Aspartyl protease inhibitors

Technical field

This invention relates to novel compounds having inhibitory activity on aspartyl proteases such as renin. It further concerns pharmaceutical compositions comprising these compounds as active ingredients as well as processes for preparing these compounds and compositions and their use in the preparation of a medicament or their use in therapy.

Background to the invention

The renin-angiotensin system (RAS) is an endocrine system involved in regulation of long-term blood pressure and fluid electrolyte balance in mammals. One of the enzymes involved in the RAS is renin, which is an aspartyl protease with a high substrate specificity, its only known substrate is angiotensinogen. Renin cleaves the N terminus of circulating angiotensinogento angiotensin I (Ang I), which thereafter is further processed to the active peptide hormone angiotensin II (Ang II) by the less specific angiotensin-converting enzyme (ACE). Ang II increases blood pressure both directly by arterial vasoconstriction and indirectly by liberating the sodium-ion-retaining hormone aldosterone. Ang II is known to work on at least two receptor subtypes called ATI and AT2. ATI seems to transmit most of the known functions of Ang II, while the role of AT2 is still unknown.

Modulation of the RAS represents a major advance in the treatment of cardiovascular diseases. Inhibition of the enzymatic activity of renin leads to a reduction in the formation of Ang I, and as a consequence, a smaller amount of Ang II is produced. The reduced concentration of that active peptide hormone is a direct cause of the hypotensive effect of renin inhibitors.

ACE inhibitors and ATI blockers have been accepted to treat hypertension and ACE inhibitors are used for renal protection in the prevention of congestive heart failure and myocardial infarction. The rationale to develop renin inhibitors is the specificity of renin. Renin inhibitors are expected to demonstrate a different pharmaceutical profile than ACE inhibitors and ATI blockers with regard to efficacy in blocking the RAS and in safety aspects.

Although several pharmaceutical companies have attempted to advance renin inhibitors to the clinic, and although potent in vitro renin inhibitors was obtained, only limited clinical experience

has been created. Insufficient pharmacokinetic properties, such as oral bioavailability, high clearance, low stability and a high cost of gods for large scale synthesis have stopped clinical development of these compounds.

To date, only one renin inhibitor has entered the market and just a few are reported to be in clinical evaluation. Thus, there is still a need for highly potent renin inhibitors with improved pharmacokinetic properties and a long duration of action. The present invention concerns inhibitors of renin which exhibit beneficial potency, selectivity and/or pharmacokinetic properties.

Drugs of the future, 2001, 26(12): 1139-1148 describes the clinical study of Aliskiren fumarate. This prior art compound differs structurally from the compounds of the present invention in the link between the phenyl ring and the peptidomimetic backbone. In the prior art compound this link is an all carbon link, whereas in the compounds of the present invention, the corresponding link includes a heteroatom, S, O or N.

In J. Med. Chem., 2006, 4544-4567, Hanessian et al describe Structure based design and synthesis of macroheterocyclic peptidomimetic inhibitors of the aspartyl protease BACE and the preparation and use of intermediates to such inhibitors. Although some of the intermediates disclosed therein have some structural similarities with the compounds of the present invention, no suggestion is provided as to alternative utilities in the field of renin inhibition.

Brief description of the Invention

In accordance with the present invention, there is provided renin inhibitors represented by the formula (I):

wherein

A is selected from the partial structures Al, A2 and A3;

Ry and Ry' are both hydrogen, or Ry and Ry' together with the nitrogen atom to which they are attached form a cyclic amine such as morpholine, piperidine, piperazine or pyrrolidine;

L is NHNH, CH ₂ NH, O or S;

Y is NH, NHNH, NHC(=O), S(=O) ₂ NH, NHS(=O) ₂ , CH ₂ , CH ₂ NH, O, S or S(=O) _P ;

Q is aryl or heterocyclyl;

Z is O, S, NRa or S(=O) _P ^; m is 0, 1 or 2; n is 0, 1, 2 or 3; p is independently 1 or 2; q is 0 or 1 ;

Ra is H or Ci-C ₄ alkyl;

R ¹ is hydrogen, Ci-Cβalkyl, C ₃ -CycycloalkylCo-C3alkyl, arylCo-Csalkyl or heterocyclylCo-

Csalkyl, wherein the Ci-Cβalkyl, C ₃ -Cycycloalkyl, aryl or the heterocyclyl moiety is optionally substituted with one, two or three substituents independently selected from: halo, haloCi-C ₄ alkyl, Ci-C ₄ alkyl, Ci-C ₄ alkoxy, hydroxy, NRaRb, N(Ra)S(=O) _p Ci-C ₄ alkyl and cyano;

R ² is hydrogen or Ci-Cβalkyl;

X' is hydrogen, fluoro, hydroxy, NRaRb or Ci-Cβalkoxy;

X" is hydrogen, or when X' is fluoro, then X" may also be fluoro;

R ³ is Ci-C ₆ alkyl;

R ^4' is Ci-C ₆ alkyl;

R ⁴ is H or Ci-Cβalkyl; or R ⁴ and R ⁴ together with the carbon atom to which they are attached define a Cs-Cβcycloalkyl;

W is Ci-Cβalkyl, C ₃ -Cycycloalkyl, aryl or heterocyclyl; wherein aryl is independently phenyl, naphthyl, or phenyl fused to Cs-Cβcycloalkyl or Cs-C ₆ cycloalkenyl;

heterocyclyl is independently a 5 or 6 membered, saturated, partially unsaturated or heteroarylic ring containing 1 to 3 heteroatoms independently selected from S, O and N, the ring being optionally fused with a benzene ring; wherein each Ci-Cβalkyl, aryl and heterocyclyl moiety above (including those in composite expressions such as alkoxy or arylalkyl), unless otherwise specified is optionally substituted with one, two or where valence allows three substituents independently selected from Ci-C4alkyl (optionally substituted with one or two substituents independently selected from aryl*Co-C ₃ alkyl, NRaRb, amido and Ci-C4alkoxyamido), C3-C7cycloalkyl, Ci-C4alkoxy, Ci-C ₄ alkoxyCi-C ₄ alkoxyCo-C3alkyl, halo, haloCi-C ₄ alkyl, hydroxy, hydroxyCi-C ₄ alkyl, NRaRb, amido, cyano, azido, Ci-C ₄ alkylcarbonyl, a cyclic amine selected from pyrrolidinyl, piperidinyl, piperazinyl and morpholinyl (any of which cyclic amines being optionally substituted with Ci- C ₄ alkyl or fluoro), aryl*Co-C ₃ alkyl or heterocyclyl*Co-C ₃ alkyl (wherein the moieties aryl* and heterocyclyl* are optionally substituted with Ci-C4alkyl, halo, hydroxy or NRaRb); or a pharmaceutically acceptable salt, hydrate or N-oxide thereof.

As stated above, A is selected from the structures Al, A2 and A3 shown below:

Consequently, the invention includes compounds having any of the structures I-Al, I-A2 and I- A3 shown below:

Q

I-A3

The compounds of general formula (I) have several centres of chirality, conveniently the compounds display at least 75%, preferably at least 90%, such as in excess of 95%, enantiomeric purity at each of the chiral centres. In typical embodiments of the invention, the chiral centre whereto the group R ³ is attached has the stereochemistry shown in structure (Ia):

R , 3 is Ci-Cβalkyl, preferably ethyl or more preferably isopropyl.

The chiral centre to which R is attached typically has the configuration shown in structure (Ib) below:

R ² is Ci-Cβalkyl such as methyl or ethyl. Preferably R ² is hydrogen.

The chiral centre to which X' and X" are attached has typically the configuration as shown in structure (Ic) below:

X' is preferably fluoro, or more preferably hydroxy.

X" is preferably hydrogen.

A further embodiment of the invention includes compounds of formula (I) or any subgroup of formula (I) wherein X' and X" are both fluoro.

In compounds of formula (I) wherein A is Al or A3 and q is 1, or A is A2, the chiral centre whereto R ⁴ and R ⁴ are attached typically has the configuration shown in the partial structure below:

R ^4' R ^4"

H R ⁴ is d-Cβalkyl, preferably isopropyl or more preferably sec. butyl.

R ⁴ " is preferably hydrogen.

According to an alternative embodiment R ⁴ and R ⁴ together define a spiro-cycloalkyl group, for example cyclopentyl or cyclobutyl or preferably cyclopropyl.

In one embodiment of the invention compounds are includes wherein A is Al, thus providing compounds of general formula (I-Al):

According to this embodiment, compounds of general formula (I) are included wherein q is 0 or 1, i.e. compounds according to structures (I-Ala) and (I-Alb) respectively.

(I-Ala) (I-Alb)

Preferred compounds according to this embodiment are those wherein q is 1, i.e. compounds of formula (I-Alb).

Preferred compounds of formula (I) wherein A is Al and q is 0 orl, are those having the stereochemistry indicated in the structure of formula (1-AIa') and (1-AIb') respectively:

(I-A1 a') (I-A1 b')

Typical embodiments of the invention where A is Al include compounds wherein q is 0 and Y is NH, NHNH, CH ₂ or CH ₂ NH, thus giving compounds having the structures shown below:

I-A1c I-A1d

I-A1e I-A1f

Preferred structures according to these embodiments are those wherein Y is NH, i.e. compounds of formula I-Alc.

Further typical embodiments of the invention where A is Al include compounds wherein q is 1 and Y is NH, NHNH, CH ₂ , or CH ₂ NH, thus giving compounds having the structures shown below:

I-A1g I-A1 h

I-A1 i I-A1j

Preferred structures according to these embodiments are those wherein Y is NH, i.e. compounds of formula I-Alg.

Further typical embodiments of the invention where A is Al and q is 1 include compounds wherein Y is NHC(=O), S(=O)2NH or NHS(=O)2, thus giving compounds having the structures shown below:

I-A1k I-A1I

I-A1 m

Preferred structures according to these embodiments are those wherein Y is NHS(=O)2, i.e. compounds of formula I-Alm.

Further typical embodiments of the invention where A is Al include compounds wherein q is 0 or 1 and Y is O or S, thus giving compounds having the structures shown below:

In a further embodiment of the invention compounds are includes wherein A is A2, thus providing compounds of general formula (I-A2):

I-A2

According to this embodiment of the invention, compounds are included wherein L is NHNH, CH ₂ NH, O or S. Preferably L is CH ₂ NH and more preferably NHNH, thus giving compounds having the structures I-A2a and I-A2b respectively as shown below.

I-A2a I-A2b

Preferably, compounds of formula I wherein A is A2, have the stereochemistry as indicated in structure I-A2' below.

I-A2'

In a further embodiment of the invention compounds are includes wherein A is A3, thus providing compounds of general formula (I -A3)

I-A3

According to this embodiment, compounds are included wherein q is 0 or 1, i.e. compounds having the structures I-A3a and I-A3b respectively.

I-A3a I-A3b

Preferably, according to embodiments where A is A3, Ry and Ry' are both hydrogen.

Alternatively, Ry and Ry' together with the nitrogen atom to which they are attached, form a cyclic amine, for example morpholine, piperidine, piperazine or pyrrolidine.

The stereochemistry of compounds of formula I wherein A is A3 and q is 0 or 1, is preferably as indicated in structure I-A3a' and I-A3b' respectively:

I-A3a" I-A3b"

As recited above, R ¹ is hydrogen, d-Cβalkyl, C3-CycycloalkylCo-C3alkyl arylCo-dalkyl or heterocyclylCo-dalkyl, wherein each Ci-Cβalkyl, cycloalkyl, aryl and heterocyclyl moiety is optionally substituted with one, two or three substituents independently selected from halo, haloCi-C ₄ alkyl, Ci-C ₄ alkyl, d-C ₄ alkoxy, hydroxy, NRaRb, N(Ra)S(=O) _p Ci-C ₄ alkyl and cyano. Suitable values for R ¹ include hydrogen, optionally substituted phenyl, optionally substituted benzyl and optionally substituted Ci-Cβalkyl.

Preferably R ¹ is hydrogen, or optionally substituted Ci-Cβalkyl or benzyl, more preferably R ¹ is hydrogen, methyl, ethyl or isopropyl.

In an alternative embodiment, R ¹ is optionally substituted heteroarylmethyl, especially where the heteroaryl is pyrid-2-yl, pyrid-3-yl or pyrid-4-yl, any of which may be substituted as defined above, such as with Ci-C ₄ alkyl (preferably methyl), Ci-C ₄ alkoxy (preferably methoxy), Ci-

C4alkoxyCi-C3alkoxyCo-C3alkyl, (preferably methoxypropoxy), cyano or halo (preferably fluoro).

Further configurations for R ¹ include phenylethyl, which is optionally substituted with a sulphonamide group such as methanesulphonamide or N-methyl methanesulphonamide.

The optional substituents to R ¹ in general are as defined above. Representative values include Ci-C ₄ alkyl such as methyl; halo such as fluoro; haloCi-C ₄ alkyl such as fluoromethyl and trifluoromethyl; and cyano.

In embodiments wherein R ¹ is arylCo-Csalkyl or heterocyclylCo^alkyl the optional substituent(s) to the aryl or heterocyclyl moiety are conveniently in the para and/or ortho position. Currently favoured configurations for R ¹ according to this embodiment are phenyl or benzyl substituted in the para position.

In a preferred embodiment Z is O.

According to another embodiment Z is NRa, wherein Ra is hydrogen or Ci-C ₃ alkyl, preferably hydrogen or methyl.

The group Q is bonded either directly to Z, i.e. n is 0, or Q is bonded via a methylene, ethylene or propylene moiety, i.e. n is 1, 2 or 3 respectively. In favoured embodiments of the invention Q is bonded directly to Z or via an ethylene moiety, i.e. n is 0 or 2. In more favoured embodiments of the invention, Q is bonded via a methylene moiety, i.e. n is 1.

As defined above, Q is aryl or heterocyclyl, optionally substituted with one, two or three substituents independently selected from Ci-C ₄ alkyl (optionally substituted with one or two substituents independently selected from Co-C3alkandiylaryl, NRaRb, amido and C ₁ - C4alkoxyamido), C3-Cycycloalkyl, Ci-C4alkoxy, Ci-C4alkoxyCi-C3alkyl, Ci-C4alkoxyCi- C ₄ alkoxyCo-C ₃ alkyl, halo, haloCi-C ₄ alkyl, hydroxy, hydroxyCi-C ₄ alkyl, cyano, azido, C ₁ - C ₄ alkylcarbonyl, NRaRb, amido, a cyclic amine selected from pyrrolidinyl, piperidinyl, piperazinyl and morpholinyl (any of which cyclic amines being optionally substituted with Ci-

C ₄ alkyl or fluoro), arylCo-Csalkyl or heterocyclylCo-Csalkyl (wherein the aryl or heterocyclyl moiety is optionally substituted with Ci-C ₄ alkyl, halo, hydroxy or NRaRb).

According to one embodiment of the invention Q is an optionally substituted bicyclic aryl or heterocyclyl moiety. Typically, the heterocyclyl moiety contain 1, 2 or 3 heteroatoms, preferably 1 or 2 heteroatoms, independently selected from nitrogen, oxygen and sulphur. Representative bicyclic rings according to this embodiment include naphthyl, quinolinyl, isoquinolinyl, indolyl, isoindolyl, indolinyl isoindolinyl each of which is optionally substituted wherein each of the bicyclic rings is optionally substituted.

A typical value for Q is naphthyl or optionally substituted naphthyl.

According to a further one embodiment of the invention Q is phenyl or 5 or 6-membered heterocyclic ring, any of which is optionally substituted with one two or three substituents. Typically, the heterocyclyl moiety contain 1, 2 or 3 heteroatoms, preferably 1 or 2 heteroatoms, independently selected from nitrogen, oxygen and sulphur. Representative monocyclic rings according to this embodiment include phenyl, pyridyl, thiazolyl, pyrimidyl, pyrazinyl, pyridazinyl, pyrrolyl, imidazolyl, triazolyl, tetrazolyl, piperidyl, piperazinyl and morpholinyl and the like.

Currently preferred heterocyclyl groups include pyrid-2-yl, pyrid-3-yl or pyrid-4-yl, any of which may be substituted as defined above, such as with 1 or 2 substituents independently selected from Ci-C4alkyl (preferably methyl), Ci-C4alkoxy (preferably methoxy), C ₁ - C4alkoxyCi-C3alkoxyCo-C3alkyl, (preferably methoxypropoxy) or halo (preferably fluoro or chloro).

Currently preferred monocyclic rings for Q are phenyl and pyridyl any of which is optionally substituted.

Optional substituents to Q are as defined above. Representative values include Ci-C4alkyl, C3- Cycycloalkyl, Ci-C4alkoxy, Ci-Csalkoxy-Ci-Cβalkoxy, halo, haloCi-C4alkyl. Currently favoured values according to these configurations for the optional substituents to Q include cyclopropyl, methoxy-ethoxy, fluoro, more favoured substituents are chloro, methyl or methoxy-propoxy.

Further representative values for the optional substituents to Q are arylCo-Csalkyl, and heterocyclylCo-Csalkyl wherein the aryl and heterocyclyl moiety is optionally substituted for example with fluoro. Typical Co-C3aryl and Co-Csheterocyclyl include, but are not limited to, phenyl, benzyl, pyrrolyl, pyrrolinyl, pyrazolyl, imidazolyl, oxazolyl, pyrimidinyl, pyrazinyl, morpholinyl and especially furyl, thienyl, thiazolyl and pyridyl.

According to one embodiment of the invention Q is 6-membered ring, which is substituted in one of the meta positions and/or in the para position. Typically, Q is meta- or para-substituted phenyl or, preferably, Q is phenyl which is substituted in one of the meta-positions and in the para-position.

Suitable configurations for Q include phenyl which is substituted in one of the meta positions with Ci-C4alkoxyCi-C4alkoxy such as 3-methoxy-propoxy, and in the para position with Ci- C4alkyl such as methyl or ethyl, cyclopropyl, halo such as fluoro or chloro, or cyano.

Further suitable configurations for Q include phenyl which is substituted in one of the meta positions with Ci-C4alkoxyCi-C4alkoxy, such as 3-methoxy-propoxy and/or in the para-position with optionally substituted phenyl, preferably p-fluorophenyl, or optionally substituted heteroaryl, preferably pyridyl, thienyl or furyl.

A further embodiment for the optional substituents to Q is benzyl which is substituted at the benzylic position. Suitable substituents for the benzylic position includes for example amino, amido or alkoxyamido such as Ci-C4alkylamino or tert-butoxycarbonylamino.

According to this embodiment, compounds wherein Q has the structure shown below are included:

wherein R ⁵ is Ci-C ₄ alkyl, Ci-C ₄ alkylcarbonyl or Ci-C ₄ alkyloxycarbonyl and R ⁵ is hydrogen, methyl or especially phenyl.

According to a further embodiment, Q is disubstituted 6-membered ring with the substituents in the two meta positions or with one substituent in the meta position and the other in the para position. Preferred substituents to Q according to this embodiment are independently chloro, methoxy-propoxy and methyl.

The group W is bonded either directly to Y, i.e. m is 0, or W is bonded via a methylene or ethylene moiety, i.e. m is 1 or 2 respectively. In favoured embodiments of the invention W is bonded via a methylene moiety, i.e. m is 1.

As stated above, W is Ci-Cβalkyl, C ₃ -Cycycloalkyl, aryl or heterocyclyl any of which is optionally substituted with one, two or three substituents.

According to one embodiment of the invention, W is optionally substituted Ci-Cβalkyl such as methyl, ethyl or isopropyl. Preferred substituents to W according to this embodiment include hydroxy and halo such as mono-, di- or trifluoro.

A favoured embodiment of the invention include compounds wherein A is Al, q is 0, Y is NH and W is optionally substituted Ci-Cβalkyl preferably butyl or isobutyl.

According to a further embodiment of the invention, W is an optionally substituted bicyclic aryl or heterocyclyl moiety. Representative bicyclic rings include naphthyl quinolinyl, isoquinolinyl, indolyl, isoindolyl, indolinyl isoindolinyl.

According to a further embodiment of the invention, W is an optionally substituted monocyclic ring, such as optionally substituted phenyl, Cs-Cβcycloalkyl or monocyclic heterocyclyl. The heterocyclic ring according to this embodiment, typically contains 1, 2 or 3 heteroatoms, preferably 1 or 2 heteroatoms, independently selected from nitrogen, oxygen and sulphur. Representative values for monocyclic heterocyclyl include pyridyl, thiazolyl, pyrimidyl, pyrazinyl, pyridazinyl, pyrrolyl, imidazolyl, triazolyl, tetrazolyl, piperidyl, piperazinyl and morpholinyl and the like, each of which is optionally substituted.

