacetic acid (3ml) and the résultant solution was 5 stirred at room température for 2 hours. The réaction mixture was thenconcentrated under reduced pressure and the residue neutralised with saturatedaqueous sodium bicarbonate solution. The résultant mixture was thenconcentrated to dryness under reduced pressure and the residue purified bycolumn chromatography on silica gel using an elution gradient of 1° dichloromethane-.methanol : 0.88 ammonia (99.8: 0 : 0.2 to 94.8 : 5 : 0.2) to givethe title compound as an oil, 250mg, 73% yield. 1H-NMR (CDCIs, 400MHz) δ: 0.87 (t, 3H), 1.39-1.84 (m, 5H), 1.90 (m, 1H), 2.60(m, 1H), 2.74 (dd, 1H), 3.13 (dd, 1H), 3.21 (m, 2H), 3.77 (t, 2H), 5.61 (br s, 1H), 6.65 (s, 1H), 7.31 (s, 1H). 15 LRMS : m/z 222 (MH+)
Anal. Found: C, 61.44; H, 8.85; N, 17.86. Ci2HigN3O*0.75H2O requires C, 61.38;H, 8.80; N, 17.89%.
[a}o = -51.6 (c 0.095, methanol) 012362 84
Example 43 (2S)-2-if2-Aminoethvnamino]-3-(1-methyÎ-1H-imidazoi-4-vl)propanoic acid
2M Sodium hydroxide solution (0.61ml, 1.22mmol) was added to a solution of theprotected amino acid from préparation 90 (200mg, 0.61 mmol) in dioxan (2ml),and the reaction stirred at room température for 18 hours. Concentratedhydrochloric acid (2ml) was carefully added, and the solution stirred for a further24 hours, then concentrated under reduced pressure. The residue was dissolvedin water, and purified by column chromatography on Amberlyst® 15 ion-exchange resin, eluting with 5% aqueous ammonia solution. The product wasobtained after freeze-drying as a gum, 80mg, 55% yield. 1H-NMR (D2O, 400MHz) δ: 2.61-2.79 (m, 4H), 2.90 (m, 2H), 3.22 (m, 1H), 3.54(s, 3H), 6.79 (s, 1H), 7.42 (s, 1H). LRMS (ES·) : m/z 211 (M-H)' [a]o s -5.83 (c 0.12, methanol)
Anal Found: C, 45.63; H, 7.68; N, 23.15. 09Η16Ν402·1.45Η2θ requires C, 45.35;H, 7.99; N, 23.50%.
Examples 44 to 47
The following examples of general structure:
were prepared from the appropriate protected amino acids (Préparations 91-94),following a similar procedure to that described in Example 43. 0^2362
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Example 48 (2S)-2-f(2-Aminoethvl)aminol·3-(1-benzvl-1H-imidazol-4-vl)propano^c acid
A solution of the compound from préparation 95 (288mg, 0.57mmol) in 4M sulphuricacid (10ml), was heated at 115°C for 36 hours. The cooled solution was neutralised 5 using 1M sodium hydroxide solution, then passed through an Amberlyst® 15 ionexchange column, eluting with 5% aqueous ammonia. The product was obtained asa gum afterfreeze-drying, 70mg, 39% yield. 1H-NMR (D2O, 400MHz) δ: 2.40 (m, 1H), 2.48 (m, 1H), 2.58 (m, 4H), 3.14 (t, 1H),5.00 (s, 2H), 6.77 (s, 1H), 7.14 (d, 2H), 7.22 (m, 3H), 7.50 (s, 1H). 10 LRMS : m/z 289 (MH+) [α]ο = +1.00 (c 0.14, methanol)
Anal. Found: C, 56.96; H, 7.17; N, 17.63. C15H2oN402.1,5H2O requires C, 57.13; H,7.35; N, 17.77%.
Example 49 15 (±)-5-Amino-2-î( 1 -isopentvl-1 H-imidazol-4-vl)methvnpentanoic acid
A solution of sodium hydroxide (192mg, 4.80mmol) in water (6ml) was added to asolution of the compound from préparation 105 (420mg, 1.20mmol) intetrahydrofuran (10ml), and the reaction stirred vîgorously for 72 hours. Concentratedhydrochloric acid (6ml) was carefully added, and the mixture stirred at room 012362 88 * température for 3 hours, then concentrated under reduced pressure. The residue » was dissolved in water (50ml), and the solution purified by column chromatographyon Amberlyst® 15 ion-exchange resin, using an elution gradient of water:0.88ammonia (100:0 to 98:2) to afford the titie compound, 120mg, 35% yield. ï 1H-NMR (D2O, 400MHz) δ : 0.72 (d, 6H), 1.23-1.40 (m, 3H), 1.46 (m, 4H), 2.30-2.43(m, 2H), 2.59 (dd, 1H), 2.79 (m, 2H), 3.80 (t, 2H), 6.76 (s, 1H), 7.42 (s, 1H). LRMS (ES') : m/z 266 (M-H)'
Anal. Found: C, 58.60; H, 9.62; N, 14.56. 014Η2δΝ302·1.0Η20 requires C, 58.92; H,9.54; N, 14.72%. 1û Example 50 (±)-2-K 1 -lsopentvl-1 B-imidazol-4-vl)methvn-5-(methvlamino)pentanoic acid
A solution of the compound from préparation 106 (170mg, 0.65mmol) in dioxan (1ml)and concentrated hydrochloric acid (2ml) was heated at reflux for 18 hours. Thecooled mixture was concentrated under reduced pressure at room température, and *5 the residue dissolved in water (50ml). The solution was purified by columnchromatography on Amberlyst® 15 ion-exchange resin, using an elution gradient ofwater:0.88 ammonia (100:0 to 98:2). Freeze drying afforded the titie compound as abrown solid, 120mg, 66% yield. 1H-NMR (D2O, 400MHz) δ: 0.75 (d, 6H), 1.25-1.42 (m, 3H), 1.50 (m, 4H), 2.34-2.44 20 (m, 2H), 2.55 (s, 3H), 2.62 (dd, 1H), 2.86 (m, 2H), 3.82 (t, 2H), 6.78 (s, 1H), 7.43 (s, 1H). LRMS: m/z 282.2 (MH+)
Anal. Found: C, 58.56; H, 9.73; N, 13.61. C15H27N3O2»1.45H2O requires C, 58.59; H,9.80; N, 13.66%. 012362 89
Example 51 (±)-5-Amino-2-lï 1 -phenyl-1 H-imidazol-4-vl)rnethvi1pentanoic acid
A solution of lithium hydroxide (2ml, 1M, 2mmol) was added to a solution of thecompound from préparation 108 (240mg, 0.68mmol) in tetrahydrofuran (2ml), and 5 the reaction stirred at room température for 5 hours. Concentrated hydrochloric acid(2ml) was added carefully, and the reaction stirred at room température for 18 hours.The solution was evaporated under reduced pressure, the residue dissolved inwater, and the solution purified by column chromatography on Amberlyst® 15 ion-exchange resin using an elution gradient of water0.88 ammonia (100.Ό to 95:5) to 1° afford the title compound as a white foam, 88mg, 45% yield. 1H-NMR (D2O, 400MHz) δ : 1.43 (m, 2H), 1.54 (m, 2H), 2.42-2.59 (m, 2H), 2.74 (dd,1H), 2.83 (m, 2H), 7.18 (s, 1H), 7.32 (m, 1H), 7.40 (m, 4H), 7.88 (s, 1H). LRMS : m/z 296 (MNa+)
Anal. Found: C, 62.21; H, 7.01; N, 14.55. Ci5H19N3O2.1 .0H2O requires C, 61.84; H, ’5 7.27; N, 14.42%. 0)2362 90
Préparation 1 (±)-Ethvl 24 ( 1 -U2-(trimethvlsilvnethoxv1methyl>-1 H-imidazoM-vOmeth yil-5-
A mixture of the alkenes from Préparation 49 (460mg, 0.77mmol) and 10% palladium5 on charcoal (100mg) in éthanol (25ml) was hydrogenated at 1.5 atm and roomtempérature for 72 hours. The reaction mixture was filtered through Arbocel™,washing through with ethano, (200ml), and the filtrate concentrated under reducedpressure. The residual oi, was purified by column chromatography on silica gel usingethyl acetate : pentane (50:50) as eluant, to give the title compound, 150mg, 33% yieid. 1H-NMR (CDCI3i 400MHz) δ: -0.02 (s, 9H), 0.95 (t, 2H), 1.18 (t, 3H), 1.46 (m, 2H), 1.45-1.70 (m, 2H), 2.09 (m, 2H), 2.64-2.79 (m, 2H), 2.90 (dd, 1H), 3.42 (t, 2H), 4.09(q, 2H), 5.18 (s, 2H), 6.75 (s, 1H), 7.17 (m, 3H), 7.22 (m, 7H), 7.42 (d, 6H). 10 012362 91
Préparation 2 (±)-Ethvl 2-K1 -n-propyl-1 H-imidazol-4-vl)rnethvil-5-(tritylamino)pentanoate
Sodium borohydride (7.2g, 190mmol) was added portionwise over 2 hours to asolution of alkenes from Préparation 50 (3.2g, 6.3mmol) and copper (I) chloride 5 (928mg, 9.5mmol) in methanol (120ml), so as to maintain the reaction température at about 45°C, and the reaction stirred at this température for 2 hours, (two additionalportions of copper (I) chloride (310mg, 3.1mmol) were added after approx 40 and 80minutes). The réaction mixture was filtered through Arbocel™ and the filtrateconcentrated under reduced pressure. The residue was partitioned between ethyl 10 acetate and water, the layers separated, and the aqueous phase extracted with ethylacetate (2x). The combined organic extracts were dried (Na2SO4) and concentratedunder reduced pressure. The crude product was purified by column chromatographyon silica gel using an elution gradient of ethyl acetate : pentane (50:50 to 100:0) togive the titte compound, 2g, 62% yield. 15 1H-NMR (CDCIs, 300MHz) δ: 0.88 (t, 3H), 1.19 (t, 3H), 1.55 (m, 4H), 1.76 (m, 2H),2.08 (m, 2H), 2.62-2.80 (m, 2H), 2.86 (dd, 1H), 3.79 (t, 2H), 4.07 (q, 2H), 6.60 (s,1H), 7.18 (m, 3H), 7.24 (m, 7H), 7.43 (d, 6H). LRMS : m/z 510 (MH*) 012362 92
Préparation 3 (±)-6-i(ferf-Butoxvcarbonv0amino1-2-r(1“n-propvl-1B-imidazol-4-vl)methynhexanoic acid
OH A solution of the compound from Préparation 4 (32mg, 0.07mmol) in tetrahydrofuran5 (2ml) and éthanol (50μΙ) was added to a cooled (-78°C) solution of sodium (20mg, 0.87mmol) in 0.88 ammonia (3ml), and the solution stirred for 15 minutes, until theblue colour disappeared. The reaction was allowed to warm to room température, theammonia evaporated off and then the remaining solution was concentrated underreduced pressure. The crude product was purified by ion exchange chromatographyon DOWEX® (50WX8-200) resin, eluting with a solvent gradient of water : 0.88ammonia (100:0 to 97:3), to give the titie compound, 17mg, 69% yield. 1H-NMR (CD3OD, 300MHz) δ: 0.90 (t, 3H), 1.42 (m, 13H), 1.61 (m, 2H), 1.80 (m,2H), 2.57-2.68 (m, 2H), 2.80-2.95 (m, 2H), 3.00 (m, 1H), 3.95 (t, 2H), 6.98 (s, 1H), 7.76 (s, 1H). 15 LRMS : m/z 354.3 (MH+) 012362 93
Préparation 4
Sodium 6-îbenzyl(tezf-butoxvcarbonvl)amino1-2-T(1 -n-propyi-1 H-imidazol-4- vümethyllhexanoate
Aqueous sodium hydroxide solution (2ml, 2N) was added to a solution of the ester 5 from Préparation 5 (50mg, 0.106mmol) in dioxane (2ml), and the reaction stirred atroom température for 18 hours. The mixture was concentrated under reducedpressure and the residue purified by column chromatography on siiica gel elutingwith dichloromethane : methanol : 0.88 ammonia (90:10:1), to give the titlecompound, 32mg, 65% yield. 10 1H-NMR (CD3OD, 300MHz) δ: 0.88 (t, 3H), 1.15-1.57 (m, 15H), 1.80 (m, 2H), 2.60(m, 2H), 2.82 (m, 1H), 3.17 (m, 2H), 3.94 (t, 2H), 4.42 (s, 2H), 6.96 (s, 1H), 7.22 (m,3H), 7.32 (m, 2H), 7.78 (br s, 1H). LRMS : m/z 444.7 (MH+) 0^2362 94
Préparation 5 (±)-Ethvl e-rbenzvl(tezf-butoxvcarbonvl)aminoÎ-2-f(1-z7-propvl-1 /7-imidazol-4- vhmethvnhexanoate
A mixture of the alkenes from Préparation 51 (620mg, 1.32mmol) and 10% palladium5 on charcoal (70mg) in methanol (50ml) was hydrogenated at 1 atm and roomtempérature for 4 hours. The reaction mixture was filtered through Arbocel™, andthe filtrate concentrated under reduced pressure to give the title compound in quantitative yield as a clear gum, which was used without further purification. 1H-NMR (CDCIs, 300MHz) δ: 0.90 (t, 3H), 1.18 (t, 3H), 1.24 (m, 2H), 1.38-1.66 (m,13H), 1.78 (m, 2H), 2.61-2.80 (m, 2H), 2.86 (dd, 1H), 3.04-3.22 (m, 2H), 3.80 (t, 2H),4.06 (q, 2H), 4.40 (br s, 2H), 6.61 (s, 1H), 7.18-7.37 (m, 6H). LRMS : m/z 472.4 (MH+) 10 012362 95
Préparation 6
Ethvl (2R)-2-K1 -n-butyl-1 H-imidazol-4-vl)methvn-5-(tritvlamino)pentanoate
and Préparation 7
Ethvl (2S)-2-f(1 -n-butyl-1 H-imidazol-4-vl)methyn-5-(tritvlamino)pentanoate
5 The racemic compound from Préparation 8 was resolved by KPLC using aChiralcel® OD 250 column (20mm), and hexane : éthanol : diethylamine (85 :15:0.45) as eluant at a rate of 10ml/minute, to afford the title compound of Préparation6, 98.3%ee, Rétention time: 13.36 minutes, 10 1H-NMR (CDCIs, 300MHz) δ: 0.92 (t, 3H), 1.20 (t, 3H), 1.28 (m, 2H), 1.45-1.78 (m,6H), 2.10 (m, 2H), 2.62-2.79 (m, 2H), 2.88 (dd, 1H), 3.81 (t, 2H), 4.08 (q, 2H), 6.60(s, 1H), 7.18 (m, 3H), 7.24 (m, 7H), 7.43 (d, 6H). and the title compound of Préparation 7, 94.2%ee, Rétention time: 14.91 minutes. 15 1H-NMR (CDCfe, 300MHz) δ: 0.92 (t, 3H), 1.20 (t, 3H), 1.28 (m, 2H), 1.45-1.78 (m,6H), 2.10 (m, 2H), 2.62-279 (m, 2H), 2.88 (dd, 1 H), 3.81 (t, 2H), 4.08 (q, 2H), 6.60(s, 1H), 7.18 (m, 3H), 7.24 (m, 7H), 7.43 (d, 6H). 012362 96
Préparation 8 (±)-Ethvl 2-f(1-n-butvl-1H-imidazol-4-vnmethvn-5-(tritv!amino)pentanoate
Sodium borohydride (871 mg, 23mmol) was added portionwise over an hour to asolution of the alkene from Préparation 52 (400mg, 0.76mmol) and copper (I) 5 chloride (112mg, 1.15mmol) in methanol (15ml). TLC analysis showed startingmaterial remaining, so additional copper (I) chloride (75mg, 0.76mmol) and sodiumborohydride (290mg, 7.7mmol) were added, and the reaction stirred at roomtempérature for a further 2 hours. The reaction mixture was filtered throughArbocel™, the filtrate concentrated under reduced pressure and the residue 10 partitioned between ethyl acetate and brine. The layers were separated, the aqueousphase extracted with ethyl acetate (2x), and the combined organic extracts dried(Na2SO4) and concentrated under reduced pressure, to give the title compound,185mg, 47% yield. 1H-NMR (CDCIs, 400MHz) δ: 0.92 (t, 3H), 1.19 (t, 3H), 1.27 (m, 2H), 1.48-1.77 (m, *5 6H), 2.10 (m, 2H), 2.62-2.79 (m, 2H), 2.88 (dd, 1H), 3.82 (t, 2H), 4.08 (q, 2H), 6.60 (s, 1H), 7.17 (m, 3H), 7.24 (m, 7H), 7.43 (d, 6H). 012362 97
