The invention relates to novel heteroaryl substituted tetrahydropyridine derivatives of the general formula (I). The invention also concerns related aspects including processes for the preparation of the compounds, pharmaceutical compositions containing one or more compounds of formula Q) and especially their use as renin inhibitors in cardiovascular events and renal insufficiency.

In the renin-angiotensin system (RAS) the biologically active angiotensin II (Ang II) is generated by a two-step mechanism. The highly specific enzyme renin cleaves angiotensinogen to angiotensin I (Ang I), which is then further processed to Ang II by the less specific angiotensin-converting enzyme (ACE). Ang II is known to work on at least two receptor subtypes called ATi and AT2. Whereas ATi seems to transmit most of the known functions of Ang II, the role of AT2 is still unknown.

Modulation of the RAS represents a major advance in the treatment of cardiovascular diseases. ACE inhibitors and ATi blockers have been accepted to treat hypertension (Waeber B. et ah, "The renin-angiotensin system: role in experimental and human hypertension", in Birkenhager W. H., Reid J. L. (eds): Hypertension, Amsterdam, Elsevier Science Publishing Co, 1986, 489-519; Weber M. A., Am. J. Hypertens., 1992, 5, 247S). In addition, ACE inhibitors are used for renal protection (Rosenberg M. E. et ctl, Kidney International, 1994, 45, 403; Breyer J. A. et ah, Kidney International, 1994, 45, S156), in the prevention of congestive heart failure (Vaughan D. E. et ah, Cardiovasc. Res., 1994, 2S ₅ 159; Fouad-Tarazi F. et ah, Am. J. Med, 1988, 84 (Suppl. 3A), 83) and myocardial infarction (Pfeffer M. A. et ah, N. Engl. J. Med, 1992, 327, 669).

The rationale to develop renin inhibitors is the specificity of renin (Kleinert H. D., Cardiovasc. Drugs, 1995, 9, 645). The only substrate known for renin is angiotensinogen, which can only be processed (under physiological conditions) by renin. In contrast, ACE can also cleave bradykinin besides Ang I and can be by-passed by chymase, a serine protease (Husain A., J. Hypertens., 1993, 11, 1155). In patients inhibition of ACE thus

leads to bradykinin accumulation causing cough (5-20%) and potentially life-threatening angioneurotic edema (0.1-0.2%) (Israili Z. H. et al, Annals of Internal Medicine, 1992, 117, 234). Chymase is not inhibited by ACE inhibitors. Therefore, the formation of Ang II is still possible in patients treated with ACE inhibitors. Blockade of the ATi receptor (e.g. by Iosartan) on the other hand overexposes other AT-receptor subtypes (e.g. AT ₂ ) to Ang II, whose concentration is significantly increased by the blockade of ATi receptors. In summary, 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.

Only limited clinical experience (Azizi M. et al, J. Hypertens., 1994, 12, 419; Neutel J. M. et al., Am. Heart, 1991, 122, 1094) has been created with renin inhibitors because of their insufficient oral activity due to their peptidomimetic character (Kleinert H. D., Cardiovasc. Drugs, 1995, 9, 645). The clinical development of several compounds has been stopped because of this problem together with the high cost of goods. Only one compound containing four chiral centers has entered clinical trials (Rahuel J. et al, Chem. Biol, 2000, 7, 493; Mealy N. E., Drugs of the Future, 2001, 26, 1139). Thus, renin inhibitors with good oral bioavailability and long duration of action are required. Recently, the first non- peptide renin inhibitors were described which show high in vitro activity (Oefher C. et al, Chem. Biol, 1999, 6, 127; Patent Application WO97/09311; Marki H. P. et al, E Farmaco, 2001, 56, 21). However, the development status of these compounds is not known.

The present invention relates to the identification of renin inhibitors of a non-peptidic nature and of low molecular weight. Described are orally active renin inhibitors of long duration of action which are active in indications beyond blood pressure regulation where the tissular renin-chymase system may be activated leading to pathophysiologically altered local functions such as renal, cardiac and vascular remodeling, atherosclerosis, and possibly restenosis. So, the present invention describes these non-peptidic renin inhibitors.

The present invention relates to novel heteroaryl substituted tetrahydropyridine derivatives of the general formula (I):

(D wherein

W represents 1,4-phenyl;

V represents a substituent of the formula -E ¹ -Z-E ² -;

E ¹ represents -0-CH ₂ -; - CH ₂ -CH ₂ -;

E ² represents a bond, -O-, -CH ₂ -;

Z represents a five-membered heteroaryl with 2 or 3 heteroatoms;

U represents phenyl or mono-, di-, tri- or terra- substituted phenyl, whereby the substituents are selected from the group consisting of halogen, -CF ₃ , C _1-7 -alkyl or C ₁ -C ₇ hydroxy-lower alkyl;

T represents -CONR ¹ -;

Q represents methylene;

M represents phenyl, mono- or di-substituted phenyl, whereby the substituents are selected from the group consisting of C ₁ . ₇ -a.kyl, C^-alkoxy, -OCF ₃ , -CF ₃ , hydroxy-Ci- ₇ -alkyl and halogen;

R ¹ represents Q- ₇ -alkyl or C ₃ -C ₆ cycloalkyl;

and optically pure enantiomers, mixtures of enantiomers such as racemates, diastereomers, mixtures of diastereomers, diastereomeric racemates, mixtures of diastereomeric

racemates, the meso-form as well as pharmaceutically acceptable salts, solvent complexes and morphological forms.

In the definitions of general formula (I) - if not otherwise stated - the term lower refers to a moiety having from one to seven carbon atoms, preferably from one to four carbon atoms.

In the definitions of general formula (I) - if not otherwise stated - the term lower alkyl, alone or in combination with other groups, means saturated, straight and branched chain groups with one to seven carbon atoms, preferably one to four carbon atoms that can be optionally substituted by halogens. Examples of lower alkyl groups are methyl, ethyl, n- propyl, iso-propyl, n-butyl, iso-butyl, sec-butyl, tert-butyl, pentyl, hexyl and heptyl. The methyl, ethyl and isopropyl groups are preferred.

The term lower alkoxy refers to a R-O group, wherein R is a lower alkyl. Examples of lower alkoxy groups are methoxy, ethoxy, propoxy, iso-propoxy, iso-butoxy, sec-butoxy and tert-butoxy.

The term hydroxy-lower alkyl alone or in combination with other groups refers to a HO-R group, wherein R is a lower alkyl. Examples of hydroxy-lower alkyl groups are HO-CH ₂ -, HO-CH ₂ CH ₂ - -CH(OH)CH ₃ , or HO-CH ₂ CH ₂ CH ₂ -.

The term halogen means fluorine, chlorine, bromine or iodine, preferably fluorine, chlorine and bromine.

The term cycloalkyl alone or in combination, means a saturated cyclic hydrocarbon ring system with 3 to 6 carbon atoms, e.g. cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl, which can be optionally mono- or multisubstituted by lower alkyl, lower alkenyl, lower alkenylene, lower alkoxy, lower alkylenoxy, lower alkylenedioxy, lower alkoxy - lower alkyl, cycloalkyl - lower alkyl, cycloalkoxy, cycloalkoxy - lower alkyl, hydroxy, halogen, -CF ₃ , and OCF ₃ .

The term cycloalkoxy refers to a R-O group, wherein R is a cycloalkyl. Examples of cycloalkoxy groups are cyclopropoxy, cyclobutoxy, cyclopentoxy.