Preferably, W is an optionally substituted monocyclic aryl, heterocyclyl or cycloalkyl, more preferably W is cyclopropyl or phenyl.

A typical embodiment for W include optionally substituted phenyl, wherein the optional substituents are selected from fluoro, chloro, methyl and cyclopropyl.

A favoured embodiment of the invention include compounds wherein A is Al, q is 1 and W is optionally substituted phenyl, pyridyl or pyrimidinyl.

In embodiments wherein W is a substituted 6-membered ring, the ring is preferably mono substituted with the substituent in the meta or para position. Preferred configurations according to this embodiment include meta or para substituted phenyl, for example p-fhioro phenyl.

In embodiments wherein the ring W is disubstituted the substituents are preferably in the two meta positions or in the meta and para positions.

Preferred optional substituents to W include one or two substituents independently selected form halo such as fluoro or chloro; C ₃ -Cycycloalkyl such as cyclopropyl; haloCi-Csalkyl such as fluoromethyl and trifluoromethyl; Ci-Cβalkyl such as methyl, ethyl and isopropyl.

It is to be understood that the above defined subgroups of compounds of formulae (Ia), (Ib), (Ib'), (Ic), (Id) and (Ie), as well as any other subgroup defined herein, are meant to also comprise any prodrugs, iV-oxides, addition salts, quaternary amines, metal complexes and stereochemical^ isomeric forms of such compounds.

As used in the foregoing and hereinafter, the scientific and technological terms and nomenclature have the same meaning as commonly understand by a person of ordinary skill in the art, in addition, the following definitions apply unless otherwise noted.

As used herein 'C _m -C _n alkyl' as a group or part of a group defines a saturated straight or branched chain hydrocarbon radical having the number of carbon atoms designated (e.g. Ci-Cβalkyl means an alkyl group having from 1 to 6 carbon atoms). Preferred alkyl groups for use in the invention are Ci-Cβalkyl groups, i.e. alkyl groups having from 1 to 6 carbon atoms. Exemplary alkyl

groups include methyl, ethyl, n-propyl, iso-propyl, n-butyl, iso-butyl, tert.butyl, pentyl, hexyl and the like. Unless otherwise indicated the alkyl group is optionally substituted with 1 or 2, or where valence permits up to 3, substituents.

The term 'C ₂ -C _n alkenyl' as a group or part of a group defines a straight or branched chain hydrocarbon radical having saturated carbon-carbon bonds and at least one carbon-carbon double bond, and having the number of carbon atoms designated, (e.g. C ₂ -Cealkenyl means an alkenyl group having from 2 to 6 carbon atoms). Preferred alkenyl groups for use in the invention are C ₂ - Cβalkenyl groups, i.e. alkenyl groups having from 2 to 6 carbon atoms. Exemplary alkenyl groups include ethenyl (or vinyl), 1-propenyl, 2-propenyl (or allyl), isopropenyl, butenyl, and the like. Unless otherwise indicated the alkenyl group is optionally substituted with 1 or 2, or where valence permits up to 3, substituents.

The term 'C ₂ -C _n alkynyl' as a group or part of a group defines a straight or branched chain hydrocarbon radical having saturated carbon-carbon bonds and at least one carbon-carbon triple bond, and having the number of carbon atoms designated, (e.g. C ₂ -Cealkynyl means an alkynyl group having from 2 to 6 carbon atoms). Preferred alkynyl groups for use in the invention are C ₂ -C ₆ alkynyl, i.e. alkynyl groups having from 2 to 6 carbon atoms. Exemplary alkynyl groups include, ethynyl, propynyl, propynyl, butynyl, and the like, especially propynyl. Unless otherwise indicated the alkynyl group is optionally substituted with 1 or 2, or where valence permits up to 3, substituents.

The term C ₃ -C _n cycloalkyl as a group or part of a group defines a saturated cyclic hydrocarbon radical having the number of carbon atoms designated, e.g. Cs-Cβcycloalkyl means a cycloalkyl group having 3, 4, 5 or 6, carbon atoms. Exemplary cycloalkyl groups include cyclopropyl, cyclobutyl cyclopentyl, cyclohexyl and the like, especially cyclopropyl. Unless otherwise indicated the cycloalkyl group is optionally substituted with 1 or 2, or where valence permits up to 3, substituents.

'C ₃ -C _n 'CycloalkylC _m -C _n alkyl' represents a C _m -C _n alkyl radical which is substituted with a C ₃ - C _n 'Cycloalkyl moiety, wherein C ₃ -C _n 'Cycloalkyl and C _m -C _n alkyl are as defined for C ₃ - C _n cycloalkyl and C _m -C _n alkyl respectively above. Preferred C ₃ -C _n 'CycloalkylC _m -C _n alkyl groups

for use in the invention are C ₃ -CycycloalkylCo-C ₃ alkyl, i.e. the cycloalkyl moiety is directly bonded (i.e. Co) or bonded through a methyl, ethyl, n-propyl or isopropyl group.

'C ₃ -C _n 'CycloalkylC ₂ -C _n alkenyl' and 'C ₃ -C _n 'CycloalkylC ₂ -C _n alkynyl' have the corresponding meanings, adjusted just for the link to the C ₃ -C _n 'Cycloalkyl moiety, as defined for 'C ₂ -C _n alkenyl' and 'C ₂ -C _n alkynyl respectively. Preferred C3-C _n 'CycloalkylC2-C _n alkenyl and C3-C _n 'CycloalkylC2- C _n alkynyl groups for use in the invention are C ₃ -C _n 'CycloalkylC ₂ -C ₃ alkenyl and C ₃ - C _n 'CycloalkylC ₂ -C ₃ alkynyl, i.e. the C ₃ -C _n 'Cycloalkyl moiety is bonded through an ethenyl, propenyl, ethynyl or propynyl group respectively.

The term C ₃ -C _n cycloalkenyl as a group or part of a group defines a cyclic hydrocarbon radical having one double bond and having the number of carbon atoms designated, e.g. C3- Cβcycloalkenyl means a cycloalkenyl group having 3, 4, 5 or 6, carbon atoms. Exemplary cycloalkenyl groups include cyclobutenyl cyclopentenyl, cyclohexenyl and the like. Unless otherwise indicated the cycloalkenyl moiety is optionally substituted with 1 or 2, or where valence permits up to 3, substituents.

'Ci-C _n alkoxy' defines a radical O-Ci-C _n alkyl wherein Ci-C _n alkyl is as defined for C _m -C _n alkyl above. Preferred alkoxy groups for use in the invention are Ci-Cβalkoxy, i.e. alkoxy groups having from 1 to 6 carbon atoms. Exemplary alkoxy groups include but are not limited to methoxy, ethoxy n-propoxy and isopropoxy, and the like.

The term 'halo' is generic to fluoro, chloro, bromo and iodo. Fluoro is typically preferred in many applications.

The term 'haloC _m -C _n alkyl' as a group or part of a group, represents a C _m -C _n alkyl radical which is substituted with one or more halogen atoms, in particular Ci-C4alkyl substituted with one, two, three, four, five, six, or more halo atoms, such as methyl or ethyl with one or more fluoro atoms, for example, difluoromethyl, trifluoromethyl, trifluoroethyl. Preferred is trifluoromethyl. In case more than one halogen atom is attached to an alkyl group within the definition of haloC _m -C _n alkyl, the halogen atoms may be the same or different.

As used herein, the term '(=0)' or 'oxo' forms a carbonyl moiety when attached to a carbon atom, a sulphoxide moiety when attached to a sulphur atom and a sulphonyl moiety when two of said terms are attached to a sulphur atom. It should be noted that an atom can only be substituted with an oxo group when the valency of that atom so permits.

'Amino' as a group or part of a group, unless the context suggests otherwise, includes NH ₂ , NHCi-Cβalkyl or N(Ci-C6-alkyl)2, wherein in the amino definitions each Ci-Cβalkyl is especially Ci-C4alkyl variants. Included are also radicals wherein the two Ci-Cβalkyl groups of the N(C ₁ - C6-alkyl)2 together with the nitrogen atom to which they are attached form a saturated cyclic amine such as pyrrolidinyl, piperidinyl, piperazinyl or morpholinyl.

'Amido' as a group or part of a group represents a radical Q=O)NH ₂ , C(=O)NHCi-C6alkyl and C(=O)N(Ci-C ₆ alkyl) ₂ , especially C(=O)NHCi-C ₄ alkyl and C(=O)N(Ci-C ₄ alkyl) ₂ . Included are also radicals wherein the two Ci-Cβalkyl groups of the dialkylcarbamoyl together with the nitrogen atom to which they are attached form a saturated cyclic amine such as pyrrolidinyl, piperidinyl, piperazinyl and morpholinyl. Also included are radicals NHC(=0)H, NH(C=O)Ci- Cβalkyl especially NH(C=O)C i-C ₄ alkyl, and N-alkyl alkanoylamino such as N(C ₁ - C ₆ alkyl)(C=O)Ci-C ₆ alkyl especially N(Ci-C ₄ alkyl)(C=O)Ci-C ₄ alkyl.The term 'alkoxyamido' is meant to include NHC(=O)Ci-Cealkoxy, such as tert-butoxycarbonylamino.

Aryl' as a group or part of a group as applied herein represents an aryl moiety such as a phenyl or naphthyl or a phenyl fused to a Cs-Cβcycloalkyl (for example indanyl), or a C ₅ - Cβcycloalkenyl. Examples of suitable aryl groups include but are not limited to phenyl, naphthyl, tetrahydronaphthyl, indenyl and indanyl. Unless otherwise indicated the aryl and/or its fused cycloalkyl moiety is optionally substituted with 1 or 2, or where valence allows up to 3 substituents.

'ArylC _m -C _n alkyl' represents a C _m -C _n alkyl radical which is substituted with an aryl moiety, wherein aryl and C _m -C _n alkyl are as defined above. Preferred arylC _m -C _n alkyl groups for use in the invention are arylCo-Csalkyl, i.e. the aryl moiety is directly bonded (i.e. Co) or bonded through a methyl, ethyl, n-propyl or isopropyl group.

'ArylC ₂ -C _n alkenyT and 'arylC ₂ -C _n alkynyl' have the corresponding meanings, adjusted just for the link to the aryl moiety as defined for 'C ₂ -C _n alkenyT and 'C ₂ -C _n alkynyl respectively. Preferred arylC2-C _n alkenyl and arylC2-C _n alkynyl groups for use in the invention are arylC ₂ - Csalkenyl and arylC ₂ -C ₃ alkynyl, i.e. the aryl moiety is bonded through an ethenyl, propenyl, ethynyl or propynyl group respectively.

ηeterocyclyl' as applied herein is meant to include a saturated, partially unsaturated or aromatic 4-7 membered monocyclic ring or a 8-12 membered bicyclic ring, which monocyclic or bicyclic ring contains 1, 2, 3 or 4 heteroatoms independently selected from S, O and N. Examples of suitable heterocyclyl groups include but are not limited to pyranyl, tetrahydropyranyl, tetrahydrothiopyranyl, thiopyranyl, furanyl, tetrahydrofuranyl, pyrazolyl, pyrazolinyl, pyrazolidinyl, imidazolyl, oxazolyl, isoxazolyl, thiazinolyl, isothiazinolyl, thiazolyl, isothiazolyl, thiazolidinyl, thiadiazolyl, oxadiazolyl, 1,2,3-triazolyl, 1,2,4-triazolyl, tetrazolyl, thienyl, pyridyl, pyrimidinyl, pyrazinyl, pyridazinyl, pyrrolyl, pyrrolinyl, pyrrolidinyl, azetidinyl, piperidinyl, piperazinyl, morpholinyl, thiomorpholinyl, triazinyl, 1 ,4-dioxanyl, quinolinyl, tetrahydroquinolinyl, isoquinolinyl, tetrahydroisoquinolinyl, quinazolinyl, tetrahydroquinazolinyl, quinoxalinyl, benzimidazolyl, benzoxazolyl, benzisoxazolyl, benzothiazinolyl, benzisothiazinolyl, benzothiazolyl, benzoxadiazolyl, benzo- 1,2,3-triazolyl, benzo- 1,2,4-triazolyl, benzotetrazolyl, benzofuranyl, benzothienyl, benzopyridyl, benzopyrimidinyl, benzopyridazinyl, benzopyrazolyl, indolyl, isoindolyl indolinyl, isoindolinyl etc. Unless otherwise indicated the heterocyclyl group is optionally substituted with one, two or where valence allows three substituents.

'HeterocylylC _m -C _n alkyl' represents a C _m -C _n alkyl radical which is substituted with a heterocyclyl moiety, wherein heterocyclyl and C _m -C _n alkyl are as defined above. Preferred heterocyclylC _m - C _n alkyl groups for use in the invention are heterocyclylCo-Csalkyl, i.e. the heterocyclyl moiety is directly bonded (i.e. Co) or bonded through a methyl, ethyl, n-propyl or isopropyl group.

'HeterocyclylC ₂ -C _n alkenyl' and 'heterocyclylC ₂ -C _n alkynyl' have the corresponding meanings, adjusted just for the link to the heterocyclyl moiety as defined for 'C ₂ -C _n alkenyl' and 'C ₂ -C _n alkynyl respectively. Preferred heterocyclylC ₂ -C _n alkenyl and heterocyclylC ₂ -C _n alkynyl groups for use in the invention are heterocyclylC ₂ -C ₃ alkenyl and heterocyclylC ₂ -C ₃ alkynyl, i.e.

the heterocyclyl moiety is bonded through an ethenyl, propenyl, ethynyl or propynyl group respectively.

'Heteroaryl' as applied herein means an aromatic heterocyclyl moiety.

Typically aryl and heterocyclyl moieties within the scope of the above definitions are thus a monocyclic ring with 5 or especially 6 ring atoms, or a bicyclic ring structure comprising a 6 membered ring fused to a 5 or 6 membered ring.

Unless otherwise indicated, the alkyl, alkenyl, alkynyl, cycloalkyl, cycloalkenyl, aryl and heterocyclyl (including those in composite expressions such as arylalkyl or heterocyclylalkyl) is independently optionally substituted with 1 or 2, or where valence permits up to 3, substituents independently selected from: Ci-C ₄ alkyl (optionally substituted with one or two substituents independently selected from aryl*Co-C ₃ alkyl, NRaRb, amido and Ci-C4alkoxyamido), C ₂ - C ₆ alkenyl, C ₂ -C ₆ alkynyl, C ₃ -C ₆ cyclolkyl, Ci-C ₄ alkoxy, Ci-C ₄ alkoxyCi-C ₃ alkyl, Ci-C ₄ alkoxyCi- C ₆ alkoxyCo-C ₃ alkyl, halo, haloCi-C ₄ alkyl, polyhaloCi-C ₄ alkyl, hydroxy, hydroxyCi-C ₄ alkyl, amino, aminoCi-C ₄ alkyl, amido, cyano, azido, nitro, Ci-Cβalkylcarbonyl, a cyclic amine selected from pyrrolidinyl, piperidinyl, piperazinyl and morpholinyl (any of which cyclic amines being optionally substituted with Ci-C ₄ alkyl or fluoro), oxo, mercapto, C ₃ -C ₇ carbocyclylCo-C ₃ alkyl, aryl*C ₀ -C ₃ alkyl, heterocyclyl*C ₀ -C ₃ alkyl, C ₃ -C ₇ carbocyclylC ₂ -C ₃ alkenyl, aryl*C ₂ -C ₃ alkenyl, heterocyclyl*C ₂ -C ₃ alkenyl, C3-C7carbocyclylC2-C ₃ alkynyl, aryl*C2-C ₃ alkynyl, heterocyclyl* C ₂ - C ₃ alkynyl, wherein the asterisked moieties aryl* and heterocyclyl* are optionally substituted with Ci-C ₄ alkyl, halo, hydroxy or NRaRb.

It should be noted that the radical positions on any molecular moiety used in the definitions may be anywhere on such a moiety as long as it is chemically stable.

Radicals used in the definitions of the variables include all possible isomers unless otherwise indicated. For instance pyridyl includes 2-pyridyl, 3-pyridyl and 4-pyridyl; pentyl includes 1- pentyl, 2-pentyl and 3-pentyl.

When any variable occurs more than one time in any constituent, each definition is independent.

Whenever used hereinafter, the term 'compounds of formula (I)', or 'the present compounds' or similar terms, it is meant to include the compounds of formula (I), their prodrugs, iV-oxides, addition salts, quaternary amines, metal complexes, and stereochemically isomeric forms.

The term 'prodrug' as used throughout this text means the pharmacologically acceptable derivatives such as esters, amides and phosphates, such that the resulting in vivo biotransformation product of the derivative is the active drug as defined in the compounds of formula (I). The reference by Goodman and Gilman (The Pharmacological Basis of Therapeutics, 8 ^th ed., McGraw-Hill, Int. Ed. 1992, "Biotransformation of Drugs", p 13-15) describing prodrugs generally is hereby incorporated. Prodrugs preferably have excellent aqueous solubility, increased bioavailability and are readily metabolized into the active inhibitors in vivo. Prodrugs of a compound of the present invention may be prepared by modifying functional groups present in the compound in such a way that the modifications are cleaved, either by routine manipulation or in vivo, to the parent compound.

Preferred are pharmaceutically acceptable ester prodrugs that are hydrolysable in vivo and are derived from those compounds of formula (I) having a hydroxy and/or a carboxyl group. An in vivo hydrolysable ester is an ester, which is hydrolysed in the human or animal body to produce the parent acid or alcohol. Suitable pharmaceutically acceptable esters for carboxy include Ci-Cβalkoxymethyl esters for example methoxymethyl, Ci-Cβalkanoyloxymethyl esters for example pivaloyloxymethyl, phthalidyl esters, Cs-CscycloalkoxycarbonyloxyCi-Cβalkyl esters for example 1-cyclohexylcarbonyloxyethyl; l,3-dioxolen-2-onylmethyl esters for example 5-methyl-l,3-dioxolen-2-onylmethyl; and Ci-Cβalkoxycarbonyloxyethyl esters for example 1-methoxycarbonyloxy ethyl which may be formed at any carboxy group in the compounds of this invention.

An in vivo hydrolysable ester of a compound of the formula (I) containing a hydroxy group includes inorganic esters such as phosphate esters and α-acyloxyalkyl ethers and related compounds which as a result of the in vivo hydrolysis of the ester breakdown will give the parent hydroxy group. Examples of α-acyloxyalkyl ethers include acetoxymethoxy and 2,2- dimethylpropionyloxy-methoxy. A selection of in vivo hydrolysable ester forming groups for hydroxy include alkanoyl, benzoyl, phenylacetyl and substituted benzoyl and phenylacetyl, alkoxycarbonyl (to give alkyl carbonate esters), dialkylcarbamoyl and N-(dialkylaminoethyl)-N-

alkylcarbamoyl (to give carbamates), dialkylamino acetyl and carboxyacetyl. Examples of substituents on benzoyl include morpholino and piperazino linked from a ring nitrogen atom via a methylene group to the 3- or 4-position of the benzoyl ring.

For therapeutic use, salts of the compounds of formula (I) or any subgroup of compounds of formula (I) are those wherein the counter-ion is pharmaceutically acceptable. However, salts of acids and bases which are non-pharmaceutically acceptable may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound. All salts, whether pharmaceutically acceptable or not are included within the ambit of the present invention.

The pharmaceutically acceptable acid and base addition salts as mentioned hereinabove are meant to comprise the therapeutically active non-toxic acid and base addition salt forms which the compounds of formula (I) are able to form. The pharmaceutically acceptable acid addition salts can conveniently be obtained by treating the base form with such appropriate acid. Appropriate acids comprise, for example, inorganic acids such as hydrohalic acids, e.g. hydrochloric or hydrobromic acid, sulphuric, nitric, phosphoric acids and the like; or organic acids such as, acetic, propanoic, hydroxyacetic, lactic, pyruvic, oxalic (i.e. ethanedioic), malonic, succinic (i.e. butanedioic acid), maleic, fumaric, malic (i.e. hydroxybutanedioic acid), tartaric, citric, methanesulfonic, ethanesulfonic, benzenesulfonic, /?-toluenesulfonic, cyclamic, salicylic, /^-aminosalicylic, pamoic acids and the like.