Préparation 9
Lithium (2R)-5-r(tezÎ-butoxvcarbonyl)amino1-2-f( 1 -n-propyl-1 B-imidazol-4- vhmethyllpentanoate
Water (2ml) and lithium hydroxide monohydrate (81 mg, 1.93mmol) were added to a 5 solution of the lactam from Préparation 10 (207mg, 0.64mmol) in tetrahydrofuran(3.5ml), and the solution stirred at room température for 23 hours. The mixture wasconcentrated under reduced pressure and the residue purified by columnchromatography on silica gel using dichloromethane : methanol : 0.88 ammonia(90:10:0 to 90:10:1) to give the title compound, 200mg, 92% yield. 10 1H-NMR (CD3OD, 300MHz) δ: 0.90 (t, 3H), 1.42 (s, 9H), 1.45-1.62 (m, 4H), 1.80 (m,2H), 2.57-2.70 (m, 2H), 2.85 (m, 1H), 3.02 (m, 2H), 3.95 (t, 2H), 6.97 (s, 1H), 7.76 (s,1H). LRMS (ES'): m/z 338 (M-H)‘
Préparation 10 15 (-)-fezf-Butyl (3R)-2-oxo-3-[n-n-propyl-1H-imidazoi-4-vBmethvll-1-
98 012362 (+)-ferf-Butvl (3S)-2-oxo-3-Ff 1 -n-propyl-1 H-imidazol-4-vl)methyri-1 - piperidinecarboxvlate
(700mg) in éthanol (120ml) was hydrogenated at 4 atm and 60°C for 18 hours. Thecooled mixture was filtered through Arbocei™, washing through with ethyl acetate,and the filtrate concentrated under reduced pressure. The crude product was purifiedby column chromatography on silica gel, eluting with dichloromethane : methanol(97:3), to afford the racemate of the title compounds as a yellow oil, 4.3g, 65% yield.This racemic compound was resolved by HPLC using a Chiralcel® OG 250 column(20 mm), and hexane : isopropanol (70:30) as eluant at a rate of 10 ml/minute, togive the title compound of Préparation 10,1.56g, 99.5% ee, Rétention time: 10.10 minutes, 1H-NMR (CDCIs, 300MHz) δ: 0.92 (t, 3H), 1.54 (s, 9H), 1.63 (m, 2H), 1.80 (m, 3H),2.00 (m, 1H), 2.65-2.88 (m, 2H), 3.18 (m, 1H), 3.58 (m, 1H), 3.70-3.90 (m, 3H), 6.72(s, 1H), 7.38 (s, 1H). LRMS : m/z 322.5 (MH+) [a]D = -34.34 (c 0.12, dichloromethane) and the title compound of Préparation 11,1.56g, 98.9% ee, Rétention time: 15.23 minutes, 1H-NMR (CDCIs, 300MHz) δ: 0.92 (t, 3H), 1.54 (s, 9H), 1.80 (m, 4H), 2.00 (m, 2H),2.63-2.85 (m, 2H), 3.19 (m, 1H), 3.58 (m, 1H), 3.90-3.98 (m, 3H), 6.72 (s, 1H), 7.37(s, 1H). LRMS : m/z 322.3 (MH+)
Md « +27.7 (c 0.22, dichloromethane) 012352 99
Préparation 12
Ethvl (2R)-2J( 1 -if2-(trimethvlsilvl)ethoxv1methvlM H-irnidazol-4-vl)methvri-5- (tritvlamino)pentanoate
and Préparation 13 5 Ethvl (2S)-2-i( 1 -ff2-(trimethvlsilvl)ethoxv1methvl}-1 H-imidazol-4-vl)methvn-5-
The compound from Préparation 1 was resolved by KPLC using a Chiraicel® OD250 column (20mm), and hexane : isopropanol : diethylamine (90 : 10 : 0.5) aseluant at 10mi/minute, to give, the title compound of Préparation 12, in 25% yield,99.4% ee, Rétention time : 16.90 minutes. 1H-NMR (CDCI3i 400MHz) δ: -0.02 (s, 9H), 0.95 (t, 2H), 1.20 (t, 3H), 1.44-1.66 (m,4H), 2.09 (m, 2H), 2.64-2.80 (m, 2H), 2.90 (dd, 1H), 3.42 (t, 2H), 4.09 (q, 2H), 5.18(s, 2H), 6.75 (s, 1H), 7.17 (m, 3H), 7.22 (m, 7H), 7.42 (d, 6H). ’5 LRMS : m/z 598.7 (MH+) and the title compound of Préparation 13, in 36% yield, 96.5% ee, Rétention time: 22.27 minutes. 012362 100 1H-NMR (CDCI3, 400MHz) δ: -0.02 (s, 9H), 0.95 (t, 2H), 1.20 (t; 3H), 1.44-1.66 (m, 4H), 2.09 (m, 2H), 2.64-2.80 (m, 2H), 2.90 (dd, 1H), 3.42 (t, 2H), 4.09 (q, 2H), 5.18 (s, 2H), 6.75 (s, 1H), 7.17 (m, 3H), 7.22 (m, 7H), 7.42 (d, 6H).
Préparation 14 5 Lithium 5-f(fert-butoxvcarbonvl)aminol-2-f(4-propyl-1 -Π2-
Lithium hydroxide monohydrate (42mg, 0.99mmol) was added to a solution of thelactam from Préparation 15 (150mg, 0.33mmol) in tetrahydrofuran (1ml) and water(1.5ml), and the reaction stirred for 4 hours at room température. The mixture was 10 concentrated under reduced pressure and the residue purified by columnchromatography on silica gel eluting with dichloromethane : methanol (90:10) aseluant to give the title compound, 108mg, 70% yield. 1H-NMR (CD3OD, 300MHz) δ: 0.00 (s, 9H), 0.96 (m, 5H), 1.42 (s, 9H), 1.54 (m, 3H), 1.63 (m, 3H), 2.58 (t, 2H), 2.80 (m, 1H), 2.88-2.98 (m, 1H), 3.02 (m, 2H), 3.16 (dd, 15 1H), 3.60 (t, 2H), 5.34 (d, 1H), 5.50 (d, 1H), 7.07 (s, 1H). LRMS : m/z 470.3 (MH+)
Préparation 15 fe/f-Butvl_2-oxo-3-r(4-n-propyl-1-(f2-(trimethvlsilvl)ethoxvÎmethvrHH-imidazol-2- vl)methvn-1 -piperidinecarboxvlate
012362 101
The title compound was obtained in 75% yield from the alkene of Préparation 54, foliowing a similar procedure to that described in Préparation 10/11. 1H-NMR (CDCIs, 300MHz) δ: -0.02 (s, 9H), 0.82-0.98 (m, 5H), 1.50 (s, 9H), 1.60 (m,3H), 1.81 (m, 2H), 2.05 (m, 1H), 2.46 (t, 2H), 2.74 (dd, 1H), 3.03 (m, 1H), 3.35 (dd, 5 1 H), 3.46 (t, 2H), 3.58 (m, 1H), 3.82 (m, 1H), 5.15 (d, 1H), 5.30 (d, 1H), 6.59 (s, 1H). LRMS : m/z 452.4 (MH+)
Préparation 16
Methvl (2S)-2“({2-f(fe/Î-butoxvcarbonvl)aminotethyl)amino)-3-( 1 H-imidazol-4- vDpropanoate
10 L-Histidine methyl ester (7.93g, 32.8mmol) and sodium acetate (10.75g, 131mmol)were added to a stirred solution of fert-butyi W-(2-oxoethyl)carbamate (5.22g,32.8mmol) in methanol (100ml). 4Â molecular sieves and sodium cyanoborohydride(4.12g, 65.6mmol) were added and the mixture was stirred at room température for17 hours. Aqueous hydrochloric acid (2N, 4ml) was added and the mixture was thenbasified with saturated aqueous sodium carbonate solution to pH=10. The mixturewas filtered to remove solid which was washed with methanol. Methanol wasremoved by évaporation under reduced pressure and the residual aqueous solutionwas extracted with ethyl acetate (2x300ml). The combined organic extracts werethen dried (MgSO4), filtered, and concentrated under reduced pressure. The 20 résultant residue was purified by column chromatography on silica gel, eluting with asolvent gradient of dichloromethane : methanol (96:4 to 92:8), to afford the titlecompound as a colourless oil, 8.07g, 79% yield. 102 0’2362 1H-NMR (CDCI3i 300 MHz) δ: 1.42 (s, 9H), 2.65 (m, 1H), 2.90 (m, 2H), 3.07 (m, 1H), 3.19 (m, 1H), 3.30 (m, 1H), 3.58 (m, 1H), 3.73 (s, 3H), 5.22 (br s, 1H), 6.97 (s, 1H), ’ 7.02 (br s, 2H), 7.91 (s, 1H). LRMS : m/z 313.1 (MH+)
Préparation 17
Methvl (2ffl-2-((2-r('terï-butoxvcarbonvl)amino1ethvl}amino)-3-d/-/-imidazoi-4- vDpropanoate
The title compound was prepared from D-histidine methyl ester according to theprocedure described in Préparation 16. 1H-NMR (CDCIs, 300 MHz) δ: 1.41 (s, 9H), 2.57 (m, 1H), 2.80 (m, 2H), 3.00 (m, 1H),3.14 (m, 1H), 3.23 (m, 1H), 3.50 (m, 1H), 3.68 (s, 3H), 6.77 (s, 1H), 7.50 (s, 1H).LRMS : m/z 313 (MH+) 15
Préparation 18 (±)-Methvl 2-({24(ferf-butoxvcarbonvl)amino1ethvïïamino)-3-( 1 H-imidazol-2- vDpropanoate
A solution of the amine from Préparation 55 (183mg, 10.8mmol) was dissolved inmethanol (7ml) and of fert-butyl /\/-(2-oxoethyl)carbamate (172mg, 10.8mmol) wasadded. Sodium acetate (354mg, 43.2mmol), 4Â molecular sieves and then sodium 012362 103 cyanoborohydride (135mg, 21.6mmol) were added, and the résultant mixture wasstirred at room température for 18 hours. Aqueous hydrochloric acid (2N, 1ml) wasthen added and the reaction mixture was stirred thoroughly and then basified withsaturated aqueous sodium carbonate solution to pH=10. The résultant mixture wasthen fiitered to remove solid and the filtrate was extracted with ethyl acetate (2x). Thecombined organic extracts were dried (MgSO4), fiitered, and then concentratedunder reduced pressure. The residue was purified by column chromatography onsilica gel elutihg with a solvent gradient of methanol : dichloromethane (1:99 to 5:95)to give the title compound, 105mg, 31% yield. 10 1H-NMR (CDsOD, 400MHz) δ: 1.42 (s, 9H), 2.58 (m, 1H), 2.74 (m, 1H), 3.11 (m, 4H), 3.67 (m, 1H), 3.70 (s, 3H), 7.10 (s, 2H).
Préparation 19
Methyl (2S)-2-({2-i(ferê-butoxvcarbonvl)arninolethvlternino)-3-( 1 -f 12- (trimethvlsitvl)ethoxvlmethvl)-1H-imidazol-2-vi)propanoate
15 A solution of the amine from Préparation 56 (120mg, 0.40mmol) was dissolved inmethanol (3.5ml) and of feri-butyl /V-(2-oxoethyl)carbamate (51 mg, 0.33mmol) wasadded. Sodium acetate (131mg, 1.60mmol), 4Â-moiecuIar sieves and then sodiumcyanoborohydride (50mg, 0.80mmol) were added, and the résultant mixture wasstirred at room température for 18 hours. Aqueous hydrochloric acid (1 N, 1ml) was 20 then added and the reaction mixture was stirred thoroughly and then basified withsaturated aqueous sodium carbonate solution to pH=10. The résultant mixture wasextracted with ethyl acetate (2x) and the combined organiç extracts were then dried(MgSC>4), fiitered, and concentrated under reduced pressure. The residue was 012362 104 purified by column chromatography on silica gel eluting with ethyl acetate : methanol : 0.88 ammonia (55 : 5 : 0.5) to give the title compound, 30mg, 21 % yield. 1H-NMR (CDCb, 300MHz) δ: -0.02 (s, 9H), 0.90 (t, 2H), 1.29 (s, 9H), 2.63 (m, 1H), 2.84 (m, 1H), 3.02 (dd, 1H), 3.13 (dd, 1H), 3.19 (m, 1H), 3.48 (t, 2H), 3.74 (s, 3H), 3.84 (m, 1H), 5.21 (dd, 2H), 5.77 (br s, 1H), 6.90 (s, 1H), 6.97 (s, 1H). LRMS : m/z 443.3 (MH+)
Préparation 20
Methvl (2S)-3-( 1 H-imidazol-5-vl)-2-f(3ff S)-pyrrolidinvlaminolpropanoate
A solution of the product from Préparation 25 (0.4g, 1.22mmol) in acetic acid (30ml) 10 was hydrogenated over palladium catalyst (10% on carbon, 50mg) at 50°C and 3.5atm for 72 hours. The solution was filtered over Arbocel™/Hyflo™ and the filtratewas concentrated under reduced pressure. The résultant oi, was dissolved indichloromethane and extracted with saturated aqueous sodium bicarbonate solution(3x20ml). The aqueous phase was concentrated under reduced pressure and the V? résultant white solid was triturated with hot ethyl acetate (2x50ml) then with hotmethanol (2x50ml). The methanol extracts were combined and evaporated underreduced pressure. The résultant residue was dissolved in dichloromethane :methanol : 0.88 ammonia (80 : 20 : 2) and purified by column chromatography onsilica gel, eluting with dichloromethane : methanol : 0.88 ammonia (80 : 20 : 5), to 20 afford the title compound as an orange oii, 200mg, 70% yield. 1H-NMR (300 MHz, D2O), mixture of diastereoisomers, δ: 1.70 (m, 1H), 2.02 (m, 1H), 2.93 (m, 3H), 3.10-3.47 (m, 4H), 3.58 (2x s, 2χ1%Η), 3.61 (m, 1H), 6.98 (2x s,2x1/2H), 8.00 (2x s, 2x%H). HRMS: m/z 239.1514 (MH+), calcd 239.1503. 012362 105
Préparations 21 and 22
Methvl (2S}-2-\((1R or S)-1-(f(tezf-butoxvcarbonvl)amino1methvl)propvl)aminol-3-(1/7- imidazol-4-vQpropanoate and
Methvl (2S}-2-l((1S or /?)-1-ff(ferf-butoxvcarbonvl)amino)methvl)propvl)amino1-3-(1H- 5 imidazol-4-vl)propanoate
L-Histidine methyl ester dihydrochloride (945mg, 3.9mmoi) and sodium acetate(1.28g, 15.6mmol) were added to a stirred solution of the product from Préparation77 (730mg, 3.9mmol) in methanol (50ml). 4Â moiecular sieves and sodiumcyanoborohydride (491mg, 7.8mmol) were added and the mixture was stirred at 10 room température for 17 hours. The mixture was filtered and the filtrate wasconcentrated to 10ml under reduced pressure. Aqueous hydrochloric acid (2N, 2ml)was added and the mixture was stirred for two minutes. Saturated aqueous sodiumhydrogen carbonate solution was added and the mixture was extracted with ethylacetate (3x150ml). The combined organic extracts were dried (Na2SO4), filtered, and *5 concentrated under reduced pressure. The residue was purified by columnchromatography on silica gel (Biotage™ column), eluting with a solvent gradient ofdichloromethane : methanol (95:5 to 90:10), to afford the title compound ofPréparation, 21,178mg, 13% yield: 1H-NMR (CDCfe ,400 MHz) δ: 0.90 (t, 3H), 1.40 (m, 2H), 1.43 (s, 9H), 2.30 (br m, 20 1H), 2.82 (dd, 1H), 2.97 (dd, 1H), 3.02 (m, 1H), 3.20 (br m, 1H), 3.65 (m, 1H), 3.72 (s, 3H), 5.21 (brs, 1H), 6.80 (s, 1H), 7.57 (s, 1H). LRMS : m/z 341.2 (MH+) TLC : dichloromethane:methanol (90:10) Rf = 0.48.