The term heteroaryl, alone or in combination, means five-membered aromatic rings containing two or three heteroatoms, preferably nitrogen, oxygen or sulfur. Preferred are five-membered aromatic rings containing one oxygen and two nitrogen atoms; five-

membered aromatic rings containing one nitrogen and one oxygen atom; five-membered aromatic rings containing two nitrogen atoms; five-membered aromatic rings containing three nitrogen atoms; five-membered rings containing two nitrogen atoms and one sulfur atom. Examples of such ring systems are triazinyl, thiazinyl, thiazolyl, isothiazolyl, oxazolyl, isoxazolyl, oxadiazolyl. Such rings may be adequatly substituted with lower alkyl, lower alkenyl, lower alkinyl, lower alkylene, lower alkenylene, lower alkylenedioxy, lower alkyleneoxy, hydroxy-lower alkyl, lower alkoxy, hydroxy, halogen, -CF ₃ , -OCF ₃ , another aryl, preferably phenyl or substituted phenyl, or another heteroaryl and the like.

The term V within the present invention V represents a substituent of the formula -E ¹ -Z- E ² - which may be connected in both possible ways to the group W and U of the compound of formula I. In a preferred embodiment of the invention the beginning part of the group -E'-Z-E ² - is linked to the group W of the compound of formula I (that means, for example, that the -E^part of -E ¹ ^-Z-E ² - is linked to the group W of the compound of formula I.

The term T within the present invention represents -CONR ¹ - which may be connected in both possible ways to the core structure of compound of formula I. In a preferred embodiment of the invention the beginning part of the group T is linked to the core structure of compound of formula I (that means, for example, that the -C(=O) part of -

CONR ¹ - is linked to the core structure of compound of formula I. Preferably, the term T within the present invention represents -CONR ¹ - ,wherein R ¹ represents C _1-7 -alkyl or C ₃ - Ce-cycloalkyl, preferably and most preferred cyclopropyl.

The expression pharmaceutically acceptable salts encompasses either salts with inorganic acids or organic acids like hydrochloric or hydrobromic acid, sulfuric acid, phosphoric acid, citric acid, formic acid, acetic acid, maleic acid, tartaric acid, benzoic acid, methanesulfonic acid, p-toluenesulfonic acid, and the like that are non toxic to living organisms or in case the compound of formula (I) is acidic in nature with an inorganic base like an alkali or earth alkali base, e.g. sodium hydroxide, potassium hydroxide, calcium hydroxide and the like.

The compounds of the general formula (I) contain two or more asymmetric carbon atoms and are prepared in form of optically pure enantiomers, mixtures of enantiomers such as racemates, diastereomers, mixtures of diastereomers, diastereomeric racemates, mixtures of diastereomeric racemates, and the meso-form and pharmaceutically acceptable salts thereof.

The present invention encompasses all these forms. Mixtures are separated in a manner known per se, i.e. by column chromatography, thin layer chromatography, HPLC or crystallization.

In a preferred embodiment M represents phenyl, mono- or di-substituted phenyl, wherein the substituents are selected from the group consisting of C _1-7 -alkyl, C _1-7 -alkoxy, and halogen.

In a further preferred embodiment M represents disubstituted phenyl, wherein the substituents are selected from the group consisting of C _1-7 -alkyl, C^-alkoxy, and chlorine.

In a further preferred embodiment R ¹ represents a cyclopropyl group.

In a further preferred embodiment Z represents an oxadiazol ring or isoxazol ring

In a further preferred embodiment U represents a mono-, di-, or trisubstituted phenyl wherein the substituents are selected from the group consisting of halogen and C _1-7 -alkyl.

In a more preferred embodiment heteroaryl substituted diazabicyclononene derivatives of general formula (I) are those selected from the group consisting of:

(rac.)-(lR*, 55'*)-7-{4-[5-(2-chloro-3,6-difluorophenyl)-[l,2,4]oxadiazol -3-ylmethoxy]- phenyl}-3,9-diazabicyclo[3.3.1]non-6-ene-6-carboxylic acid cyclopropyl-(3-methoxy-2- methylbenzyl)amide,

(røc.)-(ii?*, 55'*)-7-{4-[5-(2-chlorophenyl)-[l,2,4]oxadiazol-3-ylmethoxy] phenyl}-3,9- diazabicyclo[3.3. l]non-6-ene-6-carboxylic acid cyclopropyl-(3-methoxy-2-methyl- benzyl)amide,

(rac.)-(lR*, 5,S*)-7-(4-{2-[3-(2,3-dichlorophenyl)-[l,2,4]oxadiazol-5-yl] ethyl}phenyl)-3,9- diazabicyclo[3.3. l]non-6-ene-6-carboxylic acid cyclopropyl-(3-methoxy-2-methyl- benzyl)amide,

(rac)-(lR*, 55*)-7-(4-{2-[3-(2,6-dichlorophenyl)-[l,2,4]oxadiazol-5-yl]e thyl}phenyl)-3,9- diazabicyclo[3.3.1]non-6-ene-6-carboxylic acid cyclopropyl-(3-methoxy-2-methyl- benzyl)amide,

(rac.)-(7i?*, 55*)-7-{4-[3-(2-chloro-3,6-difluorophenyl)-[l,2,4]oxadiazol- 5-ylmethoxy]- phenyl}-3,9-diazabicyclo[3.3. l]non-6-ene-6-carboxylic acid cyclopropyl-(2,3-dichloro- benzyl)amide, and

(rac.)-(lR*, 55'*)-7-{4-[3-(2-chloro-3,6-difluorophenyl)isoxazol-5-ylmeth oxy]phenyl}- 3,9-diazabicyclo[3.3.1]non-6-ene-6-carboxylic acid cyclopropyl-(2,3-dichlorobenzyl)- amide.

The invention relates to a method for the treatment and/or prophylaxis of diseases which are 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, which method comprises administrating a compound as defined above to a human being or animal.

In another embodiment, the invention relates to a method for the treatment and/or prophylaxis of diseases which are 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, complications resulting from diabetes such as nephropathy, vasculopathy and neuropathy.

In another embodiment, the invention relates to a method for the treatment and/or prophylaxis of diseases, which are associated with a dysregulation of the renin-angiotensin system as well as for the treatment of the above-mentioned diseases.

The invention also relates to the use of compounds of formula (I) for the preparation of a medicament for the treatment and/or prophylaxis of the above-mentioned diseases.

A further aspect of the present invention is related to a pharmaceutical composition containing at least one five-membered heteroaryl derivative according to general formula (I) and pharmaceutically acceptable carrier materials or adjuvants. This pharmaceutical composition may be used for the treatment or prophylaxis of the above-mentioned disorders; as well as for the preparation of a medicament for the treatment and/or prophylaxis of the above-mentioned diseases.

Compounds of formula (I) and their pharmaceutically acceptable acid addition salts can be used as medicaments, e. g. in the form of pharmaceutical compositions containing at least one compound of formula (I) and pharmaceutically acceptable inert carrier material or adjuvants. These pharmaceutical compositions can be used for enteral, parenteral or topical administration. They can be administered, for example, perorally, e. g. in the form of tablets, coated tablets, dragees, hard and soft gelatine capsules, solutions, emulsions or suspensions, rectally, e. g. in the form of suppositories, parenterally, e. g. in the form of injection solutions or infusion solutions or topically, e. g. in the form of ointments, creams or oils.

Five-membered heteroaryl derivatives of formula (I) or the above-mentioned pharmaceutical compositions are also of use in combination with other pharmacologically active compounds comprising ACE-inhibitors, neutral endopeptidase inhibitors, angiotensin II receptor antagonists, aldosterone antagonists, endothelin receptors antagonists, vasodilators, calcium antagonists, potassium activators, diuretics, sympatholitics, beta-adrenergic antagonists, alpha-adrenergic antagonists or with other drugs beneficial for the prevention or the treatment of the above-mentioned diseases.