Acid addition salt forms can be converted to the free base form by treatment with an appropriate base.

The compounds of formula (I) containing an acidic proton may also be converted into their nontoxic metal or amine addition salt forms by treatment with an appropriate organic or inorganic base. Appropriate base salt forms comprise, for example, the ammonium salts, the alkali and earth alkaline metal salts, e.g. the lithium, sodium, potassium, magnesium, calcium salts and the like, salts with organic bases, e.g. the benzathine, JV-methyl-D-glucamine, hydrabamine salts, and salts with amino acids such as, for example, arginine, lysine and the like.

Base addition salt forms can be converted to the free acid form by treatment with an appropriate acid.

The term addition salt as used hereinabove also comprises the solvates which the compounds of formula (I) or any of the subgroups of compounds of formula (I), as well as the salts thereof, are able to form. Such solvates are for example hydrates, alcoholates and the like.

The term 'quaternary amine' as used above and hereinafter defines the quaternary ammonium salts which the compounds of formula (I) or any of the subgroups of compounds of formula (I), are able to form by reaction between a basic nitrogen of a compound of formula (I) or any of the subgroups of compounds of formula (I), and an appropriate quaternizing agent, such as, for example, an optionally substituted alkylhalide, arylhalide or arylalkylhalide, e.g. methyliodide or benzyliodide. Other reactants with good leaving groups may also be used, such as alkyl trifluoromethanesulphonates, alkyl methanesulphonates, and alkyl p-toluenesulphonates. A quaternary amine has a positively charged nitrogen. Pharmaceutically acceptable counterions include chloro, bromo, iodo, trifluoroacetate and acetate. The counterion of choice can be introduced using ion exchange resins.

The iV-oxide forms of the present compounds are meant to comprise the compounds of formula (I) wherein one or several nitrogen atoms are oxidized to the so-called iV-oxide.

The compounds according to the invention may contain one or more asymmetrically substituted carbon atoms, asymmetric or chiral centre. The presence of one or more of these asymmetric centres in compounds according to the invention can give rise to stereochemically isomeric forms, stereoisomers, and in each case the invention is to be understood to extend to all such stereoisomers, both in pure form and mixed with each others, including enantiomers and diastereomers, and mixtures including racemic mixtures thereof.

Pure stereoisomeric forms of the compounds and intermediates as mentioned herein are defined as isomers substantially free of other enantiomeric or diastereomeric forms of the same basic molecular structure of said compounds or intermediates. In particular, the term 'stereoisomerically pure' concerns compounds or intermediates having a stereoisomeric excess of at least 80% (i.e. minimum 90% of one isomer and maximum 10% of the other possible isomers) up to a stereoisomeric excess of 100% (i.e. 100% of one isomer and none of the other), more in particular, compounds or intermediates having a stereoisomeric excess of 90% up to

100%, even more in particular having a stereoisomeric excess of 94% up to 100% and most in particular having a stereoisomeric excess of 97% up to 100%. The terms 'enantiomerically pure' and 'diastereomerically pure' should be understood in a similar way, but then having regard to the enantiomeric excess, and the diastereomeric excess, respectively, of the mixture in question.

Pure stereoisomeric forms of the compounds and intermediates of this invention may be obtained by application of art-known procedures (cf. Advanced Organic Chemistry: 3rd Edition: author J March, pp 104-107). For instance, enantiomers may be separated from each other using known procedures including, for example, formation of diastereomeric mixtures by reaction with a convenient optically active auxiliary species followed by separation of the diastereomers, using for instance selective crystallisation, and finally cleavage of the auxiliary species. Examples of optically active auxiliary species are optically active acids and bases such as tartaric acid, dibenzoyltartaric acid, ditoluoyltartaric acid and camphorsulfonic acid. Alternatively, enantiomers may be separated by chromatographic techniques using chiral stationary phases. Pure stereochemically isomeric forms may also be derived from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction occurs stereospecifϊcally. When a specific stereoisomer of a compound is desired, the compound will preferably be synthesized by stereospecific methods of preparation. These methods will advantageously employ enantiomerically pure starting materials.

With reference to the instances where (R) or (S) is used to designate the absolute configuration of a chiral centre within a substituent, the designation is done taking into consideration the whole compound and not the substituent in isolation.

Where tautomers exist in the compounds of the invention, we disclose all individual tautomeric forms and combinations of these as individual specific embodiments of the invention.

It will be appreciated that the compounds of formula (I) may have metal binding, chelating or complex forming properties and therefore may exist as metal complexes or metal chelates. Such metalated derivatives of the compounds of formula (I) are intended to be included within the scope of the present invention.

The invention relates to the compounds of formula (I) or any subgroup of compounds of formula (I) per se, the prodrugs, JV-oxides, addition salts, quaternary amines, metal complexes, and stereochemically isomeric forms thereof. One embodiment comprises the compounds of formula (I) or any subgroup of compounds of formula (I) specified herein, as well as the iV-oxides, salts, as the possible stereoisomeric forms thereof.

The invention further relates to methods for the preparation of the compounds of formula (I) or any subgroup of compounds of formula (I), the prodrugs, JV-oxides, addition salts, quaternary amines, metal complexes, and stereochemically isomeric forms thereof, its intermediates, and the use of the intermediates in the preparation of the compounds of formula (I) or any subgroup of compounds of formula (I).

The invention also relates to the use of a compound of formula (I) or any subgroup of compounds of formula (I), or a prodrug, iV-oxide, addition salt, quaternary amine, metal complex, or stereochemically isomeric form thereof, for the manufacture of a medicament. Or the invention relates to the use of a of a compound of formula (I) or any subgroup of compounds of formula (I), or a prodrug, iV-oxide, addition salt, quaternary amine, metal complex, or stereochemically isomeric form thereof in therapy.

In the context of the present specification, the term 'therapy' also includes 'prophylaxis' unless there are specific indications to the contrary. The terms 'therapeutic' and 'therapeutically' should be construed accordingly.

The invention further relates to a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula (I) or a compound of any of the subgroups of formula (I) or a pharmaceutically acceptable salt thereof as specified herein, and a pharmaceutically acceptable adjuvant, diluent or carrier for administration to a subject in need thereof. A therapeutically effective amount in this context is an amount sufficient to act in a prophylactic way against, to stabilize or to reduce adverse conditions associated with RAS activity, such as or related to hypertension, heart failure, pulmonary hypertension, renal insufficiency, renal ischemia, in affected subjects or subjects being at risk of being affected.

The invention further relates to a process of preparing a pharmaceutical composition as specified herein, which comprises intimately mixing a pharmaceutically acceptable adjuvant, diluent or carrier with a therapeutically effective amount of a compound of formula (I) or any of the subgroups of formula (I) as specified herein, or a pharmaceutically acceptable salt or a solvate, prodrug, N-oxide, quaternary amine, metal complex or stereochemically isomeric form thereof as specified herein.

The compounds of formula (I) or any of the subgroups of formula (I) have enzyme inhibiting properties and are modulators of the renin-angiotensin system, in particular they are inhibitors of the natural enzyme renin and may be used in the treatment and/or prophylaxis of diseases such as or related to hypertension, congestive heart failure, pulmonary hypertension, renal insufficiency, renal ischemia, renal failure, renal fibrosis, cardiac insufficiency, cardiac hypertrophy, cardiac fibrosis, myocardial ischemia, cardiomyopathy, glomerulonephritis, renal colic, complications resulting from diabetes such as nephropathy, vasculopathy and neuropathy, glaucoma, elevated intra-ocular pressure, atherosclerosis, restenosis post angioplasty, complications following vascular or cardiac surgery, erectile dysfunction, hyperaldosteronism, lung fibrosis, scleroderma, anxiety, cognitive disorders, complications of treatments with immunosuppressive agents, and other diseases known to be related to the renin-angiotensin system. In one embodiment, the invention relates to a method for the treatment and/or prophylaxis of diseases or conditions which are associated with a dysregulation of the renin-angiotensin system, in particular to a method for the treatment or profylaxis of the above mentioned diseases, said method comprising administering to a patient a pharmaceutically active amount of a compound of formula (I) or any of the subgroups of formula (I).

The invention further provides a method of treating a disease or condition known to be related to the renin-angiotensin system which comprises administering to a patient in need thereof a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt, a solvate, prodrug, N-oxide, quaternary amine, metal complex, or stereochemically isomeric form thereof, as hereinbefore defined.

The invention also provides a method of treating diseases or conditions such as or related to the above mentioned (e.g. hypertension) which comprises administering to a patient in need thereof

a therapeutically effective amount of a compound of formula (I) or any of the subgroups of formula (I) or a pharmaceutically acceptable salt, or solvate thereof as hereinbefore defined.

For the above-mentioned therapeutic uses the dosage administered will, of course, vary with the compound employed, the mode of administration, the treatment desired and the disorder indicated. The daily dosage of the compound of formula I/salt/so lvate (active ingredient) may be in the range from 0.001 mg/kg to 75 mg/kg, in particular from 0.5 mg/kg to 30 mg/kg. This daily dose may be given in divided doses as necessary. Typically unit dosage forms will contain about 1 mg to 500 mg of a compound of this invention.

The compounds of formula (I) and pharmaceutically acceptable salts, solvates, prodrugs, TV-oxides, quaternary amines, metal complexes, or stereochemically isomeric forms thereof may be used on their own but will generally be administered in the form of a pharmaceutical composition in which the compound of formula (I) /salt/solvate (active ingredient) is in association with a pharmaceutically acceptable adjuvant, diluent or carrier. Depending on the mode of administration, the pharmaceutical composition will preferably comprise from 0.05 to 99 %w (per cent by weight), more preferably from 0.10 to 70 %w/w, of active ingredient, and, from 1 to 99.95 %w/w, more preferably from 30 to 99.90 %w/w, of a pharmaceutically acceptable adjuvant, diluent or carrier, all percentages by weight being based on total composition. A representative tablet within the scope of the pharmaceutical composition of the invention could have a mass of 500 - 1500 mg with a loading of active ingredient in the range 35 - 75%, with the balance being excipients, such as binders, disintegrants, antioxidants and the like.

The pharmaceutical compositions of this invention may be administered in standard manner for the disease or condition that it is desired to treat, for example by oral, topical, parenteral, buccal, nasal, vaginal or rectal administration or by inhalation. For these purposes the compounds of this invention may be formulated by means known in the art into the form of, for example, tablets, capsules, aqueous or oily solutions, suspensions, emulsions, creams, ointments, gels, nasal sprays, suppositories, finely divided powders or aerosols for inhalation, and for parenteral use (including intravenous, intramuscular or infusion) sterile aqueous or oily solutions or suspensions or sterile emulsions. The oral delivery route, particularly capsules or tablets is favoured.

In addition to the compounds of the present invention the pharmaceutical composition of this invention may also contain, or be co- administered (simultaneously or sequentially) with, one or more pharmacological agents of value in treating one or more of the diseases or conditions referred to hereinabove. A representative example are other pharmacologically active compounds such as ACE-inhibitors, neutral endopeptidase inhibitors, aldosterone antagonists, angiotensin II receptor antagonists, endothelin receptors antagonists, vasodilators, calcium antagonists, potassium activators, diuretics, sympatholitics, beta- adrenergic antagonists, alpha- adrenergic antagonists and/or other drugs beneficial for the prevention or the treatment of the above-mentioned diseases such as 1 ibeta-hydroxy steroid dehydrogenase type 1 inhibitors and soluble guanylate cyclase activators.

General synthetic methodology

The compounds of the present invention and intermediates useful for the synthesis of these compounds are prepared by methods and techniques known to those skilled in the art. The general schemes below illustrate typical synthetic routes to the compounds of the invention and to intermediates thereof. Alternative routes, which will be readily apparent to the ordinary skilled organic chemist, may alternatively be used to synthesize various portions of the molecules as illustrated by the general schemes and the preparative examples below.

Scheme 1 illustrates a synthetic route to a lactone which is a useful intermediate in the preparation of compounds of formula (I).

Scheme 1

The isopropylidene derivative (Ia) achieved for example as described in Tetrahedron Lett., 1987, 28, 1143, can be transferred into the methyl glucoside (Ib) by acidic hydrolysis of the acetal group effected by treatment with a suitable acid such as sulphuric acid, in the presence of methanol. The achieved free secondary hydroxy group can then be reductively removed effected for instance by transformation of the hydroxy group into a thiocarbonyl group by reaction with thiocarbonyl diimidazole (TCDI), followed by reduction of the formed thiocarbonyl group using for instance conditions such as tributyltin hydride the presence of a radical initiator like azobis- (2-methylpropyonitrile) (AIBN) or the like to give the 2,3-dideoxy glucoside (Ic). Oxidative cleavage of the methyl ether performed for example by oxidation with m-chloroperbensoesyra or the like in the presence of BF3-etherate, gives the lactone (Id). The ring substituent R ³ can then be introduced for example by treatment of the lactone with a base such as LDA or equivalent followed by reaction with a suitable alkylating agent such as an alkyl halide like an alkyl bromide or alkyl iodide or a derivative of sulphonic acid such as a mesylate, triflate or tosylate or the like, thus providing the α-alkylated lactone (Ie). Removal of the benzyl groups using any suitable conditions well known to the skilled person, such as catalytic hydrogenation, then provides the diol (If).

Compounds of formula (I) wherein Z is O can then be achieved from the diol If as generally depicted in scheme IA.

1Ad

Scheme IA

The primary hydroxy group of the lactone (If) can be selectively alkylated for example by activation of the hydroxy group with dibutyltinoxide followed by reaction with a desired

alkylating agent Q-(CH ₂ ) _D -Lg wherein Lg is a suitable leaving group such as a halide like bromide or iodide in the presence of tetrabutylammonium bromide or the like thus forming the ether derivative (IAa). Alternatively, the substituent Q-(CH ₂ )D can be introduced by using the Mitsunobu conditions (Mitsunobu, 1981, Synthesis, January, 1-28; Rano et al., Tetrahedron Lett, 1995, 36, 22, 3779-3792; Krchnak et al., Tetrahedron Lett, 1995, 36, 5, 6193-6196; Richter et al., Tetrahedron Lett., 1994, 35, 27, 4705-4706) i.e. reaction of the primary hydroxy group of the lactone (If) with an azodicarboxylate such as DIAD or the like in the presence of triphenylphosphine followed by displacement with a desired alcohol.

Replacement of the secondary hydroxy group of the alcohol (IAa) by azide may be effected by transforming the hydroxy group to a leaving group, for example a derivative of sulphonic acid like a triflate or tosylate or the like by subjecting the alcohol to sulphonylating conditions such as treatment with the appropriate anhydride or halide optionally in the presence of a base, for instance pyridine, followed by displacement of the leaving group with azide for example sodium azide, thus giving the azide derivative (IAb). Alternatively, the azide moiety can be introduced by treatment of the alcohol with diisopropyl azodicarboxylate or any other suitable azodicarboxylate, in combination with triphenylphosphine, followed by reaction with azide for example DPPA, in a solvent like THF. The linear compound (IAd) is then be achieved by opening of the lactone with a desired nucleophile (IAc) as described herein below.

Compounds of formula (I) wherein Z is O and Y is NH can be achieved by opening of the lactone (If) with an amine as shown in scheme 2.

2c 2d

Scheme 2

Opening of the lactone (IAb) with a desired amino derivative (2a) in the presence of a coupling agent for example 2-hydroxypyridine and a base like isopropyl diethylamine provides the linear compound (2b). Reduction of the azide moiety using conditions compatible with the (CH ₂ )D-Q group, for example by treatment with Ph ₃ P or by hydrogenation at atmospheric pressure in the presence of Lindlar Catalyst or equivalent then provides the corresponding amino derivative (2c). If desired, the afforded primary amine can be substituted effected by way of any convenient method for example by reaction with an alkylating agent R ¹ -Lg wherein Lg is a leaving group such a halide like chloride, bromide or iodide or a derivative of sulphonic acid such as a mesylate or a triflate or the like, optionally in the presence of a base, or alternatively an N- substituent may be introduced by reaction with an aldehyde R ¹ Q=O)H in a reductive amination reaction using any suitable reductive agent such as NaBH ₃ CN or the like.

Compounds of formula (I) wherein Z is O, Y is O or S and q is 0 may be prepared as shown in scheme 2A.

group

Scheme 2 A

Opening of the lactone (IAb) with hydroxide such as lithium hydroxide, provides the linear hydroxy acid (2Aa). Protection of the afforded secondary alcohol using any suitable hydroxy protecting group known in the art, followed by a Mitsunobu reaction with a desired alcohol HO- (CH ₂ ) _m -W or thiol HS-(CH ₂ ) _m -W and finally removal of the hydroxy protecting group, provides compounds esters and thioesters (2Ac) which subsequently can be reduced to the corresponding

amine and optionally N-alkylated as described above to yield compounds of formula (I) wherein Y is O or S respectively and q is 0.

Compounds of formula (I) wherein Z is O, Y is CH and q is 0 may be prepared as indicated in scheme 2B.

Scheme 2B

Opening of the lactone IAb with a suitable organo lithium or Grignard reagent provides the keto derivative (2Ba). Subsequent reduction of the azide moiety optionally followed by alkylation of the afforded amine then gives compounds of formula (I) wherein Y is CH and q is 0.

Amino derivatives (2a) to be used for the opening of the lactone (IAb) in scheme 2 are available commercially or they can easily be prepared by the skilled person according to literature procedures. For the preparation of compounds of the invention wherein q is 1 and Y is NH, an appropriate amine can be prepared for example as illustrated in scheme 3.

Scheme 3

The amino acid (3a), carrying the desired side chain R > 4'r R _> 4" , can be coupled to an amine W- (CH ₂ )Hi-NH ₂ using any convenient method for peptide coupling known in the art. For example, a coupling agent like HATU or isobutylchloro formate in the presence of a tertiary amine such as ethyldiisopropylamine (DIEA) or N-methylmorpholine in a solvent like dimethyl formamide can be used, thus providing the amide. Subsequent reduction of the carbonyl group using for

example Red- Al or any other suitable reducing agent followed by removal of the amino protecting group using the appropriate conditions according to the protecting group used, such as acidic treatment in the case of a Boc-group, provides the amine (3b).

Alternatively, the amine 3b can be achieved by coupling of a suitably amino protected amino alcohol (3a), carrying the desired substituents R ⁴ and R ⁴ , to the amine W-(CH ₂ )Hi-NH ₂ using for example the Mitsunobu conditions, or the amino alcohol group can be oxidized to the corresponding amino aldehyde by treatment with a suitable oxidation agent such as Dess Martin periodinane or the like, followed by reaction with the amine W-(CH ₂ ) _m -NH2 in a reductive amination reaction, thus affording the amine (3b). Subsequent removal of the N-protecting provides the amine (3b).

Similarly, amino derivatives useful for the preparation of compounds of formula (I) wherein Y is

0 or S, and q is 1 can for example be prepared by using an alcohol H0-(CH ₂ ) _m -W or thiol HS- CH ₂ ) _m -W respectively in the Mitsunobu reaction with the amino alcohol 3 a.

Compounds of formula (I) wherein Y is NHNH and q is 0 or 1 , are conveniently prepared according to the method outlined in scheme IA by using a suitable hydrazine or amino hydrazine derivative respectively in the opening of the lactone IAb. Suitable hydrazine or amino hydrazine derivatives are commercially available or they can be prepared according literature procedures. A route to amino hydrazines useful for the preparation of compounds of formula (I) wherein q is

1 and Y is NHNH is illustrated in scheme 3A.

H

Scheme 3A

Reaction of a suitably amino protected, for example boc protected, amino alcohol (3Aa) carrying the desired substituents R ⁴ and R ⁴ with a desired hydrazine derivative (3Ab) under Mitsunobu conditions i.e. treatment with an azodicarboxylate such as DIAD or the like in the presence of triphenylphosphine, provides the N,N'-dialkylated alkylated hydrazine derivative. Alternatively the amino alcohol (3Aa) can be oxidized to the corresponding amino aldehyde by treatment with a suitable oxidation agent such as Dess Martin periodinane or the like, followed by reaction with

the hydrazine derivative (3Ab) in a reductive amination reaction, to give the N,N'-dialkylated alkylated hydrazine derivative. Removal of the amino protecting group using the appropriate conditions according to the protecting group used, such as acidic treatment in the case of a boc group, then gives the hydrazine containing amine (3Ac).

A route to an amine useful for the preparation of compounds of formula (I) wherein Y is CH ₂ NH and q is 1 is illustrated in scheme 3B.