And Préparation 22,271mg, 20% yield: 012362 106 * 1H-NMR (CDCI3) 400 MHz) δ: 0.82 (t, 3H), 1.23-1.42 (m, 2H), 1.45 (s, 9H), 2.50 (brm, 1H), 2.80 (dd, 1H), 3.00 (dd, 1H), 3.03-3.18 (m, 2H), 3.60 (m, 1H), 3.73 (s, 3H), * 5.30 (br s 1H), 6.82 (s, 1H), 7.53 (s, 1H) LRMS : m/z 341.3 (MH+) 5 TLC : dichioromethane:methanol (90:10) Rf = 0.41.
Préparations 23 - 26
The compounds of the following tabuiated Préparations of the general formula:
were prepared by a similar method to that of Préparation using L-histidine methylester dihydrochloride and the appropriate aldehyde/ketone starting materiais(Products from Préparations 78-80 or commercially-available 1-benzyl-3-pyrrolidinone). 10 012362 107
108
012362 * 012362 109
Préparation 27
Methvl (2SV2-[{2-[(tezi-butoxvcarbonvi'>amino1ethviKmethvnamino1-3-(1/-/-imidazol-4- vDpropanoate
A solution of methyl (2S)-3-(4-imidazolidinyl)-2“(methylamino)propanoate (1g, 5 4.55mmol), of te/t-butyl /V-(2-oxoethyl)carbamate (833mg, 5.23mmol), sodium acetate (1.494g, 18.22mmol) and sodium cyanoborohydride (572mg, 9.10mmol) inmethanol (30ml) was stirred at 0°C under a nitrogen atmosphère. The mixture wasaltowed to warm to room température then aqueous hydrochloric acid (5ml, 1N) wasadded, followed by saturated aqueous sodium hydrogen carbonate solution. The 1° solution was filtered and the aqueous phase was extracted with ethyl acetate. Thecombined organic extracts were washed with brine, dried (MgSO4), filtered, and thenconcentrated under reduced pressure. The residue was purified by columnchromatography on silica gel, eluting with dichloromethane : methanol (100:5), toafford the title compound, 9Û0mg, 61% yield. 15 1H-NMR (CDC,3, 300 MHz) δ: 1.43 (s, 9H), 2.32 (s, 3H), 2.60 (m, 1H), 2.78 (m, 1H), 2.90 (m, 1H), 3.02 (m, 1H), 3.19 (m, 2H), 3.60 (m, 1H), 3.70 (s, 3H), 5.30 (br m, 1H), 6.80 (s, 1H), 7.55 (s, 1H). LRMS : m/z 327.1 (MH+) 012362 110
Préparation 28
Methvl (2S)-3-( 1 H-imidazol-4-vl)-2-( 1 -piperazinvDpropanoate
The productfrom Préparation 29 (200mg, 0.315mmol) was added to a suspension of4-hydroxybenzoic acid (0.22g, 1.5mmol) in hydrogen bromide solution (45% in acetic 5 acid, 5ml) at 0°C and the mixture was stirred at room température for 72 hours.Deionised water (20ml) was added to afford a suspension which was extracted withethyl acetate (3x20ml). The residual aqueous solution was then concentrated underreduced pressure. The résultant orange foam was crystallised from methanol : ethylacetate to afford the tri-hydrobromide sait of the title compound as a colourless solid, 10 82mg, 54% yield. M.p. 211-213°C. 1H-NMR (D2O, 300 MHz) δ: 2.80 (m, 2H), 2.97 (m, 2H), 3.15 (m, 6H), 3.65 (s, 3H),3.73 (t, 1H), 7.23 (s, 1H), 8.53 (s, 1H). LRMS : m/z 239.2 (MH+)
Anal. Found: C, 27.37; H, 4.45; N, 11.36. CnHi8N4O2«3HBr requires C, 27.47; H, '5 4.40; N, 11.65%.
[afo = -32.92 (c 0.11, methanol) 012362 111
Préparation 29
Methvi (2S)-2-{4-f(4-methvlphenvl)sulfonvn-1 -piperazinyl)-3-( 1 -trityl-1 /7-imidazol-4- vDpropanoate
A suspension of methyl (2S)-2-amino-3-(1-trityi-1H-imidazol-4-yl)propanoate (1g,5 2.4mmoî) in diisopropylethyiamine (5ml), was stirred at room température for 20minutes. /V,/V-Bis(2-chloroethyl)-4-methylbenzenesulfonamide (720mg, 2.4mmol)was added and the mixture was stirred at reflux for 3 hours. The mixture was allowedto cool and diluted with acetonitrile. The résultant solution was concentrated underreduced pressure and the residue was suspended in aqueous sodium carbonate solution and extracted with dichloromethane (3x20ml). The combined organicextracts were washed with brine (3x20ml), dried (Na2SO4), filtered, and thenconcentrated under reduced pressure. The residue was purified by columnchromatography on silica gel, eluting with a solvent gradient of dichloromethane :methanol (99:1). The isoiated matériel was dissolved in ether and the résultant •5 solution concentrated under reduced pressure to afford the title compound as acolourless foam, 300mg, 19% yield. 1H-NMR (CDCI3, 300 MHz) δ: 2.42 (s, 3H), 2.63 (m, 2H), 2.72 (m, 2H), 2.78 (dd, 1H), 2.97 (m, 5H), 3.57 (s, 3H), 3.60 (m, 1H), 6.50 (s, 1H), 7,07 (m, 6H), 7.50 (m, 12H). 7.62 (2x s, 2H). 012362 112 LRMS : m/z 635.3 (MH+)
Anal. Found: C, 69.51; H, 6.06; N, 8.69. C37H3sN4O4S*0.25H2O requires C, 69.51; H, *6.07; N, 8.59%.
[cc]d= -3.73 (c 0.10, dichloromethane) 5 Préparation 30 (7S)-642-((fe/f-Butoxvcarbonyl)aminotethviy-2-ethvl-7-(rnethoxvcarbonvl)-5-oxo- 5.6,7.8-tetrahvdroimidazoil .5-clpvrimidin-2-ium iodide
Ethyl iodide (99μΙ, 1.243mmol) was added to a stirred solution of the product fromPréparation 48 (200mg, 0.592mmol) in acetonitrile (5ml) and the mixture was heated 10 at reflux for 17 hours under a nitrogen atmosphère. The mixture was allowed to coolto room température and the solvent was removed by évaporation under reducedpressure. The residue was purified by column chromatography on silica gel elutingwith dichloromethane : methanol (90:10) to afford the titie compound as a whitefoam, 118mg, 40% yield. 15 1H-NMR (D2O, 300 MHz) δ: 1.27 (s, 9H), 1.42 (t, 3H), 3.22-3.47 (m, 4H), 3.58 (m,1H), 3.65 (s, 3H), 3.95 (m, 1H), 4.20 (q, 2H), 4.75 (m, 1H), 7.40 (s, 1H). LRMS : 366.9 (M+) TLC : dichloromethane : methanol : 0.88 ammonia (90 : 10 :1) Rf = 0.26. 012362
Préparations 31-46
ιη 012362 114
115 012362
116 012362
012362
Footnote: 1. Product of Préparation 81. 118 072362
Préparation 47 (7S)-6-i2-r(te/ï-butoxvcarbonyi)amino1ethvl>-7-(,methoxvcarbonvl)-2-i2-(methvlamino)- - 2-oxoethvn-5-oxo-5.6.7,8-tetrahvdroimidazol 1.5-c1pyrimidin-2-ium bromide
A mixture of the product from Préparation 48 (300mg, 0.89mmol) and 2-bro’no-/V-5 methylacetamide (Heterocycles 1995, 41, 2427) (270mg, 1.78mmol) in acetonitrile (7ml) was heated at 80°C for 72 hours. The cooled reaction was concentrated underreduced pressure and the residue purified by column chromatography on silica gelusing an elution gradient of dichloromethane : methanol (95:5 to 90:10). The productwas triturated with ether to afford the title compound as a white solid, 380mg, 87% 10 yield. 1H-NMR (D2O, 300MHz) δ: 1.30 (s, 9H), 2.71 (s, 3H), 3.23-3.47 (m, 5H), 3.60 (rn, 1H), 3.68 (s, 3H), 3.97 (m, 1H), 4.77 (m, 1H), 5.00 (br s, 2H), 7.38 (s, 1H). LRMS : m/z 410.4 (M+)
Préparation 48
Methyl (7S)-6-{2-r(ferf-butoxvcarbonvi)amino1ethvn-5-oxo-5,6.7.8-
Carbonyldiimidazole (156mg, 0.959mmol) was added to a stirred solution of theproduct from Préparation 16 (300mg, 0.959mmol) in Λ/,/V-dimethylformamide (5ml)and the mixture was heated at 60-70°C for 17 hours. The solvent was removed byévaporation under reduced pressure, the residue was .dissoived in saturated °’2362 119 aqueous sodium hydrogen carbonate solution and extracted with dichloromethane.
The combined organic extracts were dried (MgSCM, filtered and then concentrated under reduced pressure. The residue was purified by column chromatography on silica gel, eluting with dichloromethane:methanol (95:5), to afford the title compound 5 as a colourless oil, 210mg, 67% yield. 1H-NMR (D2O, 300 MHz) δ: 1.40 (s, 9H), 3.20-3.60 (m, 5H), 3.70 (s, 3H), 4.08 (m,1H), 4.33 (m, ΊΗ), 4.82 (br m, 1H), 6.80 (s, 1H), 8.13 (s, 1H). LRMS : m/z 339 (MH+) [oc]d = +39.2 (c 0.12, dichloromethane) 1° TLC : ethy, acetate:methanol (95:5) Rf - 0.79
Préparation 49
Ethvl (2E and 223-3-(1-ff2-(trimethvlsilvl)ethoxv1methvl)-1H-imidazol-4-vl)-2-i3- (tritvlamino)propyl1-2-propenoate
The géométrie isomère of the title compound were obtained in 32% and 38% yield15 respectively, from the compound from Préparation 60, and the aldéhyde from
Préparation 68, following a similar procedure to that described in Préparation 52.Isomer 1,1H-NMR (CDCb, 300MHz) δ: -0.02 (s, 9H), 0.90 (t, 2H), 1.28 (t, 3H), 1.78(m, 2H), 2.18 (t, 2H), 2.40 (br s, 1H), 2.97 (t, 2H), 3.44 (t, 2H), 4.19 (q, 2H), 5.20 (s,2H), 7.15-7.32 (m, 12H), 7.43 (d, 6H). 20 LRMS : m/z 596.5 (MH*) 012362 120 and isomer 2, 1H-NMR (CDCfe, 300MHz) δ: -0.01 (s, 9H), 0.90 (t, 2H), 1.28 (t, 3H), 1.72 (m, 2H), 2.19 (t, 2H), 2.46 (t, 2H), 3.47 (t, 2H), 4.22 (q, 2H), 5.22 (s, 2H), 6.70 (s, 1H), 7.18 (m, 3H), 7.24 (m, 6H), 7.45 (d, 6H), 7.55 (s, 1H), 7.79 (s, 1H). LRMS : m/z 596.3 (MH+) 5 Préparation 50
Ethyl (2E and 2Z)-3-(1-n-propvl-tH-imidazol-4-v0-2-i3-('tritvlamino)propvn-2- propenoate
A solution of the compound from Préparation 60 (5.9g, 11.3mmol) in tetrahydrofuran(100ml) was added to an ice-cooled solution of sodium hydride (457mg, 60% 1û dispersion in minerai oil, 11.3mmol) in tetrahydrofuran (100ml), and the mixturestirred for 45 minutes. A solution of the aldéhyde from Préparation 66 (1.56g,11.3mmol) in tetrahydrofuran (100ml) was then added. The reaction was thenallowed to warm to room température and stirred for 18 hours. The mixture wasdiluted with aqueous ammonium chloride solution, the layers separated, and the 15 aqueous phase extracted with ethyl acetate (3x). The combined organic extractswere dried (MgSO4), filtered and concentrated under reduced pressure. The residuewas purified by column chromatography on silica gel, eluting with a solvent gradientof ethyl acetate : pentane (40:60 to 60:40), to give the two géométrie isomers of thetitle compound, 1.87g, 33% yield (isomer 1): 20 1H-NMR (CDCI3, 300MHz) δ: 0.92 (t, 3H), 1.27 (t, 3H), 1.78 (m, 4H), 2.18 (t, 2H),2.52 (br s, 1H), 2.96 (t, 2H), 3.82 (t, 2H), 4.18 (q, 2H), 7.10-7.28 (m, 12H), 7.42 (d,6H). LRMS : m/z 508.2 (MH*)and 2.40g, 42% yield (isomer 2): 012362 121 1H-NMR (CDCI3i 3OOMHz) δ: 0.95 (t, 3H), 1.27 (t, 3H), 1.72 (m, 2H), 1.82 (m, 2H), 2.18 (t, 2H), 2.45 (t, 2H), 3.86 (t, 2H), 4.22 (q, 2H), 6.75 (s, 1H), 7.18 (m, 3H), 7.28 (m, 7H), 7.44 (d, 6H), 7.76 (s, 1H). LRMS : m/z 508.4 (MH+) 5 Préparation 51 Ëthvl (2E and 2Z)-2-Î4-ibenzvl(te/ï-butoxvcarbonvl)amino1butv0-3-(1-n-propyl-1H- imidazot-4-vl)-2-propenoate
The géométrie isomers of the title compound were obtained in 24% and 21% yieldrespectively, from the compound of Préparation 59, and the aldéhyde from 10 Préparation 66, following the procedure described in Préparation 52.
Isomer 1,1H-NMR (CDCfe, 300MHz) δ: 0.96 (t, 3H), 1.27 (t, 3H), 1.37-1.58 (m, 13H), 1.80 (m, 2H), 2.80 (m, 2H), 3.20 (m, 2H), 3.88 (t, 2H), 4.20 (q, 2H), 4.40 (s, 2H), 7.04(s, 1H), 7.22 (m, 5H), 7.42 (s, 1H), 7.52 (s, 1H). LRMS : m/z 470.3 (MH+) 15 Isomer 2,1H-NMR (CDCI3, 300MHz) δ: 0.94 (t, 3H), 1.28 (t, 3H), 1.38-1.58 (m, 13H), 1.80 (m, 2H), 2.38 (m, 2H), 3.18 (m, 2H), 3.85 (t, 2H), 4.22 (q, 2H), 4.40 (br s, 2H), 6.70 (s, 1H), 7.23 (m, 5H), 7.40 (s, 1H), 7.75 (s, 1H). LRMS : m/z 470.3 (MH+) 012362 122
Préparation 52
Ethvl (2E and 2Z)-3-(1-n-butvl-1H-imidazol-4-vl)-2-i3-(tritvlamino)propyri-2- propenoate
A solution of the compound from Préparation 60 (1g, 2.6mmol) in tetrahydrofuran 5 (20ml) was added to an ice-cooled solution of sodium hydride (106mg, 60% dispersion in minerai oil, 2.6mmol) in tetrahydrofuran (20ml), and the solution stirredfor 45 minutes. The aldéhyde from Préparation 67 (400mg, 2.6mmol) intetrahydrofuran (10ml) was then added, and the reaction stirred at room températurefor 18 hours. The reaction was quenched by the addition of aqueous ammonium 1C chloride solution and the mixture extracted with ethyl acetate (2x). The combinedorganic extracts were dried (Na2SO4), filtered, and concentrated under reducedpressure. The residue was dissolved in toluene, adsorbed onto silica, and purified bycolumn chromatography on silica gei, eiuting with a soivent gradient of ethyl acetate :pentane (20:80 to 40:60), to give the two géométrie isomers of the title compound, 15 390mg, 29% yield (isomer 1): 1H-NMR (CDCI3, 300MHz) δ: 0.94 (t, 3H), 1.28 (m, 5H), 1.76 (m, 4H), 2.18 (t, 2H), 2.55 (br s, 1H), 2.97 (t, 2H), 3.84 (t, 2H), 4.17 (q, 2H), 7.09-7.30 (m, 12H), 7.42 (d,6H). LRMS : m/z 522 (MH+) 20 and 400mg, 30% yield (isomer 2): 1H-NMR (CDCIs, 300MHz) δ: 0.94 (t, 3H), 1.30 (m, 5H), 1.76 (m, 4H), 2.19 (t, 2H),2.45 (t, 2H), 3.92 (t, 2H), 4.22 (q, 2H), 6.76 (s, 1H), 7.18 (m, 3H), 7.24 (m, 7H), 7.46(d, 6H), 7.75 (s, 1H). LRMS : m/z 523.1 (M+2H)+ 012362 123
Préparation 53 fe/f-Butvl (3E)-2-oxo-3-f(1 -n-propyl-1 H-imidazol-4-yl)methvlene1-1 - piperidinecarboxvlate
A solution of lithium bis(trimethylsilyl)amide in tetrahydrofuran (43.5ml, 1M, 5 43.5mmol) was added dropwise to a cooled (~78°C) solution of fert-butyl 2-oxo-1- piperidinecarboxylate (J. Org, Chem. 1983, 48, 2424) (8.7g, 43.5mmol) intetrahydrofuran (120ml) and, once addition was complété, the solution was allowedto warm to 0°C, and stirred for an hour. The solution was re-cooled to -78°C, asolution of the aldéhyde from Préparation 66 (4g, 28.9mmol) in tetrahydrofuranio (40ml) was added, and the reaction was then allowed to warm to room température.