The production of pharmaceutical preparations can be effected in a manner which will be familiar to any person skilled in the art by bringing the described compounds of formula (I) and their pharmaceutically acceptable acid addition salts, optionally in combination with other therapeutically valuable substances, into a galenical administration form together with suitable, non-toxic, inert, therapeutically compatible solid or liquid carrier materials and, if desired, usual pharmaceutical adjuvants.

Suitable carrier materials are not only inorganic carrier materials, but also organic carrier materials. Thus, for example, lactose, cornstarch or derivatives thereof, talc, stearic acid or its salts can be used as carrier materials for tablets, coated tablets, dragees and hard gelatine capsules. Suitable carrier materials for soft gelatine capsules are, for example, vegetable oils, waxes, fats and semi-solid and liquid polyols (depending on the nature of the active ingredient no carriers are, however, required in the case of soft gelatine capsules). Suitable carrier materials for the production of solutions and syrups are, for example, water, polyols, sucrose, invert sugar and the like. Suitable carrier materials for injections are, for example, water, alcohols, polyols, glycerols and vegetable oils. Suitable carrier materials for suppositories are, for example, natural or hardened oils, waxes, fats and semi-liquid or liquid polyols. Suitable carrier materials for topical preparations are glycerides, semi-synthetic and synthetic glycerides, hydrogenated oils, liquid waxes, liquid paraffins, liquid fatty alcohols, sterols, polyethylene glycols and cellulose derivatives.

Usual stabilizers, preservatives, wetting and emulsifying agents, consistency-improving agents, flavor-improving agents, salts for varying the osmotic pressure, buffer substances, solubilizers, colorants and masking agents and antioxidants come into consideration as pharmaceutical adjuvants.

Heteroaryl substituted tetrahydropyridine derivatives according to formula (I) are also used in combination with other pharmacologically active compounds comprising ACE- inhibitors, neutral endopeptidase inhibitors, angiotensin II receptor antagonists, endothelin receptors antagonists, vasodilators, calcium antagonists, potassium activators, diuretics, sympatholitics, beta-adrenergic antagonists, alpha-adrenergic antagonists or with other drugs beneficial for the prevention or the treatment of the above-mentioned diseases.

In a preferred embodiment, the amount administered to a patient of a pharmaceutically active amount of a heteroaryl substituted tetrahydropyridine derivative according to formula (I) is comprised between 2 mg and 1000 mg per day.

In a particular preferred embodiment, this amount is comprised between 1 mg and 500 mg per day.

In a more particularly preferred embodiment, this amount is comprised between 5 mg and 200 mg per day.

All forms of prodrugs leading to an active component comprised by general formula (I) above are included in the present invention.

Another aspect of the invention is related to a process for the preparation of a pharmaceutical composition comprising a heteroaryl substituted tetrahydropyridine derivative of the general formula (I). According to said process, one or more active ingredients of the general formula (I) are mixing with inert excipients in a manner known per se.

The heteroaryl substituted tetrahydropyridine derivatives of general formula (I) can be manufactured by the methods outlined below, by the methods described in the examples or by analogous methods.

Compounds mentioned below, and not including W-V-U as a specific substituent, can be prepared by the chemistry described in the patent application WO03/093267 that is incorporated by reference in this respect. A compound of type A in Scheme 1, wherein PG stands for a protecting group, can be prepared from P-methyl-T-trifluoromethanesulfonyloxy-S^-diazabicyclop.S.lj non-ό-ene- 3,6-dicarboxylic acid 3-fert-butyl ester 6-ethyl ester (WO03/093267), and a protected 4- bromophenol derivative via a Negishi coupling, or any other coupling catalyzed by a palladium catalyst. A protecting group manipulation delivers a compound of type B, then a hydrolysis of the ethyl ester yields a compound of type C. An amide coupling, for instance, leads then to a compound of type E, and a subsequent ether formation, via a Mitsunobu reaction, for instance, delivers a compound of type F. The fragment necessary for the construction of the fragment U-V is prepared in parallel, very often following literature procedures. Final deprotection yields a compound of general formula (I).

Scheme 1

Otherwise the segment U-V- can be prepared on the template directly, as described in Scheme 2. Starting from a compound of type E, a compound of type H can be prepared, wherein X stands for a precursor of the substituent V as described in general formula (I). X can be modified along the synthesis. Finally the whole segment U-V- is constructed to yield a compound of type F. Deprotection yields a compound of general formula (I).

Scheme 2

Another possibility is sketched in Scheme 3. The X-E ¹ substituent can be present from the beginning on the phenyl moiety, and be introduced on 9-methyl-7- trifluoromethanesulfonyloxy-3,9-diazabicyclo[3.3.1]non-6-ene -3,6-dicarboxylic acid 3- tert-bvLty\ ester 6-ethyl ester (WO03/093267) via aNegishi coupling to a compound of type J. Subsequent protecting group manipulations lead to a compound of type K, then hydrolysis to a compound of type L. An amide coupling, for instance, leads to a compound of type H. The synthesis can then follow the schemes 1 and 2.

Scheme 3

The following examples serve to illustrate the present invention in more detail. They are, however, not intended to limit its scope in any manner.

Chemistry

Abbreviations:

Ac Acetyl

ACE Angiotensin Converting Enzyme Ang Angiotensin aq. aqueous

Boc fert-Butyloxycarbonyl

Bu Butyl

BSA Bovine serum albumine BuLi o-Butyllithium cone. concentrated

DIPEA Diisopropylethylamine

DMAP 4-N,N-Dimethylaminopyridine

DMF N,N-Dimethylformamide DMSO Dimethylsulfoxide

EDC-HCl Ethyl-N,N-dimethylaminopropylcarbodiimide hydrochloride EIA Enzyme immunoassay Et Ethyl

EtOAc Ethyl acetate FC Flash Chromatography HOBt Hydroxybenzotriazol LC-MS Liquid chromatography - mass spectrometry MeOH Methanol org. organic PG protecting group

RAS Renin Angiotensin System rt room temperature sat. saturated sol. Solution

TBAF Tetra-M-butylammonium fluoride

TBDMS tert-Butyldimethylsilyl

Tf Trifluoromethylsulfonyl

THF Tetrahydrofuran

TLC Thin Layer Chromatography

General remarks

The compound is characterized at least by LC-MS and ¹ H-NMR. Only the LC-MS data are given here (Zorbax SB-AQ column, 5 μm, 4.6x50 mm; eluent A: 0.04% trifluoroacetic acid in water; eluent B: acetonitrile; gradient 5% -» 100% eluent B over 1.5 min, flow 1 mL/min)

Experimental part

Preparation of the aryl-heterocyclyl derivatives

2-Chloro-3,6-difluoro-benzaldehyde oxime

A mixture of 2-chloro-3,6-difluorobenzaldehyde (1.13 mmol, 200 mg) and NaHCO ₃ (3.4 mmol, 286 mg) in water (1.9 mL) was treated at it with H ₂ NOH-HCl (2.3 mmol, 160 mg) and Bu ₄ NCl (0.06 mmol, 16 mg). Acetonitrile (1.2 mL) was then added and the mixture was stirred at rt for 90 min. AcOH (0.2 mL) was then added and the mixture was stirred for 30 min at rt. The mixture was extracted with EtOAc, washed with brine, dried over MgSO ₄ , filtered, and the solvents were removed under reduced pressure. Purification of the residue by FC (EtOAc/heptane 3:7) yielded the title compound (170 mg, 78%). LC- MS: t _R = 0.84 min; ES+ + CH ₃ CN: 233.81.