Scheme 3 B

Reduction of a suitably N-protected β-amino acid (3Ba) using any convenient method such as treatment with LiAlH ₄ or the like affords the corresponding amino alcohol (3Bb). Reaction of the afforded alcohol with a desired amino derivative, H ₂ N-(CH ₂ )Hi-W, under Mitsunobu conditions, followed by removal of the amino protecting group provides the amine (3Bc).

An intermediate lactone useful for the preparation of compounds of formula (I) wherein the group Q is bound directly to the oxygen atom, i.e. Z is O and n is 0, can be prepared as shown in scheme 4.

as above

4c

Scheme 4

Treatment of the diol (If) with triphenylphosphine and an azodicarboxylate for example DIAD provides the epoxide (4a). Opening of the epoxide with a desired nucleophile Q-OH in the presence of a base, such as potassium carbonate or the like, provides the ether derivative (4b). Subsequent replacement of the secondary hydroxy group with azide, opening of the lactone and finally reduction of the azide as described above, provides the linear amine (4c).

Lactones useful for the synthesis of compounds of formula (I) wherein Z is S or NH and n is 1 or 2, can be prepared from the diol If for example by a Mitsunobu reaction with a thiol or amino derivative respectively, as illustrated in scheme 5.

Scheme 5

The primary hydroxy group of the lactone (If) can be converted to a thioether or an amine for example by transforming it into a leaving group followed by displacement of the formed leaving group with the desired group Q-(CH ₂ )D-S or Q-(CH ₂ )D-NRa. A convenient method to effect this transformation is by way of a Mitsunobu reaction, i.e. reaction of the hydroxy group with an azodicarboxylate such as DIAD or the like in the presence of triphenylphosphine or the like followed by displacement with a desired thiol or amine to provide the thioether (5a) or the amine derivative (5b) respectively. Alternatively, the amine (5b) may be achieved by using an azide derivative, such as sodium azide or DPPA in the Mitsunobu reaction with the alcohol (If), followed by reduction of the azide to the primary amine and subsequent alkylation of the amine with a suitable alkylating agent Q-CH ₂ -Lg, or by performing a reductive amination with a suitable aldehyde Q-CH(=O). A further alternative to obtain the amino derivative (5b) is to

selectively oxidize the primary hydroxy group of the alcohol (If) to the corresponding aldehyde, effected for example by treatment with Dess-Martin periodinane or by any other suitable oxidation reagent, followed by a reductive amination with the desired amino derivative Q- (CH ₂ ) _D -NHRa in the presence of a reducing agent like NaCNBH ₃ . Replacement of the secondary hydroxy group with azide, opening of the lactone and finally reduction of the azide as described above, then provides the linear compounds (5 c and 5d).

Compounds wherein the group Q is linked directly to a sulphur or nitrogen atom, i.e. an intermediate for the preparation of compounds of formula (I) wherein Z is S or NRa and n is 0, may be prepared by transformation of the primary hydroxy group of the diol (If) into a leaving group such as a derivative of sulphonic acid like a mesylate, triflate, tosylate or the like by treatment with the appropriate sulphonylating agent in a solvent like for instance pyridine or dichloromethane optionally in the presence of triethylamine or the like, followed by displacement of the leaving group with a desired thiol Q-SH or a amine Q-NHRa optionally in the presence of a base. An alternative method for the preparation of compounds wherein Z is S and n is 0 is to react the diol (If) with a desired diphenyl disulphide derivative in the presence of nBu3P. Compounds wherein Z is NRa and n is 0 may alternatively be achieved by oxidation of the primary hydroxy group of the diol (If) followed by a reductive amination with a desired aniline derivative Q-NRa in the presence of a suitable catalyst like NaCNBH ₄ or the like.

Compounds of formula (I) wherein Z is a sulphone i.e. S(=O) ₂ may be obtained by oxidation of the sulphur of corresponding thioether derivative. The oxidation can be performed either at the last step of the synthesis or on any suitable intermediate. Many suitable methods for this oxidation are described in the literature for example, a peroxyacid such as AcOOH, mCPBA can be used.

An alternative route to compounds of formula (I) which can be valuable for example when it is desired to introduce various groups Q-(CH ₂ ) _n at a late stage of the synthesis, is shown in scheme 6.

6a 6b

deprot. OH, SH or NHRa

HoN orthogonal protecting groups 6e

Scheme 6

The azide derivative (6a), prepared for example as outlined in e.g. scheme IA, wherein Pg ¹ is a hydroxy protecting group for example a benzyl group can be transformed to the corresponding amine by reduction of the azide group using any convenient reduction method such as hydrogenation in the presence of a suitable catalyst, such as Lindlars catalyst or the like in the presence Of BoC ₂ O to provide the boc protected derivative (6b). Protection of the secondary hydroxy group, using a protecting group (Pg ² ) which is orthogonal to the one used for the primary hydroxy group (Pg ¹ ), followed by removal of the primary hydroxy protecting group using the appropriate conditions according to the group used, such as for example catalytic hydrogenation in the case of a benzyl group, provides the primary alcohol (6c). Suitable protecting groups for the above route will be recognized by the skilled person and a numerous of useful protecting groups are described in Greene "Protective Groups in Organic Synthesis", John Wiley and sons, New York (1981). For example benzyl can be used as Pg ¹ and acetyl as Pg ² . The group (CH ₂ ) _n -Q can then be introduced as described above. For example, compounds wherein Z' is O and n is 1, can be prepared by reaction of the primary alcohol (6c) with an alkylating agent Q-(CH ₂ ) _n -Lg wherein Lg is a halide like a bromide, chloride or iodide or a derivative of sulphonic acid such as a triflate or mesylate or the like in the presence of a base like NaH or the like, or a trichloroacetimidate of a desired group, Q-(CH ₂ ) _n -O-C(=NH)Cl3 may be reacted with

the primary alcohol (6c) in the in the presence of a Lewis acid such as BFsOEt ₂ . Trichloroacetimidates are conveniently prepared by reaction of the corresponding alcohol with trichloroacetonitrile in the presence of a base like NaH.

Compounds wherein Z' is O, S or NRa may be prepared by a Mitsunobu reaction of the primary alcohol (6c) with a desired alcohol, Q-(CH ₂ ) _n -OH, thiol, Q-(CH ₂ ) _n -SH or amine Q-(CH ₂ ) _n - NHRa respectively as described above. Compounds wherein n is 0 and Z is O, S or N may be prepared by transforming the primary hydroxy group of the alcohol (6c) to a leaving group such as a halide like chloride or bromide or to a derivative of sulphonic acid such as a triflate, tosylate or the like which subsequently is displaced by a desired alcohol Q-OH, thiol Q-SH or amine Q- NHRa optionally in the presence of a base. An alternative method for the preparation of compounds wherein Z' is S and n is O is to react the diol (If) with a desired diphenyl disulphide derivative in the presence of nBusP. Compounds wherein Z' is NRa and n is O may alternatively be achieved by oxidation of the primary hydroxy group of the alcohol (6c) followed by a reductive amination with a desired aniline derivative Q-NRa in the presence of a suitable catalyst like NaCNBH ₄ or the like.

Substituted phenyl and heteroaryl derivatives Q-(CH ₂ ) _n - used in the schemes above are commercially available or they may be prepared according to literature procedures. A method to prepare a substituted phenyl derivative useful for the preparation of compounds of formula (I) wherein Q is phenyl substituted with aminomethyl or amidomethyl and derivatives thereof is illustrated in scheme 7.

7c 7d

Scheme 7

The hydroxy protected cyanobenzyl derivative (7a) can be prepared by protection of commercially available cyanobenzyl alcohol, illustrated herein as 3 -cyanobenzyl alcohol, with a suitable protecting group, for example a trityl or monomethoxy trityl group using standard conditions well known in the art. Subsequent alkylation of the cyano group effected for example by way of an organo metallic reaction such as a Grignard reaction or an organo lithium reaction or the like, using the suitable conditions such as treatment with the desired alkyl magnesium halide in an ethereal solvent like diethyl ether or THF or the like followed by reduction of the intermediate imine for example by LAH and finally protection of the afforded amine by treatment with the suitable agent such as di-t-butyldicarbonate, provides the carbamate (7b). The corresponding carbamate wherein the benzylic carbon is unsubstituted, i.e. R ⁵ is H can be achieved by direct reduction of the cyano group to the methyl amino group using for instance LAH or diborane or the like, followed by protection of the afforded primary amine as previously described. Removal of the hydroxy protecting group using standard conditions such as treatment with acid in the case of trityl or monomethoxy trityl group provides the alcohol (7c). The afforded alcohol (7c) can then be used in the coupling to the primary alcohol of the lactone Ig or the linear compound 4c employing for example the Mitsunobu conditions as described in scheme 2 and 4 respectively. Alternatively, the hydroxy group of the alcohol (7c) can be transformed into a leaving group such as a bromide for example by treatment with bromine or carbontetrabromide in the presence of triphenylphosphine or the like thus affording the bromoderivative (7d), or the hydroxy group can be transformed into a derivative of sulphonic acid by reaction with a suitable sulphonylating agent such as a sulphonic halide or anhydride optionally in the presence of a base for example pyridine. Subsequently, the afforded compound can be coupled to the primary alcohol of the lactone If or the linear compound 6c as described in scheme 2 and 6 respectively.

Scheme 8 illustrates an example to another substituted phenyl derivative, useful for the preparation of compounds of formula (I) wherein Q is phenyl substituted with an alkoxy-alkoxy group.

Scheme 8

Alkylation of the phenolic hydroxy group of ester (8a) using for example the Mitsunobu, such as in the presence OfPh ₃ P, an azodicarboxylate like DIAD and the suitable alcohol followed by reduction of the ester function using any convenient reduction method known in the art provides benzylic alcohol (8b). The afforded alcohol (8b) can then either be used directly in the coupling to the primary hydroxyl group of the lactone If or the linear compound 6c employing the Mitsunobu conditions, or the a hydroxy group can be transferred to a leaving group, such as a halide like bromide, and subsequently coupled to the primary hydroxyl group of the lactone If or the linear compound 6c as described above.

Even though the strategy in scheme 8 is illustrates the introduction of a methoxy-ethoxy substituent to the phenyl ring, the skilled person will easily realise that the same methodology can be applied for the introduction of other O-linked substituents, such as substituents with other chain lengths. Furthermore, despite the fact that scheme 7 and 8 are illustrated with a 1,3 substituted phenyl derivative as starting compound, the skilled person will realise that the same methodology is also applicable to other phenyl derivatives, for example the corresponding 1,2- or 1 ,4-disubstituted derivatives.

Scheme 9 shows an alternative route to compounds of the invention, starting from Garner's aldehyde.

9h

Scheme 9

α-Alkylation of the aldehyde (9a) by reaction with a suitable ester of propiolic acid, for example the methyl ester, in the presence of a base like buthyllithium followed by reduction of the triple bond for example by catalytic hydrogenation using a catalyst like palladium on carbon provides the alcohol (9c). Heating of the afforded hydroxy ester in the presence of acetic acid effects the ring closing and thus affords lactone (9d). The afforded lactone can then be alkylated at the α- carbon with a desired group R as described above, i.e. by treatment of the lactone with a base such as LDA optionally followed by addition of tripyrrolidine phosphorus oxide and finally addition of the alkylating agent or by any other suitable alkylation method which provides the alkylated lactone (9e). Cleavage of the cyclic aminal by treatment with acid such as TFA followed by reaction with BoC ₂ O in order to reprotect the amino function affords the primary alcohol (9e). The group Q-(CH ₂ ) _n can then be introduced using any suitable method such as any of those described above. For example, a trichloroimidate of the desired group Q-(CH ₂ ) _n in the

presence of TMS triflate will provide the ether derivative (9f) i.e. Z' is O. The lactone may then be opened either directly with a desired amine as described above to give the amide (9i), or alternatively, the lactone may be opened by treatment with hydroxide such as lithium hydroxide, thus affording the acid (9g). Protection of the hydroxy group, using any conventional protecting group for example a silyl group such as a tert-butyl dimethylsilyl group, followed by coupling of the acid to a suitable amine using standard peptide coupling conditions such as using a coupling agent like EDAC in the presence of HOBt and a tertiary amine like triethylamine, and finally removal of the hydroxy protecting group provide the amide (9h).

An intermediate towards compounds of formula (I) wherein X' is F and X" is H or X' and X" are both F can be prepared by replacement of the hydroxy group of compound 4a with fluoro or difluoro as exemplified in scheme 10 illustrated with Q as a phenyl group and Z as oxygen.

6a

10a

1 ) DIAD, Ph ₃ P, p-NO ₂ -benzoic acid

2) NaOMe

Scheme 10

The hydroxy group of compound (6a) can be replaced by two fluoro atoms by oxidizing the hydroxy group to a keto group using any convenient method such as using a reagent like Dess Martin periodinane or oxone® (potassium monopersulphate triple salt) or any other suitable oxidizing agent followed by treatment of the afforded keto compound with a fluorinating agent like DAST or Deoxofluor or the like in a solvent like dichloromethane as described e.g. by Singh, R. P. and Shreve, J. M. in Synthesis, 17, 1999, p. 2561-2578, or any other suitable

fluorinating conditions, to yield the difluoro compound (10a). To achieve the monofluoro compound (10c) with the desired stereochemistry, the stereochemistry at the steric centre whereto the hydroxy group is attached first has to be inverted. The inversion can be achieved for example by subjecting the alcohol to a Mitsunobu reaction with for instance p-nitrobenzoic acid and an azodicarboxylate like DIAD in the presence OfPh ₃ P followed by hydrolysis of the afforded p-nitrobenzoic ester by treatment with sodium methoxide or the like. The afforded inverted alcohol (1 Ib) is then subjected to fluorinating conditions such as treatment with DAST or Deoxofluor as described above.

Even though scheme 10 illustrates the replacement of the hydroxy group with fluoro or difluoro as the last step of the synthesis, the skilled person will realise that this transformation alternatively may be performed at any other suitable stage of the synthesis for example on any of the intermediates described above.

Compounds according to the invention wherein X' is amino may be prepared from the corresponding alcohol by replacement of the hydroxy group with an amino group, performed either as the last step of the synthesis or at any other convenient stage. A variety of methods for this transformation are described in the literature. An example is shown in scheme 11 wherein the hydroxy group is transformed into a leaving group and subsequently displaced by azide.

11a 11b

11b 11d

R1' is R1 or an N-protecting group

Scheme 11

In order to get the desired configuration at the steric centre whereto the X' is attached in the final compound, the configuration of compound (1 Ia), prepared as described above, has to be inverted, for example as described in scheme 10. The inverted alcohol (1 Ib) can then be subjected to Mitsunobu conditions, i.e. treatment with an azodicarboxylate such as DIAD or the like in the presence of triphenylphosphine followed by reaction with azide, for example diphenylphosphoryl azide (DPPA) or HN3 to give the azido derivative (1 Ic) Alternatively the azido derivative (1 Ic) can be achieved by transformation of the hydroxy group to a derivative of sulphonic acid like a mesylate, triflate, tosylate or the like by treatment with the appropriate sulphonylating agent in a solvent like for instance pyridine or dichloro methane optionally in the presence of triethylamine or the like, followed by displacement of the leaving group with sodium azide or the like. Reduction of the azide using any conventional reduction method such as hydrogenation in the presence of a suitable catalyst, or treatment with triphenylphosphine provides the corresponding amine (l id)

Even though scheme 11 illustrates the conversion of the hydroxy group to an amine as the last step of the synthesis, the skilled person will realise that this transformation is also applicable at any other suitable stage of the synthesis for example on any of the intermediates described above.

Any functional groups present on any of the constituent compounds used in the preparation of the compounds of the invention are appropriately protected where necessary. For example functionalities on the natural or non-natural amino acids are typically protected as is appropriate in peptide synthesis. Those skilled in the art will appreciate that the selection and use of appropriate protecting groups depend upon the reaction conditions. Suitable protecting groups are described in Greene, "Protective Groups in Organic Synthesis", John Wiley & Sons, New York (1981) and "The Peptides: Analysis, Synthesis, Biology", Vol. 3, Academic Press, New York (1981), the disclosure of which are hereby incorporated by reference.

Detailed Description

Various embodiments of the compounds of the invention including end compound inhibitors and key intermediates towards such compounds will now be described by way of illustration only with reference to the following non-limiting examples. Note that the exemplified intermediates,

such as the substituted lactone building blocks are readily opened with alternative amines to form additional compounds of the invention.

Example 1 Step a

5-(l,2-Bis-benzyloxyethyl)-2-methoxytetrahydrofuran-3-ol (Ia)

Sulfuric acid (13 mL, 240 mmol) was added dropwise to a solution of 5-(l,2-bis-benzyloxy- ethyl)-2,2-dimethyl-tetrahydro-furo[2,3-d][l,3]dioxole (prepared as described by Hanessian et al in Tetrahedron Lett., 1987, 28, 1142) (30.2 g, 78.6 mmol) in MeOH (180 mL) at 0 ⁰ C. The reaction mixture was then stirred at room temperature for 2 h, cooled down to 0 ⁰ C and carefully neutralized with sat. aq. NaHCOs. Methanol was distilled off under reduced pressure and the residue was taken into EtOAcZH ₂ O. The phases were separated and the aqueous phase was extracted with EtOAc. The combined organic extracts were dried (Na ₂ SO ₄ ) and the solvent evaporated to give the crude product as a yellow oil. Purification by flash chromatography on silica gel (Hex:EtOAc 10:1 to 2.5:1) afforded 18.7 g (67%) of the title compound as a colourless oil (mixture of anomers).

Step b

Imidazo Ie- 1 -carbothioic acid O- [5 -( 1 ,2-bis-benzy loxyethyl)-2-methoxytetrahydrofuran-3 -yl] ester (Ib)

A solution of the alcohol Ia (14.2 g, 39.7 mmol) and l,l '-thiocarbonyldiimidazole (7.1 g, 39.8 mmol) in THF (120 mL) was refluxed under N ₂ for 2 h. The reaction mixture was concentrated under vacuum and the residue was purified by flash chromatography on silica gel (Hex:EtOAc 2.5:1) which gave the title compound (15.8 g, 85%) (mixture of anomers).

Step c

2-(l,2-Bis-benzyloxyethyl)-5-methoxytetrahydrofuran (Ic)

A solution of compound Ib (15.8 g, 33.8 mmol) and AIBN (0.55 g, 3.3 mmol) in toluene was heated to 100 ⁰ C under N ₂ . Tributyltinhydride (13.6 rnL, 50.6 mmol) was then added carefully dropwise (gas evolution). The reaction mixture was stirred further 20 min at 100 ⁰ C after the addition was completed. Evaporation of the solvent afforded a syrup that was purified by flash chromatography on silica gel (Hex:EtOAc 20:1 to 2.5:1) which gave the title compound (8.2 g, 71%) (mixture of anomers).

Step d

5-(l,2-Bis-benzyloxyethyl)-dihydrofuran-2-one (Id)

A solution of compound Ic (10.7 g, 31.29 mmol) and m-CPBA (19.6 g, 62.5 mmol) in dichloromethane (10 mL) was treated with BFsOEt ₂ (2 mL, 15.8 mmol) at 0 ⁰ C. The reaction mixture was then allowed to warm up to room temperature and stirring was continued for 17 h. Evaporation of the solvent afforded the crude product that was purified by flash chromatography on silica gel (Hex:EtOAc 20:1 to 2.5:1) which gave the title compound (6.1 g, 60%) as a colorless oil.

Step e

5 -( 1 ,2-Bis-benzyloxyethyl)-3 -isopropyldihydrofuran-2-one ( 1 e)

A solution of compound Id (3.1 g, 9.51 mmol) in THF (30 mL) was added to a 1 M solution of

LDA in THF (20 mL, 20 mmol) at -60 ⁰ C under N ₂ . The reaction mixture was then treated with

tris(pyrrolidinophosphine) oxide (15 niL) followed by iodopropane (3.8 niL, 38.1 mmol). Stirring at -60 ⁰ C was continued for 1 h and the reaction mixture was quenched with aq. NH ₄ Cl. The phases were separated and the aqueous phase was extracted with ¹ BuOMe. The combined organic extracts were dried (Na ₂ SO ₄ ) and the solvent evaporated. The residue was purified by flash chromatography on silica gel (Hex:EtOAc 10:1) which gave the title compound (1.2 g, 34%) as a colourless oil.