The reaction mixture was stirred for 18 hours and then partitioned between waterand ethyl acetate. The phases were separated and the organic phase was dried(MgSO4), filtered, and concentrated under reduced pressure. The residue waspurified by column chromatography on silica gel, eluting with dichloromethane : 15 methanol (95:5), to give the title compound as a single géométrie isomer, 4g, 43%yield. 1H-NMR (CDCIs, 300MHz) δ: 0.89 (t, 3H), 1.50 (s, 9H), 1.78 (m, 2H), 1.86 (m, 2H), 3.00 (m, 2H), 3.70 (t, 2H), 3.85 (t, 2H), 7.07 (s, 1H), 7.46 (s, 1H), 7.62 (s, 1H). LRMS : m/z 320.3 (MH+) 20 Alternative method of svnthesis for title compound in Préparation 53
The compound from Préparation 99 (76.5g, 227mmol) was dissolved indichloromethane (300ml), the solution was cooled to 0°C, and triethylamine (57g,560mmol) was added. Methanesulphonyl chloride (23.7g, 207mmol) indichloromethane (15ml) was then added slowly to the stirred solution over 0.5 hours 25 whilst maintaining the reaction température between 0-5°C. The reaction was then 012362 124 allowed to warm to room température and was stirred for 3 hours. The reaction mixture was then quenched into water (315ml) and the organic phase separated.
The aqueous phase was then extracted with dichloromethane (1x50ml) and the combined organic extracts were washed with water (1x100ml), dried and 5 concentrated under reduced pressure to afford the title compound as a solid, 58.0g,88% yield.
Préparation 54 te/f-Butyl (3E or 3Z)-2-oxo-3-f(4-n-propvl-1-fi2-(trimethvlsilvl)ethoxvÎmethvl}-1H- imidazol-2-yl)methvlenel-1-piperidinecarboxvlate or10 fert-Butvl (3£ or 3Z)-2-oxo-3-f(5-n-propvl-1-if2-(trimethvlsilvhethoxvlmethviy-1H- imidazol-2-vl)methvlenel-1-piperidinecarboxylate
The title compound was obtained as a single stereoisomer in 10% yield from thealdéhydes from Préparation 69 and 70, and iezï-butyl 2-oxo-1-piperidinecarboxylate(J. Org. Chem. 1983, 48, 2424), following a similar procedure to that described inPréparation 53, except hexane : ether (50:50) was used as the column eluant.1H-NMR (CDCIs, 300MHz) δ: -0.03 (s, 9H), 0.88 (t, 2H), 0.98 (t, 3H), 1.56 (s, 9H), 1.66 (m, 2H), 1.92 (m, 2H), 2.58 (t, 2H), 3.22 (m, 2H), 3.48 (t, 2H), 3.77 (m, 2H), 5.30(s, 2H), 6.80 (s, 1H), 7.73 (s, 1H). LRMS : m/z 450.6 (MH*) 125
Préparation 55
Methvi (2/?S)-2-amino-3-( 1 rt-imidazol-2-vl)propanoate 012362
A mixture of the aikene from Préparation 57 (366mg, 12mmol) and 10% palladiumon charcoal (50mg) in methanol (8ml) was hydrogenated at 3.5 atm and 50°C for 18 5 hours. The cooled mixture was filtered through Arbocel™, washing through withmethanol, and the filtrate concentrated under reduced pressure to afford the titlecompound, 200mg, 98% yield. ^-NMR (CD3OD, 400MHz) δ: 3.65 (d, 2H), 3.80 (s, 3H), 4.60 (t, 1H), 7.55 (s, 2H).LRMS : m/z 170.3 (MH+) 10 Préparation 56
Methvi (,2S)-2-amino-3-(1-ff2-(trimethvlsilvl)ethoxv1methvl)-1 H-imidazol-2-
The product from Préparation 58 (950mg, 2.40mmof) was treated with aqueoushydrochloric acid (48ml, 0.25N HCl, 12.0mmol) and the résultant mixture was stirred 15 at room température for 2 hours. The reaction was then basified with 0.88 ammoniato pH=9 and extracted with ethyl acetate (2x). The combined organic extracts weredried (Na2SO4), filtered, and then concentrated under reduced pressure. Theresidue was purified by column chromatography on silica gel eluting with ethylacetate : methanol : 0.88 ammonia (95:5:0.5) to give the title compound, 600mg, 20 83% yield. 012362 126 1H-NMR (CDCI3i 300MHz) δ: -0.03 (s, 9H), 0.90 (t, 2H), 3.00 (dd, 1H), 3.20 (dd, 1H), 3.48 (t, 2H), 3.71 (s, 3H), 4.05 (m, 1H), 5.23 (dd, 2H), 6.92 (s, 1H), 6.97 (s, 1H). LRMS : m/z 300.2 (MH+)
Préparation 57 5 Methvl (2Z)-2-ff(benzvloxv)carbonvnamino)-3-( 1 /7-imidazol-2-vl)-2-propenoate
A mixture of methyl 2-{l(benzyloxy)carbonylîamino}-3-(dimethoxyphosphoryl)-propanoate (1g, 30mmol) in tetrahydrofuran (7ml) was stirred at -40°C andtetramethylguanidine (380mg, 33mmol) was added. The reaction mixture was stirredat -40°C for 20 minutes and then imidazole-2-carboxaldehyde (317mg, 33mmol) wasadded. The reaction was then allowed to warm to room température and was stirredat room température for 18h. The solvent was then removed by évaporation underreduced pressure and the residue dissolved in ethyl acetate and washed with waterand then brine. The organic phase was then dried (MgSO4), filtered, and thenconcentrated under reduced pressure. The residue was purified by columnchromatography on silica gel, eluting with a solvent gradient of ethyl acetate :pentane (30:70 to 80:20), to give the title compound, 366mg, 40% yield. 1H-NMR (CDCIs, 300MHz) δ: 3.77 (s, 3H), 5.17 (s, 2H), 6.44 (s, 1H), 7.10 (br s, 2H), 7.35 (m, 5H), 10.2(brs, 1H). LRMS : m/z 301.9 (MH+) 012362 127
Préparation 58 (2/?,5/?)-2-lsopropyl-3,6-dimethoxy-5-f( 1 -ff2-(trimethvlsilvl)ethoxy1methv0~1 H~ imidazol-2-vl)methvn-2,5-dihvdropyrazine
A solution of (2R)-2-isopropyl-3,6-dimethoxy-2,5-dihydropyrazine (111mg, 0.60mmol)5 in. tetrahydrofuran (2.5ml) was cooled to -78°C and treated with n-butyl lithium (0.388ml, 1.6M in hexanes, 0.62mmol). The reaction was stirred at -78°C for 45minutes and the organic solution from Préparation 73 was added. The reaction wasthen allowed to warm to room température and was stirred for a further 18 hours.The reaction was then quenched by the addition of methanol and then the solventio was removed by évaporation under reduced pressure. The residue was diluted withwater and ethy, acetate. The layers were separated and the aqueous phase wasextracted with further ethyl acetate (2x). The combined organic extracts were thendried (Na2SO4), filtered, and then concentrated under reduced pressure. Theresidue was purified by column chromatography on silica gel, eluting with a solvent 15 gradient of ethyl acetate : hexane (50:50 to 100:0), to give the title compound, 40mg,17% yield. 1H-NMR (CDCb, 400MHz) δ: -0.03 (s, 9H), 0.65 (d, 3H), 0.84 (t, 2H), 1.00 (d, 3H),2.16 (m, 1H), 3.03 (dd, 1H), 3.39 (dd, 1H), 3.44 (t, 2H), 3.58 (s, 3H), 3.71 (s, 3H), 3.77 (m, 1H), 4.39 (m, 1H), 5.29 (dd, 2H), 6.90 (s, 1H), 6.95 (s, 1H). 20 LRMS : m/z 394.8 (MH*) 012362 128
Préparation 59
Ethvl (2RS)-6-îbenzvl(terf-butoxvcarbonvl)amino)-2-(diethoxyphosphorvOhexanoate
Triethyl phosphonoacetate (2.6ml, 12.9mmol) was added to a solution of sodiumhydride (576mg, 14.2mmol) in tetrahydrofuran (75ml), and the solution stirred at5 room température for 30 minutes. A solution of the iodide from Préparation 64 (5.0g,12.9mmol) in tetrahydrofuran (10ml), and 18-crown-6 (40mg) were added, and thereaction heated under reflux for 18 hours. Aqueous ammonium chloride solution wasadded to the cooled reaction, and the mixture extracted with ethyl acetate (2x). Thecombined organic extracts were dried (MgSO4), filtered, and concentrated underθ reduced pressure to give a yellow oil. The crude product was purified by columnchromatography on silica gel, eluting with a solvent gradient of ethyl acetate :pentane (40:60 to 100:0), to give the title compound, 2.69g, 49% yield. 1H-NMR (CDCIs, 300MHz) δ: 1.21-1.37 (m, 9H), 1.38-1.58 (m, 13H), 1.80 (m, 1H),1.96 (m, 1H), 2.80-2.98 (m, 1H), 3.05-3.25 (m, 2H), 4.16-4.24 (m, 6H), 4.40 (s, 2H),7.18-7.37 (m, 5H). 15 012362 129
Préparation 60
Ethvi (2RS)-2-(diethoxvphosphorvD-5-(tritvlamino)pentanoate
The title compound was prepared in 34% yield from the bromide from Préparation62, following a similar procedure to that described in Préparation 59. 1H-NMR (CDCIs, 400MHz) δ: 1.28 (m, 11H), 1.84-2.02 (m, 2H), 2.15 (t, 2H), 2.93 (m,1H), 4.17 (m, 6H), 7.18 (m, 3H), 7.24 (m, 6H), 7.44 (d, 6H). LRMS : m/z 524.4 (MH+)
Préparation 61
Methvl (2R)-2-chloro-3-( 1 /7-imidazol-4-vl)propanoate
iü A cold solution of sodium nitrite (2.63g, 38mmol) in water (5ml) was added dropwiseto a stirred suspension of D-histidine (2g, 11.5mmol) in concentrated hydrochloricacid (30ml) at -5°C. The mixture was stirred at 0°C for 1 hour and then at roomtempérature for 17 hours. The mixture was cooled and basified with aqueousammonium hydroxide solution (2N) until pH = 4-5. The solvent was then removed by 15 évaporation under reduced pressure to afford (2/?)-2-chloro-3-(1/V-imidazol-4-yl)propanoic acid. 1H-NMR (D2O, 300 MHz) δ: 3.25 (m, 2H), 4.45 (t, 1H), 7.12 (s, 1H), 8.15 (s, 1H).LRMS : m/z 175.0 (MH+) 1<x)d « +13.51 (c 0.093, methanol) 'Π 236 2 130
Hydrogen chloride gas was bubbled through a stirred suspension of (2/?)-2-chloro-3-(1/7-imidazol-4-yl)propanoic acid in methanol (60ml) at 0°C for 20 minutes and thesuspension was -stirred at room température for 17 hours. The solvent was thenremoved by évaporation under reduced pressure and the chilled residue wassuspended in cold aqueous saturated sodium bicarbonate solution (20ml) andextracted with dichloromethane (4x20ml). The combined organic extracts were dried(Na2SO4), filtered, and concentrated under reduced pressure. The residue wasdissolved in diethyl ether and the résultant solution concentrated under reducedpressure to afford the title compound as an oil, 350mg, 14% yield. 1H-NMR (CDCI3) 300 MHz) δ: 3.20 (dd, 1H), 3.37 (dd, 1H), 3.75 (s, 3H), 4.59 (m, 1H), 6.90 (s, 1H), 7.57 (s, 1 H). LRMS : m/z 189.0 (MH+) [α]ο = +2.13 (c 0.16, methanol)
Triphenylphosphine (121g, 0.46mol) was added portionwise to an ice-cooled solutionof the alcohol from Préparation 63 (139g, 0.44mol) and carbon tetrabromide (153g,0.46mol) in dichloromethane (1360ml) and, once addition was complété, the reactionwas stirred at room température for 48 hours. The reaction was diluted with water,the layers separated, and the aqueous phase extracted with dichloromethane (2x).The combined organic extracts were dried (Na2SO4), filtered, and concentratedunder reduced pressure. The crude product was purified by column chromatographyon silica gel, eluting with a solvent gradient of hexane : ethyl acetate (99:1 to 95:5),to afford the title compound, 81.5g, 49% yield. 012362 131 Ή-NMR (CDCI3, 300MHz) δ: 2.02 (m, 2H), 2.28 (m, 2H), 3.58 (t, 2H), 7.19 (m, 3H), 7.27 (m, 6H), 7.46 (d, 6H).
Préparation 63 3-Hvdroxy-A/-tritvl-1 -propanamine
OH A mixture of 3-amino-1-propanol (51ml, 0.66mol), chlorotriphenylmethane (184g,5 0.66mol) and triethylamine (92ml, 0.66mol) in dichioromethane (1000ml) was stirred at room température for 18 hours. The reaction mixture was diluted with water andthe layers separated. The aqueous phase was extracted with furtherdichioromethane (2x) and the combined organic extracts were dried (Na2SO4),filtered, and concentrated under reduced pressure. The residue was triturated well 10 with diisopropyl ether, and the resulting solid was filtered and dried. This solid wasthen triturated with methanol, the suspension filtered, and the filtrate concentratedunder reduced pressure, to give the title compound as a white solid, 139.1g, 66%yield. 15 Ή-NMR (CDCfe, 300MHz) δ: 1.70 (m, 2H), 2.38 (t, 2H), 3.86 (t, 2H), 7.19 (m, 3H), 7.25 (m, 6H), 7.42 (d, 6H). LRMS : m/z 318.4 (MH+)
Préparation 64 fe/f-Butvl benzvl (4-iodobutvl)carbamate o 012362 132 A mixture of the chloride from Préparation 65 (9.3g, 31.3mmol) and sodium iodide (14.9g, lOOmmoi) in acetone (200ml) was heated under reflux for 18 hours. The cooled reaction mixture was concentrated under reduced pressure, and the residue partitioned between ether and water. The layers were separated and the aqueous 5 phase extracted with ether. The combined organic extracts were then dried(Na2SO4), filtered, and concentrated under reduced pressure to afford the titlecompound as a yellow oil, 10.5g, 87% yield. 1H-NMR (CDCfe, 300MHz) δ: 1.40-1.65 (m, 11 H), 1.79 (m, 2H), 3.19 (m, 4H), 4.42 (s,2H), 7.20-7.38 (m, 5H). 10 LRMS : m/z 390 (MH+)
Préparation 65 ferf-Butvl benzvl (4-chlorobutvl)carbamate ci
Teri-butyl benzylcarbamate (J. Org. Chem. 1993, 58, 56) (9.1g, 44mmol) was addedto a solution of sodium hydride (2.14g, 53mmol) in tetrahydrofuran (160ml), and the 15 solution stirred at room température for 20 minutes. 1-Bromo-4-chlorobutane(5.07ml, 44mmol) was then added and the reaction heated under reflux for 18 hours.The cooled reaction was quenched by the addition of aqueous ammonium chloridesolution, and the mixture extracted with ethyl acetate (2x). The combined organicextracts were dried (Na2SO4), filtered, and concentrated under reduced pressure. 20 The crude product was purified by column chromatography on silica gel, eluting withacetate.pentane (95:5), to afford the title compound as a clear oil, 6.1g, 47% yield. H-NMR (CDCb, 300MHz) δ: 1.45 (s, 9H), 1.58-1.80 (m, 4H), 3.14-3.30 (m, 2H), 3.52(t, 2H), 4.42 (s, 2H), 7.25 (m, 5H). LRMS : m/z 298.0 (MH*) - 012362 133
Préparation 66 1 -Propyl-1 H-imidazole-4-carboxaldehvde o.