5-(tert-ButyldimethyIsilanyloxymethyl)-3-(2-chloro-3,6-di fluorophenyl)isoxazole

A solution of 2-chloro-3,6-difluoro-benzaldehyde oxime (0.834 mmol, 170 mg) in DMF (0.53 mL) was added dropwise at rt to a solution of N-chlorosuccinimide (0.83 mmol, 111 mg) and pyridine (2 drops) in DMF (1.8 mL). After 1 h, fert-butyldimethylprop-2- ynyloxysilane (0.33 mmol, 0.07 mL) was added and the mixture was heated to 85 ⁰ C. Et ₃ N (0.83 mmol, 0.12 mL, 2.5 eq) in DMF (0.7 mL) was then added over 35 min. At the end of the addition, the mixture was stirred for 1 h at 85 ⁰ C. Water was added at rt and the mixture was extracted with EtOAc. The org. extracts were washed with aq. 10% KHSO ₄ , water, and brine, dried over MgSO ₄ , filtered, and the solvents were removed under reduced

pressure. Purification of the residue by FC (EtO Ac/heptane 5:95) yielded the title compound (94 mg, 79%). LC-MS: t _R = 1.16 min; ES+: 360.05.

[3-(2-ChIoro-3,6-difluorophenyl)isoxazol-5-yl]methanol

A solution of 5-(tert-butyldimethylsilanyloxymethyl)-3-(2-chloro-3,6-diflu orophenyl)- isoxazole (0.25 mmol, 94 mg) in THF (6.8 niL) at 0 ⁰ C was treated with TBAF (0.50 mmol, 0.50 mL of a 1 M solution in THF). After 1 h at rt, aq. sat. NH ₄ Cl was added. The mixture was extracted with EtOAc, dried over MgSO ₄ , filtered, and the solvents were removed under reduced pressure. Purification of the residue by FC (EtOAc/heptane 3:7) yielded the title compound (60 mg, 98%). LC-MS: t _R = 0.84 min; ES+ + CH ₃ CN: 287.14.

2-ChIoro-3,6-difluorobenzoαitriIe

2-Chloro-3,6-difluorobenzamide (0.80 g, 4.18 mmol) in POCl ₃ (12 mL) was heated for 2 h at 95 ⁰ C. The sol. was cooled and the excess of POCl ₃ was removed by distillation under reduced pressure. The residue was diluted with EtOAc and aq. 10% K ₂ CO ₃ was cautiously added. The layers were separated and the org. phase was washed with water (2x), and brine. The org. extracts were dried over MgSO ₄ , filtered, and the solvent were removed under reduced pressure to yield the title compound (0.46 g) that was not further purified.

2,3-dichIoro-iV-hydroxybenzamidine

To 2-chloro-3,6-difluorobenzonitrile (0.36 g, 2.08 mmol) in 95 % EtOH (4 mL) was added H ₂ NOH-HCl (0.32 g, 4.56 mmol) and Et ₃ N (0.66 mL, 4.77 mmol). The mixture was stirred at 75 ⁰ C for 18 h. After cooling to rt, the mixture was filtered. The filtrate was diluted with water (3 mL), and neutralized to pH 7 with aq. cone. HCl. The solvents were partially evaporated under reduced pressure, and the residue was filtered, washed with water, and dried under vacuum. The residue (0.4Ig) was used in the next step without further purification. LC-MS: R _t = 0.34 min, ES+=207.02.

(rac.)-(lR*, 5.S'*)-9-Methyl-7-(4-triisopropyIsilanyloxyphenyl)-3,9-diaza bicyclo[3.3.1]- non-6-ene-3,6-dicarboxylic acid3-tert-butyl ester 6-ethyl ester (Al)

A sol. of (4-bromophenoxy)triisopropylsilane (P. Wipf et ah, Org. Lett. 2000, 2, 26, 4213- 4216; 18.6 g; 56.4 mmol) in THF (215 mL) was cooled to -78 ⁰ C. BuLi (1.6M in hexane, 36.8 mL; 58.8 mmol) was added dropwise over 15 min. After completion of the addition, the resulting solution was stirred further at -75 ⁰ C for 30 minutes. ZnCl ₂ (1.06 M in THF; 58.8 mL, 62.3 mmol) was added dropwise over 20 min, and the reaction mixture was allowed to warm up to rt. A mixture of 9-methyl-7-trifluoromethanesulfonyloxy-3,9- diazabicyclo[3.3.1]non-6-ene-3,6-dicarboxylic acid 3-te/t-butyl ester 6-ethyl ester (WO03/093267, 10.78 g; 23.5 mmol) and Pd(PPtIs) ₄ (1.36 g; 1.18 mmol) in THF (25 mL) was added. The resulting sol. was stirred at rt for 5 min., then at 45 ⁰ C for 50 min. The reaction mixture was cooled to 0 ⁰ C, and aq. sat. NaHCO ₃ was added to this reaction mixture, followed by EtOAc. The phases were shaken, separated and the aq. phase was extracted with EtOAc (2 x 100 ml). The combined org. extracts were dried over MgSO ₄ , filtered, and the solvents were removed under reduced pressure. Purification of the residue by FC (CH ₂ C1 ₂ /CH ₃ OH 20/1) yielded the title compound (10.99 g, 84%). LC-MS: t _R = 1.03 min.

(mc.)-(-?jR* ₅ 5iS ^> *)-7-(4-TriisopropyIsiIanyloxyphenyl)-3,9-diazabicyclo [3.3.1]non-6-ene- 3,6,9-tricarboxylic acid 3,9-di-tert-butyI ester 6-ethyl ester (Bl)

A mixture compound Al (3.5 g, 6.26 mmol), NaHCO ₃ (5.26 g, 62.64 mmol) and 1- chloroethyl chloroformate (6.80 mL, 62.4 mmol) in CH ₂ ClCH ₂ Cl (69 mL) was heated to reflux for 1 h. The reaction mixture was allowed to cool to rt, and was filtered. The precipitate was washed with 1,2-dichloroethane, and the filtrate was concentrated under reduced pressure. The residue was dissolved in MeOH (69 mL), and the resulting sol. was stirred at 4O ⁰ C for Ih. The solvents were removed under reduced pressure, and the residue was dried under high vacuum for 1 h. The dried residue was dissolved in CH ₂ Cl ₂ (69 mL). The resulting sol. was cooled to 0 ⁰ C. DIPEA (6.45 mL, 37.67 mmol) was added dropwise, followed by the addition at once of Boc ₂ O (4.10 g, 18.79 mmol). After further stirring at 0 ⁰ C for 30 min., the mixture was stirred at rt for 3 h. Aq. IM HCl (35 mL) was added dropwise to the reaction mixture at 0 ⁰ C. The phases were shaken and separated. The org. phase was washed with an aq. sat. NaHCO ₃ . The combined org. extracts were dried over MgSO ₄ , filtered, and the solvents were removed under reduced pressure.

Purification by FC (CH ₂ C1 ₂ /CH ₃ OH 100/1) yielded the title product as a yellow foam (3.64 g, 90%). LC-MS : t _R = 1.30 min.