Step f

5 -( 1 ,2-Dihydroxyethyl)-3 -isopropyldihydrofuran-2-one (If)

A solution of compound Ie (2.58 g, 7.01 mmol) in MeOH (50 mL) was hydrogenated in the presence of 10% Pd/C at 3 bar H ₂ pressure. Filtration of the catalyst and evaporation of the solvent afforded the title compound (1.2 g, 92%) as a white solid.

Step g

5-(l-Hydroxy-2-benzyloxyethyl)-3-isopropyl-dihydrofuran-2 -one (Ig)

A solution of compound If (0.6 g, 3.19 mmol) and Bu ₂ SnO (1.0 g, 4.0 mmol) in toluene (50 mL) was refluxed with a Dean-Stark trap for 4 h. The reaction mixture was cooled down to 90 ⁰ C and benzyl bromide (0.43 mL, 3.66 mmol) and Bu ₄ NBr (1.18 g, 3.66 mmol) were added. Stirred further 22 h at 90 ⁰ C, concentrated under vacuum and the residue purified by flash chromatography on silica gel (Hex:EtOAc 10:1 to 2.5:1) to give the title compound (0.7 g) as a colorless oil.

Step h

5-(l-Azido-2-benzyloxyethyl)-3-isopropyldihydrofuran-2-on e (Ih)

A solution of compound Ig (278 mg, 1 mmol) and pyridine (140 μL, 1.7 mmol) in CH ₂ Cl ₂ (5 rnL) was treated with Tf ₂ O (220 μL, 1.3 mmol) at room temperature Stirred for 30 min and quenched with H ₂ O. The product was extracted into dichloromethane (DCM) and the combined extracts were dried (Na ₂ SO ₄ ) and the solvent evaporated to give a yellow oil that was dissolved in DMF (1 mL). Sodium azide (325 mg, 5 mmol) was added and the resulting suspension was stirred at 60 ⁰ C for 1 h. The reaction mixture was concentrated under vacuum and the residue was purified by flash chromatography on silica gel (Hex:EtOAc 2.5:1) which gave the title

Example 2 Step a

3-Hydroxy-4-methyl-benzoic acid methyl ester (2a)

Thionyl chloride (96 μl, 1.31 mmol) was carefully added to a solution of 3-hydroxy-4- methylbenzoic acid (200 mg, 1.31 mmol) in MeOH (2 ml). The resulting solution was then heated at 45 ⁰ C for 17 h. Evaporation of the solvent afforded a white solid that was purified by flash chromatography on silica gel (Hex:EtOAc 10:1) to give 168 mg (86%) of the title compound.

Step b

3-(3-Methoxy-propoxy)-4-methyl-benzoic acid methyl ester (2b)

Azodicarboxylic acid dipiperidide (505 mg, 2 mmol) and PPh ₃ (524 mg, 2 mmol) were added to a solution of 23a (166 mg, 1 mmol) and 3-methoxypropan-l-ol (95.7 μl, 1 mmol) in CH ₂ Cl ₂ (10

ml) and the resulting mixture was stirred under N ₂ for 17 h. The reaction mixture was then concentrated under vacuum and the residue was taken into EtOAc and washed with 10% aq. citric acid and brine. The organic phase was dried (Na ₂ SO ₄ ) and the solvent evaporated to give the crude product that was purified by flash chromatography on silica gel (Hex:EtOAc 10:1) to give 96 mg (40%) of the title compound.

Step c

[3 -(3 -Methoxy-propoxy)-4-methyl-phenyl] -methano 1 (2c)

Diisobutylaluminum hydride 1.0 M solution in hexanes (2.5 mL, 2.5 mmol) was added carefully to 2b (96 mg, 0.403 mmol) under N ₂ . The resulting solution was stirred 30 min and then carefully quenched with 1 M HCl. The product was extracted into EtOAc, and the combined organic extracts were dried (Na ₂ SO ₄ ). Evaporation of the solvent afforded 84 mg (100%) of the title compound.

Step d

4-Bromomethyl-2-(3-methoxy-propoxy)- 1 -methyl-benzene (2d)

Bromine (24 μl, 0.48 mmol) was added to a solution of PPh ₃ (126 mg, 0.48 mmol) in CH ₂ Cl ₂ (2 ml). The resulting solution was stirred for 5 min and then 2c (84 mg, 0.4 mmol) was added at once. The reaction mixture was stirred for 20 min and then concentrated under vacuum. The residue was purified by flash chromatography (Hex:EtOAc 35:1) to give 68 mg (52%) of the title compound as a white solid.

Step e

5- { 1 -Hydroxy-2-[3-(2-methoxy-ethoxy)-4-methyl-benzyloxyl-ethyl| -3-isopropyl-dihydro-furan- 2-one (2e)

The procedure described in Example 1 step g was followed but using 2d instead of benzyl bromide, which gave the title compound (56% yield).

5- { 1 -Azido-2-[3-(2-methoxy-ethoxy)-4-methyl-benzyloxyl-ethyU -3-isopropyl-dihydro-furan-2- one (2f)

Compound 2e was reacted according to the procedure described in Example 1 step g, which afforded the title compound (93%).

Ster

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid butylamide (2g)

A solution of 2f (28 mg, 0.069 mmol) in butylamine (1 mL, 10.1 mmol) was heated in a microwave cavity at 110 ⁰ C for 45 min. Evaporation of the volatiles under vacuum afforded the title compound (33 mg, 100%).

Step h

5 - Amino-4-hydroxy-2-isopropyl-6- [3 -(3 -methoxy-propoxy)-4-methyl-benzyl oxyl-hexanoic acid butylamide (2h)

A solution of the azide 2g in MeOH was hydrogenated in the presence of Pd Lindlar. The catalyst was filtered off and the solvent evaporated . The residue was purified by preparative HPLC (20% acetonitrile in H ₂ O with 0.1% TFA to 60% acetonitrile) which gave the title compound (97%) as its TFA salt.

Example 3 Step a

(3-Amino-phenyl)-acetic acid methyl ester (3a)

Thionyl chloride (4.8 mL, 66.16 mmol) was carefully added drop wise to MeOH (100 mL) at 0 ⁰ C. To the resulting solution was added a suspension of 3-amino-phenylacetic acid (5.0 g, 33.08 mmol) in MeOH (100 mL) and the reaction mixture was refluxed for 2 h. Then MeOH was distilled off and the residue was partitioned between EtOAc and sat. aq. NaHCO ₃ . The phases were separated and the organic phase was washed with sat. aq. NaHCO ₃ , dried (Na ₂ SO ₄ ) and evaporated which gave the title compound (4.77, 87%).

(3-Methanesulfonylamino-phenyl)-acetic acid methyl ester (3b)

Methanesulfonyl chloride (0.47 rnL, 6.07 mmol) was added to a solution of the amine 2i (Ig,

6.06 mmol) and pyridine (1 mL, 12.4 mmol) in CH ₂ Cl ₂ (6 mL) at 5 ⁰ C. The reaction mixture was then stirred at r.t. for 17 h. The reaction mixture was concentrated under vacuum and purified by flash chromatography on silica gel (Hep:EtOAc 2.5:1) which gave the title compound (1.1 g,

75%).

Step c

[3-(Methanesulfonyl-methyl-amino)-phenyll-acetic acid methyl ester (3 c) A suspension of compound 2j (1.1 g, 4.52 mmol), K ₂ CO ₃ (6.2 g, 45.2 mmol) and methyl iodide (0.84 mL, 13.56 mmol) in acetonitrile (50 mL) was stirred at room temperature for 17 h. The reaction mixture was then concentrated under vacuum and the residue partitioned between CH ₂ Cl ₂ and water. The organic phase was dried (Na ₂ SO ₄ ) and evaporated which gave the title compound (1.13 g, 100%).

Step d

N-Methyl-N- [3 -(2-oxo-ethyl)-phenyll -methanesulfonamide (3 d)

A 1.0 M solution of DIBAL-H in hexanes (4.3 mL, 4.3 mmol) was added dropwise to a solution of the ester 2k (1.1 g, 4.28 mmol) in CH ₂ Cl ₂ (20 mL) at -78 ⁰ C. The reaction mixture was carefully quenched with EtOAc and 2 M HCl and the product extracted into CH ₂ Cl ₂ . The

combined organic extracts were dried (Na ₂ SO ₄ ) and the solvent evaporated which gave the title compound (0.77 g, 79%).

Step e

4-Hydroxy-2-isopropyl-5 - (2- [3 -(methanesulfonyl-methyl-amino)-phenyll -ethylamino |-6-[3-(3- methoxy-propoxy)-4-methyl-benzyloxy ^" |-hexanoic acid butylamide (3e)

A solution of compound 2h (1 eq), aldehyde 21 (1 eq.) and AcOH (3 eq.) in MeOH was treated with NaBH ₃ CN (1.2 eq.) . The resulting solution was stirred at room temperature for 16 h. The solvent was evaporated under vacuum and the residue was taken into CH ₂ Cl ₂ and washed with 2M NaOH. The organic phase was dried (Na ₂ SO ₄ ) and concentrated and the residue was purified by preparative HPLC (20% acetonitrile in H ₂ O with 0.1% TFA to 47% acetonitrile) which gave the title compound (21% yield). MS [M+H] ⁺ 664.3

Example 4 Step a

2-Amino-3-methyl-pentanoic acid phenylamide (4a)

N-methylmorpholine (0.5 mL, 4.55 mmol) was added to a solution of Boc-Ile-OH (1 g, 4.32 mmol) in THF (10 mL) at -20 ⁰ C under N ₂ . The mixture was allowed to stir for 10 min. and then ώobutylchloroformate (0.59 mL, 4.55 mmol) was added dropwise keeping the temperature below -14 ⁰ C. The reaction mixture was allowed to stir for 20 min. and aniline (0.44 mL, 4.78 mmol) was then added dropwise keeping the temperature below -20 ⁰ C. After the addition was completed the reaction mixture was allowed to warm to r.t. Stirred at r.t. until LC-MS showed full conversion. The reaction mixture was diluted with toluene and washed with 2 M HCl and H ₂ O. The organic phase was dried (Na ₂ SO ₄ ) and the solvent evaporated to give a solid that was

dissolved into THF:toluene (1.5:3 niL). The resulting solution was cooled to 0 ⁰ C under N ₂ and then treated carefully with 70% w/w RedAl (6.9 mL, 24 mmol). After the addition was completed the reaction mixture was stirred at 40 ⁰ C for 21 h then cooled down to 0 ⁰ C and carefully quenched with 5 M NaOH (10 mL). Stirred 20 min at r.t. and then the product extracted into toluene. The combined organic extracts were washed with 5 M NaOH, dried (Na ₂ SO ₄ ) and evaporated to give a residue that was purified by flash chromatography on silica gel (Hep:EtOAc 10:1 to 2.5:1) to give 0.67 g of a colourless oil that was taken into TFA at r.t. and stirred for 30 min. The reaction mixture was concentrated under vacuum and the residue was partitioned between CH ₂ Cl ₂ and 2 M NaOH. The phases were separated and the organic phase was washed with 2 M NaOH, dried (Na ₂ SO ₄ ) and evaporated to give the title compound 374 mg (45%).

Step b

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid (2 -methyl- 1 -phenylaminomethyl-butyD-amide (4b)

A mixture of the lactone 2f (40 mg, 0.10 mmol), the amine 4a (77 mg, 0.40 mmol), 2- hydroxypyridine (38 mg, 0.40 mmol) and EtN ¹ Pr (17 μL, 0.10 mmol) was heated to 70 ⁰ C for 4 days. The reaction mixture was purified by preparative HPLC (40% acetonitrile in 10 mM aq. NH ₄ OAc to 90% acetonitrile) which gave 24 mg (40%) of the title compound.

Step c

5-Amino-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid

(2 -methyl- 1 -phenylaminomethyl-butyD-amide (4c)

A solution of compound 4b (24 mg, 0.040 mmol) in MeOH (20 mL) was hydrogenated in the presence of Pd Lindlar. Filtration of the catalyst and evaporation of the solvent gave 23 mg

(99%) of the title compound.

MS [M+H] ⁺ 572.4

Example 5 Step a

N-(4-Fluoro-phenyl)-3-methyl-pentane- 1 ,2-diamine (5a)

The procedure described in example 4 step a was used but using 4-fluoroaniline instead of aniline which afforded the title compound (36%).

Step b

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid { 1 -[(4-fluoro-phenylamino)-methyl]-2-methyl-butyU -amide (5b)

The procedure described in example 4 step b was followed but using the amine 5a instead of 4a which afforded the title compound (48%).

5-Amino-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid

{ 1 -[(4-fluoro-phenylamino)-methyll-2-methyl-butyl| -amide (5c)

The azido group of compound 5b was reduced according to the procedure described in example

4 step c, which afforded the title compound (94%).

MS: [M+H] ⁺ 590.3

Example 6 Step a

N-(4-tert-Butyl-phenyl)-3-methyl-pentane- 1 ,2-diamine (6a)

The procedure described in example 4 step a was used but using 4-tert-butylaniline instead of aniline which afforded the title compound (36%).

Step b

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid { 1 - r(4-tert-Butyl-phenylamino)-methyll -2-methyl-butyU -amide (6b)

The procedure described in example 4 step b was used but using 6a instead of 4a which afforded the title compound (44%).

Step c

5-Amino-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid

{ 1 - [(4-tert-butyl-phenylamino)-methyll -2-methyl-butyl| -amide (6c)

The azido group of compound 6b was reduced according to the procedure described in example

4 step c, which afforded the title compound (55%).

MS: [M+H] ⁺ 628.3

Example 7 Step a

N-(4-chloro-phenyl)-3-methyl-pentane- 1 ,2-diamine (7a)

The procedure described in example 4 step a was used but using 4-chloroaniline instead of aniline which afforded the title compound (39%).

Step b

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid { 1 -[(4-chloro-phenylamino)-methyll-2-methyl-butyl| -amide (7b)

The procedure described in example 4 step b was used but using 7a instead of 4a which afforded the title compound (28%).

Step c

5-Amino-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid

{ 1 -[(4-chloro-phenylamino)-methyll-2-methyl-butyl| -amide (7c)

The azido group of compound 7b was reduced according to the procedure described in example

4 step c, which afforded the title compound (99%).

MS: [M+H] ⁺ 606.3

Example 8 Step a

N-(3,4-dichloro-phenyl)-3-methyl-pentane-l,2-diamine (8a)

The procedure described in example 4 step a was used but using 3,4-dichloroaniline instead of aniline which afforded the title compound (44%).

Step b

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid { 1 -[(3,4-dichlorophenylamino)-methyll-2-methyl-butyl| -amide (8b)

The procedure described in example 4 step b was used but using 8a instead of 4a which afforded the title compound (31%).

Ster

5-Amino-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid

{ 1 -[(3,4-dichlorophenylamino)-methyll-2-methyl-butyl| -amide (8c)

The azido group of compound 8b was reduced according to the procedure described in example

4 step c, which afforded the title compound (71%).

MS: [M+H] ⁺ 640.4

Example 9 Step a

N-(3,5-dimethyl-phenyl)-3-methyl-pentane-l,2-diamine (9a)

The procedure described in example 4 step a was used but using 3,5-dimethylaniline instead of aniline which afforded the title compound (54%).

Step b

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid { 1 - [(3 ,5 -dimethyl-phenylamino)-methyl] -2-methyl-butyl| -amide (9b)

The procedure described in example 4 step b was used but using 9a instead of 4a which afforded the title compound (29%).

Step c

5-Amino-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid

{ 1 - [(3 ,5 -dimethyl-phenylamino)-methyl] -2-methyl-butyl| -amide (9c)

The azido group of compound 9b was reduced according to the procedure described in example

4 step c, which afforded the title compound (100%).

MS: [M+Na] ⁺ 622.5

Example 10 Step a

4-Bromo-3-(3-methoxy-propoxy)-benzoic acid methyl ester (IQa)

4-Bromo-3-hydroxy-benzoic acid methyl ester [4.2 g, 18 mmol (prepared according J. Med. Chem. 2005, pp. 1596-1609)] and 3-methoxy-l-propanol (9.5 g, 36.4 mmol) were dissolved in DCM (200 ml). Triphenylphosphine (9.5 g, 36.4 mmol) and l,l '-(azidocarbonyl) dipiperidine (9.2 g, 36.4 mmol) were added and the reaction mixture was stirred at room temperature over night. The reaction mixture was filtered. Evaporation of the solvent afforded a residue that was purified on several silica gel columns eluting with hexanes/ethyl acetate 9:1 to give 4.15 g (75%) of the title compound with a GC purity of 80%.

Step b

3-(3-Methoxy-propoxy)-4-vinyl-benzoic acid methyl ester (IQb) 4-Bromo-3-(3-methoxy-propoxy)-benzoic acid methyl ester (455 mg, 1.5 mmol), dichlorobis(triphenylphosphine)-palladium(II) (105 mg), lithium chloride (190 mg) and 2,6-di- te/t-butyl-4-methylphenol (10 mg) were dissolved n DMF (8 ml). The air was carefully removed and replaced by argon. Tributyl(vinyl)tin (0.48 ml, 1.65 mmol) was added and the reaction mixture was stirred at +60 ⁰ C over night. Thereafter, ethyl acetate and sat. potassium fluoride were added and the mixture was stirred at room temperature for 1 h. The organic phase was evaporated and the residue purified on 2 silica gel columns eluting with hexanes 9 and ethyl acetate 1 to give 224 mg (75%) of title compound.

¹ H-NMR (400MHz, CDCl ₃ ): δ 7.62-7.50 (m, 3H), 7.06 (dd, IH), 5.87 (d, IH), 5.37 (d, IH), 4.15 (t, 2H), 3.91 (s, 3H), 3.58 (t, 2H), 3.36 (s, 3H), 2.13-2.07 (m, 2H).

Step c

4-Ethyl-3-(3-methoxy-propoxy)-benzoic acid methyl ester (IQc)

3-(3-Methoxy-propoxy)-4-vinyl-benzoic acid methyl ester (0.5 g, 2 mmol) was dissolved in 95% ethanol (25 ml). 10% Pd on carbon (100 mg) was added and the reaction mixture was stirred under hydrogen at atmospheric pressure for 4 h. Filtration and evaporation of solvents gave about 0.5 g of the title compound. LC/MS confirmed the correct structure with a M+l of 253.

Step d

[4-Ethyl-3-(3-methoxy-propoxy)-phenyll methanol (IQd)

4-Ethyl-3-(3-methoxy-propoxy)-benzoic acid methyl ester ( 0.5 g, 2 mmol) was dissolved in DCM (20 ml) and the solution cooled to +0° C. DIBAL-H [1.0 M solution in DCM (7 ml, 7 mmol)] was added and the reaction mixture stirred for Ih at room temperature. Sat. ammonium chloride was carefully added followed by 1.5 M HCl and DCM. The organic phase was evaporated and the residue purified on silica gel with ethyl acetate 2 and hexanes 3 as the eluent to give 0.45 g (100%) of the title compound.

¹ H-NMR (400MHz, CDCl ₃ ): δ 7.12 (d, IH), 6.88-6.85 (m, 2H), 4.64 (s, 2H), 4.07 (t, 2H), 3.58 (t, 2H), 3.35 (s, 3H), 2.63 (q, 2H), 2.10-2.03 (m, 2H), 1.19 (t, 3H).

Step e

4-Bromomethyl- 1 -ethyl-2-(3-methoxy-propoxy)-benzene ( 1 Oe)

Triphenylphosphine (0.44 g, 1.7 mmol) was dissolved in DCM (10 ml). Bromine (85 μl, 1.7 mmol) was added and the reaction mixture stirred for 5 min and then the obtained solution was added to [4-ethyl-3-(3-methoxy-propoxy)-phenyl] methanol (0.4 g, 1.8 mmol). The reaction mixture was stirred at room temperature for 30 min. The solvent was evaporated and the residue purified on silica gel using hexanes 9 and ethyl acetate 1 as eluent to give 0.45 (87%) of the title compound. LC/MS confirmed the correct structure with characteristic double peaks at 289 (M+l) and 306 (M+NH ₄ ).

Step f

5 - (2- [4-Ethyl-3 -(3 -methoxy-propoxy)-benzyloxy] - 1 -hydroxy-ethyl-3 -isopropyl-dihydro-furan-2- one (IQf)

The procedure described in Example Ih was followed using 4-bro mo methyl- l-ethyl-2-(3- methoxy-propoxy)-benzene (0.45 g, 1.6 mmol) instead of benzyl bromide, which gave the title compound (0.48 g, 78%).