H
N CH. lmidazoie-4-carboxaldehyde (30g, 0.31 mol) was added portionwise to a solution ofsodium hydride (13.9g, 60% dispersion in minerai oii, 0.348mol) in tetrahydrofuran 5 (450ml), and the solution stirred for 45 minutes. n-Propyl bromide (31.2ml, 0.344mol) was then added portionwise, followed by 18-crown-6 (150mg), and the reactionheated under reflux for 18 hours. Aqueous ammonium chloride solution was addedto the cooled reaction, and the mixture extracted with ethyl acetate (2x) anddichloromethane (2x). The combined organic extracts were dried (MgSO4), filtered,iO and concentrated under reduced pressure. The crude product was purified bycolumn chromatography on siiica gel, eluting with ethyl acetate : pentane (40:60), to give the title compound, 20.2g, 47% yield. 15 Ή-NMR (DMSO-de, 400MHz) 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 277.3 (2M+H)+
Préparation 67 1 -n-Butvl-1 B-imidazole-4-carboxaldehvde O.
H
HaC 012362 134 lmidazole-4-carboxaldehyde (10g, 104mmol) was added portionwise to a solution ofsodium hydride (4.56g, 60% dispersion in minerai oil, 114mmol) in tetrahydrofuran *(150ml), and the solution stirred for 30 minutes. n-Butyl bromide (15.7g, 114mmol)was added portionwise, foilowed by 18-crown-6 (50mg), and the reaction heated 5 under reflux for 18 hours. Aqueous ammonium chloride solution was added to thecooled reaction and the mixture extracted with ethyl acetate (2x) anddichloromethane (2x). The combined organic extracts were then dried (MgSO4),filtered, and concentrated under reduced pressure. The residue was purifïed bycolumn chromatography on siiica gel, eluting with a solvent gradient of pentane : io ethyl acetate (50:50 to 25:75), to give the title compound, 4.45g, 28% yield. 1H-NMR (CDCfe, 300MHz) 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 153.3 (MH*)
Préparation 68 1 ~ff 2-fT rimethvlsilvl)ethoxv1methvft-1 H-imidazole-4-carboxaldehvde
H3C CHa lmidazole-4-carboxaldehyde (1g, 10.4mmol) was added portionwise to a solution ofsodium hydride (463mg, 60% dispersion in minerai oil, 11.4mmol) in N,N-dimethylformamide (15ml), and the solution stirred for 30 minutes at roomtempérature. 2-(Trimethylsilyl)ethoxymethyl chloride (2.03ml, 11.4mmol) was addedand the reaction stirred at room température for 18 hours. The reaction wasquenched by the addition of aqueous ammonium chloride solution, and the mixtureextracted with ethyl acetate (2x). The combined organic extracts were dried(Na2SO4), filtered, and concentrated under reduced pressure. The residue waspurified by column chromatography on siiica gel, eluting with methanokethyl acetate(3:97), to give the title compound, 1.8g, 77% yield.
135 1H-NMR (CDCI3, 300MHz) δ: -0.02 (s, 9H), 0.92 (t, 2H), 3.52 (t, 2H), 5.33 (s, 2H), 7.68 (s, 1H), 7.72 (s, 1H), 9.92 (s, 1H).
Préparations 69 and 70 4- Propvl-t-{f2-(trimethvlsilvr)ethoxvlmethvl}-1/-y-imidazole-2-carboxaldehvde and 5- Propvl-1-ff2-(trimethvisilvi)ethoxvlmethvl)-1H-imidazole-2-carboxaldehvde
n-Butyl lithium (11.9ml, 1.6M in hexanes, 19.14mmol) was added dropwise to acooled (-40°C) solution of the imidazoles from Préparations 71 and 72 (4.6g,19.14mmol) in tetrahydrofuran (75ml) and, once addition was complété, the resulting 10 red solution was stirred for 20 minutes. /V,A/-Dimethylforrnamide (1.36ml, 19.14mmoi)was added dropwise over 15 minutes, and the reaction then allowed to warm to roomtempérature and stirred for 18 hours. The reaction was quenched by the addition ofaqueous ammonium chloride, extracted with ether and the combined organic extracts were concentrated under reduced pressure. The crude product was purified 15 by column chromatography on silica gel, eluting with hexane:ethyt acetate (75:25), togive the tïtle compounds of Préparations 69 and 70 respectively in a 3:1regioisomeric mixture, 3.4g, 66% yield. 1H-NMR (CDCI3i 300MHz) δ: -0.02 (s, 9H), 0.84-1.02 (m, 3H), 1.74 (m, 4H), 2.61 (m,2H), 3.57 (m, 2H), 5.75 (s, 1.5H), 5.80 (s, 0.5H), 6.98 (s, 0.25H), 7.10 (s, 0.75H), 20 9.75 (s, 0.25H), 9.77 (s, 0.75H). LRMS : m/z 269.0 (MH+) 012362 136
Préparations 71 and 72 4- n-Propyi-1 -([2-(trimethvlsilv0ethoxv1methyl)-1 B-imidazole and 5- n-Propyl-1 -ff2-(trimethylsilv0ethoxv1methvl)-1 B-imidazole
10 15 A solution of the imidazole from Préparation 76 (4.9g, 44.6mmol) in tetrahydrofuran(20ml) was added dropwise to a solution of sodium hydride (1.96g, 60% dispersion inminerai oil, 49.1mmol) in tetrahydrofuran (20ml) and, once addition was complété,the solution was stirred for an hour. The solution was cooled to 0°C and 2-(trimethylsilyl)ethoxymethyl chloride (8.28ml, 46.8mmol) was added dropwise over 20minutes. The reaction mixture was stirred at room température for 18 hours, thenconcentrated under reduced pressure. The residue was partitioned between etherand water, the layers separated, and the aqueous phase extracted with ether. Thecombined organic extracts were washed with brine, dried (MgSO4), filtered, andconcentrated under reduced pressure. The residual brown oil was purified by columnchromatography on silica gel, eluting with dichloromethane : methanol (95:5), toafford the title compounds of Préparation 71 and 72 respectively in a regioisomericmixture of 3:1,7g, 65% yield. 1H-NMR (CDCI3, 300MHz) δ: 0.0 (s, 9H), 0.90 (m, 3H), 1.65 (m, 4H), 2.58 (m, 2H),3.45 (m, 2H), 5.20 (s, 2H), 6.74 (s, 0.75H), 6.80 (s, 0.25H), 7.28 (s, 1H). LRMS : m/z 241.1 (MH+) 20 012362 137
Préparation 73 2-(ChloromethvO-1-(i2-(trimethvlsilvnethoxvlmethvi}-1B-imidazole
A solution of the alcohol (150mg, 0.66mmol) from Préparation 74 in dichioromethane(3.7m!) wastreated with triethylamine (0.138mt, 0.99mmol). Methanesulfonyl chloride(0.061ml, 1.79mmol) was then added and the réaction mixture was stirred for 1 hour.The reaction was then diluted with water and extracted with dichioromethane (2x).The combined organic extracts were dried (Na2SO4) and filtered. A small aliquot ofthe résultant solution was concentrated under reduced pressure to provide a sampleof the title compound for characterisation. The remaining organic solution wasconcentrated to a small volume (0.5ml) and diluted with tetrahydrofuran (5ml). Thisorganic solution was used directly in Préparation 58. 1H-NMR (CDCIs, 400MKz) δ: 0.00 (s, 9H), 0.94 (t, 2H), 3.52 (t, 2H), 4.72 (s, 2H), 5.37(s, 2H), 7.01 (s, 2H). LRMS : m/z 247 (MH+)
Préparation 74 ( 1 -(Î2-(T rimethvtsiM)ethoxvlmethvB-1 B-imidazol-2-vhmethanol
A solution of the aldéhyde (2.3g, 10.2mmol) from Préparation 75 in methanol (30ml)was cooled to -20°C. Sodium borohydride (462mg, 12.2mmol) was addedportionwise to the stirred solution and the reaction was allowed to warm to roomtempérature over 1 hour. The reaction was quenched by the addition of aqueous 012362 138 ammonium chloride solution and the résultant mixture was extracted with dichloromethane (2x). The combined organic extracts were dried (Na2SO4), filtered, and concentrated under reduced pressure to give the title compound as a beige solid, 2.15g, 93% yield. 1H-NMR (CDCIs, 400MHz) δ: -0.03 (s, 9H), 0.90 (t, 2H), 3.52 (t, 2H), 4.71 (s, 5.35 (s, 2H), 6.94 (s, 1H), 6.97 (s, 1H).
Préparation 75 1-ÏÏ2-ÎT rimethvlsilvDethoxv1methvl}-1 H-imidazole-2-carboxaldehvde
Sodium hydride (463mg, 60% dispersion in minerai oil, 11.4mmol) was washed withhexane under an atmosphère of dry nitrogen. /V,A/-Dimethylformamide (15 ml) wasadded, the résultant mixture was stirred at room température and imidazole-2-carboxaldehyde (1g, 1Û.4mmol) was added portionwise. The reaction was thenstirred for 1.5 hours, 2-(trimethylsilyl)ethoxy methyl chloride (2.03ml, 11.4mmol) wasadded, and the résultant mixture was then stirred at room température for 18 hours.The reaction was quenched by the addition of aqueous ammonium chloride solutionand the résultant mixture then extracted with ethyl acetate (2x). The combinedorganic extracts were dried (Na2SO4), filtered, concentrated under reduced pressureand then azeotroped with xylene to give the title compound, 2.3g, 98% yield. 1H-NMR (CDCIs, 400MHz) δ: -0.03 (s, 9H), 0.90 (t, 2H), 3.55 (t, 2H), 5.77 (s, 2H), 7.32 (s, 1H), 7.35 (s, 1H), 9.84 (s, 1H). 012362 139
Préparation 764-Propvl-1 H-imidazole
A mixture of 2-bromopentanal (15g, 91mmol) (Bull. Chim. Soc. Fr. 1973, 1465) andformamide (32ml, 806mmol) were heated at 180°C for 8 hours, then allowed to cool.Excess formamide was removed by vacuum distillation, and the residue partitionedbetween aqueous sodium bicarbonate solution and ethyl acetate. The layers wereseparated, and the organic phase was concentrated under reduced pressure. Thecrude product was purified by column chromatography on silica gel, eluting with asolvent gradient of dichloromethane : methanol (93:7 to 90:10), to give the titlecompound, 9g, 90% yield. 1H-NMR (CDCI3, 300MHz) δ: 0.98 (t, 3H), 1.67 (m, 2H), 2.60 (t, 2H), 6.79 (s, 1H), 7.25 (s, 1H), 7.58 (s, 1H). LRMS : m/z 221 (2M+H)*
Préparation 77 fert-Butvl N-(2-oxobutvl)carbamate
o
Ethy, magnésium bromide (1M solution in tetrahydrofuran, 13.7ml, 13.7mmol)) wasadded to a stirred solution of ferf-butyl 2-[methoxy(methyl)amino]-2-oxoethylcarbamate (Synth. Commun. 1988, 18, 2273) (1g, 4.58mmol) intetrahydrofuran (25ml) at 0°C then stirred at 0°C for 15 minutes. The solution wasallowed to warm to room température and was stirred for 45 minutes. Ethyl acetate(5ml) wàs added, followed by saturated ammonium chloride solution. The aqueousphase was extracted with ethyl acetate. The combined organic extracts were washedwith saturated aqueous sodium hydrogen carbonate solution and farine. The organicphase was then dried (NazSCU), filtered, and coiicentrated under reduced pressure. 312362 140
The residue was purified by column chromatography on silica gel, eluting with asolvent gradient of hexane:ethyl acetate (85:15 to 70:30), to afford the title *compound as a colourless oil, 730mg, 84% yield. 1H-NMR (CDCIs, 300 MHz) δ: 1.10 (t, 3H), 1.43 (s, 9H), 2.45 (q, 2H), 4.01 (m, 2H), 5.22 (br s, 1H). LRMS : m/z 187.9 (MH+), 204.9 (MNH?) TLC: hexane : ethyl acetate (70:30) Rf = 0.41
Préparations 78 and 79
The compounds of the following tabulated Préparations of the general formula: o
were prepared by a similar method to that of Préparation 77 using ferf-butyl 2-[methoxy(methyl)amino]-2-oxoethylcarbamate (Synth. Commun. 1988,18, 2273 andthe appropriate Grignard starting materials. 012362 141
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Préparation 80 ferf-Butvl (1 S)-1-methvi-2-oxopropylcarbamate
Methyl magnésium bromide (3.0M solution in diethyl ether, 4.3ml, 12.9mmol)was added to a stirred solution of ferf-butyl (1S)-2-[methoxy(methyl)amino]-1- 5 'nethyl-2-oxoethyfcarbamate (Tetrahedron: Asymmetry 1996, 7, 985) (1g,4.3mmol) in anhydrous tetrahydrofuran (20ml) at -60 °C under a nitrogenatmosphère. The mixture was allowed to warm to 0°C and to then roomtempérature and was stirred at room température for 1 hour. Aqueous saturatedammonium çhloride was added and aqueous phase was extracted with diethyl 10 ather (2x 76ml). The combined organic extracts were then washed with saturatedaqueous ammonium çhloride solution and brine. The organic phase was thendried (MgSO4), filtered, and concentrated under reduced pressure. The residuewas purified by column chromatography on silica gel, eluting with a solventgradient of dichloromethane : methanol (99:1 to 98:2), to afford the title 15 compound as a colourless solid, 412mg, 51% yield. 1H-NMR (CDCb, 300 MHz) δ: 1.35 (d, 3H), 1.45 (s, 9H), 2.20 (s, 3H), 4.30 (m,1H), 5.22 (brs, 1H). 012362 143
Préparation 81 (±)-2-Methoxv-1 -methvîethvl 4-methvlbenzenesulfonate
A solution of 1-methoxy~2-propanol in dichloromethane (2.3g, 25.5mmol) (25ml)and pyridine (5ml) was cooled to between -5 and 0°C. 4-Methylbenzenesulfonyl 5 chloride (5.35g, 28.1mmol) was added dropwise and the mixture was stirred at0°C for 15 minutes. The mixture was then stirred at room température for 18hours. Ice was added and the mixture was stirred for 1 hour. The organic phasewas separated, washed with 10% aqueous sulfuric acid (4x) and water (1x), andthen dried (MgSO4) and filtered. The filtrate was purified by column 10 chromatography on silica gel eluting with dichloromethane. The solution obtainedwas dried (MgSO4), filtered, and concentrated under reduced pressure to affordthe title compound as a colourless oil, 4.3g, 69% yield. 1H-NMR (CDCIs, 400 MHz) δ: 1.27 (d, 3H), 2.43 (s, 3H), 3.23 (s, 3H), 3.37 (m,2H), 4.70 (m, 1H), 7.32 (d, 2H), 7.80 (d, 2H). 15 LRMS : m/z 262.0 (MNH?) 012362 144
Préparation 82
Methvl (2S)-2-r(terf-butoxvcarbonvi)amino1-3-ri-(4,4,4-trifiuorobutv0-W-imidazol- 4-vnpropanoate
Césium carbonate (1.95g, 6mmol) and 1-bromo-4,4,4-trifluorobutane (954mg,5mmol) were added to a solution of methyl (2S)-2-[(te/ï-butoxycarbonyl)amino]-3- 5 (1H-imidazol-4-yl)propanoate (1.08g, 4mmol) in Ν,Ν-dimethylformamide (5ml),and the reaction stirred at 70°C for 3 hours. The cooled mixture wasconcentrated under reduced pressure and the residue partitioned between ethylacetate (150ml) and water (50ml). The layers were separated, the organic phasedried (MgSO4), filtered, and concentrated under reduced pressure. The crudeio product was purified by column chromatography on silica gel using an elutiongradient of cyclohexane:ethyl acetate (100:0 to 0:100) to afford the title compound as an oil, 840mg, 55% yield. 1H-NMR (CDCIs, 400MHz) δ: 1.41 (s, 9H), 2.01 (m, 4H), 3.01 (m, 2H), 3.68 (s,3H), 3.98 (t, 2H), 4.57 (m, 1H), 5.84 (m, 1H), 6.66 (s, 1H), 7.38 (s, 1H). 15 LRMS : m/z 380.3 (MH+) [ajo = —0.81 (o 0.148, methanol) 012362 145
Préparation 83
Methvl (2S)-2-i(tezi-butoxvcarbonvBamino1-3-ri-f1,3-thiazol-5-vlmethvl)-1 H- imidazol-4-vllpropanoate