(rac.)-(lR*, 55 ^r *)-7-[4-(tert-Butyldimethylsilanyloxy)phenyl]-3,9-diaz abicyclo[3.3.1]- non-6-ene-3,6,9-tricarboxylic acid 3,9-di-tert-bntyl ester (Cl)

To a sol. of compound Bl (8.78 g, 13.6 mmol) in EtOH (100 mL) was added aq. IM NaOH (41 mL, 41 mmol). The resulting mixture was stirred at 80 ⁰ C for 20 h. The solvents were partially removed under reduced pressure. EtOAc, then aq. IM HCl were added. The mixture was shaken, and the phases were separated. The aq. phase was extracted with EtOAc (2x). The combined org. extracts were dried over MgSO ₄ , filtered, and the solvents were removed under reduced pressure. The residue was dried under high vacuum, and dissolved in DMF (155 mL). Imidazole (4.51 g; 66.2 mmol) and TBDMS-Cl (6.24 g, 41.4 mmol) were added. The reaction mixture was stirred at rt for 5 h. The reaction mixture was cooled to 0 ⁰ C, and aq. sat. NH ₄ Cl (155 ml) was slowly added. The resulting mixture was extracted with heptane/Et ₂ O (1/1, 5x), and the combined org. extracts were dried over MgSO ₄ , filtered, and the solvents were removed under reduced pressure. The resulting residue was dissolved in THF (315 mL). MeOH (32 mL), water (32 mL) and K ₂ CO ₃ (1.56 g) were added. The reaction mixture was stirred at rt for 30 min. The reaction mixture was cooled to 0 ⁰ C, and aq. sat. NH ₄ Cl (300 mL) was slowly added. The phases were separated and the aq. phase was extracted with Et ₂ O. The combined org. extracts were dried over MgSO ₄ , filtered, and the solvents were removed under reduced pressure. Purification of the residue by FC (CH ₂ Cl ₂ ZCH ₃ OH 40/1) yielded a mixture of the title compound with (rac.)-(lR*, 55' ^5|! )-7-[4-(tert-butyldimethylsilanyloxy)phenyl]-3,9- diazabicyclo[3.3.1]-non-7-ene-3,6,9-tricarboxylic acid 3,9-di-tert-butyl ester (5.57 g, 71%). LC-MS: t _R - 1.14 min. and 1.15 min. (minor isomer).

(rac.)-(lR*, 55 ^r *)-7-[4-(tert-Butyldimethylsilanyloxy)phenyl]-6-[cyclo propyl-(3-meth- oxy-2-methyIbenzyl)carbamoyl]-3,9-diazabicydo[3.3.1]non-6-en e-3,9-dicarboxyIic acid di-tert-butyl ester (Dl)

To a sol. of compound Cl with its regioisomer (1.64 g; 2.853 mmol; 1 equiv.) in CH ₂ Cl ₂ (32 ml) were added at rt cyclopropyl-(3-methoxy-2-methylbenzyl)amine (WO2004/096366; 1.64 g, 8.56 mmol), DIPEA (1.95 mL, 11.39 mmol) DMAP (87 mg,

0.71 mmol), HOBt (578 mg, 4.28 mmol), and EDC-HCl (2.19 g, 11.413 mmol). The mixture was stirred at rt for 3 days. CH ₂ Cl ₂ (50 mL) was added to the reaction mixture, and the sol. was washed with aq. IM HCl (3x). The combined aq. layers were extracted with CH ₂ Cl ₂ (25 mL). The combined org. extracts were washed with aq. sat. NaHCO ₃ (25 ml). The org. extracts were dried over MgSO ₄ , filtered, and the solvents were removed under reduced pressure. Purification of the residue by FC (CH2C1 ₂ /CH ₃ OH 100/3) yielded the title compound (1.45 g, 68%). LC-MS: t _R = 1.29 min.

(rac.)-(lR*, 55 ^r *)-7-[4-(tert-ButyldimethylsilanyIoxy)pheiiyl]-6-[cycl opropyI-(2,3- dichlorobenzyI)carbamoyI]-3,9-diazabicyclo[3.3.1]non-6-ene-3 ,9-dicarboxyIic acid di- tert-bntyl ester (D2)

A solution of compound Cl (3.48 mmol, 2.0 g) in CH ₂ Cl ₂ (28 mL) was treated with

DMAP (0.87 mmol, 106 mg), HOBt (4.18 mmol, 0.56 g), EDC-HCl (8.70 mmol, 1.67 g), and DIPEA (13.92 mmol, 2.38 mL). After 30 min at rt, cyclopropyl-(2,3-dichlorobenzyl)- amine (WO2004096366; 10.4 mmol, 2.30 g) was added and the mixture was stirred for 5 days at rt. The reaction was then diluted with CH ₂ Cl ₂ , washed with aq. IM HCl, dried over MgSO4, filtered, and the solvents were removed under reduced pressure. Purification of the residue by FC (EtOAc/heptane 30:70) yielded the title compound (1.2 g, 45%). LC- MS: t _R = 1.31 min; ES+: 772.55.

(rac.)-(lR*, 55'*)-6-[Cyclopropyl-(3-methoxy-2-methylbenzyl)carbamoyI]-7- (4- hydroxyphenyl)-3,9-diazabicyclo[3.3.1]non-6-ene-3,9-dicarbox yIic acid di-tert-butyl ester (El)

To a sol. of compound Dl (2.13 g, 2.85 mmol) in THF (32 mL) at 0 ⁰ C was added dropwise a sol. of TBAF (899 mg 2.85 mmol) in THF (9 mL). The reaction mixture was stirred at O ⁰ C for 40 min. Aq. sat. NH ₄ Cl was added. EtOAc was then added and the phases were shaken before being separated. The org. phase was washed with aq. sat. NH ₄ CI and with brine. The org. extracts were dried over MgSO ₄ , filtered, and the solvents were reduced under reduced pressure. Purification of the residue by FC (CH ₂ Cl ₂ ZCH ₃ OH 100/3) yielded the title compound (1.73 g, 96%). LC-MS: t _R = 1.09 min.

(rac.)-(lR*, 5£*)-6-[CyclopropyI-(2,3-dichIorobenzyl)carbamoyl]-7-(4-hyd roxy- phenyI)-3,9-diazabicycIo[3.3.1]non-6-ene-3,9-dicarboxylic acid di-tert-butyl ester (E2)

A solution of compound D2 (1.55 mmol, 1.2 g) in THF (31 mL) at 0 ⁰ C was treated with TBAF (3.3 mmol, 3.3 mL of a IM solution in THF). After 4 h at rt, aq. sat. NH ₄ Cl was added. The mixture was extracted with EtOAc, dried over MgSO ₄ , filtered, and the solvents were removed under reduced pressure. Purification of the residue by FC (EtOAc/heptane 50:50) yielded the title compound (0.85 g, 83%). LC-MS: t _R = 1.12 min; ES+: 658.47.

(rac.)-(lR *, 5^*)-7-{4-[5-(2-ChIoro-3,6-difluorophenyl)-[l,2,4]oxadiazoI- 3-ylmethoxy]- phenyl}-6-[cyclopropyl-(3-methoxy-2-methyIbenzyl)carbamoyl]- 3,9-diazabicyclo- [3.3.1]non-6-ene-3,9-dicarboxyIic acid di-tert-butyl ester (Fl)

To a sol. of compound H2 (0.07 g, 0.1 mmol) in pyridine (1 mL) was added 2-chloro-3,6- difluorobenzoyl chloride (0.025 g, 0.12 mmol) and the reaction mixture was refluxed for 4 h. EtOAc was added and the mixture was washed with aq. IM HCl (3x), water and brine. The org. phase was dried over Na ₂ SO ₄ , filtered, and the solvents were removed under reduced pressure. The residue was used in the next step without further purification. LC- MS: t _R = 1.23 min, ES+=862.51.