¹ H-NMR (400MHz, CDCl ₃ ): δ 7.11 (d, IH), 6.83-6.81 (m, 2H), 4.50 (d, 2H), 4.44-4.40 (m, IH),

4.06 (t, 2H), 3.88-3.82 (m, IH), 3.63-3.51 (m, 4H), 3.36 (s, 3H), 2.66-2.58 (m, 3H), 2.46 (d, IH),

2.35-2.28 (m, IH), 2.18-2.03 (m, 4H), 1.19 (t, 3H), 1.02 (d, 3H), 0.93 (d, 3H).

Ster

5- { 1 -Azido-2-[4-etyl-3-(3-methoxy-propoxy)-benzyloxy|-ethyU -3-isopropyl-dihydro-furan-2- one (IQg)

Compound 1Oh (0.48 g, 1.2 mmol) was treated according to the procedure described in Example liwhich gave the title compound was in a yield of 64%. LC/MS confirmed the correct structure with a M+NH ₄ ion at 537.

5-Azido-6-[4-ethyl-3-(3-methoxy-propoxy)-benzyloxy-4-hydr oxy-2-isopropyl-hexanoic acid { 1 - [(4-fluoro-phenylamino)-methyl-butyl| -amide (1 Oh)

A mixture of 5- { 1 -azido-2-[4-etyl-3-(3-methoxy-propoxy)-benzyloxy]-ethyl} -3-isopropyl- dihydro-furan-2-one (130 mg, 0.31 mmol), the compound of example 5a (100 mg, 0.5 mmol), 2- hydroxypyridine (200 mg) and DIPEA (200 μl) was heated at +70° C for 4 days. The reaction mixture was evaporated and the residue purified on silica gel using DCM 95 and methanol 5 as eluent to give 77 mg of the title compound. The purity was 92% according HPLC and LC/MS confirmed the structure with a M+l ion at 630.

Step i

5-Amino-6-[4-ethyl-3-(3-methoxy-propoxy)-benzyloxyl-4-hyd roxy-2-isopropyl-hexanoic acid { 1 - [(4-fluoro-phenylamino)-methyll -2-methyl-butyl| amide ( 1 Oi)

A solution of 5-azido-6-[4-ethyl-3-(3-methoxy-propoxy)-benzyloxy-4-hydroxy -2-isopropyl- hexanoic acid {l-[(4-fluoro-phenylamino)-methyl-butyl} -amide (77 mg, 0.122 mmol) in methanol (75 ml) was hydrogenated in a H-cube instrument using Lindlars catalyst as the catalyst. The reaction mixture was filtered and the solvent evaporated to give 70 mg of a crude product. 20 mg was purified on preparative LC (ammonium acetate and acetonitrile) to give 17 mg of the title compound. HPLC purity >95% and LC/MS confirmed the structure with a M+l ion at 587.

Example 11 Step a

3-Methyl-N*l*-pyridin-3-yl-pentane-l,2-diamine (1 Ia)

The procedure described in example 4 step a was used but using 3-aminopyridine instead of aniline which afforded the title compound (12%).

Step b

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid [2-methyl-l-(pyridin-3-ylaminomethyl)-butyll -amide (1 Ib)

The lactone 2f was opened according to the procedure described in example 4 step b but using the amine 11a instead of 4a which afforded the title compound (28%).

Step c

5-Amino-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid

[2-methyl- 1 -(pyridin-3 -ylaminomethyD-butyl] -amide (l ie)

The azido group of compound 1 Ib was reduced according to the procedure described in example

4 step c, which afforded the title compound (44%).

MS: [M+H] ⁺ 573.9

Example 12 Step a

3 -Methyl-N* 1 * -pyridin-2-yl-pentane- 1 ,2-diamine (12a)

The procedure described in example 4 step a was used but using 2-aminopyridine instead of aniline which afforded the title compound (25%).

Step b

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid [2-methyl- 1 -(pyridin-2-ylaminomethyl)-butyll -amide ( 12b)

The lactone 2f was opened according to the procedure described in example 4 step b but using the amine 12a instead of 4a which afforded the title compound (30%).

Step c

5-Amino-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid [2-methyl- 1 -(pyridin-2-ylaminomethyl)-butyll -amide (12c)

The azido group of compound 12b was reduced according to the procedure described in example 4 step c, which afforded the title compound (94%).

MS: [M+H] ⁺ 574.9

Example 13 Step a

3-Methyl-N* 1 *-pyridin-4-yl-pentane- 1 ,2-diamine (13a)

The procedure described in example 4 step a was used but using 4-aminopyridine instead of aniline which afforded the title compound (37%).

Step b

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid [2-methyl- 1 -(pyridin-4-ylaminomethyl)-butyll -amide (13b)

The lactone 2f was opened according to the procedure described in example 4 step b but using the amine 13a instead of 4a which afforded the title compound (20%).

Step c

5-Amino-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid [2-methyl- 1 -(pyridin-4- ylaminomethyD-butyll -amide (13c)

The azido group of compound 13b was reduced according to the procedure described in example 4 step c, which afforded the title compound (15%). MS: [M+H] ⁺ 573.2

Example 14 Step a

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid ( 1 -hydroxymethyl-2-methyl-propyl)-amide (14a)

The lactone 2f was opened according to the procedure described in example 4 step b but using (S)-valinol instead of 4a, which afforded the title compound (16%).

Step b

5 - Amino-4-hydroxy-2-isopropyl-6- [3 -(3 -methoxy-propoxy)-4-methyl-benzyloxyl -hexanoic acid

( 1 -hydroxymethyl-2-methyl-propyl)-amide ( 14b)

The azido group of compound 14a was reduced according to the procedure described in example

4 step c, which afforded the title compound (100%).

MS: [M+Na] ⁺ 505.2

Example 15 Step a

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyl oxyl-hexanoic acid (2-hydroxy-l-methyl-2-phenyl-ethyl)-amide (15a)

The lactone 2f was opened according to the procedure described in example 4 step b but using (7i?-2iS)-norephedrine instead of 4a which afforded the title compound (25%).

Step b

5 - Amino-4-hydroxy-2-isopropyl-6- [3 -(3 -methoxy-propoxy)-4-methyl-benzyl oxyl-hexanoic acid (2-hydroxy-l-methyl-2-phenyl-ethyl)-amide (15b)

The azido group of compound 15a was reduced according to the procedure described in example

4 step c, which afforded the title compound (96%).

MS: [M+Na] ⁺ 553.2

Example 16 Step a

4-(2-Amino-3-methyl-pentylamino)-phenol (16a)

The procedure described in example 4 step a was followed but using 4-aminophenol instead of aniline which afforded the title compound (43%).

Step b

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid { 1 -[(4-hydroxy-phenylamino)-methyll-2-methyl-butyl| -amide (16b)

The lactone 2f was opened according to the procedure described in example 4 step b but using 16a instead of 4a, which afforded the title compound (15%).

Step c

5-Amino-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid

{ 1 -[(4-hydroxy-phenylamino)-methyll-2-methyl-butyl| -amide (16c)

The azido group of compound 16b was reduced according to the procedure described in example

4 step c, which afforded the title compound (42%).

MS: [M+H] ⁺ 588.0

Example 17 Step a

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid ( 1 -benzyl-2-hydroxy-ethyl)-amide (17a)

The lactone 2f was opened according to the procedure described in example 4 step b but using (S)-phenylalaninol instead of 4a, which afforded the title compound (13%).

Step b

5-Amino-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid

( 1 -benzyl-2-hydroxy-ethyl)-amide ( 17b)

The azido group of compound 17a was reduced according to the procedure described in example

4 step c, which afforded the title compound (100%).

MS: [M+Na] ⁺ 531.4

Example 18 Step a

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyl oxyl-hexanoic acid (l-carbamoyl-2-methyl-butyl)-amide (18a)

The lactone 2f was opened according to the procedure described in example 4 step b but using

(IS-S^-l-amino-S-methyl-pentanoic acid amide instead of 4a which afforded the title compound

(51%).

Step b

5 - Amino-4-hydroxy-2-isopropyl-6- [3 -(3 -methoxy-propoxy)-4-methyl-benzyl oxyl-hexanoic acid (l-carbamoyl-2-methyl-butyl)-amide (18b)

The azido group of compound 18a was reduced according to the procedure described in example

4 step c, which afforded the title compound (78%).

MS: [M+Na] ⁺ 510.3

Example 19 Step a

N* 1 *-(4-Fluoro-benzyl)-3-methyl-pentane- 1 ,2-diamine (19a)

A solution of (5)-isoleucinal (2.2 g, 10.2 mmol), 4-fluorobenzylamine (1.1 mL, 9.62 mmol), sodium cyanoborohydride (0.63 g, 10 mmol) and acetic acid (0.060 mL, 1.05 mmol) in MeOH (50 mL) was stirred at r.t. for 16 h. The reaction mixture was concentrated under vacuum and the residue was partitioned between 2 M NaOH and CH ₂ Cl ₂ . The phases were separated and the organic phase was washed with 2 M NaOH, dried (Na ₂ SO ₄ ) and evaporated to give 2.5 g. The residue was purified by RP-HPLC (60% acetonitrile in 10 mM aq. NH ₄ OAc to 80% acetonitrile) which gave 0.96 g (29%) of the Boc protected amine. Deprotection was achieved by stirring a solution of the Boc protected amine in HCl in MeOH. The reaction mixture was concentrated to

dryness and the residue was taken into 2 M NaOH. The product was extracted into CH ₂ Cl ₂ and the combined extracts were dried (Na ₂ SO ₄ ) and the solvent evaporated which gave the title compound in 62% yield.

Step b

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyl oxyl-hexanoic acid {l-[(4-fluoro-benzylamino)-methyll-2-methyl-butyl| -amide (19b) The lactone 2f was opened according to the procedure described in example 4 step b but usin^ 19a instead of 4a, which afforded the title compound (21%).

Step c

5-Amino-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid

{ 1 -[(4-fluoro-benzylamino)-methyll-2-methyl-butyl| -amide (19c)

The azido group of compound 19b was reduced according to the procedure described in example

4 step c, which afforded the title compound (43%).

MS: [M+H] ⁺ 604.5

Example 20 Step a

4-Fluoro-3-hydroxy-benzoic acid methyl ester (20a)

4-Fluoro-3-hydroxybenzoic acid (0.90 g, 5.77 mmol) was dissolved in 9 rnL of MeOH and the mixture was cooled to 0 ⁰ C and stirred. Thionyl chloride (0.45 mL, 6.23 mmol) was carefully added dropwise, after which the mixture was heated to reflux for 30 min and then cooled to room temperature. The solvent volume was reduced to approx 4 mL by rotary evaporation, and the residue was cooled on ice. 15 mL of water was added, and the mixture was allowed to stand for 15 min with ice cooling. A white precipitate was formed which was filtered off, washed with cold water (10 mL) and air dried. The product was dissolved in toluene (20 mL) which was evaporated in vacuo to yield 0.73 g of the title compound (74%). LC/MS [M-H] ^" 169.

Step b

4-Fluoro-3-(3-methoxy-propoxy)-benzoic acid methyl ester (20b)

The procedure described in Example 10 step a was followed but using 4-fluoro-3-hydroxy- benzoic acid methyl ester (0.69 g, 4.06 mmol) instead of 4-bromo-3-hydroxy-benzoic acid methyl ester, which gave the title compound (0.76 g, 77%) after silica gel column chromatography (gradient elution with hexanes/ethyl acetate 20:1 - 6:1).

Step c

[4-Fluoro-3-(3-methoxy-propoxy)-phenyll-methanol (20c)

The methyl ester 20b (0.73 g, 3.01 mmol) was dissolved in heptane (10 mL) and the solution was cooled to 0 ⁰ C and stirred. DIBAL-H (9.0 mL of a 1.0 M solution in hexane, 9.0 mmol) was

added dropwise. The reaction was quenched after 30 min by careful dropwise addition of 3 M HCl (10 mL). The mixture was diluted with Et ₂ O (40 mL) and 1 M HCl (20 rnL) and the layers were separated. The aqueous phase was extracted with Et ₂ O (40 mL) and the combined organic phases were washed with 1 M HCl (30 mL) and brine (30 mL), dried (MgSO4), filtered and evaporated to yield the title compound (0.50 g, 78%).

Step d

4-Bromomethyl- 1 -fluoro-2-(3-methoxy-propoxy)-benzene (2Od)

The benzyl alcohol 20c (0.50 g, 2.33 mmol) and triphenylphosphine (0.64 g, 2.45 mmol) was dissolved in DCM (7 mL) and the solution was stirred and cooled to 0 ⁰ C on an ice bath. A solution of tetrabromo methane (0.85 g, 2.57 mmol) in DCM (7 mL) was added dropwise after which the cooling bath was removed and stirring was continued for 2 h. Additional triphenylphosphine (64 mg) was added and stirring was continued for 2 h. 1.5 g of SiO2 was added and the solvent was evaporated. The product was purified by column chromatography (gradient elution with hexanes/ethyl acetate 50:1 - 20:1) which gave the title compound (0.54 g,

%).

Step e

5- {2-[4-Fluoro-3-(3-methoxy-propoxy)-benzyloxy]- 1 -hydroxy-ethyl} -3-isopropyl-dihydro- furan-2-one (2Oe)

The procedure described in Example 1 step g was followed but using 4-bromomethyl-l-fluoro-2- (3-methoxy-propoxy)-benzene (0.50 g, 1.80 mmol) instead of benzyl bromide, which gave 0.60

g (96%) of the title compound after column chromatography (elution with hexanes/ethyl acetate 10:1 - 2:1 with 1% MeOH throughout). LC/MS [M+H] ⁺ 385, [M+Na] ⁺ 407.

Step f

5- { 1 -Azido-2-[4-fluoro-3-(3-methoxy-propoxy)-benzyloxyl-ethyl| -3-isopropyl-dihydro-furan-2- one (2Of)

The alcohol 2Oe (0.59 g, 1.53 mmol) was reacted according to the procedure described in Example 1 step g, which gave the title compound (0.47 g, 73%).

Ster

5-Azido-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid (l-carbamoyl-2-methyl-butyl)-amide (2Og)

The lactone 2Of was opened according to the procedure described in example 4 step b but using the amine (25'-35)-2-amino-3-methyl-pentanoic acid amide instead of 4a, which afforded the title compound (57%).

5-Amino-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid (l-carbamoyl-2-methyl-butyl)-amide (2Oh)

The azido group of compound 2Og was reduced according to the procedure described in example

4 step c, which afforded the title compound (89%).

MS: [M+H] ⁺ 590.3

Example 21 Step a

N-(2-Amino-3-methyl-pentyl)-4-methyl-benzenesulfonamide (21 a)

A solution of l-aminomethyl-2-methyl-butyl)-carbamic acid tert-butyl ester (0.5g, 2.31 mmol), [prepared as described by Islam, I. et al. in J. Med. Chem. 1994, 37, 293-304] and pyridine (0.37 mL, 4.62 mmol) in CH ₂ Cl ₂ (10 mL) was reacted with TsCl (0.53 g, 2.78 mmol) at r.t. for 18 h. The reaction mixture was quenched with 2 M HCl and the phases were separated. The organic phase was washed with sat. aq. NaHCO ₃ and dried (Na ₂ SO ₄ ). The product was purified by flash chromatography on SiO ₂ gel (eluent heptane:EtOAc 10:1 to 2.5:1) to give 0.26 g of the protected amine which was taken into a 1 M HCl solution in MeOH and stirred at r.t. until full deprotection was achieved. The reaction mixture was concentrated under vacuum and the residue was partitioned between 2 M NaOH and CH ₂ Cl ₂ . The phases were separated and the aqueous phase was extracted with CH ₂ Cl ₂ . The combined organic extracts were dried (Na ₂ SO ₄ ) and the solvent evaporated to give 152 mg (24%) of the title compound.

Step b

5-Azido-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid {2-methyl-l-[(toluene-4-sulfonylamino)-methyl] -butyl) -amide (21b) The lactone 2Of was opened according to the procedure described in example 4 step b but using the amine 21a instead of 4a, which afforded the title compound (51%).

Step c

5-Amino-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid {2-methyl-l-[(toluene-4-sulfonylamino)-methyll -butyl) -amide (21c)

The azido group of compound 21b was reduced according to the procedure described in example

4 step c, which afforded the title compound (45%).

MS: [M+H] ⁺ 730.3

Example 22 Step a

N-(2-Amino-3-methyl-pentyl)-4-fluoro-benzamide (22a)

4-fluorobenzoyl chloride (0.34 mL, 2.87 mmol) was added slowly dropwise to a solution of compound 21a (0.62 g, 2.87 mmol) and Et ₃ N (0.8 mL, 5.74 mmol) in CH ₂ Cl ₂ (10 mL). The

reaction mixture was stirred at r.t. for 1 h and then extracted with 2 M HCl (x2). The organic phase was dried (Na ₂ SO ₄ ) and the solvent evaporated. The product was purified by flash chromatography on SiO ₂ gel (heptane: EtOAc 10:1 to 2.5:1) to give 0.56 g of the protected amine which then was taken into a 1 M HCl solution in MeOH and stirred at r.t. until full deprotection was achieved. The reaction mixture was concentrated under vacuum and the residue was partitioned between 2 M NaOH and CH ₂ Cl ₂ . The phases were separated and the aqueous phase was extracted with CH ₂ Cl ₂ . The combined organic extracts were dried (Na ₂ SO ₄ ) and the solvent evaporated which gave 393 mg (58%) of the title compound.

Step b

N-(2-{5-Azido-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl -4-ylmethoxyl-4-hydroxy-2- isopropyl-hexanoylamino|-3-methyl-pentyl)-4-fluoro-benzamide (22b)

The lactone 2Of was opened according to the procedure described in example 4 step b but using the amine 22a instead of 4a, which afforded the title compound (31%).

Step c

N-(2-{5-Amino-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl -4-ylmethoxyl-4-hydroxy-2- isopropyl-hexanoylamino|-3-methyl-pentyl)-4-fluoro-benzamide (22c)

The azido group of compound 22b was reduced according to the procedure described in example 4 step c, which afforded the title compound (100%).

MS: [M+H] ⁺ 698.3

Example 23 Step a

5-Azido-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid (l-hydroxymethyl-2-methyl-butyl)-amide (23a)

The lactone 2Of was opened with the amine (5)-isoleucinol according to the procedure described in example 4 step b, which afforded the title compound (27%).

Step b

5-Amino-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid (l-hydroxymethyl-2-methyl-butyl)-amide (23b)

The azido group of compound 23 a was reduced according to the procedure described in example

4 step c, which afforded the title compound (97%).

MS: [M+H] ⁺ 577.3

Example 24 Step a

5-Azido-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid isobutyl- amide (24a)

A solution of the lactone 2Of (23 mg, 0.047 mmol) and isobutylamine (1 niL, 10 mmol) was heated at 70 ⁰ C for 16 h. The reaction mixture was concentrated under vacuum and the residue was purified by preparative HPLC (60% acetonitrile in 10 mM aq. NH ₄ OAc to 95% acetonitrile) which gave the title compound (12 mg, 46%).

Step b

5-Amino-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid isobutyl- amide (24b)

The azido group of compound 24a was reduced according to the procedure described in example

4 step c, which afforded the title compound (100%).

MS: [M+H] ⁺ 533.3

Example 25 Step a

N-(2-Amino-3-methyl-pentyl)-methanesulfonamide (25a)

The procedure described in Example 21, step b was followed but using mehanesulfonyl chloride instead of TsCl, which gave the title compound.

Step b

5-Azido-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylmeth oxyl-4-hydroxy-2-isopropyl- hexanoic acid [l-(methanesulfonylamino-methyl)-2-methyl-butyl ^" |-amide (25b) The lactone 2Of was opened according to the procedure described in example 4 step b but using the amine 25a instead of 4a, which afforded the title compound (38%).

Step c

5-Amino-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid [l-(methanesulfonylamino-methyl)-2-methyl-butyll-amide (25c)

The azido group of compound 25b was reduced according to the procedure described in example

4 step c, which afforded the title compound (100%).

MS: [M+H] ⁺ 654.3

Example 26 Step a

N-(2-Amino-3-methyl-pentyl)-4-chloro-benzenesulfonamide (26a)

The procedure described in Example 21, step b was followed but using 4-chlorobenzenesulfonyl chloride instead of TsCl, which gave the title compound.