The title compound was obtained as an oil in 20% yield, from methyl (2S)-2-[(tert-butoxycarbonyI)amino]-3'(1H-imidazol-4-yî)propanoate and 5-(chloromethyl)-l ,3-thiazole hydrochloride (EP 373891), following a similarprocedure to that described in préparation 82, except methanohethyl acetate(10:90) was used as the coiumn eluant. 1H-NMR (CDCIs, 400MHz) δ: 1.41 (s, 9H), 3.03 (m, 2H), 3.65 (s, 3H), 4.55 (m,1H), 5.22 (s, 2H), 5.86 (m, 1H), 6.78 (s, 1H), 7.01 (s, 1H), 7.50 (s, 1H), 8.80 (s,1H). LRMS : m/z 367.1 (MH+)
Préparation 84
Methvl (2S)-2-f(fert-butoxvcarbonvnamino1-3-{ 142-(2-pvridinvl)ethvn-1 H-imidazoi- 4-vl>propanoate
The title compound was obtained in 16% yield, from methyl (2S)-2-[(tertbutoxycarbonyl)amino]-3-(1/-/-imidazol-4-yI)propanoate and 2-(2 012362 146 )romoethyl)pyridine hydrobromide (J. Ket. Chem. 1973, 10, 39) following a îimilar procedure to that described in préparation 82, except methanokethyl acetate was used as the column eluant. H-NMR (CDCI3, 400MHz) δ: 1.41 (s, 9H), 2.95 (m, 1H), 3.03 (m, 1H), 3.18 (t, 5 >H), 3.65 (s, 3H), 4.32 (t, 2H), 4.50 (m, 1H), 5.80 (m, 1H), 6.58 (s, 1H), 6.95 (d, 1H), 7.15 (m, 1H), 7.20 (s, 1H), 7.58 (m, 1H), 8.58 (d, 1H). -RMS : m/z 375.2 (MH+)
Préparation 85
Methvl (2S)-24(tert-butoxycarbonvi)aminol-3-( 1 -phenvl-1 H-imidazol-0 4-vl)propanoate
Phenyiboronic acid (2.44g, 20mmol), copper acetate (2.72g, 15mmol), 4Âmolecular sieves (3g) and pyridine (1.62ml, 20mmol) were added to a solution ofmethyi (2S)-2-[(ferf-butoxycarbonyl)amino]-3-(1/7-imidazoI-4-y!)propanoate(2.69g, 10mmol) in dichloromethane (60ml), and the reaction mixture stirred atroom température whilst bubbling through compressed air, for 2 days. A solutionof ethyienediaminetetraacetic acid (5g, 17mmol) in saturated sodium bicarbonatesolution (200ml) was added and the mixture stirred at room température for 20minutes. The phases were separated, the aqueous iayer extracted withdichloromethane (2x100ml), and the combined organic extracts dried (MgSO4),fîltered, and concentrated under reduced pressure. ie residue was azeotropedwith toluene (300ml), and then purified by column uiromatography on silica gelusing an elution gradient of pentanerethyl acetate (100:0 to 40:60), to afford thetitle compound as a yeilow gum, 1.87g, 52% yield. 012362 147 H-NMR (CDCfe, 400MHz) δ: 1.42 (s, 9H), 3.05-3.19 (m, 2H), 3.72 (s, 3H), 4.60 m, 1H), 5.84 (m, 1H), 7.04 (s, 1H), 7.36 (m, 3H), 7.46 (m, 2H), 7.78 (s, 1H). .RMS: m/z346.1 (MH+) \nal. Found: C, 60.59; H, 6.56; N, 11.57. C^HasNsO^OJôHsO requires C, 5 >0.24; H, 6.88; N, 11.71%. a]o = +10.64 (c 0.126, methanol)
Préparation 86
Methvl (,2S)-2-amino-3-f1-(4.4.4-trifluorobutvl)-1H-imidazol-4-vnpropanoate
IM Hydrochloric acid in dioxan (5ml) was added to the protected amine from^réparation 82 (830mg, 2.19mmol), in an ice-cooled flask. The solution wasallowed to warm to room température, and stirred for 3 hours. The mixture was:oncentrated under reduced pressure, the residue azeotroped with ethyl acetate3x100ml), then dried in vacuo, to afford the title compound as a white foam inquantitative yield. 'H-NMR (D2O, 400MHz) δ: 2.00-2.19 (m, 4H), 3.28 (m, 2H), 3.70 (s, 3H), 4.17 (t,2H), 4.37 (t, 1H), 7.40 (s, 1H), 8.62 (s, 1H). .RMS: m/z280.1 (MH+) = +14.60 (c 0.1, methanol) 012362 148
Préparation 87
Methyl (2S)-2-amino-3-f 1 -phenvl-1 H-imidazol-4-vllpropanoate dihvdrochioride
The title compound was obtained in 90% yield as a yellow solid, after triturationrom diethyl ether, from the protected amine from préparation 85, foliowing a 5 similar procedure to that described in préparation 86. H-NMR (D2O, 400MHz) δ: 3.40 (m, 2H), 3.77 (s, 3H), 4.42 (t, 1H), 7.50 (m, 5H), 7.77 (s, 1H), 9.00 (s, 1H). _RMS : m/z 246 (MH+) \nal. Found: C, 47.86; H, 5.51; N, 12.61. C^HnNaOaC^I.OHzO requires C, 10 17.72; H, 5.54; N, 12.84%. a]D - +12.55 (c 0.11, methanol)
Préparation 88
Methyl (2S)-2-amino-3-i1 -(1,3-thiazoi-5-vlmethvl)-1 B-imidazol-4-vnpropanoate
4M Hydrochloric acid in dioxan (6ml) was added to the protected amine fromoreparation 83 (1.3g, 3.5mmol) in an ice-cooled flask. Water (5ml) followed bysoncentrated hydrochloric acid were then added, and the solution stirred at room 012362 149 smperature for 18 hours. The mixture was concentrated under reduced pressure ind azeotroped with éthanol to afford the title compound, 1.2g, 100% yield. H-NMR (CD3OD, 400MHz) δ: 3.30-3.46 (m, 2H), 3.81 (s, 3H), 4.43 (m, 1H), 5.62 s, 2H), 7.63 (s, 1H), 7.95 (s, 1H), 9.10 (s, 1H), 9.18 (s, 1H). .RMS : m/z 267.0 (MH*) α]ο = +14.60 (c 0.1, methanol)
Préparation 89
Methvi (2S)-2-amino-3-f 1 -T2-(2-pyridinvl)ethvn-1 H-imidazol-4-vDpropanoate dihydrochloride
rhe title compound was obtained as a gum in 95% yield, from the protectedamine from préparation 84, foliowing the procedure described in préparation 88.H-NMR (D2O, 400MHz) δ: 3.30 (m, 2H), 3.58 (m, 4H), 3.70 (s, 3H), 4.36 (m,ΙΗ), 7.40 (s, 1H), 7.78 (d, 1H), 7.85 (dd, 1H), 8.41 (dd, 1H), 8.61 (m, 2H). -RMS: m/z 275.1 (MH*) 012362 150
Préparation 90
Methvl (2S)-2-({2-f(fer^butoxvcarbonv0aminolethyl)arnino)-3-(1-methvl-1/·/- ·
Methyl (2S)-2-amino-3-(1 -methyl-1 H-imidazo!-4-yl)propanoate dihydrochloride(1.06g, 4mmol), sodium acetate (1.3g, 16mmoi) and 4Â molecular sieves 5 (500mg) were added to a solution of fe/ï-butyl N-(2-oxoethyi)carbamate (637mg, 4mmol) in methanol (10ml), and the solution stirred for 10 minutes. Sodiumcyanoborohydride (1.3g, 16mmol) was then added, and the reaction stirred atroom température for 72 hours. 2M Hydrochloric acid (2ml) and water (50mi)were added, and the solution then basified using saturated sodium bicarbonateio solution. The mixture was extracted with ethyl acetate (5x100ml), the combinedorganic extracts dried (MgSO4), filtered, and concentrated under reducedpressure. The crude product was purified by column chromatography on silicagel using an elution gradient of ethyl acetate:methanol:diethylamine (100:0:0 to 96:2:2) to afford the title compound as a colourless oil, 220mg, 17% yield. 15 1H-NMR (CDCIs, 400MHz) δ: 1.41 (s, 9H), 2.62 (m, 1H), 2.77-2.86 (m, 2H), 2.98(dd, 1H), 3.18 (m, 2H), 3.60 (m, 4H), 3.70 (s, 3H), 5.38 (m, 1H), 6.63 (s, 1H),7.34 (s, 1H). LRMS : m/z 327.2 (MH+) [a]o = -1.48 (c 0.108, methanol) 012362 151
Préparation 91
Methyl (2S)-2-({2-r(tert-butoxvcarbonvnamino1ethvl)amino)-3-F 1 -(4,4.4- trifluorobutvl)-1H-imidazol-4-vnpropanoate
4Â Molecular sieves (500mg) and fe/f-buty, N-(2-oxoethyl)carbamate (350mg, 5 2.2mmol) were added to a solution of the amine from préparation 86 (780mg, 2.2mmoi) in methanol (5ml), and the mixture stirred for 20 minutes. Sodiumcyanoborohydride (276mg, 4.4mmol) was added, and the reaction stirred atroom température for 18 hours. 2M Hydrochloric acid (5ml) was added, themixture then neutralised using sodium bicarbonate solution, and filtered throughio ^rbocel®. The filtrate was concentrated under reduced pressure and the residuepartitioned between ethyl acetate (100ml) and water (20ml). The layers wereseparated and the organic layer was dried (MgSO4), filtered, and concentrated under reduced pressure. The crude product was purified by columnchromatography on silica gel using an elution gradient of ethyl acetate.methanol 15 (100:0 to 90:10) to afford the title compound as a colourless oil, 300mg, 32% yield. ’H-NMR (CDCb, 400MHz) δ: 1.42 (s, 9H), 2.02 (m, 4H), 2.62 (m, 1H), 2.78-2.92(m, 2H), 2.98 (dd, 1H), 3.18 (m, 2H), 3.60 (t, 1H), 3.68 (s, 3H), 3.98 (t, 2H), 5.40(m, 1 H), 6.70 (s, 1 H), 7.38 (s, 1 H). 20 LRMS : m/z 423.2 (MH+) [a]o - +2.0 (c 0.1, methanol) c1236 2 152
Préparations 92 to 94
The following compounds of general structure:
were prepared from the appropriate amines (préparations 87-89) and fert-butylN-(2-oxoethyl)carbamate, following a similar procedure to that described in 5 préparation 91. 012362 153
1 = the product was further purified by column chromatography on silica gel, using ethyl acetate:methanol:diethylamine(90:5:5) as eluant 2 = the product was additionally purified by column chromatography on reverse phase polystyrène gel usingwater:methanol (100:0 to 0:100) as eluant ,236 2 154
Préparation 95 (7SV2-Benzvl-6-{2-f(ferf-butoxvcarbonyl')aminoiethvi'i-7-(methoxvcarbonvl')-5-oxo- 5,6.7.8-tetrahvdroimidazof1.5-cipyrimidin-2-ium bromide
Benzyl bromide (119μΙ, 1mmol) was added to a solution of the compound from 5 préparation 48 (270mg, 0.8mmol) in acetonitrile (5ml), and the mixture heated at60°C for 18 hours. The cooled mixture was concentrated under reduced pressureand the residue purified by column chromatography on silica gel using an elutiongradient of dichloromethane:methanol (100:0 to 90:10) to afford the trtle compound,299mg, 59% yield. Lù 1H-NMR (DMSOds, 400MHz) δ: 1.28 (s, 9H), 3.18 (m, 3H), 3.42 (m, 2H), 3.61 (s, 3K),3.95 (m, 1H), 4.85 (m, 1H), 5.42 (dd, 2H), 6.94 (m, 1H), 7.38-7.48 (m, 5H), 7.64 (s,1H), 10.08 (s, 1H). LRMS : m/z430 (M+) [<x]D - +42.09 (c 0.096, methanol) 15 Préparation 96 1 -lsopentvl-1 B-imidazole-4-carboxaldehvde
A mixture of sodium hydride (20g, 60% dispersion in minerai oil, 0.5mol) intetrahydrofuran (300ml) was cooléd to 0°C, and 2-imidazolecarboxaldehyde (45g,0.47mol) was added portionwise over 30 minutes. Once addition was complété, the 012362 155 reaction was stirred at 0°C for 30 minutes, then allowed to warm to room température. 1-Bromo-3-methylbutane (60.8ml, 0.5moi) and 18-crown-6 (140mg) were added, and the reaction was heated at reflux for 18 hours. The cooled reaction was quenched by the addition of water (400mi), and the resulting mixture extracted 5 with dichloromethane (800ml in total). The combined organic extracts were dried(MgSO4) and evaporated under reduced pressure. The residual orange oil waspurified by column chromatography on silica gel using an elution gradient of ethyiacetate:pentane:methanol (40:60:0 to 100:0:0 to 98:0:2) to afford the title compound,19.6g. w Further purification of impure fractions using a Biotage® silica gel column, and ethyiacetate:cyclohexane (40:60) as eluant afforded a further 11.4g of the title compound.Combination of the two batches provided 31g of the title compound, 41% yield.1H-NMR (CDCfe, 400MHz) δ: 0.90 (d, 6H), 1.52 (m, 1H), 1.63 (dt, 2H), 3.97 (t, 2H), 7.47 (s, 1H), 7.58 (s, 1H), 9.80 (s, 1H). 15 LRMS : m/z 189 (MNa+)
Anal. Found: C, 63.73; H, 8.43; N, 16.36. C9Hi4N2O;0.2H2O requires C, 63.65; ,8.55; N, 16.50%.
Préparation 97 ferf-Butvl 3-f hvdroxvi 1 -isopentvl-1 H-imidazol-4-vl)methvn-2-oxo-1 - -G piperidinecarboxvlate
Lithium diisopropylamide (6.5mi, 2M in heptaneAetrahydrofuran/ethylbenzene,13mmol) was added dropwise over 5 minutes to a cooled (-78°C) solution of tert-butyl 2-oxo-1-piperidinecarboxylate (J. Org. Chem. 1983, 48, 2424; J. Chem. Soc. I,1989, 721) (2.6g, 13mmol) in tetrahydrofuran (25ml), so as to maintain a 25 température below -70°C. Once addition was complété, the solution was stirred for30 minutes, then allowed to warm to -10°C, and stirred for a further 30 minutes, 012362 156 before recooling to -78°C. A solution of the aldéhyde from préparation 96 (1.66g, 10mmol) in tetrahydrofuran (5ml) was added dropwise so as to maintain the température below -70°C, and once addition was complété, the reaction was stirred for 30 minutes. Saturated ammonium chloride solution (30ml) was added, the 5 mixture allowed to warm to room température and then partitioned between waterand ethyl acetate. The layers were separated, the aqueous phase extracted withethyl acetate, and the combined organic extracts dried (MgSO4), filtered andconcentrated under reduced pressure. The resulting yellow oil was purified bycolumn chromatography on silica gel using an eiution gradient of ethylίο acetate:diethylamine:methanol (100:0:0 to 88:6:6) to afford the title compound, 1.1g, 30% yield. 1H-NMR (CDCI3, 400MHz) (mixture of diastereoisomers) δ: 0.90 (d, 6H), 1.46-1.64(m, 13H), 1.76 (m, 3H), 2.98 (m, 1H), 3.52 (m, 1H), 3.74 (m, 1H), 3.84 (t, 2H), 4.08, 4.90 (2xm, 1H), 4.58, 5.34 (2xm, 1H), 6.85 (2xs, 1H), 7.35 (2xs, 1H). 15 LRMS : m/z 388 (MNa+)
Préparation 98 3-rHvdroxv(1-isopentvl-1H-imidazol-4-vl)methvn-1-methvl-2-piperidinone
The title compound was obtained in 67% yield from the aldéhyde from préparation 96 and 1-methyl-2-piperidinone, following the procedure described in préparation 97.zü 1H-NMR (CDCI3, 400MHz) (mixture of diastereoisomers) δ: 0.88 (2xd, 6H), 1.35-1.82 (m, 7H), 2.67, 2.81 (m, 1H), 2.88, 2.94 (2xs, 3H), 3.18, 3.22 (m, 2H), 3.84 (t, 2H), 4.78 (m, 1H), 5.04 (m, 1H), 6.83 (2xs, 1H), 7.32 (2xs, 1H). LRMS : m/z 302 (MNa+) 157 012362
Préparation 99 ferf-Butyl 3-ihvdroxy( 1 -propyl-1 H-imidazol-4-yl)methvn-2-oxo-1 -piperidinecarboxylate
10 15
Lithium bis(trimethylsiiyl)amide (244ml, 1M in tetrahydrofuran, 244mmol) was addeddropwise over an hour to a cooled (-75°C) solution of tert-buty! 2-oxo-1-piperidinecarboxylate (J. Org. Chem. 1983, 48, 2424; J. Chem. Soc. I, 1989, 721)(48.7g, 244mmol) in tetrahydrofuran (200ml) under nitrogen, so as to maintain thetempérature below -70°C. The mixture was warmed to 0°C, stirred for 90 minutes,then re-cooled to -75°C. A solution of the imidazole from préparation 66 (26.0g,188mmoi) in tetrahydrofuran (86ml) was added dropwise over 30 minutes, and onceaddition was complété, the reaction was stirred for 2 hours at -75°C. The mixturewas poured into 15% aqueous citric acid solution (650ml), and extracted with ethylacetate (3x250ml). The aqueous solution was basified to pH 8 using 10% sodiumhydroxide, and extracted with dichloromethane (3x250ml). These organic extractswere dried and concentrated under reduced pressure to give the title compound as apale yellow solid, 54.1g.