(rac.)-(lR*, 55*)-7-{4-[5-(2-Chlorophenyl)-[l,2,4]oxadiazoI-3-ylmethoxy]p henyI}-6- [cyclopropyI-(3-methoxy-2-methylbenzyl)carbamoyI]-3,9-diazab icycIo[3.3.1]non-6- ene-3,9-dicarboxylic acid di-tert-butyl ester (F2)

To a sol. of compound H2 (0.07 g, 0.1 mmol) in pyridine (1 mL) was added 2-chloro- benzoyl chloride (0.025 g, 0.12 mmol) and the reaction mixture was refluxed for 4 h. EtOAc was added and the mixture was washed with aq. IM HCl (3x), water and brine. The org. phase was dried over Na ₂ SO ₄ , filtered, and the solvents were removed under reduced pressure. The residue was used in the next step without further purification. LC- MS: t _R = 1.23 min, ES+=826.46.

(rac.)-(lR*, 55*)-6-[Cyclopropyl-(3-methoxy-2-methyIbenzyl)carbamoyl]-7-( 4-{2-[3- (ijS-dichlorophenylJ-tl^^loxadiazoI-S-yllethylJpheny^-S^-dia zabicycIop.S.llnon-δ- ene-3,9-dicarboxylic acid di-tert-bntyl ester (F3)

A sol. of compound H5 (0.52 g, 0.05 mmol), 2,3-dichloro-N-hydroxybenzamidine (0.011 g, 0.055 mmol), DMAP (0.002 g, 0.012 mmol), DIPEA (0.035 mL, 0.2 mmol), HOBt

(0.042 g, 0.062 mmol) and EDC-HCl (0.014 g, 0.075 mmol) in toluene (2 mL) was stirred for 72 h. The mixture was then heated at HO ⁰ C for 6h. After being cooled, the mixture was diluted with CH ₂ Cl ₂ and poured on a syringe containing diatomaceous earth pretreated with aq. IM HCl (Isolute Sorbent Technology, Johnson, C.R., et ah, Tetrahedron, 1998, 54, 4097). The product was eluted with CH ₂ Cl ₂ and the solvent was removed under reduced pressure. The residue was used in the next step without further purification. LC- MS: t _R = 1.25 min, ES-l-858.41.

(rac)-(lR *, 55*)-7-(4-{2-[3-(2-Chlorophenyl)-[l,2,4]oxadiazol-5-yl]ethyl }phenyl)-6- [cyclopropyl-(3-methoxy-2-methyIbenzyI)carbamoyI]-3,9-diazab icycIo[3.3.1]noii-6- ene-3,9-dicarboxylic acid di-tert-bntyl ester (F4)

A sol. of compound H5 (0.52 g, 0.05 mmol), 2-chloro-N-hydroxybenzamidine (0.011 g, 0.055 mmol), DMAP (0.002 g, 0.012 mmol), DIPEA (0.035 mL, 0.2 mmol), HOBt (0.042 g, 0.062 mmol) and EDC-HCl (0.014 g, 0.075 mmol) in toluene (2 mL) was stirred for 72 h. The mixture was then heated at 110 ⁰ C for 6h. After being cooled, the mixture was diluted with CH ₂ Cl ₂ and poured on a syringe containing diatomaceous earth pretreated with aq. IM HCl (Isolute Sorbent Technology, Johnson, C.R., et ah, Tetrahedron, 1998, 54, 4097). The product was eluted with CH ₂ Cl ₂ and the solvent was removed under reduced pressure. The residue was used in the next step without further purification. LC-MS: tR = 1.24 min, ES-H=858.50.

(rac.)-(lR*, 5.S ^r *)-7-{4-[3-(2-Chloro-3,6-difluorophenyl)isoxazol-5-ylm ethoxy]phenyl}- 6-[cyclopropyI-(2,3-dichlorobenzyl)carbamoyl]-3,9-diazabicyc Io[3.3.1]non-6-ene-3,9- dicarboxylic acid di-tert-butyl ester (FS)

A solution of compound E2 (0.08 mmol, 50 mg) in toluene (1 mL) was treated with [3-(2- chloro-3,6-difluorophenyl)isoxazol-5-yl]methanol (0.11 mmol, 28 mg) followed by azodicarboxylic dipiperidide (0.15 mmol, 38 mg) and tributylphosphine (0.27 mmol, 0.06 mL). The mixture was stirred at 110 ⁰ C for 1 h, cooled to rt, and filtered on IsoluteO. The solvents were removed under reduced pressure. Purification of the residue by HPLC yielded the title compound (33 mg, 47%). LC-MS: t _R = 1.24 min; ES+: 887.25.

(rac.)-(lR*, 5S*)-7-{4-[3-(2-Chloro-3,6-difluorophenyl)-[l,2,4]oxadiazoI- 5-ylmethoxy]- phenyl}-6-[cyclopropyl-(2,3-dichlorobenzyl-carbamoyI]-3,9-di azabicyclo[3.3.1]non-6- ene-3,9-dicarboxylic acid di-tert-bntyl ester (F6)

A sol. of compound H6 (0.30 g, 0.43 mmol), 2-chloro3,6-difluoro-N-hydroxybenzamidine (0.17 g, 0.85 mmol), DMAP (0.005 g, 0.04 mmol), DIPEA (0.22 mL, 1.28 mmol), HOBt

(0.065 g, 0.43 mmol) and EDC-HCl (0.12 g, 0.64 mmol) in CH ₂ Cl ₂ (4 mL) was stirred for 48 h. The mixture was diluted with CH ₂ Cl ₂ , washed with aq. 1 HCl, water, and finally brine. The organic layer was dried over MgSO ₄ , filtered, and the solvent was evaporated under reduced pressure. Pyridine (1 mL) was added to the residue and the mixture was then heated at HO ⁰ C for 4 h. After being cooled, the mixture was diluted with CH ₂ Cl ₂ , washed with aq. IM HCl, water, and brine. The org. extracts were evaporated under reduced pressure. The residue (0.28 g) was used in the next step without further purification. LC-MS: R _t = 1.24 min, ES+=888.38.

{rac)-(lR*, 5.y*)-7-(4-Cyanomethoxyphenyl)-6-[cyclopropyI-(3-methoxy-2-m ethyl- benzyl)carbamoyl]-3,9-diazabicycIo[3.3.1]non-6-ene-3,9-dicar boxyIic acid di-tert- butyl ester (Hl)

To a sol. of compound El (1.40 g 2.20 mmol) in anhydrous acetone (16 mL) were added successively K ₂ CO ₃ (381 mg, 2.76 mmol), NaI (63 mg, 0.42 mmol), and chloroacetonitrile (208 mg, 2.75 mmol). The mixture was heated to reflux for 4h. The crude reaction mixture was allowed to cool to rt, and was filtered. The filtrate was concentrated under reduced pressure, and was purified by FC (CH ₂ C1 ₂ /CH ₃ OH 100/3). This yielded the title compound (1.35 g, 91%). LC-MS: t _R = 1.14 min.

{rac.)-(lR*, 5.S'*)-6-[CycIopropyl-(3-methoxy-2-methylbenzyl)carbamoyl]-7 -[4-(iV- hydroxycarbamimidoylmethoxyJphenyll-S^-diazabicycIop.S.ljnon -ό-ene-SjP- dicarboxylic acid di-fert-butyl ester (H2)

To a sol. of compound Hl (1.39 g, 2.07 mmol) in EtOH (25 mL) were added successively water (6 mL), H ₂ NOH-HCl (531 mg, 7.64 mmol) and K ₂ CO ₃ (485 mg, 3.51 mmol). The mixture was heated to reflux for 3.5 h. The mixture was allowed to cool to it, and the solvents were removed under reduced pressure. Water and CH ₂ Cl ₂ were added: The layers were shaken and separated, and the aq. layer was extracted with CH ₂ Cl ₂ , until the aq. layer did not contain any product. The combined org. layers were evaporated under reduced pressure. Drying the solid residue under high vacuum yielded the title compound (1.45 g), which was used without further purification. LC-MS tR = 0.92 min.

(rac.)-(lR*, 5.S*)-7-{4-[2-(fert-Butyldimethylsilanyloxy)propyl]phenyl}-6 -[cyclopropyI- (S-methoxy^-methylbenzylJcarbamoylJ-S^-diazabicycIop.S.ll-no n-β-ene-S^- dicarboxylic acid di-te/tf-butyl ester (H3)

To a stirred sol. of the compound L2 (12.0 g, 19.4 mmol) in CH ₂ Cl ₂ (158 mL) were added EDC'HCl (9.34 g, 48.7 mmol), HOBt (3.18 g, 23.4 mmol), DMAP (0.596 g, 4.87 mmol), cyclopropyl-(3-methoxy-2-methylbenzyl)amine (11.2 g, 58.4 mmol) and DIPEA (13.4 mL, 77.9 mmol). The mixture was stirred at rt for 5 days. The reaction mixture was partitioned between aq. IM HCl and CH ₂ Cl ₂ , and the org. phase was washed with aq. sat. NaHCO ₃ . The org. extracts were dried over MgSO ₄ , filtered, and the solvents were removed in vacuo. Purification by FC (heptane/EtOAc 8/2) yielded the title compound (8.20 g, 52%).

(rac.)-(lR*, 55*)-6-[Cyclopropyl-(3-methoxy-2-methylbenzyl)carbamoyl]-7-[ 4-(2- hydroxypropyl)phenyl]-3,9-diazabicycIo[3.3.1]non-6-ene-3,9-d i-carboxylic acid di- tert-buty\ ester (H4)

To a sol. of the compound H3 (10.0 g, 12.7 mmol) in THF (225 mL) was added TBAF (IM in THF, 25.3 mL, 25.3 mmol) and the mixture was stirred at 0 °C for 4 h. The reaction mixture was partitioned between aq. sat. NH ₄ Cl and EtOAc, the org. phase washed with brine, dried over MgSO ₄ , filtered, and the solvents were removed in vacuo.

Purification by FC (0-10% MeOH in CH ₂ Cl ₂ ) yielded the title compound (7.00 g, 82 %) as a yellow foam.

(rac.)-{lR*, 55*)-6-[CyclopropyI-(2,3-dichlorobenzyI)carbamoyl]-7-(4-meth oxy- carbonylmethoxyphenyl)-3,9-diazabicyclo[3.3.1]non-6-ene-3,9- dicarboxyIic acid di- tert-hvάyl ester (SS)

To a sol. of compound E2 (0.33 g, 0.5 mmol) in anhydrous CH ₃ CN (3 mL) was added K ₂ CO ₃ (0.15 g, 1.1 mmol) and methyl bromoacetate (0.046 mL, 0.5 mmol). The reaction mixture was refluxed for 18 h. After cooling to rt, CH ₂ Cl ₂ was added, the solid was filtered, and washed with CH ₂ Cl ₂ . The solvents were removed under reduced pressure, and the residue was dissolved in CH ₂ Cl ₂ , washed with water, and brine. The org. extracts were dried over MgSO ₄ , filtered, and the solvents were evaporated under reduced pressure to yield the title compound (0.35 g) that was not further purified. LC-MS: R _t = 1.17 min, ES+=730.29.

(rac.)-(lR*, 55*)-7-(4-Carboxymethoxyphenyl)-6-[cyclopropyl-(2,3-dichloro benzyI)- carbamoyl]-3,9-diazabicyclo[3.3.1]non-6-ene-3,9-dicarboxylic acid di-tert-buty\ ester (H6)

To a sol. of compound H5 (0.35 g, 0.48 mmol) in THF (1 mL) was added a solution of NaOH IM (1 mL), and the reaction mixture was stirred for 1 h. CH ₂ Cl ₂ (20 mL) was added, as well as aq. 0.5M HCl (6 mL). The organic layer was separated and dried over MgSO ₄ , filtered, and the solvents were evaporated under reduced pressure. The obtained title compound(0.31 g) was not further purified. LC-MS: R ₁ = 1.10 min, ES-H=716.40.

(rac.)-(lR*, 5.?*)-7-[4-(2-Hydroxypropyl)phenyl]-3,9-diazabicyclo[3.3.1]n on-6-ene- 3,6,9-tricarboxylic acid 3,9-di-tert-butyl ester 6-ethyl ester (Kl)

To a suspension of NaHCO ₃ (15.5 g, 184 mmol) and 7-{4-[2-(tert-butyl- dimethylsilanyloxy)propyl]phenyl}-9-methyl-3,9-diazabicyclo[ 3.3. l]non-6-ene-3,6- dicarboxylic acid 3-tert-butyl ester 6-ethyl ester WO03/093267, 10.3 g, 18.4 mmol) in CH ₂ ClCH ₂ Cl (190 mL) was added 1-chloroethyl chloroformate (20.0 mL, 55.1 mmol). The mixture was heated and stirred to 80 ⁰ C. After 3 h, the reaction mixture was allowed

to cool to rt, filtered, and the solvents were thoroughly removed in vacuo. The residue was dried under high vacuum for 15 min. MeOH (130 mL) was added and the mixture was stirred at 50 ⁰ C for 20 min. The sol. was allowed to cool to rt, and the solvents were removed in vacuo. The resdidue was dried under high vacuum. The residue was dissolved in CH ₂ Cl ₂ (190 mL), DIPEA (15.7 mL, 91.8 mmol) and Boc ₂ O (12.0 g, 55.1 mmol) were added, and the mixture was stirred at rt for 30 min. The mixture was washed with aq. IM HCl (Ix), and aq. sat. NaHCO ₃ (Ix). The org. phase was dried over MgSO ₄ , filtered, and the solvents were removed in vacuo. Purification by FC (50% EtOAc in heptane) yielded the title compound (8.86 g, 91%).

(rac.)-(lR*, 55 ^r *)-7-[4-(2-Hydroxypropyl)phenyl]-3,9-diazabicycIo[3.3. 1]non-6-ene- 3,6,9-tricarboxylic acid 3,9-di-tert-butyl ester (Ll)

1 M NaOH (198 mL) was added to a sol. of compound Kl (15.0 g, 28.3 mmol) in EtOH (396 mL). The resulting mixture was stirred at 80 ⁰ C for 3 h, cooled to rt and the solvents were removed in vacuo. The crude mixture was partitioned between EtOAc and aq. IM HCl. The aq. layer was extracted once more with EtOAc. The combined org. extracts were dried over MgSO ₄ , filtered, and the solvents were removed under reduced pressure to yield the title compound (13.3 g, 94%).

(rac.)-(lR*, 5-y*)-7-{4-[2-(te/t-ButyIdimethyIsilanyloxy)propyl]phenyl}-3 ,9- diazabicycIo[3.3.1]non-6-ene-3,6,9-tricarboxyIic acid 3,9-di-tert-butyl ester (L2)

TBDMS-Cl (7.38 g, 48.7 mmol) and imidazole (5.32 g, 77.9 mmol) were added to a stirred sol. of compound Ll (9.79 g, 19.5 mmol) in DMF (100 mL). The reaction mixture was stirred at rt over 15 h. The solvents were was removed in vacuo, the crude mixture partitioned between Et ₂ O and aq. sat. NH ₄ Cl. The org. extracts were dried over MgSO ₄ , filtered, and the solvents were removed under reduced pressure. To a sol. of the reaction mixture in THF (190 mL) were added MeOH (63 mL), water (33 mL) and K ₂ CO ₃ (1.46 g), and the mixture stirred at rt for 30 min. The mixture was then partitioned between Et ₂ O and aq. sat. NH ₄ Cl and the aq. layer extracted once more with Et ₂ O. The combined org. extracts were dried over MgSO ₄ , filtered, and the solvents were removed in vacuo to yield the title compound (12.2 g, quantitative yield) as a yellow foam.

Examples

Example 1

(rαc.)-(^* _» -5 ₁ S'*)-7-{4-[5-(2-Chloro-3,6-difluorophenyl)-[l,2,4]oxad iazol-3-yImethoxy]- phenyl}-3,9-diazabicycIo[3.3.1]non-6-ene-6-carboxylic acid cycIopropyl-(3-methoxy-2- methylbenzyl)amide

To a sol. of the compound Fl (0.1 mmol) in CH ₂ Cl ₂ (1 mL), cooled to 0 ⁰ C was added 4M HCl/dioxane (1 mL). The ice bath was removed and the sol. was stirred for 90 min. The solvents were evaporated under reduced pressure without heating. Purification of the residue by HPLC (H ₂ O, MeOH, NH ₄ OH) yielded the title compound (14 mg). LC-MS: t _R = 0.83 min, ES+=662.47.

Example 2

(rac.)-(lR*, 55 ^r *)-7-{4-[5-(2-ChlorophenyI)-[l,2,4]oxadiazol-3-ylmetho xy]phenyl}-3,9- diazabicyclo[3.3.1]non-6-ene-6-carboxylic acid cyclopropyl-(3-methoxy-2-methyl- benzyl)amide

To a sol. of the compound F2 (0.1 mmol) in CH ₂ Cl ₂ (1 mL), cooled to 0 °C was added 4M HCl/dioxane (1 mL). The ice bath was removed and the sol. was stirred for 90 min. The solvents were evaporated under reduced pressure without heating. Purification of the residue by HPLC (H ₂ O, MeOH, NH ₄ OH) yielded the title compound (14 mg). LC-MS: t _R = 0.82 min, ES+=626.50.

Example 3

(rac.)-(lR*, 55'*)-7-(4-{2-[3-(2,3-Dichlorophenyl)-[l,2,4]oxadiazol-5-yl] ethyl}phenyl)- 3,9-diazabicyclo[3.3.1]non-6-ene-6-carboxylic acid cycIopropyl-(3-methoxy-2-methyl- benzyl)amide

To a sol. of compound F3 (0.05 mmol) in CH ₂ Cl ₂ (0.5 mL), cooled to 0 ⁰ C, was added 4M HCl/dioxane (0.5 mL). The ice bath was removed and the sol. was stirred for 90 min. The

solvents were evaporated under reduced pressure without heating. Purification of the residue by HPLC (H ₂ O, MeOH, NH ₄ OH) yielded the title compound (9 mg). LC-MS: t _R = 0.85 min, ES-l-=658.38.

Example 4

(mc.)-(lR*, 5S*)-7-(4-{2-[3-(2,6-Dichlorophenyl)-[l,2,4]oxadiazoI-5-yl]e thyI}phenyl)- 3,9-diazabicycIo[3.3.1]non-6-ene-6-carboxyIic acid cyclopropyl-(3-methoxy-2-methyI- benzyl)amide

To a sol. of compound F4 (0.05 mmol) in CH ₂ Cl ₂ (0.5 rnL), cooled to 0 ⁰ C, was added 4M HCl/dioxane (0.5 mL). The ice bath was removed and the sol. was stirred for 90 min. The solvents were evaporated under reduced pressure without heating. Purification of the residue by HPLC (H ₂ O, MeOH, NH ₄ OH) yielded the title compound (9 mg). LC-MS: t _R = 0.83 min, ESH-=658.38.

Example 5

(rac.)-(lR*, 55*)-7-{4-[3-(2-ChIoro-3,6-difluorophenyl)-[l,2,4]oxadiazol- 5-yImethoxy]- phenyl}-3,9-diazabicyclo[3.3.1]non-6-ene-6-carboxyIic acid cyc!opropyl-(2,3-dichloro- benzyl)amide

To a sol. of the compound F6 (0.074 g, 0.08 mmol) in CH ₂ Cl ₂ (1 mL), cooled to 0 ⁰ C, was added 4M HCl in dioxane (1 mL). The ice bath was removed and the sol. was stirred for 2 h. The reaction mixture was quenched with aq. 2M NaOH (2 mL) and poured on a syringe containing diatomaceous earth (4 g, Isolute Sorbent Technology, Johnson, C.R., et al., Tetrahedron, 1998, 54, 4097). The product was eluted with CH ₂ Cl ₂ , and the solvents were removed under reduced pressure. The residue was purified by HPLC (H ₂ O, MeOH, NH ₄ OH) to yield the title product (0.026g). LC-MS: R _t = 0.86 min, ES+=688.28.

Example 6

(rac.)-(lR*, 5S*)-7-{4-[3-(2-Chloro-3,6-difluorophenyl)isoxazol-5-ylmetho xy]phenyl}-

3,9-diazabicyclo[3.3.1]non-6-ene-6-carboxylic acid cyclopropyl-(2,3-dicblorobenzyl)- amide

To a sol. of compound F5 (0.05 mmol) in CH ₂ Cl ₂ (0.5 mL), cooled to 0 ⁰ C, was added 4M HCl/dioxane (0.5 mL). The ice bath was removed and the sol. was stirred for 90 min. The solvents were evaporated under reduced pressure without heating. Purification of the residue by HPLC (H ₂ O, MeOH, NH ₄ OH) yielded the title compound (15 mg). LC-MS: t _R = 0.86 min; ES+: 685.29.

Biological Assays

In vitro assay:

Inhibition of human recombinant renin by the compounds of the invention

The enzymatic in vitro assay was performed in 384-well polypropylene plates (Nunc). The assay buffer consisted of 10 mM PBS (Gibco BRL) including 1 niM EDTA and 0.1% BSA. The incubates were composed of 50 μL per well of an enzyme mix and 2.5 μL of renin inhibitors in DMSO. The enzyme mix was premixed at 4 ⁰ C and consists of the following components:

• human recombinant renin (0.16 ng/mL)

• synthetic human angiotensin(l-14) (0.5 μM)

• hydroxyquinoline sulfate (1 mM)

The mixtures were then incubated at 37 ⁰ C for 3 h.

To determine the enzymatic activity and its inhibition, the accumulated Ang I was detected by an enzyme immunoassay (EIA) in 384-well plates (Nunc). 5 μL of the incubates or standards were transferred to immuno plates which were previously coated with a covalent complex of Ang I and bovine serum albumin (Ang I - BSA). 75 μL of Ang I-antibodies in essaybuffer above including 0.01% Tween 20 were added and a primary incubation made at 4 ⁰ C overnight. The plates were washed 3 times with PBS including 0.01% Tween 20, and then incubated for 2 h at rt with an antirabbit-peroxidase coupled antibody (WA 934,

Amersham). After washing the plates 3 times, the peroxidase substrate ABTS (2.2'-azino- di-(3-ethyl-benzthiazolinsulfonate), was added and the plates incubated for 60 min at room temperature. After stopping the reaction with 0.1 M citric acid pH 4.3 the plate was evaluated in a microplate reader at 405 nm. The percentage of inhibition was calculated of each concentration point and the concentration of renin inhibition was determined that inhibited the enzyme activity by 50% (IC50). The IC50 values of all compounds tested are below 100 nM. However selected compounds exhibit a very good bioavailability and are metabolically more stable than prior art compounds.

Examples of inhibition:

Example 1 : IC ₅₀ = 4 nM

Example 5 : IC ₅₀ = 3.2 nM

Example 6: IC ₅₀ = 0.8 nM