Step b

5-Azido-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid {l-[(4-chloro-benzenesulfonylamino)-methyll-2-methyl-butyl| -amide (26b) The lactone 2Of was opened according to the procedure described in example 4 step b but using the amine 26a instead of 4a, which afforded the title compound (36%).

Ster

5-Amino-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylmeth oxyl-4-hydroxy-2-isopropyl- hexanoic acid {l-[(4-chloro-benzenesulfonylamino)-methyll-2-methyl-butyl| -amide (26c) The azido group of compound 26b was reduced according to the procedure described in example 4 step c, which afforded the title compound (50%). MS: [M+H] ⁺ 750.3

Example 27 Step a

5-Azido-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid (2-methyl-butyl)-amide (27a)

The lactone 2Of was opened according to the procedure described in example 4 step b but using the amine (5)-2-methyl-l-butylamine instead of 4a, which afforded the title compound (40%).

Step b

5-Amino-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid (2-methyl-butyl)-amide (27b)

The azido group of compound 27a was reduced according to the procedure described in example

4 step c, which afforded the title compound (100%).

MS: [M+H] ⁺ 547.3

Example 28 Step a

5-Azido-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid 4-methoxy-benzylamide (28a)

The lactone 2Of was opened according to the procedure described in example 4 step b, but using 4-methoxybenzylamine instead of 4a which afforded the title compound (52%).

Step b

5-Amino-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid 4-methoxy-benzylamide (28b)

The azido group of compound 28a was reduced according to the procedure described in example

4 step c, which afforded the title compound (100%).

MS: [M+H] ⁺ 597.3

Example 29 Step a

5-Azido-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid amide (29a)

A solution of the lactone 2Of (79 mg, 0.163 mmol) in THF (0.1 niL) was added to a pressurized vessel containing liquid ammonia (2 mL) at r.t. The reaction mixture was stirred at r.t. for 18 h. Ammonia was carefully removed by evaporation and the resulting residue was purified by flash chromatography on SiO ₂ (5% MeOH in CH ₂ Cl ₂ ) to give 28 mg (34%) of the title compound.

Step b

5-Amino-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid amide (29b)

The azido group of compound 29a was reduced according to the procedure described in example

4 step c, which afforded the title compound (71%).

MS: [M+H] ⁺ 477.2

Example 30 Step a

5-Azido-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid methylamide (30a)

A solution of the lactone 2Of (43 mg, 0.09 mmol) and methylamine in THF (0.1 mL) was heated at 70 ⁰ C for 18 h. The reaction mixture was concentrated under vacuum and the residue was

purified by preparative HPLC (40% acetonitrile in 10 rnM aq. NH ₄ OAc to 90% acetonitrile) which gave the title compound (39 mg, 85%).

Step b

5-Amino-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid methylamide (30b)

The azido group of compound 30a was reduced according to the procedure described in example

4 step c, which afforded the title compound (83%).

MS: [M+H] ⁺ 491.2

Example 31 Step a

5-Azido-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid ethylamide (31a)

The lactone 2Of was opened according to the procedure described in example 30 step a but using ethylamine instead of methylamine which afforded the title compound (55%).

5-Amino-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid ethylamide (31b)

The azido group of compound 31a was reduced according to the procedure described in example

4 step c, which afforded the title compound (86%).

MS: [M+H] ⁺ 505.3

Example 32 Step a

5-Azido-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid {l-[(4-fluoro-phenylamino)-methyll-2-methyl-butyl|-amide (32a) The lactone 2Of was opened according to the procedure described in example 4 step b but using the amine 5 a instead of 4a which afforded the title compound (27%).

Step b

5-Amino-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid {14(4-fluoro-phenylamino)-methyl]-2-methyl-butyU -amide (32b)

The azido group of compound 32a was reduced according to the procedure described in example

4 step c, which afforded the title compound (100%).

MS: [M+H] ⁺ 670.37

Example 33 Step a

5-Azido-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid {l-[(4-chloro-phenylamino)-methyll-2-methyl-butyl|-amide (33a) The lactone 2Of was opened according to the method described in example 4 step b but using the amine 7a instead of 4a, which afforded the title compound (27%).

Step b

5-Amino-6-[4'-fluoro-2-(3-methoxy-propoxy)-biphenyl-4-ylm ethoxyl-4-hydroxy-2-isopropyl- hexanoic acid {l-[(4-chloro-phenylamino)-methyll-2-methyl-butyl|-amide (33b)

The azido group of compound 33a was reduced according to the procedure described in example

4 step c, which afforded the title compound (100%).

MS: [M+H] ⁺ 686.36

Example 34 Step a

1 -Chloro-2-(3-methoxy-propoxy)-4-methyl-benzene (34a)

Triphenylphosphine (5.25 g, 20 mmol) and azodicarboxylic dipiperidine (5.04, 20 mmol) were added to a solution of 2-chloro-5-methyl-phenol (1.43 g, 10 mmol) and 3-methoxy-propan-l-ol (1 mL, 10 mmol) in CH ₂ Cl ₂ (100 mL). The reaction mixture was stirred at r.t. 16 h and the evaporated into SiO ₂ gel. Purification of the afforded residue by flash chromatography on silica gel (Hep:EtOAc 20:1) gave the title compound (2.0 g, 80%).

Step b

4-Bromomethyl- 1 -chloro-2-(3-methoxy-propoxy)-benzene (34b)

N-Bromosuccinimide (284 mg, 1.6 mmol) and benzoyl peroxide (11 mg, 0.034 mmol) were added to a solution of 34a in CCU (6 mL) and the resulting suspension was refluxed for 1 h. The reaction mixture was filtered through SiO ₂ using Hep:EtOAc 10:1 as eluent. Evaporation of the solvent afforded 304 mg of the title compound (78% yield).

Step c

5- {2-[4-Chloro-3-(3-methoxy-propoxy)-benzyloxyl- 1 -hydroxy-ethyll -3-isopropyl-dihydro- furan-2-one (34c)

The procedure described in Example 1 step g was followed but using 34b instead of benzyl bromide, which gave the title compound (54% yield).

Step d

5- { 1 -Azido-2-[4-chloro-3-(3-methoxy-propoxy)-benzyloxy ^" |-ethyl| -3-isopropyl-dihydro-furan-2- one (34d)

Compound 34c was reacted according to the procedure described in Example 1 step g, which afforded the title compound (72%).

Step e

5-Azido-6-[4-chloro-3-(3-ethoxy-propoxy)-benzyloxyl-4-hyd roxy-2-isopropyl-hexanoic acid (2- methyl- 1 -phenylaminomethyl-butyD-amide (34e)

The lactone 34d was opened according to the procedure described in example 4 step b, which afforded the title compound (6%).

Step f

5-Amino-6-[4-chloro-3-(3-ethoxy-propoxy)-benzyloxyl-4-hyd roxy-2-isopropyl-hexanoic acid

(2 -methyl- 1 -phenylaminomethyl-butyD-amide (34f)

The azido group of compound 34e was reduced according to the procedure described in example

4 step c, which afforded the title compound (100%).

MS: [M+H] ⁺ 606.3

Example 35 Step a

2,2-Dimethyl-oxazolidine-3,4-dicarboxylic acid 3-tert-butyl ester 4-methyl ester (35a) 2,2-Dimethoxypropane (50 ml, 400 mmol) and boron trifluoride etherate (0.35 ml, 2.8 mmol) was added to a solution of N-Boc serine methyl ester (1, 10.0 g, 45.6 mmol) in acetone (165 ml). The resulting orange solution was stirred at room temperature for 2.5 h until disappearance of starting material. 0.9 ml triethylamine was then added to the reaction mixture whereafter the solvent was removed under reduced pressure. The residue was partitioned between diethyl ether and sat. NaHCOs, the organics extracts were combined, dried and concentrated which gave the title compound (10.5 g, 90%) which was used without further purification.

Step b

4-Hvdroxymethyl-2,2-dimethyl-oxazolidine-3-carboxylic acid tert-butyl ester (35b)

A 250-ml two-necked flask was equipped with a magnetic stirring bar, reflux condenser bearing a drying tube and a dropping funnel. The flask was charged with tetrahydrofuran (100 ml) and lithium aluminium hydride (2.16 g, 57.0 mmol). While the suspension in the flask was stirred, a solution of the ester 12a (9.90 g, 38.2 mmol) in THF (50 ml) was added dropwise during 20 min. The reaction was monitored by thin layer chromatography. When the reaction was finished, the mixture was cooled in an ice bath and a solution of 10% potassium hydroxide (20 ml) was added dropwise during 10 min. The mixture was stirred for 2 h at room temperature, whereafter the white precipitate was removed by filtration through celite. The combined organic filtrates were washed with 100 ml of aqueous phosphate buffer (pH 7), and the aqueous layer was extracted with ether. The combined organic phases were dried and concentrated which gave the title compound (8.3 g, 94%). The residue was used without further purification.

Step c

4-Formyl-2,2-dimethyl-oxazolidine-3-carboxylic acid tert-butyl ester (35c) A solution of dimethylsulfoxide (8.10 g, 103.71 mmol) in dichloromethane (10 ml) was added dropwise during 25 min to a solution of oxalyl chloride (6.58 g, 51.9 mmol) in dichloromethane (80 ml) at -78 ⁰ C. At the end of the addition the reaction solution was warmed up to -60 ⁰ C, and a solution of the alcohol 12b (8.0 g, 34.6 mmol) in dichloromethane (60 ml) was added dropwise during 50 min. N,N-diisopropylethyl amine (36 ml, 200 mmol) in dichloromethane (5 ml) was then added to the reaction mixture -45 ⁰ C during 30 min whereafter the reaction mixture was allowed to warm to 0 ⁰ C during 10 min. The reaction mixture was then transferred to a separation funnel charged with ice-cold 1 M HCl solution (130 ml). The two phases were separated and the aqueous phase was extracted with dichloromethane. The combined organic extracts were dried and concentrated which gave the title compound (7.89 g, 99%). The residue was used in the next step without further purification.

Step d

4-(l-Hydroxy-3-methoxycarbonyl-prop-2-ynyl)-2,2-dimethyl- oxazolidine-3-carboxylic acid tert- butyl ester (35d)

BuLi (71.5 ml, 114 mmol, 1.6 M in hexanes) was added dropwise to a solution of methyl propiolate (10.5 ml, 125 mmol) in ether (300 ml) at -78 ⁰ C. The solution was stirred at this temperature for 1 h, then a solution OfZnBr ₂ (31.6 g, 140 mmol) in ether (200 ml) was added via canula. The mixture was stirred at room temperature for ~2 h, then cooled down to -20 ⁰ C and of the aldehyde 12c (10.4 g, 45.7 mmol) in ether (80 ml) was added. The mixture was stirred at room temperature overnight, then the reaction was quenched by addition of sturated NH ₄ Cl at 0 ⁰ C. The mixture was extracted with ether, dried and concentrated. The residue was purified by column chromatography, eluted with ethyl acetate - iso-hexane 1 :4, which gave the title compound (5.8 g, 20.4 mmol, 45%).

Step e

4-(l-Hydroxy-3-methoxycarbonyl-propyl)-2,2-dimethyl-oxazo lidine-3-carboxylic acid tert-butyl ester (35e)

Compound 12d (4.6 g, 14.70 mmol) was dissolved ethyl acetate (50 ml) and a catalytic amount of Pd/C was added. The mixture was stirred under an atmosphere of hydrogen for ~3 h, then filtered through celite and concentrated which gave the title compound (4.5 g, 14.20 mmol, 97%). The residue was used in the next step without further purification.

Step f

Acetic acid (420 μl) was added to a solution of compound 12e (4.5 g, 14.20 mmol) in toluene (100 ml). The reaction mixture was refluxed for 4-5 h, until the starting material disappeared. The solvent was removed under vacuum and the residue was purified by column chromatography (Rf 0.40, toluene - acetone 6:1) which gave the title compound (3.7 g, 13.2 mmol, 93%).

Ster

4-(4-Isopropyl-5-oxo-tetrahydro-furan-2-yl)-2,2-dimethyl- oxazolidine-3-carboxylic acid tert- butyl ester (35g)

A solution of BuLi in hexanes (1.6 M, 2.3 ml, 3.67 mmol) was added dropwise to a solution of DIPA (494 μl, 3.50 mmol) in THF (3 ml) at -78 ⁰ C under an atmosphere of argon whereafter the mixture was stirred at this temperature for 30 min. A solution of the lactone 12f (500 mg, 1.75 mmol) in THF (3 ml) was then added to the solution followed by TPPO (tripyrrolidine phosphorus oxide, 3.0 ml) under vigorous stirring. ¹ PrI (700 μl, 7.0 mmol) was added dropwise and the reaction mixture was stirred at this temperature for 2 h, followed by another 2 h at room temperature. The reaction was then quenched by addition of sat. NH ₄ Cl, extracted with TBME, dried and concentrated. The residue was purified by column chromatography which gave the title compound (380 mg, 1.16 mmol, 66%).

Step h

[2-Benzyloxy-l-(4-isopropyl-5-oxo-tetrahydro-furan-2-yl)- ethyll-carbamic acid tert-butyl ester (35h)

The isopropyl lactone 12g (204 mg, 0.624 mmol) was treated with TFA - H ₂ O (5 ml, 9:1), stirred at room temperature for 1 h, and then concentrated under vacuum. The residue was dissolved in a mixture of dioxane and a 10% solution of sodium carbonate (10 ml, 1 :1, pH ~8). BoC ₂ O (204 mg, 0.936 mmol) was added in portions and the mixture was stirred at room temperature for 2 h. Dioxane was then removed under vacuum and the aqueous phase was extracted with dichloro methane, dried and concentrated. The residue was dissolved in dichloromethane and cooled to 0 ⁰ C, and benzyl 2,2,2-trichloroacetimidate (174 μl, 0.936 mmol) and TMS-triflate (22 μl) was added. The reaction mixture was stirred at room temperature until the reaction was complete and then washed with sat. sodium bicarbonate, dried and concentrated. The residue was purified by column chromatography (Rf 0.60 ethyl acetate - iso-hexane 1 :2) which gave the title compound (143 mg, 0.379 mmol, 61%).

6-Benzyloxy-5-tert-butoxycarbonylamino-4-(tert-butyl-dime thyl-silanyloxy)-2-isopropyl- hexanoic acid (35i)

LiOH (1 M sol in water, 2.98 ml, 2.98 mmol) was added to a solution of the lactone 12h (750 mg, 1.99 mmol) in dioxane-water (12 ml, 2:1). The reaction mixture was stirred at room temperature until the starting material had disappeared (~1 h). The solvents were removed under vacuum and the residue partitioned between ether and 10% citric acid, the organic phase was dried and concentrated. The residue was dissolved in DMF (20 ml) whereafter imidazole (3.0 g, 44 mmol) and TBDMSCl (3.0 g, 20 mmol) was added and the solution was stirred at room temperature for 2 days. The reaction was quenched by addition of methanol, whereafter the mixture was stirred at room temperature for 2 h, and then concentrated. The residue was purified by column chromatography (Rf 0.65 ethyl acetate - iso-hexane 1:2) which gave the title compound (502 mg, 0.98 mmol, 50%).

Ster

[l-Benzyloxymethyl-2-(tert-butyl-dimethyl-silanyloxy)-5-m ethyl-4-phenylcarbamoyl-hexyll- carbamic acid tert-butyl ester (35j)

A solution of 35a (200 mg, 0.39 mmol), HATU (149 mg, 0.39 mmol) and ethyldiisopropylamine (0.07 ml, 0.40 mmol) in DMF (4 mL) was stirred at 0 ⁰ C for 5 min. Aniline (0.036 mL, 0.40 mmol) was then added at once and the reaction mixture was further stirred 30 min at 0 ⁰ C. The reaction mixture was concentrated into SiO ₂ and the product was purified by flash chromatography in SiO ₂ (heptane: EtOAc 5:1 to 2.5:1) to give 155 mg (68%) of the title compound.

Step k

5-Amino-6-benzyloxy-4-hydroxy-2-isopropyl-hexanoic acid phenylamide (35k)

A solution of compound 35j (125 mg, 0.21 mmol) in THF (3 mL) was treated with TBAF (1 mmol) at r.t. for 2 h. The reaction mixture was concentrated under vacuum and redisolved in

TFA, stirred at r.t for 30 min and concentrated under vacuum. The residue was purified by preparative HPLC (40% acetonitrile in 10 mM aq. NH ₄ OAc to 75% acetonitrile) which gave 2.6 mg (3%) of the title compound.

MS: [M+H] ⁺ 371.32

Example 36 Step a

5-Azido-6-[4-fluoro-3-(3-methoxy-propoxy)-benzyloxyl-4-hy droxy-2-isopropyl-hexanoic acid { 1 -[(4-fluoro-phenylamino)-methyll-2-methyl-butyl| amide (36a)

Azide 2Of (102 mg, 0.249 mmol), amine 5a (210 mg, 1.00 mmol), 2-hydroxypyridine (95 mg, 1.00 mmol) and DIPEA (48 μL, 0.275 mmol) were stirred and heated to 70 ⁰ C for 3 days. Purification by preparative HPLC (40% MeCN in H ₂ O with 10 mM NH ₄ Ac to 80% MeCN) gave the title compound after freeze drying (73 mg, 47%). LC/MS confirmed the structure with a M+l (H ⁺ ) ion at 620.3.

Step b

5 - Amino-6- [4-fluoro-3 -(3 -methoxy-propoxyVbenzyloxy] -4-hydroxy-2-isopropyl-hexanoic acid { 1 - [(4-fluoro-phenylamino)-methyll -2-methyl-butyl| amide (36b)

A solution of azide 36a (73 mg, 0,12 mmol) in MeOH (35 mL) was hydrogenated at atmospheric pressure in the presence of Lindlars catalyst. The mixture was filtered and the solvent evaporated, and the residue redissolved in 1,4-dioxane (3 mL) and freeze dried to yield 69 mg (99%) of the title compound. LC/MS confirmed the structure with a M+l (H ⁺ ) ion at 594.8 and a M+23 (Na ⁺ ) ion at 616.1. The purity was determined by HPLC with a DAD detector to be 94%.

Example 37 Step a

l-(t-butoxycarbonylamino)-l-(3-(4-methoxytrityloxymethyl)phe nyl)-(li?,61-l-phenylmethane (37a)

3-(4-Methoxytrityloxymethyl)benzonitrile (1.6 g, 3.95 mmol) was dissolved in dry tetrahydrofuran (THF, 18 ml) and added dropwise over 20 min under nitrogen to a cooled (0 °C) 3M diethyl ether solution of phenylmagnesiumbromide., followed by reduction with lithium aluminium hydride, hydrolysis and treatment of the crude amine with di-t-butyldicarbonate, according to the method described in Example 2, step b. The mixture was then stirred at room temperature for 30 min, and then at 60 ⁰ C for 2 h. It was cooled to 0 ⁰ C and lithium aluminium hydride was added as a slurry in THF, and the suspension was stirred at room temperature for Ih and then refluxed for 1 h. It was cooled to 0 ⁰ C and water, 10% NaOH and water were added successively. The resulting suspension was filtered through Celite, and the solid was washed with THF and DCM. The filtrate was evaporated and the residue partitioned between DCM and water. The organic extract was dried thorough sodium sulphate and evaporated. The residue was dissolved in EtOAc and di-tert-butyl dicarbonate was added and the mixture was stirred at room temperature over night. The reaction was quenched with MeOH, evaporated and the residue partitioned between DCM and saturated aqueous sodium bicarbonate. The organic extract was dried by sodium sulphate and evaporated. Silica gel column chromatography (gradient 50% DCM / hexane - 75% DCM - 100% DCM), which gave the title compound (2.04 g, 88%). ¹ H-NMR (400 MHz, CDCl ₃ ): δ 7.47-7.18 (m, 21H), 6.82 (d, 2H), 5.90 (br, s, IH) CH, 5.12 (br, s, IH) NH, 4.13 (s, 2H), 3.79 (s, 3H), 1.43 (br, s, 9H) t-but.