The ethyl acetate extracts from above were combined, evaporated under reducedpressure and the residue re-suspended in 10% aqueous citric acid solution (100ml).This was extracted with ethyl acetate (3x50ml), and the aqueous basified to pH 8using 10% sodium hydroxide solution. The aqueous solution was extracted withdichloromethane (3x50ml), and these organic extracts dried and evaporated underreduced pressure to give additional product as a pale yellow solid, 22.4g. Overallyield of the title compound was thus 76.5g, 93% yield. 1H-NMR (CDCI3, 300MHz) (mixture of diastereoisomers) δ: 0.88 (t, 3H), 1.52 (s, 9H), 1.78 (m, 6H), 3.00 (m, 1H), 3.58 (m, 2H), 3.74 (m, 1H), 3.82 (t, 2H), 5.38 (d, 1H), 6.87 (s, 1 H), 7.38 (s, 1 H).- 20 012362 158
Préparation 100 terf-Butvl 3-ïhvdroxv(1-tritvl-1H-imidazol-4-vDmethvn-2-oxo-1-piperidinecarboxylate
Lithium diisopropylamide (8ml, 1.5M in cyclohexane, 12mmol) was added dropwiseover 5 minutes to a cooled (-78°C) solution of ferf-butyl 2-oxo-1- , 5 piperidinecarboxylate (J. Org. Chem. 1983, 48, 2424; J. Chem. Soc. I, 1989, 721)(1.99g, 10mmol) in tetrahydrofuran (40mi), so as to maintain a température below-70°C. Once addition was complété, the solution was stirred for 20 minutes. Asolution of 1-tritylimidazole-4-carboxaldehyde (J. Med. Chem. 1977, 20, 721) (4.06g,12mmol) in tetrahydrofuran (60ml) was added slowly, and once addition was 10 complété, the reaction was stirred at -78°C for 2 hours. Saturated aqueousammonium chloride solution (50mJ) was added, the mixture allowed to warm to roomtempérature and then partitioned between water (50ml) and ethyl acetate (300ml).The phases were separated, the organic layer dried (MgSO4), filtered, andconcentrated under reduced pressure to give the title compound, 5.3g, 99% yield. 15 1H-NMR (CDCb, 400MHz) (mixture of diastereoisomers) δ: 1.50 (2xs, 9H), 1.60-1.81(m, 4H), 3.00 (m, 1H), 3.58 (m, 1H), 3.74 (m, 1H), 4.10, 4.90 (2xm, 1H), 4.62, 5.40(2xm, 1H), 6.80 (2xs, 1H), 7.14 (m, 6H), 7.25-7.40 (m, 10H). LRMS : m/z 538 (MH+) 072362 159
Préparation 101 ferf-Butyl (3£)-3-i(1-isopentvl-1H-imidazol-4-vl)methyienel-2-oxo-1-
5 were added to a solution of the compound from préparation 97 (1.1g, 3.0mmol) indichloromethane (15ml), and the reaction stirred at room température for 18 hours.The solution was poured into water (200ml), and extracted with ethyl acetate(300ml). The organic extract was dried (MgSO4), filtered, and concentrated underreduced pressure. The crude product was purified by column chromatography on 10 silica gel using an elution gradient of pentane:ethyl acetate (25:75 to 0:100) to affordthe title compound as a white solid, 430mg, 41% yield. 1H-NMR (CDCI3, 400MHz) δ: 0.92 (d, 6H), 1.52 (s, 9H), 1.56 (m, 1H), 1.64 (m, 2H), 1.88 (m, 2H), 3.03 (t, 2H), 3.73 (dd, 2H), 3.92 (t, 2H), 7.05 (s, 1H), 7.45 (s, 1H), 7.62(s, 1H). 15 LRMS : m/z 348.1 (MH+)
Anal. Found: C, 65.47; H, 8.49; N, 12.05. C19H29N3O3 requires C, 65.68; H, 8.41; N,12.09%. 012362 160
Préparation 102 i3E)-3-K1-lsopentvl-1H-imidazol-4-vl)methvlene1-1-methvl-2-piperidinone and (3ZM4(1-isopentvMH4midazol-4-vl)methvlene1-1-methvl-2-piperidinone
5 The title compound was obtained as a yellow solid in 46% yield, from the compoundfrom préparation 98, following a similar procedure to that described in préparation101, except ethyl acetate:diethylamine:methanol (100:0:0 to 96:2:2) was used as thecolumn eluant. 1H-NMR (CDCI3, 400MHz) δ (mixture of isomers): 0.94 (d, 6H), 1.58 (m, 1H), 1.70 (m, 10 2H), 1.92 (m, 2H), 3.03 (s, 3H), 3.12 (m, 2H), 3.40 (t, 2H), 3.97 (t, 2H), 7.02 (s, 1H), 7.48 (s, 1H), 7.58 (s, 1H). LRMS : m/z262 (MH+)
Préparation 103 ferf-Butvl (3E)-2-oxo-3-i(1-tritvl-1/-/-imidazol-4-vl)methvlene1-1-piperidinecarboxviate and 15 ferf-Butvl (3Z)-2-oxo-3-f( 1 -trityl-1 H-imidazol-4-vl)methvlenel-1 -piperidinecarboxvlate
Triethyiamine (2.78ml, 20.0mmol) and methanesulphonyl chloride (773μΙ, lO.Ommol)were added to an ice-cooled solution of the compound from préparation 100 (5.3g,lO.Ommol) in dichloromethane (50ml), and the reaction stirred at room température 012362 161 for 18 hours, and a further 4 hours at reflux. The cooled solution was concentrated under reduced pressure and the residue purified by column chromatography on silica gel using an elution gradient of toluenerethyl acetate (100:0 to 20:80) to afford the title compound, 2.6g, 50% yield. ’H-NMR (CDCI3, 400MHz) δ (mixture of isomers): 1.54 (2xs, 9H), 1.85 (m, 2H), 3.00(t, 2H), 3.68 (t, 2H), 6.99 (s, 1H), 7.10 (m, 6H), 7.30 (m, 9H), 7.44 (s, 1H), 7.58 (s,1H). LRMS : m/z 520.1 (MH+)
Anal. Found: C, 76.40; H, 6.51; N, 7.85. C33H33N3O3 reqires C, 76.28; H, 6.40; N,8.09%.
Préparation 104 (2E)-2-(3-i(terf-Butoxvcarbonvl)amino1propvD-3-( 1 -propyl-1 H-imidazol-4-vl)-2- propenoic acid P«3
A solution of sodium hydroxide (171.3g, 4.28M) in water (4.55L) was added to asolution of the compound from préparation 53 (455g, 1.42M) in tetrahydrofuran(2.275L), and the reaction stirred at room température for 18 hours. The mixture wasconcentrated under reduced pressure to remove the tetrahydrofuran and theremaining aqueous solution was adjusted to pH 5 using glacial acetic acid. Theresulting precipitate was granulated in an ice-bath for 1 hour, then filtered, washedwith water and dried in vacuo. This solid was recrystallised from isopropanol andwater to afford the title compound as a white solid, 304g, 63% yield. 1H-NMR (DMSOde, 400MHz) δ: 0.81 (t, 3H), 1.38 (s, 9H), 1.56 (m, 2H), 1.74 (m,2H), 2.75 (t, 2H), 2.93 (m; 2H), 3.95 (t, 2H), 6.97 (bs, 1 H), 7.37 (s, 1H), 7.52 (s, 1H), 7.76 (s, 1H), 12.02 (bs, 1H). 0123q2 162
Préparation 105 (±)-fe/f-Butyl 3-f(1-isopentyl-1 tf-imidazoM-yl)methvn-2-oxo-1-piperidinecarboxylate
The alkene from préparation 101 (430mg, 1.25mmol), and 10% palladium oncharcoal (Degussa® 101) (100mg) in éthanol (10ml) was hydrogenated at 60psî and 5 room température for 18 hours. The mixture was filtered through Arbocel®, washingthrough with éthanol. The filtrate was concentrated under reduced pressure to givethe title compound as a colourless oil, 420mg, 97% yield. 1H-NMR (CDCIs, 400MHz) δ : 0.89 (d, 6H), 1.50 (m, 10H), 1.62 (m, 4H), 1.78 (m, 1H), 1.98 (m, 1H), 2.63 (dd, 1H), 2.77 (m, 1H), 3.15 (dd, 1H), 3.54 (m, 1H), 3.70 (m, 1H), 10 3.81 (t, 2H), 6.68 (s, 1 H), 7.30 (s, 1 H). LRMS : m/z 350 (MH*)
Préparation 106 (+)-3-17 1 -lsopentvl-1 H-imidazol-4-vi)methvn-1-methvl-2-piperidinone
The title compound was obtained as a colourless oil in 24% yield, from the alkenesfrom préparation 102, following a similar procedure to that described in préparation105, except the product was additionally purified by column chromatography on silicagel using an elution gradient of ethyl acetate:diethylamine:methanol (100:0:0 to90:5:5). 163 1H-NMR (CDCI3i 400MHz) δ : 0.94 (d, 6H), 1.55 (m, 1R), 1.62 (m, 3H), 1.75 (m, 2H), 1.86 (m, 1H), 2.60 (m, 1H), 2.73 (dd, 1H), 2.94 (s, 3H), 3.22 (m, 3H), 3.85 (t, 2H), 6.69 (s, 1H), 7.35 (s, 1H). LRMS : m/z 264 (MH+) °’23e2
Préparation 107 (±)-fe/Î-Butvl 3-(1 /-/-imidazol-4-yimethvl)-2-oxo-1 -piperidinecarboxvlate
on charcoal (Degussa® 101) (200mg) in éthanol (400ml) was hydrogenated at 50°Cand 60 psi for 18 hours. TLC analysis showed starting material remaining, so 10 additional 10% palladium on charcoal (Degussa® 101) (100mg) was added, and themixture hydrogenated for a further 72 hours. The mixture was filtered throughArbocel®, and the filtrate concentrated under reduced pressure. The crude productwas purified by coiumn chromatography on silica gel using an elution gradient ofdichloromethane:ethyl acetate:methanol (100:0:0 to 0:100:0 to 0:90:10) to afford the 15 title compound as a solid, 1.2g, 93% yield. 1H-NMR (CDCI3i 400MHz) δ: 1.46-1.62 (m, 10H), 1.81 (m, 2H), 1.98 (m, 1H), 2.66(m, 1H), 2.95 (m, 2H), 3.55 (m, 1H), 3.78 (m, 1H), 6.80 (s, 1H), 7.24 (s, 1H), 7.50 (s,1H). LRMS : m/z 280 (MH+) 4* 012362 164
Préparation 108 (±)-ferf-Butvl 2-oxo-3-i( 1 -phenvl-1 H-imidazol-4-yl)methvn-1 -piperidinecarboxylate
Phenylboronic acid (366mg, 3mmol), 4À molecular sieves (1g), copper acetate(408mg, 2.25mmol) and pyridine (243μΙ, 3mmol) were added to a solution of the 5 imidazole from préparation 107 (419mg, 1.5mmol) in dichloromethane (10ml), andthe reaction mixture stirred at room température for 4 hours in the presence of a slowstream of compressed air. The air flow was then stopped, and the reaction wasstirred for a further 18 hours at room température. A solution ofethylenediaminetetraacetic acid (2g) in aqueous sodium bicarbonate solution (10ml)θ was added,. the mixture stirred for 10 minutes, then diluted with dichloromethane(100ml). The layers were separated, the organic phase dried (MgS04) andconcentrated under reduced pressure. The residue was purified by columnchromatography on silica gel using an ëlution gradient of ethyi acetate:pentane (50:50 to 80:20) to afford the title compound as a gum, 253mg, 47% yield. 15 1H-NMR (CDCIs, 400MHz) δ: 1.52 (s, 9H), 1.81 (m, 2H), 2.05 (m, 1H), 2.78-2.90 (m,2H), 3.22 (dd, 1H), 3.58 (m, 1H), 3.77 (m, 2H), 7.11 (s, 1H), 7.36 (m, 3H), 7.42 (m,2H), 7.77 (s, 1H). LRMS : m/z 356.1 (MH+) 0123S2 165
Préparation 109 (±)-5-i(terf-Butoxvcarbonvl)aminol-2-[(1 -propyl-1 H-imidazol-4-yi)methvnpentanQic acid
A mixture of the compound from préparation 104 (302g, 0.895M) and 5% palladiumon charcoal (30g) in éthanol (3.0L) was hydrogenated at 60 psi and 60°C for 18 5 hours. The cooled reaction was filtered through Arbocel® and the fiitrate evaporatedunder reduced pressure to give a colourless oil. This was crystallised from ethylacetate and pentane, to afford the title compound as a white solid, 291.7g, 96%yield. 1H-NMR (CDCIs, 300MHz) δ: 0.90 (t, 3H), 1.42 (m, 10H), 1.58 (m, 2H), 1.66-1.86 (m, 10 3H), 2.70 (m, 1H), 2.83 (d, 2H), 3.10 (m, 2H), 3.84 (t, 2H), 4.63 (bs, 1H), 6.68 (s, 1H), 7.49 (s, 1H).
Préparation 110 (2S)-5-f ( fe/f-Butoxvcarbonvl)arninol-2-f( 1 -propyl-1 H-irnidazol-4-vl)methvnpentanoic acid with quinidine
15 A mixture of the acid from préparation 104 (20g, 59mmol), quinidine (19.23g,59mmol) and methanoi (160ml) in.a pressure vessel was purged with nitrogen, andthen hydrogen to a pressure of 3 psi. The vessél was heated to 60°C, a solution of 012362 166
A
[(R)-iPrFerroLANE Rh(C0D)]BF4 (Chirotech Technology Limited) (9.8mg, 0.012mmol) in deoxygenated methanol (1ml) was added, and the reaction mixture hydrogenated at 145 psi for 40 hours. The cooled solution was concentrated under reduced pressure and the crude product dissolved in ethyl acetate, with warming to 5 60°C. On cooling to room température with stirring, précipitation occurred, and the solid was filtered and dried in vacuo to afford the title compound, 29.8g, 76% yîeid(94% ee determined by CE).
Alternative method of svnthesis for title compound in préparation 110 A mixture of the acid from préparation 109 (50g, 147mmol) and quinidine (47.8g, ' 147mmol) in ethyl acetate (1.75L) was heated at 50°C on a steam bath, until asolution was obtained. The solution was warmed to 60°C, the heat removed and thesolution allowed to cool, then stirred at room température for 18 hours. The resultingprecipitate was filtered, washed with ethyl acetate and dried at 80°C in vacuo toafford the title compound as a white solid, 45.1g, 46% yield. ’ 1H-NMR (CD3OD, 400MHz) δ: 0.83 (t, 3H), 1.10-1.20 (m, 1H), 1.40 (s, 9H), 1.45-1.62(m, 5H), 1.65-1.80 (m, 4H), 1.88 (m, 1H), 2.37 (m, 1H), 2.50-2.64 (m, 3H), 2.84 (m, 1H), 3.00-3.14 (m, 3H), 2.21 (m, 1H), 3.39 (m, 1H), 3.80 (m, 2H), 3.96 (m, 4H), 5.17- 5.25 (m, 2H), 5.91 (m, 1H), 6.07-6.18 (m, 1H), 6.89 (s, 1H), 7.38 (d, 1H), 7.43 (dd, 1H), 7.57 (s, 1H), 7.76 (d, 1H), 7.98 (d, 1H), 8.72 (d, 1H). 3 LRMS : m/z 340 (MH+), 325 (quinidineH*)
Anal. Found: C, 65.82; H, 8.17; N, 10.32. 037^3^06*0.5^0 requires 66.05; H,8.09; N, 10.41%.
[a]D = +121.36 (c 0,15, methanol)

Claims (46)

167 012362 Claims 1) Compounds of formula (!) R9
O) Where: X is N or CHn is 0 to 3 5 R1 is: a) C-t-e alkyl, straight Chain or branched chain, b) Ci-6 alkenyl, straight chain or branched chain, c) Ci.6 alkynyl, straight chain or branched chain, d) Heterocycle, l0 e) Aromatic heterocycle, f) Aryl; g) hydrogen; said groups (a), (b) and (c) optionaiiy further substituted by: C3.7 cycloalkyl, aryl,aromatic heterocycle, heterocycle, OR11, N R11 R12, S(O)PR11, OC(0)R11, CO2R11, 15 CONR11R12, SO2NR11R12, halo and NHSO2R11, where R1 may be attached at any position on the imidazole ring, R2 and R3 are each independentty selected from hydrogen, C-i-s alkyl, optionaiiyfurther substituted by OR11, halo; or wherein R2 and R3 may be joined to form a link, said link is C2-e alkylene,
012362 168 R4 is hydrogen, alkyl, optionally further substituted by C3.7 cycloalkyl, aryl, OR11, halo and R11; or wherein R4 and R10 may be joined to form a link, wherein said link is Cm alkylene, optionally further substituted by OR11, halo and R11, R5 and R6 are seiected from: hydrogen, aryl, C-i-S alkyl, said alkyl optionally further substituted by C3-7 cycloalkyl,aromatic hetérocycle, heterocycle, aryl, OR11, R11 and halo; orwherein R10 and either of R5 or R6 may be joined to form a link, wherein said iink is aC1.3 alkylene, optionally further substituted by OR11, halo, R11 and aryl; orwherein R5 and R6 may be joined to form a link, wherein said link is C2-e alkylene, R7 and R8 are independently seiected from: hydrogen, Ci-6 alkyl, optionally further substituted by OR11, halo, aryl and R11; orwherein R7 and R8 may be joined to form a iink, wherein said iink is C2.6 alkylene, R9 and R10 are independently seiected from: Hydrogen, C(NR11)NR11R12, Cm alkyl, said alkyl optionally substituted by OR11, halo,aryl and R11; or wherein R9 and R10 may be joined to form a link, wherein said link is C2.s alkylene, R11 and R12 are each independently seiected from hydrogen or Cm alkyl; or whenforming a NR11R12 moiety, R11 and R12 may aiso be joined to form a iink whereinsaid link is C2-6 alkylene, p is 0,1 or
2 Wherein: Aryl is defined as a 6-14 membered aromatic carbocycle, optionally furthersubstituted by R11, halo, OR11, NR11R12, NR11CO2R12, CO2R11, NR11SO2R12, CN,haloalkyl, O(haloalkyl), S(O)PR11, OC(O)R11, SO2NR11R12, C(O)NR11R12, Aromatic heterocycle is defined as a 5 to 7 membered ring; containing from 1 to 3heteroatoms, each independently seiected from O, S and N, said heterocycle group 0 169 optionally substituted by OR11, NR11R12, CO2R11, NR11CO2R12, R11, halo, CN,haloalkyl, O(haloalkyl), S(O)PR11, OC(O)R11, NR11SO2R12, SO2NR11R12, C(O)NR11R12, Heterocycle is defined as a 3-8 membered ring containing from 1-3 heieroatoms,each independently selected from O, S and N, said ring being saturated or partiallysaturated, said heterocycle group optionally substituted by OR11, NR11R12, CO2R11,NR11CO2R12, R11, halo, CN, haloalkyl, O(haloalkyl), S(O)PR11, OC(O)R11, NR11SO2R12, SO2NR11R12, C(O)NR11R12, Or a pharmaceutically acceptable sait, solvaté or prodrug thereof. f 10 2) A compound of formula (1) as claimed in daim 1 wherein the compoundpossesses the stereochemistry of a compound of formulae (IA) or (IB) RS | RS
3) A compound of formula (IA) as claimed in daim 2.
4) A compound of formula (I) as claimed in any one of daims 1-3 wherein theimidazole is 1,4 disubstituted wherein the R1 group is attached to N1.
5) A compound of formula (I) as claimed in any one of daims 1-3 wherein the15 imidazole is 2,4 disubstituted wherein the R1 group is attached to C4.
6) A compound of formula (I) as claimed in any one of daims 1 -5 wherein R1 is anaryl group, Ci.5 alkenyl group or a alkyl group, wherein said alkyl group is 012362 170 optionally substituted by one or more groups selected from CO2R11, OR11, aryl, C3.7cycioalkyl, NHSO2R11, halo and aromatic heterocycte.
7) A compound of formula (I) as claimed in any one of ciaims 1-6 wherein R1 is Ci. 3 alkyl. 5
8) A compound of formula (I) as claimed in any one of ciaims 1-7 wherein R2 and R3 are hydrogen.
9) A compound of formula (l) as claimed in any one of ciaims 1-8 wherein R4 isindependently selected from hydrogen and C-,.3 alkyl; or wherein R4 and R10 may bejoined to form a link, said iink is C2-3 alkylene. 16
10) A compound of formula (I) as claimed in any one of ciaims 1-9 wherein R4 is hydrogen.
11) A compound of formula (I) as claimed in any one of ciaims 1-10 wherein R5 andR6 are preferably independently selected from hydrogen and Ci-e alkyl, said alkylgroup optionally substituted by phenyi; or wherein R5 and R10 may be joined to form 15 a link, said iink is C1.3 alkylene.
12) A compound of formula (I) as claimed in any one of ciaims 1-11 wherein R5 andR6 are hydrogen.
13) A compound of formula (!) as claimed in any one of ciaims 1-12 wherein R7 andR8 are independently selected from hydrogen and C-|.6 alkyl. 20
14) A compound of formula (l) as claimed in any one of ciaims 1-13 wherein R7 and R8 are hydrogen. <tk
15) A compound of formula (!) as claimed in any one of ciaims 1-14 wherein R8 andR10 are independently selected from hydrogen and Cv3 alkyl; or wherein R10 and R4may be joined to form a link, said link is C2-3 alkylene. 171 °T2362
16) A compound of formula (I) as claimed in any one of ciaims 1-15 wherein R9 andR10 are hydrogen.
17) A compound of formula (I) as claimed in any one of ciaims 1-16 wherein R11and R12 are independently selected from hydrogen and C-i-e alkyl. 5
18) A compound of formula (ί) as claimed in any one of ciaims 1-17 wherein R11 and R12 are independently selected from hydrogen and CH3.
19) A compound of formula (I) as claimed in any one of ciaims 1-18 wherein X isCH.
20) A compound of formula (I) as claimed in any one of ciaims 1-19 wherein n is 0.
21) A compound of formula (I) as claimed in any one of ciaims 1-20 wherein aryl isphenyl.
22) A compound of formula (l) as claimed in any one of ciaims 1-21 whereinAromatic heterocycie is defined as a 5 to 6 membered ring, containing from 1 to 2heteroatoms, each independently selected from O, S and N.
23) Compounds of formula (il) and (IIS)Rs | R8r10/N'X[ZLr7
(lli) 172 012362 * Wherein R1, R2, R3, R4, R5, R6, R7, R8 and X are as described in any one of claims 1- il 22, R9 and R10 are as described in any one of daims 1-22 or additionally one or bothgroups may be a suifabie nitrogen proteciing group and R13 is an appropriate oxygenproteciing group.
24) Compounds of formula (XXIII) and (XXIV)
(XXIII)
(XXIV) Where R1, R3, R5, R6, R7, R8, R10 and Z are as described in any one of daims 1-22,R4 is hydrogen, X is CH and R9 is as described in any one of daims 1-22, or is anappropriate nitrogen proteciing group. ιυ
25) A process for the préparation of a compound of formulae (IA) or (IB) accordingto any one of daims 2-22 which comprises the steps of a) hydrolysis of a compound of formula (XXIII)
Where R1, R3, R5, R6, R7 and R8 are as described in any one of daims 1-22, R4 ishydrogen, n is 0, X is CH and R9 is as described in any one of daims 1-22, or is anappropriate nitrogen proteciing group; To give a compound of formula (XXIV) 15 012362 173
Wherein R1, R3, R4, R5, R6, R7, R8, η, X and R9 are as hereinbefore defined and R10is hydrogen; b) hydrogenating the compound of formula (XXIV) so obtained; then! c) resolving the enantiomeric mix to give compounds of formulae (IA) and (IB); 5 then d) optionally removing the nitrogen protecting group when R9 is a nitrogenprotecting group; and e) optionally converting said compound of formulae (IA) or (IB) to apharmaceuticaliy acceptable sait thereof. 10
26) The process as claimed in daim 25 wherein said hydrogénation is an asymmetric hydrogénation.
27) A composition comprising a compound of formula (I) or a pharmaceuticaliyacceptable sait, solvaté or prodrug thereof as claimed in any one of daims 1-22 anda pharmaceuticaliy acceptable diluent or carrier. 15
28) A compound of formula (I) or a pharmaceuticaliy acceptable sait, solvaté or prodrug thereof as claimed in any one of daims 1-22 for use as a médicament.
29) The use of a TAFIa inhibitor in the préparation of a médicament for thetreatment or prévention of a condition selected from thrombosis, atherosclerosis,adhesions, dermal scarring, cancer, fibrotic conditions, inflammatory diseases and 20 those conditions which benefit from maintaining or enhancing bradykinin leveis in thebody. 174 012362
30) The use as claimed in claim 29 wherein the TAFIa inhibitor is a compound of formula (l) or a pharmaceuticaliy acceptable sait, solvaté or prodrug thereof as claimed in any one of claims 1-22.
31 ) The use as claimed in any one of claims 29 or 30 where the condition is a5 thrombotic condition seiected from myocardial infarction, deep vein thrombosis, stroke, young stroke, cérébral infarction, cérébral thrombosis, cérébral embolism,peripheral vascular disease, angina and other forms of acute coronary syndromes,disseminating intravascular coagulation, sepsis, pulmonary embolism, embolieevents secondary to cardiac arrhythmias and the prévention of cardiovascular events θ following surgical revascularisation or intervention.
32) The use as claimed in any one of claims 29 and 30 wherein the condition isatheroscierosis.
33) The use as claimed in any one of claims 29 and 30 wherein the condition isadhesions or dermal scarring.
* 34) The use as claimed in any one of claims 29 and 30 wherein the condition is cancer.
35) The use as claimed in any one of daims 29 and 30 wherein the condition is afibrotic condition seiected from cystic fibrosis, pulmonary fibrotic diseases, chronicobstructive pulmonary disease (COPD), adult respiratory distress syndrome (ARDS),fibromuscular dysplasia, fibrotic lung disease, fibrin deposits in the eye duringopthalmic surgery and arthritis.
36) The use as claimed in any one of daims 29 and 30 wherein the condition is aninflammatory disease seiected from asthma, endometriosis, infiammatory boweldiseases, psoriasis and atopie dermatitis and neurodegenerative diseases,Alzheimers and Parkinsons. 25 175 /2362
37) The use as claimed in any one of claims 29 and 30 wherein the condition is one which benefits from maintaining or enhancing bradykinin levels in the body selected from hypertension, angina, heart failure, pulmonary hypertension, rénal failure and organ failure.
> 38) The use of a TAFIa inhibitor in the préparation of a médicament in combination with an antithrombotic for the treatment of thrombosis.
39) The use as claimed in claim 38 wherein the TAFIa inhibitor is a compound offormula (I) or a pharmaceutically acceptable sait, solvaté or prodrug thereof asclaimed in any one of claims 1-22. 10
40) The use as claimed in any one of claims 38 and 39 where the antithrombotic is a profibrinolytic.
41) The use as claimed in any one of claims 38, 39 and 40 where theantithrombotic is recombinant tissue plasminogen activator (tPA).
42) The use of TAFIa inhibitors and/or TAFl inhibitors as a coating on intravascularis devices.
43) The use as claimed in claim 42 where the TAFIa inhibitor is a compound offormula (I) as claimed in any one of claims 1-22.
44) An intravascular device coated with a TAFIa inhibitor.
45) An intravascular device as claimed in claim 44 where the TAFIa inhibitor is a2G compound of formula (!) as claimed in any one of claims 1-22.
46) A kit comprising: a) a composition comprising a compound of formula (I) or a pharmaceuticallyacceptable sait, solvaté or prodrug thereof as claimed in any one of claims1-22 and a pharmaceutically acceptable diluent or carrier; b) a composition comprising an antithrombotic and a pharmaceuticallyacceptable diluent or carrier; and c) a container 25
OA1200300039A 2000-08-17 2001-08-08 Substituted imidazoles as tafia inhibitors. OA12362A (en)

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HRP20040659A2 (en) * 2002-01-22 2004-10-31 Pfizer 3-(imidazolyl)-2-alkoxypropanoic acids as tafia inhibitors
US6713496B2 (en) 2002-01-22 2004-03-30 Pfizer Inc 3-(imidazolyl)-2-alkoxypropanoic acids
WO2003061653A1 (fr) * 2002-01-22 2003-07-31 Pfizer Limited Acides 3-(imidazolyl)-2-aminopropanoiques servant d'inhibiteurs tafia pour traiter des maladies thrombotiques
CN100507563C (zh) 2003-04-29 2009-07-01 苏黎世大学 借助通过fac配位作用高效标记[M(OH2)3(CO)3]+而用于偶联生物分子的Nε和/或Nα衍生化的、金属和有机保护的L-组氨酸
SE0302853D0 (sv) * 2003-10-29 2003-10-29 Astrazeneca Ab Chemical compounds
CA2544573A1 (fr) * 2003-11-03 2005-06-02 Probiodrug Ag Combinaisons utiles au traitement de troubles neuronaux
WO2005051384A1 (fr) * 2003-11-25 2005-06-09 Pfizer Limited Compositions pharmaceutiques stabilisees
EP1874322A1 (fr) * 2005-04-18 2008-01-09 Bayer Schering Pharma Aktiengesellschaft Utilisation d'inhibiteurs de tafi pour ameliorer la reperfusion myocardique et faciliter la pci
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HRP20150514T1 (hr) * 2006-12-06 2015-09-11 Sanofi DERIVATI SULFAMIDA KAO INHIBITORI TAFIa
WO2009103432A2 (fr) * 2008-02-21 2009-08-27 Sanofi-Aventis Sondes d’imagerie à liaison covalente
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CN105263928A (zh) 2013-06-10 2016-01-20 赛诺菲 作为TAFIa的抑制剂的大环脲衍生物、其制备及其作为药物的用途
MX392524B (es) 2015-10-07 2025-03-24 Salk Inst Biological Studies Compuestos, composiciones farmaceuticas y metodos de uso de agonistas de ppar
FR3046793B1 (fr) * 2016-01-14 2018-01-05 Les Laboratoires Servier Nouveaux derives de phosphinanes et azaphosphinanes, leur procede de preparation et les compositions pharmaceutiques qui les contiennent
EP3437641A4 (fr) 2016-03-29 2019-11-20 Daiichi Sankyo Company, Limited Agent thérapeutique contre les maladies inflammatoires de l'intestin
BR112018069930B1 (pt) * 2016-04-13 2024-02-20 Mitobridge, Inc Compostos agonistas de ppar, uso dos mesmos e composição farmacêutica
CN113943255B (zh) * 2020-07-17 2024-08-06 深圳信立泰药业股份有限公司 一种手性3-(4-环丙基-2,5-二氧杂咪唑啉-4-基)丙酸的制备方法

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