Step b

1 -(t-butoxycarbonylamino)- 1 -(3 -(hvdroxymethyl)phenyl)-( Ii?, S)- 1 -phenylmethane (37b)

l-(t-butoxycarbonylamino)-l-(3-(4-methoxytrityloxymethyl)phe nyl)-(li?,5)-l-phenylmethane (2.04 g, 3.48 mmol) was treated with 80% acetic acid and dioxane and was stirred at room temperature for 1 h. The solvents were removed by evaporation on rotavapor and the residue was co-evaporated once from acetonitrile and once from toluene. Silica gel column chromatography (gradient 0 - ¹ A - 2% EtOH / DCM) gave the title compound (1.06 g, 97%). 1H-NMR (400 MHz, CDCl ₃ ): δ 7.33-7.16 (m, 9H), 5.90 (br, s, IH) CH, 5.15 (br, s, IH) NH, 4.65 (d, 2H), 1.69 (t, IH) OH, 1.43 (br, s, 9H) t-but.

Step c

1 -(3 -(bromomethyl)phenyl)- 1 -(t-butoxycarbonylamino)-( Ii?, S)- 1 -phenylmethane (37c) Carbontetrabromide (3.6 mmol) dissolved in DCM (3 ml) was added dropwise to a DCM solution (8 ml) of l-(t-butoxycarbonylamino)-l-(3-(hydroxymethyl)phenyl)-(li?,5 )-l- phenylmethane (1.04 g, 3.40 mmol) and triphenylphosphine (3.7 mmol) and the resulting solution was stirred at room temperature over night. The solvent was removed by evaporation on rotavapor and the afforded residue was purified by silica gel column chromatography (DCM) which gave the title compound (0.75 g, 58%).

¹ H-NMR (400 MHz, CDCl ₃ ): δ 7.34-7.15 (m, 9H), 5.89 (br, s, IH) CH, 5.13 (br, s, IH) NH, 4.45 (s, 2H), 1.43 (br, s, 9H) t-but.

Step d

2,3-dideoxy-6-O-(3-(l-(t-butoxycarbonylamino)-(li?,61-l-p henylmethyl) benzyl)-2-isopropyl-D- glucono-l,4-lactone (37d)

Compound If(IOO mg, 0.53 mmol) was alkylated with compound 37c according to the method described in Example 1, step g, which gave the title compound (180 mg, 70%). MS mlz 542.2

(M+AcO) ^" .

¹ H-NMR (400 MHz, CDCl ₃ ): δ 7.31-7.13 (m, 9H), 5.90 (br, s, IH) CH, 5.12 (br, s, IH) NH, 4.52

(s, 2H), 4.53 (s, 2H), 4.39 (m, IH), 3.82 (m, IH), 3.60 (dd, IH), 3.53 (dd, IH), 2.61 (m, IH),

2.50 (br, s, IH) OH, 2.30 (m, IH), 2.17-2.01 (m, 2H), 1.42 (s, 9H) t-but.,1.01 (d, 3H), 0.92 (d,

3H).

Step e

5-azido-2,3,5-trideoxy-6-O-(3-(l-(t-butoxycarbonylamino)- (li?,5)-l-phenylmethyl)benzyl)-2- isopropyl-D-glucono- 1 ,4-lactone (37e)

2,3-Dideoxy-6-O-(3-(l-(t-butoxycarbonylamino)-(li?,5)-l-phen ylmethyl)benzyl)-2-isopropyl-D- glucono-l,4-lactone (180 mg, 0.37 mmol) was treated with triflic anhydride, followed by sodium azide according to the method described in Example 1, step h, which gave 120 mg of the title compound (63%). MS mlz 567 '.1 (M+AcO) ^" .

¹ H-NMR (400 MHz, CDCl ₃ ): δ 7.34-7.18 (m, 9H), 5.91 (br, s, IH) CH, 5.20 (br, s, IH) NH, 4.54 (s, 2H), 4.51 (m, 2H), 3.74 (d, IH), 3.72 (s, IH), 3.62 (m, IH), 2.71 (m, IH), 2.21-2.10 (m, 2H), 1.43 (s, 9H) t-but.,1.01 (d, 3H), 0.92 (d, 3H).

Step f

N-isobutyl-{(2R.4S.5iS)-5-azido-6-(3-(l-(t-butoxycarbonyl amino)-(lR.iS)-l- phenylmethyl)benzyloxy)-4-hydroxy-2-isopropyl| -hexanoic amide (37f)

The lactone 37e (40 mg, 0.079 mmol) was dissolved in isobutylamine (1 ml) and stirred at 70 °C for 16 hours. The volatile matter was evaporated on rotavapor and the residue was partitioned between DCM and 5% citric acid. The organic extract was dried through sodium sulphate and evaporated. Silica gel column chromatography (gradient 0 - Vi - 1 - 1 /4EtOH/DCM) gave the title compound (43 mg, 93%). MS mlz 582.3 (M+H) ⁺ .

¹ H-NMR (400 MHz, CDCl ₃ ): δ 7.36-7.14 (m, 9H), 5.92 (br, s, IH) CH, 5.78 (br, s, IH) NH, 5.31

(br, s, IH) NH, 4.50 (s, 2H), 3.79-3.60 (m, 3H), 3.40 (m, IH), 3.15 (m, IH), 2.96 (m, IH), 2.90

(br, s, IH) OH, 2.04 (m, IH), 1.75 (m, 2H), 1.42 (s, 9H) t-but, 0.95-0.84 (m, 12H).

Ster

N-isobutyl-((2i?.4 _t S'.5 _t S)-5-amino-6-(3-(l-(t-butoxycarbonylamino)-(li?. _t S)-l- phenylmethyl)benzyloxy)-4-hydroxy-2-isopropyl| -hexanoic amide (37g) The azide 37e (43 mg, 0.074 mmol) was hydrogenated according to the method described in Example 4, step c, which gave the title compound (39 mg, 95%). MS mlz 556.3 (M+H) ⁺ . 1H-NMR (400 MHz, CDCl ₃ ): δ 7.37-7.14 (m, 9H), 5.96-5.80 (br, s, 2H) NH, CH, 5.40 (br, s, IH) NH, 4.48 (s, 2H), 3.50 (m, IH), 3.40 (m, 2H), 3.18 (m, IH), 3.01 (m, IH), 2.78 (br, s, IH) OH, 2.10 (br, s, 2H) NH ₂ , 2.10(m, IH), 1.90-1.50 (m, 4H), 1.42 (s, 9H) t-but., 0.98-0.84 (m, 12H).

Example 38 Step a

5-Azido-6-[4-ethyl-3-(3-methoxy-propoxy)-benzyloxyl-4-hyd roxy-2-isopropyl-hexanoic acid (2-morpholin-4-yl-ethyl)-amide (38a)

A mixture of the lactone 1Og (70 mg, 0.16 mmol)], 2-(4-morpholino)ethylamine (45 μl, 0.32 mmol), 2-hydroxypyridine (100 mg) and DIPEA (100 μl) was heated at + 70 ⁰ C for 24 hours. The reaction mixture was evaporated and the residue purified by silica gel chromatography (dichloromethane and increasing amount of methanol finally 4%) to afford 47 mg of the title compound. LC/MS confirmed the structure with a M+l ion at 550.3.

Step b

5-Amino-6-[4-ethyl-3-(3-methoxy-propoxy)-benzyloxyl-4-hyd roxy-2-isopropyl-hexanoic acid (2-morpholin-4-yl-ethyl)-amide (38b)

A solution of example 38a (47 mg) in methanol (50 ml) was hydrogenated at atmospheric pressure in an H-Cube instrument using Lindlars catalyst as the catalyst. The reaction mixture was evaporated and 50 mg of the title product was obtained with a LC purity at 230 nm of 94.5%. LC/MS confirmed the structure with a M+l ion at 524.0.

Example 39 Step a

2-Amino-3-methyl-pentanoic acid N'-(4-fluoro-phenyl)-hydrazide (39a)

A solution of Boc-Ile-OH (1 g, 4.32 mmol), EtN ¹ Pr (1.7 mL, 9.74 mmol) and HATU (1.6 g, 4.32 mmol) in DMF (5 mL) was stirred for 5 min at 0 ⁰ C before adding 4-fluorophenylhydrazine (0.7 g, 4.32 mmol). The reaction mixture was stirred for 3 h and then concentrated under vacuum. The residue was purified by flash chromatography on silica gel (Hep:EtOAc 2.5:1) to give 1.1 g of the Boc protected amine. The solid was dissolved in TFA and the resulting solution was

stirred at r.t. for Ih. Evaporation of the solvent afforded an oil that was taken into CH ₂ Cl ₂ and washed with 1 M NaOH. The organic phase was dried (Na ₂ SO ₄ ) and concentrated to give the title compound (99%).

Step b

5-Azido-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid { 1 - [N'-(4-fluoro-phenyl)-hydrazinocarbonyll -2-methyl-butyl| -amide (39b) The lactone 2f was opened with the amine 39a according to the procedure described in example 1 step g which gave the title compound (9%).

Step c

5-Amino-4-hydroxy-2-isopropyl-6-[3-(3-methoxy-propoxy)-4- methyl-benzyloxyl-hexanoic acid

{ 1 - [N'-(4-fluoro-phenyl)-hydrazinocarbonyll -2-methyl-butyl| -amide (39c)

A solution of the azide 39b in MeOH was hydrogenated in the presence of Pd Lindlar. Filtration of the catalyst and evaporation of the solvent gave the title compound (99%).

MS: [M+H] ⁺ 619.5

Example 40 Step a

5- { 1 -Hydroxy-2-[3-(3-methoxypropoxy)-benzyloxyl-ethyU -3-isopropyldihydrofuran-2-one

(40a)

The procedure described in Example 1 step g was followed but using 3-(3-methoxy-propoxy)- benzyl bromide instead of benzyl bromide, which gave the title compound (67% yield).

Step b

5- { 1 -Azido-2-[3-(3-methoxypropoxy)-benzyloxy ^" |-ethyU -3-isopropyldihydrofuran-2-one (40b) Compound 40a was reacted according to the procedure described in Example 1 step h, which gave the title compound (10% yield).

Example 41 Step a

5-[ 1 -Hydroxy-2-(naphthalen- 1 -ylmethoxy)-ethvH-3-isopropyldihydrofuran-2-one (41 a)

The procedure described in Example 1 step g was followed but using 1-bromomethyl- naphthalene instead of benzyl bromide, which gave the title compound (90% yield).

Step b

5-[ 1 -Azido-2-(naphthalen- 1 -ylmethoxy)-ethyll-3-isopropyldihydrofuran-2-one (4 Ib) Compound 41a was reacted according to the procedure described in Example 1 step h, which gave the title compound (32% yield).

Example 42 Step a

5-[ 1 -Hydroxy-2-(naphthalen-2-ylmethoxy)-ethyll-3-isopropyldihydr ofuran-2-one (42a) The procedure described in Example 1 step g was followed but using 1-bromomethyl- naphthalene instead of benzyl bromide, which gave the title compound (67% yield).

Step b

5-[ 1 -Azido-2-(naphthalen-2-ylmethoxy)-ethyl]-3-isopropyldihydrof uran-2-one (42b) Compound 42a was reacted according to the procedure described in Example 1 step h, which gave the title compound (67% yield).

Example 43 Step a

3-(4-Methoxytrityloxymethyl)benzonitrile (43a)

3-(hydroxymethyl)benzonitrile (2 g, 15 mmol) was dissolved in dry pyridine (120 ml). 4- methoxytrityl chloride (6 g, 19.5 mmol) was added and the mixture was stirred over night at room temperature (r.t.). The reaction was quenched with ethanol (EtOH), evaporated on rotavapor and the residue partitioned between dichloromethane (DCM) and saturated aqueous sodium bicarbonate. The organic phase was dried by sodium sulfate and evaporated. Silica gel column chromatography (gradient 30%DCM / hexane - DCM) gave the title compound (5.98 g,

98%).

¹ H-NMR (400MHz, CDCl ₃ ): δ 7.70-7.25 (m, 16H), 6.85 (d, 2H), 4.23 (s, 2H), 3.81 (s, 3H).

Step b

(IR,S)- 1 -(t-Butoxycarbonylamino)- 1 -(3-(4-methoxytrityloxymethyl)phenyl) ethane (43b) The cyano derivative 43a (2.4 g, 5.92 mmol) was dissolved in dry THF (18 ml) and added dropwise over 20 min under nitrogen to a cooled (0 ⁰ C) 3M diethylether solution of methylmagnesiumbromide (3.95 ml). The mixture was then stirred at r.t. for 30 min, and then at 60 ⁰ C for 2 h. The reaction mixture was cooled to 0 ⁰ C and lithium aluminium hydride (450 mg) was added as a slurry in THF (18 ml), and the suspension was stirred at r.t. for Ih and then refluxed for 1 h. The reaction mixture was cooled to 0 ⁰ C and water (0.55 ml), 10%NaOH (0.55

ml) and water (1.84 ml) were added successively. The resulting suspension was filtered through Celite, and the solid was washed with THF and DCM. The filtrate was evaporated and the residue partitioned between DCM and water. The organic extract was dried by sodium sulfate and then evaporated. The residue was dissolved in ethyl acetate (15 ml) and di-t- butyldicarbonate was added (2.6 g, 11.85 mmol) and the mixture was stirred at r.t. over night. The reaction was quenched with MeOH, evaporated and the residue was partitioned between DCM and saturated aqueous sodium bicarbonate. The organic extract was dried by sodium sulfate and evaporated. Purification by silica gel column chromatography (gradient 50% DCM / hexane - 75%DCM - DCM) gave the title compound (2.81 g, 91%).

Step c

(IR,S)- 1 -(t-Butoxycarbonylamino)- 1 -(3-(hydroxymethyl)phenyl)ethane (43c) Compound 43b was dissolved in a mixture of 80% acetic acid (150 ml) and dioxane (20 ml) and was stirred at r.t. for 1 h. The solvents were removed by evaporation and the residue was co- evaporated once from acetonitrile and once from toluene. Silica gel column chromatography (gradient 0 - ¹ A - 2% EtOH / DCM) gave the title compound (1.50 g, quant.).

Step d

(IR, S)-I -(3 -(Bromomethyl)phenyl)- 1 -(t-butoxycarbonylamino)ethane (43d) Carbontetrabromide (0.66 g, 2.51 mmol) dissolved in DCM (3 ml) was added dropwise to a solution of the alcohol 43c (0.6 g, 2.39 mmol) and triphenylphosphine (0.87 g, 2.63 mmol) in DCM (8 ml) and the resulting solution was stirred at r.t. over night. The solvent was removed by evaporation on rotavapor and silica gel column chromatography (DCM) gave the title compound (0.48 g, 64%).

Step e

2,3-Dideoxy-6-O-(3-((li?,y)-l-(t-butoxycarbonylamino)ethy l)benzyl)-2-isopropyl-D-glucono-

1.4-lactone (43e)

The lactone If (185 mg, 0.98 mmol) and dibutyltin oxide (319 mg, 1.28 mmol) were slurried in benzene (30 ml) and the mixture was refluxed through a Dean-Stark tube for 3 h. Then the temperature was settled to 80 ⁰ C and compound 43d (355 mg, 1.13 mmol) and tetrabutylammonium bromide (365 mg, 1.13 mmol) were added and the mixture was stirred over night. The suspension was filtered and the filtrate evaporated. Silica gel column chromatography

(gradient Q - ¹ A - I - 2%EtOH/DCM) gave the title compound (281 mg, 68%). MS mlz 480.3

(M+NH ₄ +MeCN) ⁺ .

¹ H-NMR (CDCl ₃ ): δ 7.31-7.16 (m, 4H), 4.78 (br, s, 2H) CH, NH, 4.52 (s, 2H), 4.40 (m, IH),

3.81 (m, IH), 3.60 (dd, IH), 3.51 (dd, IH), 2.58 (m, IH), 2.51 (br, s, IH) OH, 2.29 (m, IH),

2.17-2.01 (m, 2H), 1.42-1.36 (m, 12H) Me, t-but.,0.99 (d, 3H), 0.90 (d, 3H).

Step f

5-Azido-2,3,5-trideoxy-6-O-(3-((li?,61-l-(t-butoxycarbony lamino)ethyl) benzyl)-2-isopropyl-D- glucono-l,4-lactone (43f)

Triflic anhydride (224 μl) dissolved in DCM (1.2 ml) was added dropwise under nitrogen to a stirred DCM (5.8 ml) solution cooled at 0 ⁰ C of the alcohol 43e (281 mg, 0.67 mmol) and pyridine (0.16 ml). The mixture was stirred at 0 ⁰ C for 1 h. The mixture was poured into a chilled 5% NaHSO ₄ solution and extracted with DCM. The organic extract was dried through sodium sulfate and evaporated on rotavapor below r.t. The residue was dissolved in dimethylformamide (DMF, 5.8 ml) and then sodium azide (173 mg, 2.67 mmol) was added and

the mixture was stirred at 70 ⁰ C for 1 h. The solvent was removed by evaporation and the residue was partitioned between DCM and water. The organic extract was dried by sodium sulfate and evaporated. Silica gel column chromatography (gradient 10% EtAc / hexane - 30% EtAc / hexane) gave the title compound (178 mg, 60%). MS mlz 505.1 (M+NH ₄ +MeCN) ⁺ . 1H-NMR (CDCl ₃ ): δ 7.34-7.20 (m, 4H), 4.79 (br, s, 2H) CH, NH, 4.57, 4.56 (2xs, 2H), 4.50 (m, IH), 3.78 (m, 2H), 3.72 (m, IH), 2.72 (m, IH), 2.19-2.11 (m, 3H), 1.46-1.36 (m, 12H) Me, t- but, 1.01 (d, 3H), 0.91 (d, 3H).

Example 44 Step a

5-[2-(3,4-Dichloro-benzyloxy)-l-hydroxy-ethyll-3-isopropy l-dihydro-furan-2-one (44a) The procedure described in Example 1 step g was followed but using 3,4-dichlorobenzyl bromide instead of benzyl bromide, which gave the title compound (56% yield).

Step b

5-[l-Azido-2-(3,4-dichloro-benzyloxy)-ethyll-3-isopropyl- dihydro-furan-2-one (44b) Compound 44a was reacted according to the procedure described in Example 1 step h, which afforded the title compound (79%).

Biological Examples

To evaluate the enzymatic inhibition of renin exhibited by the compounds of the invention, an assay using Fluorescence Resonance Energy Transfer (FRET) to generate a spectroscopic

response to peptidase cleavage was used. The activity was measured by a continuous detection of increased fluorescence intensity exhibited by the cleavage product (peptide-EVANS). The enzyme used in the assay was recombinant human renin (supplied by Proteos), the substrate consisted of a peptide which in one end is linked to a fluorophore, 5-

(aminoethyl)aminonaphtalene sulphonate (EDANS), and in the other end to a non-fluorescent chromophore, 4'-dimethylaminoazobenzene (Dabcyl), typically Arg-Glu(ED ANS)-Ile-His-Pro- Phe-His-Leu-Val-Ile-His-Thr-Lys(DABCYL)-Arg (Sigma- Aldrich). The cleavage site by human renin is the peptide bond between Leu and VaI. The compounds were tested at a range of concentrations whereas the enzyme and substrate concentrations were fixed. The assay used employs the enzyme at a concentration of 6.25nM in an assay buffer consisting of of 0.1 mM Tris-HCl, 0.05 M NaCl, 0.5 mM EDTA, 0.05% CHAPS at pH=7.4. The substrate was prepared at a 20 μM stock solution in DMSO. To each well of a 96-well polypropylene plate was added the enzyme containing assay buffer (90.0 μl) and inhibitor of different concentrations (1 μl). To control wells were added DMSO (1 μl) instead of inhibitor. The renin enzyme was preactivated by incubation at 37 ⁰ C for 20 min whereafter the reactions were started by addition of substrate, 10 μl/well, thus giving a total volume of 100 μl/well and a substrate concentration of 2 μM. The assay was performed during 20 min at 37 ⁰ C. The total concentration of DMSO was not above 1 %. Product fluorescence (emission filter 340 nM, excitation filter 500 nM) was monitored with a Thermo Labsystems Fluoroskan Ascent plate reader. The Ki was determined by Prism Software. Activity of the inhibitors was determined by measuring the fluorescence at λe _X 340nm and λ _em 500nm. Percent inhibition is calculated as follows: % Inhibition is equal to the (Fluorescence^ inhibitor - Fluorescence _δωC £gro»«rf); divided by the (Fluorescence _mmus inhibitor - Fluorescence _δωC £ _g ro»«rf);

For example, Table 1 shows the Ki- value expressed in nM for a representative selection of compounds according to the invention when tested in a renin enzyme assay such as the one described above. Category A indicates < 50 nM inhibition, category B indicates 51 - 200 nM inhibition and category C indicates > 200 nM: