CZ388799A3 - Protease inhibitors - Google Patents

Abstract

The present invention relates to 3-hydroxy- and 3-ketocyclohetero-substituted leucine compounds which are inhibitors cysteine proteases, especially cathepsinKs, are useful in treating diseases in which factor is inhibiting bone loss. 3- Hydroxy- or 3-keto groups attached to tetrahydrothiophene, tetrahydrothiopyran, tetrahydrofuran or tetrahydropyran ring.

Description

Technical field

The present invention relates to novel protease inhibitors, in particular cysteine and serine protease inhibitors, in particular compounds which inhibit cysteine protease. In particular, the compounds of the invention relate to compounds that inhibit cysteine proteases from the papain superclass, and particularly proteases from the cathepsin class. In a most preferred embodiment, the present invention relates to compounds that inhibit cathepsin K. Such compounds are particularly useful for the treatment of diseases in which cysteine protease is involved, especially diseases characterized by excessive bone or cartilage loss, such as osteoporosis, periodontitis and arthritis.

BACKGROUND OF THE INVENTION 1

Kathepsin K is a member of a class of enzymes that are part of the papain superclass cysteine proteas. Cathepsins Β, H, L, N and S have been described in the literature. Recently, cathepsin K polypeptide and cDNA encoding such a polypeptide have been disclosed in U.S. Patent No. 5,501,969 (referred to herein as cathepsin 0). Kathepsin K is currently expressed, purified and characterized. Bossard M. J. et al., J. Biol. Chem., 271, 12517, 12524 (1996), Drake F. H. et al., J. Biol. Chem., 271,

12511-12516 (1996), Bromme D. et al., J. Biol. Chem.,

271, 2126-2132 (1996).

Kathepsin K has been referred to differently in the literature as cathepsin 0, cathepsin X, or cathepsin 02.

- 2 thepsin K is considered more appropriate (designation given by the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology).

The papain supernatants of the cysteine protease cathepsins function in the normal physiological process of protein degradation in animals, including humans, e.g. in the degradation of connective tissues. However, elevated levels of these enzymes in the body may result in pathological conditions leading to disease. Thus, Kathepsins are involved in various disease states, including but not limited to infections caused by Pneumocystis carinii, Trypsanoma cruzi, Trypsanoma brucei brucei and Crithidia fusiculata, as well as schistosomiasis, malaria, tumor metastasis, metachromatic leukodystrophy, muscular dystrophy, muscular dystrophy, muscular dystrophy. International Publication Number WO 94/04172, published Mar. 3, 1994, and references cited therein, see also European Patent Application EP 0 603 873 A1 and references cited therein. Two bacterial cysteine proteases from P. gingivallis, called gingipains, have been involved in the pathogenesis of gingivitis, Potempa J et al., Perspectives in Drug Discovery and Design, 2, 445-458 (1994).

Cathepsin K is believed to play a causal role in diseases with excessive bone or cartilage loss. Bone consists of a protein matrix into which needle or flat hydroxyapatite crystals are incorporated. Type I collagen is the major bone structural protein comprising approximately 90% of the structural proteins. The remaining 10% of the matrix consists of many non-collagenous proteins, including osteocalcin, proteoglycans, osteopontin, osteonectin, thrombospondin, fibronectin and bone sialoprotein. Skeletal bone undergoes remodeling during individual life ·

- 3 ci. These sites, or remodeling units, undergo a cycle consisting of a bone resorption phase followed by a bone replacement phase.

Bone resorption is performed by osteoclasts, which are multinucleated hematopoietic lineage cells. The osteoclasts adhere to the bone surface and form a tightly closed zone, followed by a strong wrinkling of the membranes on their apical (i.e., resorbing) surface. This creates a closed extracellular compartment on the bone surface, which is acidified by proton pumps from the puckered membrane, and into which the osteoclast secretes proteolytic enzymes. The low pH of the compartment dissolves hydroxyapatite crystals on the bone surface, while proteolytic enzymes digest the protein matrix. In this way, a resorption lacquer is formed. At the end of this phase of the cycle, he lays down a new osteoblast protein matrix, which is then mineralized. In several disease states such as osteoporosis, Paget's disease, the normal balance between bone resorption and formation is disrupted, and bone loss is ultimately lost in each cycle. This ultimately leads to bone weakness and may result in an increased risk of fracture with minimal injury.

The over-selective expression of cathepsin K in osteoclasts strongly suggests that this enzyme is essential for bone resorption. Thus, selective inhibition of cathepsin K can provide effective treatment of diseases with excessive bone loss including, but not limited to, osteoporosis, gum disease such as gingivitis and periodontitis, Paget's disease, hypercalcaemia in cancer and metabolic bone disease. Elevated cathepsin K levels have also been demonstrated in osteoarthritic synovium chondroclasts. Thus, selective inhibition of cathepsin K may be useful in the treatment of non-cathepsin K.

4

- 4 powers with excessive cartilage or matrix degradation, including but not limited to osteoarthritis and rheumatoid arthritis. Metastatic neoplastic cells also typically exhibit high levels of proteolytic enzymes that degrade the surrounding matrix. Thus, selective inhibition of cathepsin K may be useful in the treatment of certain neoplastic diseases.

It has now been found that the new class of compounds are protease inhibitors, most specifically cathepsin K inhibitors, and these compounds are useful in the treatment of diseases where inhibition of bone resorption such as osteoporosis and periodontal disease is indicated.

SUMMARY OF THE INVENTION

It is an object of the present invention to provide protease inhibitors such as cysteine and serine protease inhibitors. In particular, this invention relates to compounds that inhibit cysteine protease, particularly cysteine protease from the papain superclass. Preferably, the present invention relates to compounds that inhibit the cysteine protease of the cathepsin class, in particular compounds that inhibit cathepsin K. The compounds of the present invention are useful in the treatment of diseases that can be therapeutically influenced by disrupting the activity of such proteases.

Thus, in a first aspect, the present invention provides a compound of formula (I):

• «• · • ·. · · ·

... · ··

In another aspect, the invention provides a pharmaceutical composition comprising a compound of formula (I) and a pharmaceutically acceptable carrier.

In yet another aspect, the invention provides a method of treating diseases in which the pathology of the disease can be therapeutically modified by inhibiting proteases such as cysteine and serine proteases. In particular, the method comprises treating diseases by inhibiting cysteine proteases, particularly cysteine proteases from the papain superclass. In particular, inhibition of cathepsin class cysteine proteases such as cathepsin K is described.

In another aspect, the compounds of the invention are particularly useful in the treatment of diseases characterized by bone loss such as osteoporosis, gum disease such as gingivitis and periodontitis, or excessive degradation of cartilage or matrix such as osteoarthritis and rheumatoid arthritis.

Detailed description of the invention

The present invention provides compounds of formula (I)

in which R, RC (O), RC (S), RSO ₂ , ROC (O), RR'NC (O)

- 6 or ROC (O) NR'CH (R ⁶ ) C (O),

R ² is H, C 1-4 alkyl, C ₂ . C ₆ -C ₆ alkenyl, Ar-C _0-6 alkyl or C ₁ -C ₆ alkyl;

Het-C ₀ _ ₆ alkyl,

R ³ is H, C ^ alkyl, C ₂ _ ₆ alkenyl, C ₂ _ ₆ alkynyl, C ₃ _ ₆ cycloalkyl-Co_ ₆ alkyl, Ar-C ₀ _ _alkyl or Het-C ₀ _ _g alkyl,

R ⁴ is H, C ^ alkyl, C ₂ _ ₆ alkenyl, Ar-C ₀ _ ₆ alkyl or

Het-C ₀ . ₆ alkyl, each R ⁵ is independently H, C ₁ _ ₆ alkyl, C ₂ _ ₆ alkenyl, Ar-C _{0th g} alkyl or Het-C ₀ _ ₆ alkyl,

R ⁶ is H, C 1-4 alkyl, C ₂ . ₆ alkenyl, C _{3rd 6} cycloalkyl-C ₀ . _g _

alkyl, Ar-C ₀ _ _g alkyl or Het-C ₀ _ ₆ alkyl,

R 1 is H, C 1-4 alkyl, C ₂ . ₆ alkenyl, Ar-C ₀ . ₆ alkyl, or

Het-C ₀ . _g alkyl,

R is C ^ alkyl, Ar-C ₀ _ ₆ alkyl, Het-C ₀ _ ₆ alkyl, Ar-C _{2nd g} alkenyl or Het-C ₂ _ _g alkenyl,

X is O or S, and n is 1, 2 or 3, or a pharmaceutically acceptable salt thereof.

The present invention includes all hydrates, solvates, complexes and prodrugs of the compounds of the invention. Drug precursors are any covalently bound compounds which in vivo release the active parent compound of formula (I). When a chiral center or other form of isomeric center is present in a compound of the present invention, it is intended to protect all forms of such isomer or isomers, including enantiomers and diastereoisomers. The inventive compounds containing the chiral center can be used as a racemic mixture, as an enantiomerically enriched mixture, or the racemic mixture can be separated using well known techniques and the single enantiomer itself can be used. According to the present invention, the S-form for the furan ring coupling is preferred for compounds of formula (I).

Where the compounds have unsaturated double bonds between two carbon atoms, both cis (Z) and trans (E) isomers are within the scope of the invention. In cases where the compounds may exist in tautomeric forms, such as keto-enol tautomers, each tautomeric form is intended to be included in the invention whether they exist in equilibrium or predominantly in one of the forms.

The meaning of any substituent at any occurrence in formula (I) or any sub-formula thereof is independent of its meaning or the meaning of any other substituent at any other occurrence, unless otherwise specified.

With regard to general formula (I):

Suitably, R ² and R ⁴ are H and R ³ is C _x _ ₆ alkyl or C ₂ _ ₆ alkenyl. Preferably R ^{3 is} isobutyl.

Suitably each R ⁵ is H.

• φ

Suitably, R ¹ is R "OC (O), RSO ₂ or RC (O) wherein R is Ar-C ₀ _ ₆ alkyl or Het-C ₀ _ ₆ alkyl, and most preferably it is R

wherein B ₂ is OH, CN, OCF ₃ , OC 1-4 alkyl, OAr, SO 1-4 alkyl, C 1-4 alkyl or halogen.

Suitably n is 1 or 2. Preferably n is 1.

Suitably X is O.

In one particular embodiment, the present invention is a compound of formula (II):

Preferably, the compound of formula (I) is as defined in claim 1

- 9 of the invention by a compound of formula (IIa):

(Ha).

Alternatively, the compound of formula (I) of the present invention is a compound of formula (IIb):

In another embodiment of Formula (IIc):

the present invention is a compound

(Hc).

Specific representative compounds of the invention are:

4- (R, S) -amino-N - [(3,4-methylenedioxybenzoyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(3,4-dichlorobenzoyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(2-quinolinecarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(8-quinolinecarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] -2,2-dibenzyltetrahydrofuran-3-one,

4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(3,4-dimethoxybenzoyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(indol-6-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(benzofuran-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(5-aminobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(5-chlorobenzofuran-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(5-methoxybenzofuran-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(3-bromobenzoyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(4-bromobenzoyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(5-chlorobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(4-fluorobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N- [(4-phenoxybenzoyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(4-phenylbenzoyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(6-trifluoromethylbenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(4-ethylbenzoyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [{4- (tert-butyl) benzoyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(5-methoxybenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(4-nitrobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one;

• φ · ⁴ · • · • «« ⁴

- 12 4- (R, S) -amino-N - [(β-bromobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(5-bromobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N- ((6-methoxybenzo [b] thiophen-2-ylcarbonyl) -S-leucine) tetrahydrofuran-3-one,

4-S-amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4-R-amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4-S-amino-N - [(2-naphthoyl) -S-leucine] tetrahydrofuran-3-one,

4-R-amino-N - [(2-naphthoyl) -S-leucine] tetrahydrofuran-3-one,

4-S-amino-N - [(quinoline-2-carbonyl) -S-leucine] tetrahydrofuran-3-one,

4-R-amino-N - [(quinoline-2-carbonyl) -S-leucine] tetrahydrofuran-3-one,

4-S-amino-N - [(S-methoxybenzofuran-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4-S-amino-N - [((4-pyrid-3-yl) benzoyl) -S-leucine) -tetrahydrofuran-3-one,

4-S-amino-N - [((4-pyrid-2-yl) benzoyl) -S-leucine] tetrahydrofuran-3-one, ···· · ·

• ·

- 13 *· ·· "· · ··

4-S-amino-N - [(benzyloxycarbonyl) -S-leucine] tetrahydrofuran-3-one,

4-S-amino-N - [(3,4-dimethoxybenzoyl) -S-leucine] tetrahydrofuran-3-one,

4-S-amino-N - [(benzofuran-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one;

4-S-amino-N - [(4- [6-methylpyrid-3-yl] benzoyl) -S-leucine] tetrahydrofuran-3-one,

4-S-amino-N - [(5-chlorobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4-S-amino-N - [((4-pyrid-4-yl) benzoyl) -S-leucine] tetrahydrofuran-3-one,

4-S-amino-N - [(2-chlorobenzoyl) -S-leucine] tetrahydrofuran-3-one,

4-S-amino-N - [(4-bromobenzoyl) -S-leucine] tetrahydrofuran-3-one,

4-S-amino-N - [(4-chlorobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4-S-amino-N - [(4-benzylpiperidin-1-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one,

4-S-amino-N - [(3,4-dichlorobenzoyl) -S-leucine] tetrahydrofuran-3-one;

• · ···

4-S-amino-N - [(3-chlorobenzoyl) -S-leucine] tetrahydrofuran-3-one,

4- (R, S) -amino-N - [(3,4-dimethoxybenzoyl) -S-leucine] tetrahydropyran-3-one,

4- (R, S) -amino-N - [(4-phenoxybenzoyl) -S-leucine] tetrahydropyran-3-one,

4- (R, S) -amino-N - [(quinolin-2-ylcarbonyl) -S-leucine] tetrahydropyran-3-one,

4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] tetrahydropyran-3-one,

4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydropyran-3-one and

4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrothiophen-3-one or a pharmaceutically acceptable salt thereof.

In yet another aspect, the present invention provides novel intermediates useful in the preparation of compounds of formula (I) represented by formulas (III), (IV) and (V):

, 3 .4

Ό (ΙΠ)

- 15 99 9 9999 9 99 9 β 999 999 999

9,999 9 «9999

999 9994

9999 99 999 99 99 in which

R ¹ is R, RC (O), RC (S), RSO ₂ , ROC (O), RR'NC (O) or ROC (O) NR'CH (R ⁶ ) C (O),

R ² is H, C ^ alkyl, C _2-6 alkenyl, Ar-C _{0th 6} alkyl, or

Het-C ₀ _ ₆ alkyl,

R ³ is H, C 1-4 alkyl, C ₂ . ₆ alkenyl, C ₂ _ ₆ alkynyl, C _{3rd 6} cycloalkyl-C ₀ _ ₆ alkyl, Ar-C ₀ _ ₆ alkyl or Het-C ₀ _ ₆ alkyl,

R ⁴ is H, C ^ alkyl, C ₂ _ ₆ alkenyl, Ar-C _{0 alkyl,} or _

Het-C ₀ _ ₆ alkyl, each R ⁵ is independently H, C ^ alkyl, c _{2. 6} alkenyl, Ar-C ₀ . _g alkyl or Het-C ₀ _ ₆ alkyl,

R ⁶ is H, C ^ -alkyl, C _2-6 -alkenyl, C ₃ _ _{cycloalkyl-C 0} _ ₆ alkyl,

Ar-C ₀ _ ₆ alkyl, Het-C ₀ _ ₆ alkyl,

R 1 is H, C _x . ₆ alkyl, c _2. C ₆ -C ₆ alkenyl, Ar-C _0-6 alkyl or C ₁ -C ₆ alkyl;

Het-C ₀ . ₆ alkyl,

R is a _{first 6} alkyl, Ar-C ₀ _ _g alkyl, Het-C ₀ _ ₆ alkyl, Ar-C _2-6 alkenyl or Het-C ₂ _ ₆ alkenyl, and n is 1, 2 or 3, or a pharmaceutically acceptable salt or

Φ

9- ·

in which

R ¹ is R, RC (O), RC (S), RSO ₂ , ROC (O), RR'NC (O) or ROC (O) NR'CH (R ⁶ ) C (O),

R ² is H, Ci.galkyl, C ₂ _ ₆ alkenyl, Ar-C ₀ _ ₆ alkyl or

Het-C ₀ _ ₆ alkyl,

R ³ is H, C 1-4 alkyl, C ₂ . ₆ alkenyl, C ₂ _ ₆ alkynyl, C _{3rd 6} cycloalkyl-C ₀ _ ₆ alkyl, Ar-C _{0th 6} alkyl, or Het-C ₀ _ ₆ alkyl,

R "is H, C ₁ _ ₆ alkyl, C _{2nd 6} alkenyl, Ar-C ₀ . ₆ alkyl, or

Het-C ₀ _ ₆ alkyl, each R ⁵ is independently H, C ^ alkyl, C ₂ _ ₆ alkenyl, Ar-C _{0th 6} alkyl, or Het-C ₀ _ ₆ alkyl;

R ⁶ is H, C _n -alkyl, C ₂ _ ₆ alkenyl, C ₃ _ ₆ cycloalkyl-C ₀ _ ₆ alkyl,

Ar-C ₀ _ ₆ alkyl, Het-C ₀ _ ₆ alkyl,

R 'is H, C ^ alkyl, C ₂ _ ₆ alkenyl, Ar-C ₀ _ ₆ alkyl or

Het-C ₀ _ ₆ alkyl,

R is Cj.galkyl, Ar-C ₀ _ _g alkyl, Het-C ₀ _ _g alkyl, Ar-C ₂ _ ₆ alkenyl or Het-C _{2nd 6} alkenyl and n is 1, 2 or 3, in € 9. · * ······ Or a pharmaceutically acceptable salt thereof; or

in which

R ¹ is R, RC (O), RC (S), RSO ₂ , ROC (O), RR'NC (O) or ROC (O) NR'CH (R ⁶ ) C (O),

R ³ is H, C ^ alkyl, C ₂ _ ₆ alkenyl, C ₂ _ ₆ alkynyl, C ₃ _ ₆ cycloalkyl-C ₀ _ ₆ alkyl, Ar-C ₀ _ ₆ alkyl or Het-C ₀ _ ₆ alkyl,

R ⁴ is H, C ₁ . ₆ alkyl, C ₂ _ ₆ alkenyl, Ar-C _{0th 6} alkyl, or

Het-C ₀ _ ₆ alkyl, each R ⁵ is independently H, C ^ alkyl, C ₂ _ ₆ alkenyl, Ar-C ₀ _ ₆ alkyl or Het-C ₀ _ ₆ alkyl,

R ⁶ is H, C ^ alkyl, C ₂ _ ₆ alkenyl, C ₃ _ ₆ cycloalkyl-C _{0th 6-} alkyl,

Ar-C ₀ _ ₆ alkyl, Het-C ₀ _ ₆ alkyl,

R 'is H, C ^ alkyl, C ₂ _ ₆ alkenyl, Ar-C ₀ _ ₆ alkyl or

Het-C ₀ _ ₆ alkyl,

R is Ci.galkyl, Ar-C ₀ _ _g alkyl, Het-C ₀ _ ₆ alkyl, Ar-C ₂ _ ₆ alkenyl or Het-C _{2nd 6} alkenyl;

X is O or S, and

r 9 9 9 9 9 9

999 β · 99

N is 1, 2 or 3, or a pharmaceutically acceptable salt thereof.

Representative intermediates of the invention are:

trans-4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] -3-hydroxytetrahydrofuran, trans-4- (R, S) -amino-N- [(tert-butoxycarbonyl) - S-leucine) -3-hydroxytetrahydrofuran, trans-4- (R, S) -amino-N- (S-leucine) -3-hydroxytetrahydrofuran, trans-4- (R, S) -amino-N- [(3 4-methylenedioxybenzoyl) -S-leucine] -3-hydroxytetrahydrofuran, trans-4- (R, S) -amino-N - [(3,4-dichlorobenzoyl) -S-leucine] -3-hydroxytetrahydrofuran, trans-4 - (R, S) -amino-N - [(2-quinolinecarbonyl) -S-leucine] -3-hydroxytetrahydrofuran, trans-4- (R, S) -amino-N - ((benzyloxycarbonyl) -S-leucine] -3-hydroxytetrahydropyran, trans -4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] -3-hydroxytetrahydropyran;

trans-4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S]

* · · · · ·

9 9 9

0 99

19-leucine] -3-hydroxytetrahydrofuran, trans-4- (R, S) -amino-N - [(indole-6-ylcarbonyl) -S-leucine] -3-hydroxytetrahydrofuran, trans-4- (R, S) -amino-N- [(5-aminobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] -3-hydroxytetrahydrofuran, trans-4-amino-3-hydroxytetrahydrofuran, trans-3-hydroxy-4-benzyloxycarbonylaminotetrahydrofuran,

4-benzyloxycarbonylaminotetrahydrofuran-3-one,

3.3-dimethoxy-4-benzyloxycarbonylaminotetrahydrofuran,

3.3-dimethoxy-4-aminotetrahydrofuran, trans-4-S-amino-3-R-hydroxytetrahydrofuran, trans-4-S-amino-N - [(benzyloxycarbonyl] -S-leucine] -3-R-hydroxytetrahydrofuran, trans- 4-S-amino-N- (S-leucine) -3-R-hydroxytetrahydrofuran, trans-4-S-amino-N - [(3,4-dichlorobenzoyl) -S-leucine] -3-R-hydrexytetrahydrofuran, trans-4-S-amino-N - [(2-quinolinecarbonyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans-4-S-amino-N - [(benzo (b) thiophen-2-ylcarbonyl)] -S-leucine] -3-R-hydroxytetrahydrofuran, also known as < - &gt; Trans-4-S-amino-N - [(benzofuran-2-ylcarbonyl) -S-leucine] trans-4-S-amino-N - [(benzofuran-2-ylcarbonyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans-4-S-amino-N - [(2-naphthoyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans-4-S-amino-N - [(5-methoxybenzofuran) -2-ylcarbonyl) -S-leucine] -3-R-hydroxytecrahydrofuran, trans-4-S-amino-N - [(5-chlorobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] -3-R -hydroxytetrahydrofuran, trans -4-S-amino-N - [(4-chloro benzo [b] thiophen-2-ylcarbonyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans -4-S-amino-N - [(4-bromobenzoyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans-4-S-amino-N - [(4- (pyrid-2-yl) benzoyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans-4-S-amino-N - [(4- ( pyrid-3-yl) benzoyl-5-leucine] -3-R-hydroxytetrahydrofuran, trans -4-S-amino-N - [(3,4-dimethoxybenzoyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans-4-amino-3-hydroxytetrahydropyran, trans-4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] -3-hydroxytetrahydropyran,

9 9900 99 * 9 «« 99 9900

9,999,999 9 9

999 9999

9999 99 999 99 99 trans-4- (R, S) -amino-N- (S-leucine) -3-hydroxytetrahydropyran, trans-4- (R, S) -amino-N- [(benzo [b] thiophene] 2-ylcarbonyl) -5-leucine] -3-hydroxytetrahydropyran, N-benzo [b] thiophen-2-ylcarbonyl-L-leucine methyl ester,

N-benzo [b] thiophen-2-ylcarbonyl-L-leucine, N-benzo [b] thiophen-2-ylcarbonyl-L-leucine-S- (methoxycarbonylmethyl) -L, D-cysteine ethyl ester and

2-methoxycarbonyl-4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrothiophen-3-one or its salts.

These intermediates are prepared using methods analogous to those described in Schemes 1-4 and in the Examples described below.

The prodrugs of the compounds of the present invention may be a prodrug of a ketone functional group of compounds of formula (I), specifically ketals or hemiketals of formula (VI):

X (VI) • 4 · 9 · 9 9 9 9 9 9 9 9 9 9

- 22 in which

R ¹ is R, RC (O), RC (S), RSO ₂ , ROC (O), RR'NC (O) or ROC (O) NR'CH (R ⁶ ) C (O),

R ² is H, C ^ alkyl, C ₂ - _g alkenyl, Ar-C _{0th g} alkyl or

Het-C ₀ _ ₆ alkyl,

R ³ is H, C ^ alkyl, C ₂ _ ₆ alkenyl, c _{2. 6} alkynyl, C ₃ _ _g cycloalkyl-C ₀ _ ₆ alkyl, Ar-C ₀ _ _g alkyl or Het-C ₀ _ _g alkyl,

R ⁴ is H, C ^ alkyl, C ₂ _ ₆ alkenyl, Ar-C ₀ _ _g alkyl or

Het-C ₀ _ ₆ alkyl, each R ⁵ is independently H, C ^ alkyl, C ₂ _ ₆ alkenyl, Ar-C ₀ _ ₆ alkyl or Het-C ₀ _ _g alkyl,

R ⁶ is H, C ^ alkyl, C ₂ _ ₆ alkenyl, C ₃ _ ₆ cycloalkyl-C ₀ _ ₆ alkyl,

Ar-C ₀ _ _g alkyl or Het-C _{0th 6} alkyl,

R 1 is H, C 1-4 alkyl, C ₂ . _g alkenyl, Ar-C ₀ _ _g alkyl or

Het-C ₀ _ ₆ alkyl,

R is C ^ alkyl, Ar-C ₀ _ _g alkyl, Het-C ₀ _ ₆ alkyl, Ar-C ₂ _ ₆ alkenyl or Het-C ₂ _ ₆ alkenyl,

X is O or S, and n is 1, 2 or 3,

R ^a and R ^a 'are independently H or C 1-6 alkyl, provided that when one of R ^a or R ^a ' is H, the other is C 1-6 alkyl or together are (CH ₂ ) ₂ - ₃ forming 5- or 6 -member circle

or a pharmaceutically acceptable salt thereof.

Abbreviations and symbols commonly used in the chemical and peptide arts are used to describe the compounds of the present invention. Amino acid abbreviations are generally governed by the IUPAC-IUB Joint Commission on Biochemical Nomenclature nomenclature, which is described in Eur. J. Biochem., 9, 158 (1984). The term amino acid as used herein refers to the D- or L-isomers of alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine. , tryptophan, tyrosine, and valine.

C1-6alkyl as used herein is intended to include substituted and unsubstituted methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl and tert-butyl, pentyl, n-pentyl, isopentyl, neopentyl and hexyl and its simple aliphatic isomers . Any C 1-4 alkyl group may be optionally substituted independently with one or two halogens, SR ', OR', N (R ') ₂ , C (O) N (R') ₂ , carbamyl or Coalkyl, wherein R 'is H or Coalkyl . Alkyl _of C means that the section is not present no alkyl group. _Ar-C alkyl is therefore equivalent to Ar.

C _3-6 cycloalkyl, as used herein, is intended to include substituted (i.e. alkyl, OR, SR or halogen) and unsubstituted cyclopropane, cyclobutane, cyclopentane and cyclohexane.

C ₂ _ ₆ alkenyl A1 as used herein means an alkyl group with 2-6 C atoms, where a single

the bond between two carbon atoms is replaced by a double bond between two carbon atoms. C _2-6 alkenyl includes ethylene, 1-propene, 2-propene, 1-butene, 2-butene, isobutene and the several isomeric pentenes and hexenes. Both cis and trans isomers are included.

C 6-6 alkynyl means an alkyl group of 2 to 6 carbon atoms, wherein the single bond between two carbon atoms is replaced by a triple bond between two carbon atoms. C ₂ _ ₆ A1 "kinyl includes acetylene, 1-propyne, 2-propyne, 1-butyne, 2-butyne, 3-butyne and the simple isomers of pentyne and hexyne.

Halogen or halo means F, Cl, Br, and I.

Ar or aryl means unsubstituted phenyl or naphthyl, or phenyl or naphthyl substituted with one or more Ph-C ₀ . ₆ al kyl groups, Het-C ₀ . ₆ alkyl groups, C ^ alkoxy, Ph-C ₀ _ ₆ alkoxy, Het-C ₀ _ ₆ alkoxy, a group of the formula OH _(CH2) ^ NR '(CH _2)! - ₆ NR'R', wherein each R 'is independently H, C ^ alkyl, Ar-C ₀ _ ₆ alkyl or Het-C ₀ _ ₆ alkyl, or phenyl or naphthyl substituted by one to three groups selected from C ^ alkyl groups, groups of formula oR', N (R ') ₂ , SR', CF ₃ , NO ₂ , CN, CO ₂ R ', CON (R') ₂ , F, Cl, Br and I, or substituted with methylenedioxy.

As used herein, Het or heterocyclic represents a stable 5- to 7-membered monocyclic or stable 7- to 10-membered bicyclic heterocyclic ring, which is either saturated or unsaturated, and which consists of carbon atoms and 1 to 4 heteroatoms selected from the group of consisting of N, O and S and wherein the nitrogen and sulfur heteroatoms may optionally be oxidized and the nitrogen heteroatom may be optionally

optionally quaternized, and including any bicyclic group in which any of the heterocyclic rings defined above is linked to a benzene ring. The heterocyclic ring may be attached at any heteroatom or carbon atom which will lead to the creation of a stable structure, and may optionally be substituted with one or two groups selected from groups of formula C _{1st 4} alkyl, OR ', N (R') ₂ , SR ', CF ₃ , NO ₂ , CN, CO ₂ R', CON (R ') ₂ , F, Cl, Br and I, where R' is as defined above. Examples of such heterocycles include piperidinyl, piperazinyl, 2-oxopiperazinyl, 2-oxopiperidinyl, 2-oxopyrrolidinyl, 2-oxoazepinyl, azepinyl, thienyl, pyrrolyl, 4-piperidonyl, pyrrolidinyl, pyrazolyl, pyrazolidinyl, imidazolyl, pyridyl, pyrazinyl, oxazolidinyl, oxazolidinyl, oxazolidinyl, oxazolidinyl, oxazolidinyl, oxazolidinyl oxazolyl, isoxazolyl, morpholinyl, thiazolidinyl, thiazolinyl, isothiazolyl, thiazolyl, quinuclidinyl, indolyl, quinolyl, isoquinolyl, benzimidazolyl, benzothienyl, benzopyranyl, benzoxazolyl, benzofuranyl, furyl, pyranyl, tetrahydrofuryl, tetrahydrofuryl, tetrahydrofuryl, tetrahydrofuryl, tetrahydrofuryl, tetrahydrofuryl, tetrahydrofuryl, thiol benzothiazolyl, benzoisothiazolyl, benzisoxazolyl, pyrimidinyl, cinnolyl, quinazolyl, quinoxalyl, 1,5-naphthyridinyl, 1,6-naphthyridinyl, 1,7-naphthyridinyl, 1,8-naphthyridinyl, tetrazolyl, 1,2,3-triazolyl and 1, 2,4-triazolyl.

Some radical groups are abbreviated herein. t-Bu refers to a tertiary butyl group, Boc or BOC refers to a tert-butyloxycarbonyl group, Fmoc refers to a fluorophenylmethoxycarbonyl group, Ph refers to a phenyl group, Cbz or CBZ refers to a benzyloxycarbonyl group.

Some agents are sometimes abbreviated herein.

· 9 · 9 9 9 9 9 9 9 9 9 9

9 9 9 9 9

9 9 9 9 99 99 9 9 9 9

DCC refers to dicyclohexylcarbodiimide, EDC or EDCI refers to N-ethyl-N (dimethylaminopropyl) carboimide. HOBT or HOBt refers to 1-hydroxybenzotriazole, DMF refers to dimethylformamide, DIEA refers to diisopropylethylamine, Lawesson's reagent is 2,4-bis (4-methoxyphenyl) -1,3-dithia-2,4-diphosphethane-2,4-disulfide , TFA refers to trifluoroacetic acid and THF refers to tetrahydrofuran.

Compounds of formula (I) are generally prepared by:

(i) reaction of a compound of formula (III):

(III) wherein

R ¹ , R ² , R ³ , R ⁴ , R ⁵ and ⁿ are as defined in formula (I) wherein any reactive functional groups are protected, with an oxidizing agent or (ii) decarboxylating a compound of formula (IV):

(IV) wherein

R ¹ , R ² , R ³ , R ⁴ , R ⁵ and ⁿ are as defined in formula (I), wherein any reactive functional groups are protected or (iii) by reaction of a compound of formula (V):

(V) in which

R ¹ , R ² , R ³ , R ⁴ , R ⁵ and ⁿ are as defined in formula (I), wherein any reactive functional groups are protected, with an acid, followed by removal of each of the protecting groups and optionally forming a pharmaceutically acceptable salt.

Compounds of formula (I) are prepared by procedures analogous to those described in the synthetic solution of Scheme 1 or the solid process of Scheme 2 or the synthetic solution in Scheme 1.

3.

Scheme 1

α ·· ·

Ν \\. Ν Λ

ΟΗ

HCl H + N + _χ ΟΗ

>> - / c

ZI

(a) NaN ₃ , ammonium chloride, methanol: water,

b) 10% Pd / C, EtOH, H ₂ , ethanolic HCl,

c) trimethylacetyl chloride, N-BOC-leucine, DIEA, CH ₂ C1 ₂

d) TFA, CH ₂ C1 ₂

(e) RCOC1, sodium bicarbonate, 1,4-dioxane,

f) Dess-Martin periodinane, CH ₂ C1 ₂ 9 9 9 9 9

I 9 9 I> · 4

Compounds of formula (I) wherein n is 1 and R ^x is R ¹ 'C (O) are prepared according to the procedures outlined in Scheme 1. Treatment of the known epoxide 1 from sodium azide and ammonium chloride in aqueous methanol at elevated temperatures. Scheme 1 yields the azide 2 of Scheme 1. Reduction of the azide 2 of Scheme 1 using techniques known in the art, such as palladium-on-carbon reduction in ethanol in a hydrogen atmosphere, gives ammonium salt 3 of Scheme 1 after treatment with ethanolic hydrogen chloride. salt 3 of Scheme 1 can be condensed with a carboxylic acid by methods known in the art, such as acid chloride acylation or acid condensation in the presence of EDC and HOBT to obtain amide 4 of Scheme 1. The tert-butoxycarbonyl group can be removed by treating with a strong acid such as TFA in an aprotic solvent such as dichloromethane to obtain the salt 5 of Scheme 1. The salt 5 of Scheme 1 can be acylated with acid chloride in 1,4-dioxane in the presence of an aqueous base such as saturated sodium bicarbonate solution to obtain alcohol 6 of Scheme 1. The alcohol 6 of Scheme 1 can be oxidized by methods known in the art, such as Dess-Martin periodinane in an aprotic solvent , such as dichloromethane.

Scheme 2

(d)

9999

99

9 9 9

9 9 9

9 9 9 • 9 9

999

R‘COOH

EDC, HOAt

(4) ft ft ft ft ft ft ft ft ft ft ft ft

Compounds of formula (I) wherein R ¹ is C (O) R 1, R ² is H, R ⁴ is H, R ⁵ is Η, X is 0 and n is 1, are prepared by the solid synthesis (SPS) procedure shown in Scheme 2. In detail, in a first step, sodium triacetoxyborohydride is added to a stirred solution of Ellman resin in DMF containing 1% HOAc, and then the alpha-amino acid methyl ester is added to give compound 2 of Scheme 2. The amino group is condensed with a carboxylic acid using known procedures such as of the carboxylic acid with EDC to form amide 2 of Scheme 2. Then the ester group is hydrolyzed, for example with potassium trimethylsilanoate in THF, and the liberated acid group is condensed with 3,3-dimethoxy-4-amino-tetrahydrofuran using, for example, EDC in NMP to The blocking group is then removed by known cleavage procedures such as the addition of 7: 2: 1 TFA / CH ₂ Cl ₂ / H ₂ O to give the desired compound 5 of Scheme 2.

Compounds of formula (I) wherein R ¹ is C (O) R 1, R ² is H, R ⁴ is H, R ⁵ is Η, X is 0 and n is 2, are prepared by a solid synthesis process (SPS) ) as shown in Scheme 2, except that 3,3-dimethoxy-4-aminotetrahydropyran is used instead of 3,3-dimethoxy-4-aminotetrahydrofuran.

Scheme 3

COCI

• 4

4 4 4 •

9 • ·

4 4 4 · 4 4 4 441

443 4 4 4 4

COOH

a) L-Leu, ⁱ Pr ₂ NEt, CH ₂ C1 ₂

b) LiOH, THF / H, O,

c) (i) ClCO / Pr, Et ₃ N, CH ₂ Cl ₂ , (ii) L-Cys ethyl ester, (iii) BrCH ₂ CO ₂ Me,

d) (i) NaOMe, (ii) AcOH / HCl, H ₂ O • φ!

φφφ ♦ * «·

Ο ♦ <* φ φ

· 9

Φ Φ Φ Φ

Scheme 4 below shows the preparation of certain compounds of formula (I) which are S-diastereomers at the furan ring linkage.

Scheme 4

as in Scheme 1

- 34 φ »φ · · · · ·

The R-diastereoisomer is prepared in an identical manner from the opposite enantiomer of 3-azido-4-hydroxytetrahydrofuran of formula:

Intermediates of the present invention, such as 3,3-dimethoxy-4-aminotetrahydrofuran, can be prepared according to the procedure of Scheme 5.

Scheme 5

CBzCI ca 70%

CbzHN \

OH /

bleach

TEMPO ca 90%

CbzHN (MeO) ₃ CH pTSA ca 70%

H ₂ N

H ₂ , Pd / C ca 95%

About me

Í ^ OMe ·

4 ·· ·

J * ’• * • · t

«» 4 4 f>

• 4 4 4

Jr * 4 4 4 4

4 4 4

4 4 · 4 4

The steps in Scheme 5 are presented as follows.

Firstly, a nitrogen protecting group is introduced by reacting 3-hydroxy-4-aminotetrahydrofuran with benzyl chloroformate in dioxane containing aqueous sodium carbonate. Thereafter, an alcohol group is oxidized to obtain the ketone by known procedures, such as the addition of sodium bicarbonate bleach in the presence of sodium bromide and TEMPO in EtOAc, toluene and water. The ketone is converted to dimethyl ketal by the addition of trimethyl orthoformate in the presence of para-toluenesulfonic acid in methanol. Finally, the protecting group is removed by hydrogenation using, for example, palladium on carbon in the presence of ethanol under a hydrogen atmosphere.

The starting materials used herein are commercially available amino acids or are prepared by routine methods well known to those of ordinary skill in the art and can be found in standard reference books such as COMPENDIUM OF ORGANIC SYNTHETIC METHODS, Volumes I through IV (published by Wiley-Interscience) .

The coupling procedures to form amide bonds herein are generally known in the art. Peptide synthesis procedures generally designed by Bodansky et al., THE PRACTICE OF PEPTIDE SYNTHESIS, Springer-Verlag, Berlin, 1984, E.

Gross and J. Meienhofer, THE PEPTIDES, Vol. 1, 1-284 (1979) and J. M. Stewart and J. D. Young, SOLID PHASE PEPTIDE SYNTHESIS, 2nd Edition, Pierce Chemical Co.,

Rockford, 111. (1984) generally illustrate the technique and are incorporated herein by reference.

Synthetic processes for preparing the compounds of this invention often utilize protecting groups to conceal reaction functionalities or minimize undesirable side effects.

»• 1 1 1 1 1 1 1 1 1 9 9 9 9

- 36 more responsive. Such protecting groups are generally described in Green T. W., PROTECTIVE GROUPS IN ORGANIC SYNTHESIS, John Wiley & Sons, New York (1981). The term amino protecting group generally refers to a Boc, acetyl, benzoyl, Fmoc and Cbz group and derivatives thereof as known in the art. Methods for protecting and removing protection and replacing an amino protecting group with another moiety are well known in the art.

Acid addition salts of compounds of formula (I) are prepared in standard manner in a suitable solvent from the parent compound and in excess of an acid such as hydrochloric, hydrobromic, hydrofluoric, sulfuric, phosphoric, acetic, trifluoroacetic, maleic, succinic and methanesulfonic acid. Certain of these compounds form internal salts or zwitterions, which may be acceptable. Cationic salts are prepared by treating the parent compound with an excess of a basic reagent, such as a hydroxide, carbonate or alkoxide, containing the appropriate cation, or by treating with the appropriate organic amine. Cations such as Li ⁺ , Na ⁺ , K ⁺ , Ca ⁺⁺ , Mg ⁺⁺ and NH 3 are specific examples of cations present in pharmaceutically acceptable salts. Halides, sulfate, phosphate, alkanoates (such as acetate and trifluoroacetate), benzoates and sulfonates (such as mesylate) are examples of anions present in pharmaceutically acceptable salts.

The invention also provides a pharmaceutical composition comprising a compound of formula (I) and a pharmaceutically acceptable carrier, diluent or excipient. Accordingly, the compound of formula (I) may be used in the manufacture of a medicament. A pharmaceutical preparation of a compound of formula (I) prepared as described herein may be formulated as solutions or lyophilized.

·. · Φ • · · · 9 9 9 9 9 9 9 9 9 9 9 9

- 37 powders for parenteral administration. The powders may be reconstituted prior to use by addition of a suitable diluent or other pharmaceutically acceptable carrier. Liquid formulations may be buffered, isotonized aqueous solutions. Examples of suitable diluents are normal isotonic sodium chloride solution, standard 5% dextrose in water or buffered sodium or ammonium acetate solution. Such a formulation is particularly suitable for parenteral administration, but may also be used for oral administration or contained in a metered dose inhaler or nebulizer for inhalation. It may be desirable to add excipients such as polyvinylpyrrolidone, gelatin, hydroxycellulose, acacia, polyethylene glycol, mannitol, sodium chloride or sodium citrate.

Alternatively, the compounds may be encapsulated, tableted, or prepared into an emulsion or syrup for oral administration. Pharmaceutically acceptable solid or liquid carriers may be added to aid in or stabilize the formulation or to facilitate formulation. Solid carriers include starch, lactose, calcium sulfate dihydrate, white clay, magnesium stearate or stearic acid, talc, pectin, acacia, agar or gelatin. Liquid carriers include syrup, peanut oil, olive oil, sodium chloride solution and water. The carrier may also include a slow release material, such as glyceryl monostearate or glyceryl distearate, alone or with a wax. The amount of solid carrier varies but will preferably be between about 20 mg to about 1 g per dosage unit. The pharmaceutical preparations are manufactured according to conventional pharmacy techniques including milling, mixing, granulating and compressing, if necessary, for tablet forms or milling, mixing and filling for solid gelatin capsule forms. If a liquid carrier is used, the preparation will be made in the form of a syrup, elir · 4 4 · 4 4 4 4 4 4 4 4 4 4 I 4 I 4 · »♦» * * *

- 38 x, emulsion or aqueous or non-aqueous suspension. Such a liquid formulation may be administered directly orally or filled into a soft gelatin capsule.

For rectal administration, the compounds of the invention may also be combined with excipients such as cocoa butter, glycerin, gelatin or polyethylene glycols and formed into a suppository.

Use of the present invention

The compounds of formula (I) are useful as protease inhibitors, in particular as cysteine and protease inhibitors, more specifically as cysteine protease inhibitors, even more specifically as papain superclass cysteine protease inhibitors, and even more specifically as cathepsin class cysteine protease inhibitors, most specifically as cathepsin K inhibitors. The present invention also provides useful compositions and formulations of said compounds, including pharmaceutical compositions and formulations of said compounds.

The present compounds are useful in the treatment of diseases in which cysteine protease is involved, including infections of Pneumocystis carinii, Trypsanoma cruzi, Trypsanoma brucei and Crithidia fusiculata, as well as in schistosomiasis, malaria, tumor metastasis, metachromatic leukodystrophy, and especially muscular dystrophy, muscular dystrophy is involved in cathepsin K, most specifically diseases with excessive bone or cartilage loss including osteoporosis, gum disease including gingivitis and periodontitis, arthritis, more specifically osteoarthritis and rheumatoid arthritis, Paget's disease, hypercalcaemia in cancer and metabolic bone diseases.

Metastatic neoplastic cells also typically exhibit high levels of proteolytic enzymes that degrade the surrounding matrix, thus certain tumors and metastatic neoplasms can be effectively treated with the compounds of the invention.

The present invention also provides methods of treating diseases caused by pathological levels of proteases, particularly cysteine and serine proteases, more specifically cysteine proteases, even more specifically cysteine proteases from the papain class, and even more specifically cysteine proteases from the cathepsin class, the method comprising administering a compound of the present invention to an animal. especially mammals, most specifically human, in need thereof. In particular, the present invention provides methods of treating diseases caused by pathological levels of cathepsin K, which methods comprise administering a cathepsin K inhibitor including a compound of the invention to an animal, especially a mammal, most specifically a human, in need thereof. In particular, the present invention provides methods of treating diseases in which cysteine protease is involved, including infections of Pneumocystis carinii, Trypsanoma cruzi, Trypsanoma brucei and Crithidia fusiculata, as well as in schistosomiasis, malaria, tumor metastasis, metachromatic leukodystrophy, and especially muscular dystrophy. in which cathepsin K is involved, most specifically diseases with excessive bone or cartilage loss including osteoporosis, gum diseases including gingivitis and periodontitis, arthritis, more specifically osteoarthritis and rheumatoid arthritis, Paget's disease, cancer hypercalcemia, and bone metabolic diseases.

• · · φ * 9 φ · · ·

9 9 9 •

9

9 ·

* Φ ·

The invention further provides a method of treating osteoporosis or inhibiting bone loss which comprises administering to a patient an effective amount of a compound of formula (I), alone or in combination with other bone resorption inhibitors such as bisphosphonates (ie alendronate), hormone replacement therapy, antiestrogens. or calcitonin. In addition, treatment with a compound of the invention and an anabolic agent such as bone morphogenetic protein, iproflavone, may be used to prevent bone loss or increase bone mass.

In accordance with the present invention, an effective amount of a compound of formula (I) is administered to inhibit a protease associated with a particular condition or disease. The dosage amount will of course be further modified according to the type of administration of the compound. For example, an effective amount for acute therapy prefers parenteral administration of a compound of formula (I). Intravenous infusion of this compound in 5% dextrose in water or normal saline or a similar formulation with a suitable excipient is most effective, although intramuscular bolus injection is also useful. Typically, a parenteral dose will be about 0.01 to about 100 mg / kg, more preferably between 0.1 and 20 mg / kg, administered in such a way as to maintain the plasma concentration of the drug to a concentration effective to inhibit cathepsin K. four times a day at a level to achieve a total daily dosage of about 0.4 to about 400 mg / kg per day. The precise amount of the inventive compound, which is therapeutically effective and the most appropriate route of administration, will be readily determined by one of ordinary skill in the art by comparing the blood levels of the compound with the concentrations required to achieve a therapeutic effect.

• ·

Drug precursors of the compounds of the present invention may be prepared by any suitable method. For those compounds where the drug precursor group is a ketone function, specifically ketals and / or hemiketals, the conversion may be performed according to conventional procedures.

The compounds of the invention may also be administered orally to a patient in a manner such that the concentration of the drug is sufficient to inhibit bone resorption or any other therapeutic indication as disclosed herein. Typically, the pharmaceutical composition comprising the compound is administered orally at a dosage of about 0.1 to about 50 mg / kg in a manner consistent with the patient's condition. Preferably, the oral dose should be about 0.5 to about 20 mg / kg.

No unacceptable toxicological effects are expected when the compounds of the present invention are administered in accordance with the present invention.

The compounds of the invention can be tested in one of several biological assays to determine the concentration of compound that is required to achieve a given pharmacological effect.

Determination of cathepsin K proteolytic catalytic activity

All cathepsin K assays were performed using human recombinant enzyme. Standard assay conditions used a fluorogenic peptide substrate, typically Cbz-Phe-Arg-AMC, to determine kinetic constants and were determined in 100 mM sodium acetate pH 5.5 containing 20 mM cysteine and 5 mM EDTA. Substrate stock solutions were prepared at concentrations of 10 or 20 mM in DMSO with final •

»· · 9 · 9 • • 9 9 « • • • • * • • « • * « « • ♦ 9 • • • 9 9 • · ·  * ·· * «

- 42 substrate concentrations in the 20 μΜ test. All assays contained 10% DMSO. Independent experiments revealed that this level of DMSO had no effect on ezymic activity or kinetic constants. All assays were performed at room temperature. Product fluorescence (excitation at 360 nm, emission at 460 nm) was monitored using a Perceptive Biosystems Cytofluor II fluorescence plate reader. Product development curves were generated within 20 to 30 minutes after AMC product formation.

Inhibition studies

Potential inhibitors were evaluated using the development curve method. Assays were performed in the presence of various concentrations of the test compound.

Reactions were initiated by the addition of enzyme to buffered inhibitor and substrate solutions. Data analysis was performed according to one of two procedures depending on the appearance of development curves in the presence of inhibitors. For those compounds whose development curves were linear, the apparent inhibition constants (K _{i (app} ) were calculated according to formula 1 (Brand et al., Biochemistry, 28, 140 (1989)):

v = V ^ A / tKJ1 + I / K _{i (app} ) + A] (1) where v is the reaction rate with the maximum reaction rate V _m ,

A is the concentration of substrate with Michaelis constant of K _a, and I is the inhibitor concentration.

For those compounds whose development curves showed a descending curvature of the time-dependent inhibition characteristic, the data from each of the kits was analyzed to provide the _obs according to relationship 2:

• · · · · · · t «t

- 43 [AMC] = v _ss t + (v0 - v _3S ) [1 - exp (-k _obs t)] / k _ob3 (2) where [AMC] is the concentration of product formed over time t, vO is the initial reaction rate and in _3S , the velocity at the final equilibrium state. The values for k _obs were then analyzed as a linear function of inhibitor concentration to obtain an apparent second order inhibition constant (k _obs / inhibitor concentration or K _obs / [I]) describing the time-dependent inhibition. A complete discussion of this kinetic processing is fully described (Morrison et al., Adv. Enzymol. Relat. Areas Mol. Biol., 61, 201 (1988)).

One of ordinary skill in the art would consider any compound with a K _{b of} less than 50 μΜ as a potential lead compound. Preferably, the compounds used in the process of the present invention have a K value of less than 1 micromolar. Most preferably said compounds have a Kimene value of greater than 100 nanomolar. 4- (R, S) -amino-N - [(8-quinolinesulfonyl) -S-leucine] -3-tetrahydrofuran-3-one, a compound of formula (I), has a K _i value of greater than 10 micromolar.

Determination of resorption of human osteoclasts

Aliquots of the osteoclastoma-derived cell suspension are removed from storage in liquid nitrogen, rapidly heated to 37 ° C and washed once in RPMI-1640 medium by centrifugation (1000 rpm, 5 minutes at 4 ° C). The medium is aspirated and replaced with murine anti-HLA-DR antibodies diluted 1: 3 in RPMI-1640 medium and incubated for 30 minutes on ice. The cell suspension is often stirred.

· Φ φ φ · · · · · · ·

Φ 9 9 9 9 9 ΦΦΦ * φ 9 9 9 9 9 9 9 9 9 9

9 φ · Φ 9 9 9 9

999 9 ΦΦ 99 9 Φ * ··

Cells are washed 2 times by cold RPMI-1640 centrifugation (1000 rpm, 5 minutes at 4 ° C) and then transferred to a sterile 15 ml centrifuge tube. The number of mononuclear cells was counted in an improved Neubauer chamber counter.

Sufficient magnetic beads (5 per 1 mononuclear cell) coated with goat anti-mouse IgG are removed from the storage bottle and placed in 5 ml of fresh medium (thereby eluting the toxic azide preservative). The medium was removed by immobilizing the beads on the magnet and replacing it with fresh medium.

The beads are mixed with the cells and the suspension is incubated for 30 minutes on ice. The suspension is often stirred. The bead-coated cells are immobilized on a magnet and the remaining cells (osteoclast-rich fraction) are pooled into a sterile 50 ml centrifuge tube. Fresh medium is added to the bead-coated cells to release any trapped osteoclasts. This washing process is repeated 10 times. The bead-coated cells are removed.

The osteoclasts are counted in a chamber counter using a plastic disposable pasteur pipette with a large opening to fill the chamber with sample. Cells are centrifuged into spheres and the osteoclast density is adjusted to 1.5x10 ⁴ / ml in EMEM medium supplemented with 10% fetal calf serum and sodium bicarbonate at a concentration of 1.7 g / L. 3 ml aliquots of cell suspension (per assay) are poured into tubes in a 15 ml centrifuge. Cells are centrifuged into pellets. Add 3 ml of the appropriate formulation (diluted to 50 μΜ in EMEM medium) to each tube. Controls of the respective vehicles were also included,

4 · · 44

444 ♦ ♦ · · 4 4 4 4 4 4 4 • 9

4 4

4 4 4 4 4

The positive control (87MEM1 diluted to 100 pg / ml) isotope control (IgG2a, diluted to 100 pg / ml) is incubated for 30 minutes at 37 ° C.

and

Test tubes

0.5 ml aliquots of cells are plated on sterile dentine sections on a 48-well plate and incubated for 2 hours at 37 ° C. Each embodiment is monitored four times. Sections are washed in six exchanges of warm PBS (10 ml per well in a 6-well plate) and then placed in fresh composition or control and incubated for 48 hours at 37 ° C. The sections are then washed with phosphate buffered saline and fixed in 2% glutaraldehyde (0.2 M sodium cacodylate) for 5 minutes, then washed in water and incubated in buffer at 37 ° C for 5 minutes. The sections are then washed in cold water and incubated for 5 minutes in cold acetate buffer / solid garnet red at 4 ° C. The excess buffer is aspirated and after washing with water the sections are air-dried.

TRAP positive osteoclasts are counted with a light microscope and then removed from the dentine surface by ultrasound. Well volumes were determined using a Nikon / Lasertec ILM21W confocal microscope.

DETAILED DESCRIPTION OF THE INVENTION

In the following synthesis examples, unless otherwise noted, all starting materials were obtained from commercial sources. It is believed that without further elaboration, one skilled in the art can, using the preceding description, utilize the present invention to its fullest extent. These examples are given to illustrate the invention but not to limit its scope. Reference is made to the claims in what is reserved below for the inventor.

Example 1

Preparation of 4- (R, S) -amino-N - [(3,4-methylenedioxybenzoyl) -S-leucine] tetrahydrofuran-3-one

(a) trans -4-Azido-3-hydroxytetrahydrofuran

3,4-Epoxytetrahydrofuran (9 g, 105 mmol) was added to a stirred solution of sodium azide (27 g, 415 mmol) and ammonium chloride (9 g, 159 mmol) in aqueous methanol (95%, 200 mL). The reaction mixture was heated to 75 ° C for 20 h and stirred. The reaction mixture was cooled, filtered and evaporated under reduced pressure. The residue was diluted with water and extracted with ethyl acetate, dried and evaporated under reduced pressure to give the title compound as a colorless oil, 10 g, 74% yield.

1 H NMR δ (CDCl ₃ ) 4.32 (m, 1H), 4.09 (dd, 1H, J = 4.8,

9.9 Hz), 3.99 (dd, 1H, J = 4.3, 10.1 Hz) 3.94 (m, 1H), 3.81 (dd, 1H, J = 2.1, 9.9 Hz), 3.73 (dd, 1 H, J = 1,8, 10.1 Hz), 2.72 (d, 1H, J = 4.6Hz).

b) Trans-4-amino-3-hydroxytetrahydrofuran hydrochloride

A mixture of trans-4-azido-3-hydroxytetrahydrofuran (10 g, 77 mmol) and 10% palladium on carbon (1 g) in ethanol (150 mL) was stirred under a hydrogen atmosphere (12 psi) for 12 hours. The mixture was filtered and treated with 100 mL of ethanolic HCl to give, after evaporation under reduced pressure, the title compound as a brown solid, 10.5 g, 97% yield, mp 132 ° C.

9 · * 99 8

9 9 ·

1 H NMR δ (d ₆ DMSO) 8.37 (s, 3H), 4.13 (m, 1H), 3.84 (dd, 1H, J = 4.9 and 14.3 Hz), 3.76 ( dd, 1H, J = 5.5, 10.0 Hz), 3.58 (dd, 1H, J = 2.7, 10.0 Hz), 3.34 (m, 3H).

c) trans-4- (R, S) -amino-N - [(tert-butoxycarbonyl) -S-leucine] -3-hydroxytetrahydrofuran

Trimethylacetyl chloride (3.5 mL, 29 mmol) was added to a stirred solution of N-Boc-L-leucine (7.3 g, 31 mmol) and diisopropylethylamine (9 mL, 52 mmol) in dichloromethane (200 mL). After 1 hour trans-4-amino-3-hydroxytetrahydrofuran hydrochloride (4 g, 28 mmol) was added and the mixture was allowed to stir overnight. The reaction mixture was poured into water and extracted with dichloromethane. The combined organic layers were washed with 0.5N HCl, saturated sodium carbonate solution, brine and dried. Evaporation under reduced pressure gave the title compound as a yellow foam, 5 g, 44% yield.

Δ NMR (CDCl ₃ ) 8.08 (d, 0.5 H, J = 4.8 Hz), 7.89 (d, 0.5 H, J = 7.4 Hz), 6.20 (d, 0.5H, J = 8.3 Hz), 6.09 (delta, 0.5H, J = 8.7 Hz), 4.81 (d, 1H, J = 16.0 Hz), 4.40 ( m, 2H), 4.20 (m, 2H), 3.77 (m, 2H), 1.60 (m, 3H), 1.50 (s, 9H), 0.92 (m, 6H).

d) Trans-4- (R, S) -amino-N- (S-leucine) -3-hydroxytetrahydrofuran salt with TFA, trans-4-Amino-N - [(tert-butoxycarbonyl) -S-leucine] - 3-Hydroxytetrahydrofuran (2.5 g, 8.0 mmol) was added to a stirred solution of 20% trifluoroacetic acid in dichloromethane (100 mL). After 2 hours, the reaction mixture was evaporated in vacuo.

9 9 9 9999 ·· 999 9999

999 9 99 999 99 99 under reduced pressure to give the title compound as a white gum 2.6 g, 100% yield.

¹ H NMR δ (MeOD) 4.18 (m, 2H), 4.08 (m, 2H), 3.97 (m,

2H), 3.86 (apparent t, 2H, J = 7.1 Hz) 3.69 (dd, 2H, J = 1.6, 7.4 Hz), 1.68 (m, 3H), O, 99 (d, 6H, J = 2.1Hz).

e) trans-4- (R, S) -Amino-N - [(3,4-methylenedioxybenzoyl) -S-leucine] -3-hydroxytetrahydrofuran

Piperonyloyl chloride (400 mg, 2.2 mmol) was added to a stirred solution of trans-4-amino-N- (S-leucine) -3-hydroxytetrahydrofuran salt with TFA (380 mg, 1.1 mmol) in saturated sodium bicarbonate solution ( 10 mL) and 1,4-dioxane (10 mL). The reaction mixture was allowed to stir for 1 hour and then diluted with ether. The organic layer was washed with 1N hydrochloric acid, sodium bicarbonate, brine and dried. Evaporation of the solvent gave the title compound as a white foam, 250 mg, 60% yield.

.Delta. NMR (CDCl3 ₎ ) 8.23 (d, 0, 5H, J = 4.8 Hz), 8.15 (d, 0,5H, J = 7.4 Hz) , 7.85 (d, 0, 5H, J = 7.6 Hz), 7.3 (m, 2H), 6.66 (dd, 1 H, J = 8.1, 11, 5 Hz), 5.92 (d, 2H, J = 6.2 HZ), 4.78 (m, 1 H), 4.50 (s, IH) 4.2 (s, 1H) , 08-3 .75 (m 4H), 3.74-3.48 (m, 3H), 1.82-1, 48 (m, 3 H), 0, 90 (m, 6H). MS you 'calculated for (C ₁₈ H ₂₄ N ₂ O ₅ + H) ⁺ : 365, find eno: 365.

f) 4- (R, S) -amino-N - [(3,4-methylenedioxybenzoyl) -S-leucine] tetrahydrofuran-3-one

Dess-Martin periodinane (500 mg, 1.2 mmol) was added

added to a stirred solution of trans-4- (R, S) -amino-N - [(3,4-methylenedioxybenzoyl) -S-leucine] -3-hydroxytetrahydrofuran (220 mg, 0.60 mmol) in dichloromethane (10 mL) . After 1 h, ether was added followed by sodium thiosulfate (570 mg, 3.6 mmol). After a further 15 minutes, the reaction mixture was washed with saturated sodium bicarbonate solution, brine and dried. Evaporation of the solvent gave the title compound as a white foam, 200 mg, 100% yield.

^X H NMR delta (CDC1 ₃₎ 8.14 (d, 0.5H, J = 6.2) Hz), 7.90 (d, O, 5H, J = 5.9Hz) 7.54 (d, 0.5H, J = 7.3) Hz), 7.46 (d, O, 5H, J = 5.1 Hz), 7 23 (d, 1H, J = 6.6Hz) 7, 14 (s, 1H), 6, 67 (m, 1H). 5.93 (s, 2H), 4.73 (m, IH), 4.37 -3.71 (m, 5H 1.68 (m, 3H); 0.85 (m, 6 H). MS calculated for (C ₁₈ H ₂₂ N ₂ O ₆ + H) ⁺ : 363, found eno: 363.

Example 2

Preparation of 4- (R, S) -amino-N - [(3,4-dichlorobenzoyl) -S-leucine] tetrahydrofuran-3-one

Following the procedure of Example 1 (e-f) except that 3,4-dichlorobenzoyl chloride was used in place of piperonyloyl chloride, the title compound was prepared.

¹ H NMR δ (MeOD) (d, 1 H, J = 2.0 Hz), 7.75 (dd, 1 H, J = 2.0, 8.3 Hz), 7.61 (d, 1 H, J = 8.3 Hz), 4.60 (m, 1H), 4.50-3.84 (m, 5H), 1.66 (m, 3H), 1.00 (m, 6H).

• 9

- 50 MS calculated for (C ₁₇ Cl ₂ H ₂₀ N ₂ O ₄ + H) ⁺ : 387 and 389, found: 387 and 389.

Example 3

Preparation of 4- (R, S) -amino-N - [(2-quinolinecarbonyl) -S-leucine] tetrahydrofuran-3-one

Following the procedure of Example 1 (e-f), except that 2-quinolinecarbonyl chloride was used in place of piperonyloyl chloride, the title compound was prepared as a white foam.

1 H NMR δ (CDCl ₃ ) 8.57 (dd, 1H, J = 1.5, 5.1 Hz), 8.20 (m, 3H), 7.77 (m, 2H), 7.62 (m 1 H, 7.40 (d, 1 H, = 6.2 Hz), 4.80 (m, 1 H), 4.57 (m, 1 H), 4.30 (m, 1 H), 4.26- 3.87 (m, 3H), 1.96-1.71 (m, 3H), 1.0 (m, 6H).

MS calculated for (C ₂₀ H ₂₃ N ₃ O ₄ + H) ⁺ : 370, found 370.

Example 4

Preparation of 4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] -tetrahydrofuran-3-one

Following the procedure of Example 1, except that N-CBZ-leucine was used instead of N-BOC-leucine, the title compound was prepared as a white foam.

¹ H NMR δ (CDCl ₃ ) 7.31 (m, 5H), 5.71 (apparently dd, 1H, J = 8.4, 14.8 Hz), 5.06 (m, 2H), 4.50 (dd, 1H, J = 8.9, 18.0 Hz), 4.30-3.88 (m, 4H), 3.80 (apparently t, 1H, J = 9.6 Hz),

9 9

9 9 99 9 9 9 99 99 99 99 99 99

51 1.64 (m, 3H), 0.91 (m, 6H).

MS calculated for (C ₁₈ H ₂₄ N ₂ O ₅ + H) ⁺ : 349, found: 349.

Example 5

Preparation of 4- (R, S) -amino-N - [(3,4-methylenedioxybenzoyl) -S-leucine] tetrahydrofuran-3-one

Following the procedure of Example 1, except that 3,4-methylenedioxybenzoyl chloride was used in place of piperonyloyl chloride, the title compound was prepared.

Example 6

Preparation of 4- (R, S) -amino-N - [(8-quinolinecarbonyl) -S-leucine] tetrahydrofuran-3-one

Following the procedure of Example 1 (e-f), except that 8-quinolinecarbonyl chloride was used in place of piperonyloyl chloride, the title compound was prepared as a white foam. Melting point: 123 ° C (HCl salt).

¹ H NMR δ (CDCl ₃ ) 11.54 (m, 1H), 8.84 (d, 1H, J = 1.5 Hz), 8.65 (d, 1H, J = 7.2 Hz), δ 17 (d, 1H, J = 8.0Hz),

7.86 (d, 1H, J = 8.0 Hz), 7.7 (s, 1H), 7.54 (t, 1H, J =

7.7 Hz), 7.40 (dd, 1H, J = 3.8, 7.7 Hz), 4.78 (dd, IH, J =

7.5, 13.6 Hz), 4.47 (dd, 1H, J = 8.6, 13.6 Hz), 4.18 (m,

1H), 4.16-3.79 (m, 3H), 1.88 (m, 3H), 0.88 (m, 6H).

·

- 52 MS calculated for (C ₂₀ H ₂₃ N ₃ O ₄ + H) ⁺ : 370, found 370.

Example 7

Preparation of 4- (R, S) -amino-N - [(8-quinolinesulfonyl) -S-leucine] tetrahydrofuran-3-one

Following the procedure of Example 1 (e-f), except that 8-quinolinesulfonyl chloride was used in place of piperonyloyl chloride, the title compound was prepared as a brown solid. Melting point: 98 ° C (HCl salt).

¹ H NMR δ (CDCl ₃ ) 9.07 (dd, 1H, J = 1.7 and 4.3 Hz), 8.40 (m, 1H), 8.30 (m, 1H), 8.00 ( m, 1H), 7.66 (m, 2H), 7.11 (d, 0.5H, J = 5.9 Hz), 6.96 (d, 0.5H, J = 5.7 Hz), 4.51 (t, 0.5H, d = 8.8 Hz), 4.39 (t, 0.5H, J = 8.7 Hz), 4.12-3.71 (m, 4.5H) 3.53 (dd, 0.5H, J = 1.0, 5.9 Hz), 1.43 (m, 2H), 0.64 (m, 3H), 0.33 (m, 3H).

MS calcd for (C ₁₉ H ₂₃ N ₃ 0 ₅ S + Η) ^+: 406, Found: 406th

Example 8

Preparation of 4- (R, S) -amino-N - [((4-methyl-3-pyridyl) carbonyl) -S-leucine] tetrahydrofuran-3-one

Following the procedure of Example 1 (e-f), except that (4-methyl-3-pyridyl) carbonyl chloride was used in place of piperonyloyl chloride, the title compound was prepared.

* «··· ·

- 53 Example 9

Preparation of 4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] -2,2-dibenzyltetrahydrofuran-3-one

To a stirred solution of 4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] tetrahydrofuran-3-one (300 mg, 0.9 mmol) and benzyl bromide (0.4 mL, 3.4 mmol) ) in methanol (5 mL) was added sodium methoxide (140 mg, 2.6 mmol). After 12 hours, the reaction mixture was poured into ether (100 mL), washed with water, brine and dried. Evaporation under reduced pressure and purification by flash column chromatography (30% ethyl acetate-hexane) gave the title compound as a colorless gum, 250 mg, 55% yield.

^X H NMR .delta (CDC1 ₃₎ 7.34-7.25 (m, 10H) 7.24-7.0 (wide p 5H), 6.56 (s, 0.5H), 6.37 (s, 0.5H), 5.18 (d, 1 H, J = 11.5 Hz), 5.07 (m, 2H), 4.48-4.42 (m, 1 H), 4, 20-3,98 (m, 4H) 3.12 (dd, 1 H, J = 12.5 and 1.25 Hz), 2.88 (dd, 1 H, J = 12.5 and 12.5 Hz), 1 , 72-1.32 (m. 3H) , 0.92-0.76 (m, 7H)

F

MS calculated for (C ₃ H ₂₁ N ₂ O ₅ = C ₇ H ₇ ) ⁺ : 439, found: 439.

Example 10

Preparation of 4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one

Following the procedure of Example 1 (e-f), with the difference that

• ··· · · «· · « ·· • · • • • ·· • • • • • • • • • • • • • • • 4 • • • • • • • • E • ··· • ·· • · · • · • ·

If benzo [b] thiophen-2-ylcarbonyl chloride is used instead of piperonyloyl chloride, the title compound is prepared.

¹ H NMR δ (CDCl ₃ ) 8.33 (d, 1H), 7.78 (m, 4H), 7.38 (m, (m, 5H), 1.88 (m, 3H), 1.00

0.5H, J = 6.6 Hz], 8.00 (m, 2.5H), 4.87 (m, 1H), 4.63-3.88 (m, 6H).

MS calcd for (c ₁₉ h ₂₂ n ₂ o ₄ s)

H) ⁺ : 373, found: 373.

Example 11

Preparation of 4- (R, S) -amino-N - [(3,4-dimethoxybenzoyl) -S-leucine] tetrahydrofuran-3-one according to the procedure of Example 1 (ef) except that instead of piperonyloyl chloride, 3 4-dimethoxybenzoyl chloride, the title compound is prepared.

Δ NMR δ (CDCl ₃ ) 7.58 (d, 0.5H, J = 6.7 Hz), 7.47 (d,

0.5H, J = 6.2Hz), 7.33 (m, 2H), 7.04 (d, 0.5H, J = 8.0Hz), 6.92 (d, 0.5H, J = 8.0 Hz), 6.83 (d, 0.5H, J = 8.4 Hz),

4.77 (m, 1H), 4.53-3.67 (m, 5H), 1.68 (m, 3H), 0.85 (m, 6H).

MS calculated for (C ₁₉ H ₂₆ N ₂ O ₆ - H) ⁺ : 377, found: 377.

Example 12

Preparation of 4- (R, S) -amino-N - [(indol-6-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one. · · <<<<• • • • • • • <<<<<<<<<

Following the procedure of Example 1 (e-f), except that indol-6-ylcarbonyl chloride was used in place of piperonyloyl chloride, the title compound was prepared.

¹ H NMR δ (d ₆ DMSO) 8.57-7.48 (m, 7H), 6.43 (s, 1H), 4.48-3.55 (m, 6H), 1.80-1, 48 (m, 3H), 0.98-0.82 (m, 6H).

MS calculated for (C ₁₉ H ₂₃ N ₃ O ₄ - H) ⁺ : 356, found: 356.

Example 13

Preparation of 4- (R, S) -amino-N - [(benzofuran-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one

Following the procedure of Example 1 (e-f), except that benzofuran-2-ylcarbonyl chloride was used in place of piperonyloyl chloride, the title compound was prepared.

Ή NMR delta (CDC1 ₃₎ 8.00-7.25 (m, 7H), 4.90-4.78 (m, 1H), 4, 53-4.48 (m, IH), 4.38-4.21 (m, 1H), 4.25-3.92 (m, 3H), 1, 88 (m, 3H), 1.68 (m, 3H) 0.97 (m, 6 H).

MS calculated for (C ₁₉ H ₂₂ N ₂ O ₅ - H) ⁺ : 357, found: 357.

Example 14

Preparation of 4- (R, S) -amino-N - [(5-aminobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one

According to the procedure of Example 1 (e-f), except that φ · φ · φ · »

56 that 5-aminobenzo [b] thiophen-2-ylcarbonyl chloride is used instead of piperonyloyl chloride, the title compound is prepared.

Ή NMR delta (CDC1 ₃ ) 8.00-7.71 (m, 1H); 7.60 (d, 1H, J = Hz), 7.53 (dd ., 1H, J = 8.5 and 4.0 Hz), 7.47-6.74 (m, 3H), 4.77 (m, 1 H), 4.43 (m, 1H), 4.38-3.50 (m, 4H), 1.77 (m, 3H), 0.85 (m, 6H).

MS calculated for (C ₁₉ H ₂₃ N ₃ O ₄ S + H) ⁺ : 390, found: 390.

Example 15

Preparation of cyclic alkoxy ketones by solid synthesis

(a) 3-trans-Hydroxy-4-benzyloxycarbonylaminotetrahydrofuran

Benzyl chloroformate (20 mL) was added dropwise to a stirred solution of trans-4-amino-3-hydroxytetrahydrofuran (5 g, 20.6 mmol) in dioxane (100 mL) containing 10% aqueous sodium carbonate (200 mL). After 3 hours, the mixture was evaporated to remove dioxane and then extracted with EtOAc. The combined organic layers were washed with saturated sodium bicarbonate solution, brine and dried (MgSO 4). Evaporation under reduced pressure and purification of the residue by column chromatography gave the title compound as white crystals, 8.00 g, 70% yield.

1 H NMR δ (CDCl ₃ ) 7.35 (s, 5H), 5.30 (s, 2H), 4.91 (broad s, 1H), 4.32 (broad s, 1H), 4.12-4 .00 (m, 3H),

3.71-3.62 (m, 2H); 2.72 (s, 1H).

- 57 (b) 4-Benzyloxycarbonylaminotetrahydrofuran-3-one

A bleaching solution (100 mL) containing sodium bicarbonate (7.34 g) was added dropwise to a rapidly stirred mixture of 3-hydroxy-4-benzyloxycarbonylaminotetrahydrofuran (21 g, 88 mmol), sodium bromide (9.4 g), TEMPO ( 50 mg) in EtOAc (140 mL), toluene (140 mL) and water (40 mL). After developing a persistent orange color, the mixture was extracted with EtOAc and the combined organic layers were washed with saturated sodium bicarbonate solution, brine and dried (MgSO 4. Evaporation under reduced pressure and purification of the residue by column chromatography gave the title compound as white crystals, 18 g, 87% yield.

¹ H NMR δ (CDCl ₃ ) 7.35 (s, 5H), 5.30 (s, 1H), 5.11 (s,

2H), 4.70 (apparent t, 1H, J = 8.9 Hz), 4.36-4.17 (m, 2H), 3.96-3.76 (2H).

c) 3,3-Dimethoxy-4-benzyloxycarbonylaminotetrahydrofuran

Trimethylorthoformate (29 mL) was added dropwise to a solution of 4-benzyloxycarbonylaminotetrahydrofuran-3-one (18g, 78 mmol) and PTSA (500 mg) in MeOH (100 mL) maintained at reflux. After 3 hours, the reaction mixture was filtered and evaporated to give the title compound as a yellow oil, 16.2 g, 76% yield after column chromatography.

¹ H NMR δ (CDCl ₃ ) 7.35 (s, 5H), 5.30 (s, 1H), 5.11 (s,

2H), 4.70 (apparent t, 1H, J = 8.9 Hz), 4.36-4.17 (m, 2H), 3.96-3.76 (2H).

* • t t t t · · · ·

58 d) 3,3-Dimethoxy-4-amino-tetrahydrofuran

A mixture of 3,3-dimethoxy-4-benzyloxycarbonylaminotetrahydrofuran (16 g, 57 mmol) and 10% palladium on carbon (2 g) in ethanol (200 mL) was stirred under a hydrogen atmosphere (351.85 kPa) for 12 hours. The mixture was filtered and evaporated to give the title compound as a yellow oil, 8 g, 100% yield.

¹ H NMR δ (CDCl ₃ ) 7.04 (s, 5H), 4.22-4.04 (m, 5H), 3.83-3.69 (m, 2H), 3.36 (s, 3H) 1.33 (s, 3H).

e) SPS using an Ellman linker

Step A:

Sodium triacetoxyborohydride (10 equivalents) is added to a stirred solution of Ellman resin (ref: CG Boojamra, KM Burow, LA Thompson and JA Ellman, J. Org. Chem., 62, 1240 (1997)) in DMF containing 1% HOAc. After 5 minutes, alpha-amino acid methyl ester (10 equivalents) was added and the mixture was shaken for 1 hour. The resin was then washed with DMF (7x), CH ₂ Cl ₂ (7x), ether (2x) and dried to constant weight.

Step B:

To a mixture of the above resin, carboxylic acid (10 equivalents) and EDC (10 equivalents) was added NMP. The mixture was then shaken for 3 hours and then washed with DMF (3x), CH ₂ Cl ₂ (3x), MeOH (2x) and ether (2x). The resin is then re-exposed to the above reaction conditions, and the resin is re-exposed.

- 59 is washed again after 3 hours.

Step C:

Potassium trimethylsilanoate (10 equivalents) was added to the shaken mixture of the above resin in THF.

After 18 hours, the resin was washed with 5% citric acid in THF (2x), THF (2x), THF-H, O (2x), H ₂ O (2x), THF-H ₂ O (2x) and finally THF (2x ).

Step D:

To a mixture of the above resin and EDC (3 equivalents) was added 3,3-dimethoxy-4-aminotetrahydrofuran (3 equivalents). After 3 hours the resin was washed with DMF (7x), CH ₂ Cl ₂ (7x) and ether (2x). The resin is then re-exposed to the reaction conditions for 3 hours and then washed again as above.

Step E: Cleavage

A 7: 2: 1 mixture of TFA / CH ₂ Cl ₂ / H ₂ O is added to the above resin. After 2 hours, the mixture was filtered and the resin was further washed with CH ₂ Cl ₂ . Removal of the solvent gave the desired tetrahydrofuran-3-one.

Example 16

Preparation of 4- (R, S) -amino-N - [(5-chlorobenzofuran-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one

According to the general details of Example 15, using •

- 60 of the appropriate amino acid and carboxylic acid methyl ester as reagents consistent with the end product, the title compound is prepared.

¹ H NMR δ (CDCl ₃ ) 7.62-7.05 (m, 6H), 4.81-4.64 (m, 1H), 4.62-4.54 (m, 1H), 4.43 -3.81 (m, 4H), 1.90-1.60 (m, 3H), 1.08-0.81 (m, 6H).

MS calculated for (C ₁₉ H ₂₁ N ₂ O ₅ Cl + H) ⁺ : 393, found: 393.

Example 17

Preparation of 4- (R, S) -amino-N - [(5-methoxybenzofuran-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one

Following the general details of Example 15, using the appropriate amino acid and carboxylic acid methyl ester as reagents consistent with the end product, the title compound is prepared.

1 H NMR δ (CDCl ₃ ) 7.65-6, 84 (m, 6H), 4.83-4.54 (m, 1H), 4.53-4.48 (m, 1H), 4.46- 3.68 (m, 4H), 1.90-1.62 (m, 3H),

1.08-0.81 (m, 6H).

MS calculated for (C ₂₀ H ₂₄ N ₂ O ₆ + H) ⁺ : 389, found: 389.

Example 18

Preparation of 4- (R, S) -amino-N - [(4-bromobenzoyl) -S-leucine] tetrahydrofuran-3-one

According to the general details of Example 15, using

61 of the corresponding amino acid and carboxylic acid methyl ester as reagents consistent with the final product, the title compound is prepared.

1 H NMR δ (CDCl ₃ ) 7.64 (apparent d, 2H, J (apparent d, 2H, J = 8.5 Hz), 4.81-4.67 (m, (m, 1H), 3.36) -4.19 (m, 1H), 4.18-3.78 (m, (m, 3H), 1.05-0.80 (m, 6H).

= 8.5Hz), 7.60 (1H), 4.62-4.48 (3H), 1.81-1.59

MS calcd for (C 1 H 11 N 3 O 4 Br)

397, found: 397.

Example 19

Preparation of 4- (R, S) -amino-N - [(4-bromobenzoyl) -S-leucine] tetrahydrofuran-3-one

Following the general details of Example 15, using the appropriate amino acid and carboxylic acid methyl ester as reagents consistent with the end product, the title compound is prepared.

Ή NMR .delta (CDC1 ₃ ) 7.91 (m, 1H) , 7.76-7.59 (m, 2H) / 7, 39-7,18 (m, 1H), 7.04-6.89 (m, 1H), 7.84-6.68 (m, 1H), 4, 89-4.68 (m, 1H), 4,66-4,5 6 (m, 1H), 4.27-3.76 (m, 4H), 1, 88-1.68 (m, 3H), 1,03-0,78 (m, 6H).

MS calculated for (C ₁₇ H ₂₁ N ₂ O ₄ Br) ⁺ : 397, found: 397.

Example 20

Preparation of 4- (R, S) -amino-N - [(5-chlorobenzo [b] thiophene-2)]

• ·

62-ylcarbonyl-5-leucine] tetrahydrofuran-3-one

Following the general details of Example 15, using the appropriate amino acid and carboxylic acid methyl ester as reagents consistent with the end product, the title compound is prepared.

1 H NMR δ (CDCl ₃ ) 4.81 (broad d, 1H, 8.8 Hz), 4.43-4.30 (m, 3H), 1.05-0.82

7.88-7.67 (m, 5H), 7.48-7.38 (m, 1H), J = 6.7 Hz), 4.60 (apparent t, 1H, J = (m, 1H)) 4.28-3.84 (m, 3H), 1.86-1.62 (m, 6H).

MS calculated for (C ₁₉ H ₂₁ N ₂ O ₄ SC 1 - H) ⁺ : 408, found: 408.

Example 21

Preparation of 4- (R, S) -amino-N - [(4-fluorobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one

Following the general details of Example 15, using the appropriate amino acid and carboxylic acid methyl ester as reagents consistent with the end product, the title compound is prepared.

1 H NMR δ (CDCl ₃ ) 7.96 (s, 1H), 7.68-7.58 (m, 2H), 7.48-7.32 (m, 1H), 7.06 (dd, 1H, J = 9.0 Hz and 9.0 Hz), 4.89-4.72 (m, 1H), 4.65-4.55 (apparent t, 1H, J = 8.8 Hz), 4.35 (apparent q, 1H, J = 8.2 Hz), 4.28-3.82 (m, 3H), 1.88-1.62 (m, 3H), 1.05-0.82 (m, 6H).

MS calculated for (C ₁₉ H ₂₁ N ₂ O ₄ SF + H) ⁺ : 393, found: 393.

• 4

63 Examples 22 to 66

Preparation of cyclic alkoxy ketones

By procedures analogous to those described in either Example 1 or Example 15 using the appropriate amino acid and acid or acid chloride as reagents consistent with the end products, the compound of Table 1 was prepared. Ή NMR spectra and / or mass spectra were consistent with structures in Table 1.

Table 1

Example R ³ R procedure synthesis 22nd Λ 3,4-difluorophenyl solution 23 1 »1 4-Benzyl-piperidin-1-yl solution 24 H 4-Benzyl-piperazin-1-yl solution 25 rr 4- (3,4-methylenedioxybenzyl) piperazin-1-yl solution 26 π 4- (tert-Butoxycarbonyl) - piperazin-1-yl solution 27 Mar: in* piperazin-1-yl solution 28 TO benzimidazol-5-yl solution 29 IV 6-quinolyl solution 30 ff 5-indolyl solution

Φ Φ 9 · 9

9 9 9 • ··

- 64 Table 1 - continued

Example R ³ R procedure synthesis 31 II 2-Naphthyl solution 32 VI 2-pyridyl solution 33 II 4-Benzyloxyphenyl solution 34 IV 3-Benzyloxyphenyl solution 35 II 4-hydroxyphenyl solution 36 II 5-nitrobenzo [b] thiophen-2-yl solution 37 II 2- (thien-2-yl) ethen-1-yl solution 38 II 4-methoxyphenyl SPS 39 II 3-methoxyphenyl SPS 40 IV 7-Ethoxybenzofuran-1-yl SPS 41 II 5-nitrobenzofuran-1-yl SPS 42 II 4- (2-methoxyphenyl) phenyl SPS 43 IV 3- (2-methoxyphenyl) phenyl SPS 44 II 4-cyanophenyl SPS 45 II 3-nitrophenyl SPS 46 IV 3- (dimethylaminoethyl) -4- -methoxyphenyl SPS 47 th 2- (2-chlorophenyl) ethen-1-yl SPS 48 II 4-trifluoromethoxyphenyl SPS 49 IV 4-methanesulfonylphenyl SPS 50 II 4-iodophenyl SPS 51 II 4-chlorobenzo [b] thiophen-2-yl SPS 52 11 5,6-dimethoxybenzo [b] thiophen-2-yl SPS 53 IV 5,6-methylenedioxybenzo [b] - thiophen-2-yl SPS 54 11 7-chlorobenzo [b] thiophen-2-yl SPS

- 65 Table 1 - Continuation

Example R ³ R synthesis procedure Ph 55 benzo [b] thiophen-2-yl SPS 56 II 2-thienyl SPS 57 II 3,4-dimethoxyphenyl SPS 58 II 4-bromophenyl SPS 59 II quinolin-2-yl SPS 60 / 2-thienyl SPS 61 WITH II ch ₃ SPS 62 \ λ benzo [b] thiophen-2-yl SPS 63 II 2-thienyl SPS 64 II 3,4-dimethoxyphenyl SPS 65 II 4-bromophenyl SPS 66 II quinolin-2-yl SPS

Example 67

Preparation of 4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] tetrahydropyran-3-one

a) Trans-4-amino-3-hydroxytetrahydropyran hydrochloride

Following the procedure of Examples 1 (a) and 1 (b), •

66 instead of using 3,4-epoxytetrahydropyran instead of 3,4-epoxytetrahydrofuran, the title compound is prepared.

b) trans-4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] -3-hydroxytetrahydropyran

Pivaloyl chloride (5.28 mL, 43 mmol) was added to a solution of N-carbobenzyloxy-L-leucine (12.48 g, 47 mmol) in dichloromethane (300 mL). After 1 hour, a mixture of trans-4-amino-3-hydroxytetrahydropyran hydrochloride (6 g, 39 mmol) and triethylamine (10.8 mL, 79 mmol) in dichloromethane (100 mL) was added and the mixture was allowed to stir overnight. The reaction mixture was washed with 1N HCl, saturated sodium bicarbonate solution and dried. Evaporation under reduced pressure gave a light oil. Purification by chromatography (eluent ethyl acetate / hexane) gave trans-4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] -3-hydroxytetrahydropyran as a white paste, 5.5 g, 39% yield.

^; 1 H NMR δ (CDCl ₃ ) 7.35 (s, 5H), 5.0 (m, 3H), 4.16-3.89 (m, 4H), 3.44-3.36 (m, 2H) 3.13 (t, 1H) 1.84-1.53 (m,

2H), 1.28-1.22 (m, 3H), 0.93 (m, 6H).

c) 4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] tetrahydropyran-3-one

Following the procedure of Example 1 (f) except that 4- (R, S) was used in place of 4- (R, S) -amino-N - [(3,4-methylenedioxybenzoyl) -Sleucine] -3-hydroxytetrahydrofuran. 1-amino-N - [(benzyloxycarbonyl) -S-leucine] -3-hydroxytetrahydropyran, the title compound is prepared.

• · • ·· ·

- 67 ^X H NMR delta _(d6 -DMSO) 8.15 (d, IH), 7.39-7.29 (m, 5H), 5.02 (d, 2H), 4.63 (m, IH) 4.14-4.10 (m, 2H); 3.97-3.83 (m,

3H), 2.10 (m, 1H), 1.92 (m, 1H), 1.61 (m, 1H), 1.45 (m, 1H);

1H), 0.95 (m, 6H).

MS calcd for (C ₁₉ H ₂₆ N ₂ O ₅ + H) ⁺ : 363, found 363.

Example 68

Preparation of 4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydropyran-3-one

a) Trans-4- (R, S) -amino-N- (S-leucine) -3-hydroxytetrahydropyran hydrochloride

A mixture of 4- (R, S) -amino-N - [(benzyloxycarbonyl-S-leucine) -3-hydroxytetrahydropyran (2.8 g, 7.75 mmol) and 10% palladium on carbon (300 mg) in ethanol (100 g). The mixture was filtered and treated with ethereal HCl to give, after evaporation under reduced pressure, the title compound as a brown solid, 1.40 g, 68% yield. .

¹ H NMR δ (CDCl ₃ ) 8.30 (m, 2H), 8.02 (m, 1H), 4.06-3.92 (m, 4H), 3.50-3.35 (m, 2H) 3.12 (t, 1H), 1.89-1.54 (m,

2H), 1.23 (m, 3H), 0.93 (m, 6H).

b) trans-4- (R, S) -Amino-N - [(benzothiophene-2-carbonyl) -S-leucine] -3-hydroxytetrahydropyran

Benzothiophene-2-carbonyl chloride (442 mg, 2.25 mmol) was added to a solution of trans-4- (R, S) -amino-N- (S • ♦).

68 -leucine) -3-hydroxytetrahydropyran (133 mg, 0.5 mmol) in dioxane (7 mL) and saturated sodium bicarbonate solution (7 mL). After 30 minutes, the reaction mixture was diluted with ethyl acetate, the organic layer was washed with saturated sodium bicarbonate solution, dried and evaporated under reduced pressure to give a white solid. Purification by chromatography (ethyl acetate / hexane eluent) gave trans-4- (R, S) -amino-N- [(benzothiophene-2-carbonyl) -S-leucine] -3-hydroxytetrahydropyran as a white solid, 160 mg, 84 % yield.

c) 4- (R, S) -Amino-N - [(benzothiophene-2-carbonyl) -S-leucine] -tetrahydropyran-3-one

Following the procedure of Example 1 (f), except that 4- (R, S) -amino-N - [(3,4-methylenedioxybenzoyl) -S-leucine] -3-hydroxytetrahydrofuran was replaced by trans-4- (R, S) -Amino-N - [(benzothiophene-2-carbonyl) -S-leucine] -3-hydroxytetrahydropyran, the title compound is prepared.

¹ H NMR δ (CDCl ₃ ) 7.84-7.76 (m, 2H), 7.41 (m, 2H), 7.05 (m, 1H), 4.83-4.61 (m, 2H) 4.18-3.77 (m, 4H), 2.73-2.53 (m, 1H), 1.98-1.75 (m, 2H), 1.26 (m, 3H), 0.92 (m, 6H).

MS calcd for (C ₂₀ H ₂₄ N ₂ O ₄ S + H) ⁺ : 389, found 389.

Example 69

Preparation of 4- (R, S) -amino-N - [(4-phenoxybenzoyl) -S-leucine] tetrahydrofuran-3-one

A procedure analogous to that detailed in Example 15 provides the title compound.

Ή NMR δ (CDCl ₃ , 250 MHz) 7.74 (d, 2H, J = 10.9 Hz),

7.37 (dd, 2H, J = 7.7 and 7.7 Hz), 7.20 (dd, 1H, J = 7.5 and

7.5 Hz), 7.03 (d, 2H, J = 7.7 Hz), 7.0 (d, 2H, J = 7.7 Hz), 6.98-6.82 (m, 2H) 4.83-4.68 (m, 1H); 4.66-4.45 (m,

1H), 4.34-3.70 (m, 4H), 1.98-1.54 (m, 3H), 1.08-0.78 (m,

6H).

MS calcd for (C ₂₃ H ₂₆ N ₂ O ₅ - H) ⁺ : 409, found: 409.

Example 70

Preparation of 4- (R, S) -amino-N - [(4-phenylbenzoyl) -S-leucine] tetrahydrofuran-3-one

In a manner analogous to that detailed in Example 15, the title compound was prepared.

1 H NMR δ (CDCl ₃ , 250 MHz) 7.83 (d, 2H, J = 8.2 Hz), 7.70-50 (m, 4H), 7.48-7.32 (m, 4H), 7.30-7.12 (m, 1 H),

6.88-6.72 (m, 1H), 4.88-4.70 (m, 1H), 4.65-52 (m, 1H), 4.38-3.78 (m, 4H), 1.92-1.60 (m, 3H), 1.08-0.78 (m, 6H).

MS calcd for (C ₂₃ H ₂₆ N ₂ 0 ₄

H) ⁺

393, found: 393.

Example 71 • · · ::; ····.:. : ·· .......

70 Preparation of 4- (R, S) -amino-N - [(6-trifluoromethyl-benzo [b] thiophen-2-yicarbonyl) -S-leucine] tetrahydrofuran-3-one

In a manner analogous to that detailed in Example 15, the title compound was prepared.

¹ H NMR δ (DMSO, d ₆ , 250 MHz) 8.82 (d, 1H, J = 6.0 Hz),

8.43 (s, 1H), 8.48 -8.44 (m, 0.5H), .delta , 34 (d, 0.5 H, J = 7.8 Hz), 8.23 (with, 1H), 8.04 (d, 1 H, J = 8.4 Hz), 7, 60 (dd, 1 H, J = 1.5 and 8.5 Hz), 4.49-4.38 (m, IH) , 4.26-3.68 (m, 5H) 1.70-1.36 (m, 3H), 0.80 (d, 3 H, J = 6 Hz), 0.76 (d, 3H, J) 6.0 Hz) MS will calculate tané for (C ₂₀ H ₂₁ F ₃ N ₂ O ₄ S - H) ⁺ : 443, finds 443.

Example 72

Preparation of 4- (R, S) -amino-N - [(4-ethylbenzoyl) -S-leucine] tetrahydrofuran-3-one

In a manner analogous to that detailed in Example 15, the title compound was prepared.

¹ H NMR δ (CDCl ₃ , 250 MHz) 7, 76 (d, 2H, J = 8.0Hz) 7.62-7.42 (m, 1 H), 7.22 (d, 2H) J = 7 8 Hz), 7.04-6.80 (m, 1H), 4.91-4.73 (m, 1H), 4,61-4, 45 (m, 1H), 4.36-3.72 (m, 4H), 2.68 (q, 2H, J = 7.6 Hz), 1, 88- 1.58 (m, 3H); 1.23 (t,

3H, J = 7.6 Hz), 0.98-0.88 (m, 6H).

· <«« · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · · ·

- 71 MS calcd for (C ₁₉ H ₂₆ N ₂ O ₄ - H) ⁺ : 345, found: 345.

Example 73

Preparation of 4- (R, S) -amino-N - [(4- (tert-butyl) benzoyl) -S-leucine] tetrahydrofuran-3-one

In a manner analogous to that detailed in Example 15, the title compound was prepared.

7.76 (broad d, 2H, J = 7.5 7.5 Hz), 6.92-6.76 (m, 2H), (m, 1H), 4.37-3.71 (m, 4H) (9H), 1.00-0.82 (m, 6H).

Έ NMR δ (CDCl ₃ , 250 MHz) Hz), 7.46 (broad d, 2H, J = 4.88-4.68 (m, 1H), 4.58-4.43 1.82-1, 57 (m, 3 H), 1.32 (s,

MS calculated for (C ₂₁ H ₃₀ N ₂ O ₄ + H) ⁺ : 375, found 375.

Example 74

Preparation of 4- (R, S) -amino-N - [(5-methoxybenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one

In a manner analogous to that detailed in Example 15, the title compound was prepared.

₁ H NMR δ (CDCl ₃ , 250 MHz) 7.84-7.54 (m, 2H), 7.20 (d, 1H, J = 2.0 Hz), 7.10 (d, 1H, J = 2) 3 Hz), 7.06 (d, 1H, J = 2.3 Hz), 6.78 (d, 1H, J = 8.1 Hz), 4.82-4.68 (m, 1H, 4 59 (apparent t, 1H, J = 8.8 Hz), 4.49-3.61 (m, 4H),

-72 3.84 (s, 3H), 1.82-1.58 (m, 3H), 1.08-0.72 (m, 6H).

MS calcd for (C ₂₀ H ₂₄ N ₂ O ₅ S - H) ⁺ : 403, found: 403.

Example 75

Preparation of 4- (R, S) -amino-N - [(4-nitrobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one

In a manner analogous to that detailed in Example 15, the title compound was prepared.

₁ H NMR δ (CDCl ₃ , 250 MHz) 8.31 (s, 1H), 7.96 (d, 1H, J = 7.2 Hz), 7.62-7.18 (m, 4H), 4, 84 (apparent d, 1H, J = 7.5 Hz), 4.70-3.72 (m, 5H), 1.94-1.60 (m, 3H), 1.11-0.80 (m ,

6H).

MS calculated for (C ₁₉ H ₂₁ N ₃ O ₆ S - NO ₂ ) ⁺ : 373, found: 373.

Example 76

Preparation of 4- (R, S) -amino-N - [(6-bromobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one

In a manner analogous to that detailed in Example 15, the title compound is prepared.

₁ H NMR δ (CDCl ₃ , 250 MHz) 8.03 (s, 1H), 7.81-7.58 (m, 2H), 7.54-7.40 (m, 1H), 7.22-6 , 98 (m, 2H), 4.74 (apparent d, β · · · 9 · 9 · 9 · 9 · · ·

9 9 9 9 9

9 »» 9 9

- 73 1Η, J = 7.5 Hz), 4.59 (apparent q, 1H, J = 7.5 and 14.0 Hz), 4.48-3.74 (m, 4H), 1.85- 1.58 (m, 3H); 1.10-0.78 (m, 6H).

MS calcd for (C ₁₉ H ₂₁ BrN ₂ O ₄ S - H) ⁺ : 452, found: 452 '

Example 77

Preparation of 4- (R, S) -amino-N - [(5-bromobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one

In a manner analogous to that detailed in Example 15, the title compound was prepared.

1 H NMR δ (CDCl ₃ , 250 MHz) 8.04-7.90 (m, 1H), 7.88-7.18 (m, 5H), 4.88-4.68 (m, 1H), 4 , 68-4.52 (m, 1H), 4.46-3.88 (m, 4H), 1.92-1.52 (m, 3H), 1.08-0.80 (m, 6H) .

MS calculated for (C ₁₉ H ₂₁ BrN ₂ O ₄ S - H) ⁺ : 452, found: 452.

Example 78

Preparation

4- (R, S) -amino-N - [(6-methoxybenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one

In a manner analogous to that detailed in Example 15, the title compound was prepared.

Ή NMR δ (CDCl ₃ , 250 MHz) 7.79-7.61 (m, 2H), 7.32-7.14 · · · V · i · V · V · i · Φφ φφφ · * * φ

- 74 (m, 2Η), 7.00 (dd, 1Η, J = 1.5 and 8.8 Hz), 6.89-6.67 (m,

1H), 4.75 (apparent q, 1H, J = 8.2 and 16.2 Hz), 4.57 (apparent t, 1H, J = 8.6 and 17.4 Hz), 4.49-3 71 (m, 4H), 3.85 (s, 3H), 1.92-1.52 (m, 3H), 1.08-0.78 (m, 6H).

MS calcd for (C ₂₀ H ₂₄ N ₂ O ₅ S - H) ⁺ : 403, found: 403.

Examples 79 to 93

By procedures analogous to those described in Example 15 using the appropriate amino acid and an acid chloride as reagents consistent with the final product was also prepared from Table 2. ^X H NMR spectra and / or mass spectra were consistent with the structures in Table 2.

Table 2

'0

4 4 4 4 · · ·

4 4 4444 4444

Λ 4,444 44 «·

44 44 44 44 44 «44 4444 • 44 44 44 4 4 44 44

Example R ³ R procedure synthesis 79 AND phenyl SPS 80 1 at 3-chlorophenyl SPS 81 IV 2-Phenylethylen-1-yl SPS 82 lf 3-fluorophenyl SPS 83 lf 3-hydroxyphenyl SPS 84 lf 4-methylphenyl SPS 85 ff 4-isopropylphenyl SPS 86 lf 4-trifluoromethylphenyl SPS 87 ff 4-methylthiophenyl SPS 88 ff 4- (benzylsulfonylamino) phenyl SPS 89 ff 4- (diethylaminosilphonyl) phenyl SPS 90 ff 4- (acetylamino) phenyl SPS 91 ff 4-Benzoylphenyl SPS 92 lf 4-acetylphenyl SPS 93 ff 4- (4-Oxopent-1-yl) phenyl SPS

Example 94

Preparation of 4-S-amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one. · • ··· ·

• · · · · · · · · · · · · ·

a) Trans-4-S-amino-3-R-hydroxytetrahydrofuran hydrochloride

A mixture of trans-4-S-azido-3-R-hydroxytetrahydrofuran (LE Martinez reference, Leighton JL, DE Carsten and EN Jacobsen, J. Amer. Chem. Soc., 117, 5897 (1995)) (10 g, 77 mmol) ) and 10% palladium on carbon (1 g) in ethanol (150 ml) were stirred under a hydrogen atmosphere (351.85 kPa) for 12 hours. The mixture was filtered and treated with 100 mL of ethanolic HCl to give, after evaporation under reduced pressure, the title compound as a brown solid, 10.5 g, 97% yield, mp 132 ° C.

1 H NMR δ (D ₂ O) 4.54-4.52 (m, 1H), 4.24-4.13 (m, 2H), 3.98-3.61 (m, 3H).

(b) trans -4-S-amino-N - [(benzyloxycarbonyl) -S-leucine] -3-R-hydroxytetrahydrofuran

Trimethylacetyl chloride (5.4 mL, 44 mmol) was added to a stirred solution of N-Cbz-L-leucine (12.7 g, 48 mmol) and triethylamine (14 mL, 52 mmol) in THF (200 mL). After 1 hour trans-4-S-amino-3-R-hydroxytetrahydrofuran hydrochloride (5.58 g, 40 mmol) was added and the mixture was allowed to stir at reflux for 16 hours. The reaction mixture was filtered and evaporated under reduced pressure. Flash column chromatography (80% ethyl acetate-hexane) gave the title compound as a white foam, 10.6 g, 76% yield.

¹ H NMR δ (CDCl ₃ ) 7.76 (d, 1H, J = 5.3 Hz), 7.33-7.20 (m, 5,), 6.43 (d, 1H, J = 8, 9 Hz), 5.01 (apparent d, 2H, J =

3.0 Hz), 4.60-3.65 (m, 6H), 1.61-1.42 (m, 3H), 0.93-0.88 (m, 6H)

(c) Trans-4-S-amino-N- (S-leucine) -3-R-hydroxytetrahydrofuran hydrochloride

A mixture of trans-4-S-amino-N - [(carbobenzyloxy) -S-leucine] -3-R-hydroxytetrahydrofuran (2.0 g, 5.7 mmol) and 10% palladium on carbon (500 mg) in ethanol ( 100 ml) was stirred under a hydrogen atmosphere (351.85 kPa) for 12 hours. The reaction mixture was then filtered, diluted with ethereal HCl (100 mL, 1 M) and evaporated under reduced pressure to give the title compound as a white foam, 1.5 g, 100% yield.

¹ H NMR δ (D ₂ O) 4.20-4.05 (m, 2H), 4.06-4.00 (m, 1H), 3.90-3.83 (m, 2H), 3, 68-3.52 (m, 2H), 1.65-1.47 (m, 3H), 0.86-0.78 (m, 6H).

(d) trans -4-S-amino-N - [(2-benzo (b) thiophenecarbonyl) -S-leucine-3-R-hydroxytetrahydrofuran

N, N-Diisopropylethylamine (0.4 mL, 2.0 mmol) was added to a stirred solution of trans-4-S-amino-N- (S-leucine) -3-R-hydroxytetrahydrofuran hydrochloride (380 mg,

1.1 mmol) in dichloromethane (10 mL). After 5 minutes, benzo [b] thiophene-2-carbonyl chloride (196 mg, 1.0 mmol) was added and the mixture was allowed to stir for 1 hour then evaporated under reduced pressure. Flash column chromatography (40% acetone / hexane) gave the title compound as a white foam, 271 mg, 75% yield.

¹ H NMR δ (d ₆ DMSO, 400 MHz) 8.72 (d, 1H, J = 8.5 Hz),

8.25 (s, 1 H), 8.04 (d, 1 H, J = 8.5 Hz), 7.95 (d, 1 H, J =

8.5 Hz), 7.46-7.43 (m, 2H), 5.23 (d, 1H, J = 3.94 Hz), 4.544.47 (m, 1H), 4.03-4, Δ (m, 2H), 3.90 (dd, 1H, J = 5.4 and 8.9 Hz), 3.82 (dd, 1H, J = 4.4 and 9.3 Hz), 3.53 - 3.48 (m, · · ·)

- 78 2H), 1.72-1.66 (m, 2Η), 1.52-1.48 (m, 1Η), 0.91 (d, 3H, J = 6.4 Hz), 0.88 (d, 3H, J = 6.4Hz).

MS calculated for (C ₁₈ H ₂₄ N ₂ O ₅ + H) ⁺ : 365, found: 365.

(e) 4-S-amino-N - [(2-benzo (b) thiophenecarbonyl) -S-leucine] tetrahydrofuran-3-one

Dess-Martin periodinane (200 mg, 0.5 mmol) was added to a stirred solution of trans-4-S-amino-N - [(2-benzo (b) thiophenecarbonyl) -S-leucine-3-R-hydroxytetrahydrofuran (150 mg, 0.40 mmol) in dichloromethane (10 mL). After 1 hour, ether (20 mL) was added, followed by sodium thiosulfate (1 g). After a further 15 minutes, the reaction mixture was washed with saturated sodium bicarbonate solution, brine and dried. Evaporation of the solvent gave the title compound as a white foam, 147 mg, 100% yield.

Ή NMR δ (d ₆ DMSO, 400 MHz) 8.82 8, 46 (d, OH, J = 4.0 Hz), 8.28 (s, 4, 0 Hz), 7.98 (d, 1 H, J = 4.0 Hz), 4, 54-4,51 (m, ÍH), 4.33-4.18 (m, 3H) 1, 74-1.67 (m, 2H), 1.58-1.56 (m, 1H) Hz), 0.88 (d, 3H, J = 6.4 Hz).

(d, 1H, J = 4.0 Hz),

1 H), 8.05 (d, 1 H, J = 7.47-7.44 (m, 2H), 4.08-3.80 (m, 3H), 0.92 (d, 3H, J) = 6.4

MS calculated for (C ₁₉ H ₂₂ N ₂ O ₄ - H) ⁺ :

373, found: 373.

Example 95

Preparation of 4-R-amino-N - [(2-benzo [b] thiophenecarbonyl) -S-leucinl] tetrahydrofuran-3-one

According to the procedure of Example 94 (ae), but starting from the procedure of Example 94 (a) starting from trans-4-R-azido-3-S-hydroxytetrahydrofuran (ref. LE Martinez, JL Leighton, DE). Carsten and EN Jacobsen, J. Amer. Chem. Soc. 117. 5897 (1995)), was prepared upon the title compound ^X H NMR delta _(d6 DMSO, 400 MHz) 8.82 (d, H, J = 4 , 0 Hz)

8.52 (d, 1 H, J = 4.0 Hz), 8.28 (s, 1 H), 8.01 (d, 1 H, J =

4.0 Hz), 7.95 (d, 1H, J = 4.0 Hz), 7.47-7.44 (m, 2H), 4.54-4.51 (m, 1H), 4, 33-4.18 (m, 3H), 4.33 (apparent t, 1H, J = 8.5 Hz), 4.22 (dd, 1H, J = 8.7 and 16.0 Hz), 4, 07 (apparent d, 1H, J = 16.6 Hz), 3.90 (apparent d, 1H, J = 16.6 Hz), 3.81 (apparent t, 1H, J = 8.5 Hz), 1 74-1.67 (m, 2H), 1.58-1.56 (m, 1H), 0.92 (d, 3H, J = 6.4 Hz), 0.88 (d, 3H, J = 6.4 Hz).

MS calculated for (C ₁₉ H ₂₂ N ₂ O ₄ - H) ⁺ : 373, found: 373.

Example 96

Preparation of 4-S-amino-N - [(2-naphthoyl) -S-leucine] tetrahydrofuran-3-one

Following the procedure of Example 94, using the appropriate carbyxylic acid chloride and 4-azido-3-hydroxytetrahydrofuran to produce the desired stereochemistry consistent with the final product, the following compound was prepared.

¹ H NMR δ (d ₆ DMSO, 400 MHz) 8.64 (d, 1H, J = 3.8 Hz),

9 9 9

-80 8.54 (s, 1H), 8.42 (d, 1H, J = 3.8 Hz), 8.03 (d, 1H, J =

3.8 Hz), 8.02-7.95 (m, 3H), 7.62-7.58 (m, 2H), 4.63-4.60 (m, 1H), 4.35-4 27 (m, 2H), 4.08 (apparent d, 1H, J = 16.8 Hz), 3.86 (apparent d, 1H, J = 16.8Hz), 3.82 (apparent t, 1H, J = 8.0 Hz), 1.75-1.64 (m, 2H), 1.62-1.55 (m, 1H), 0.92 (d, 3H, J = 6.0 Hz), 0.88 (d, 3H, J = 6.0 Hz).

MS calcd for (C ₂₁ H ₂₄ N ₂ O ₄ - H) ⁺ : 367, found: 367,

Example 97

Preparation of 4-R-N - [(2-naphthoyl) -S-leucine] tetrahydrofuran-3-one

Following the procedure of Example 94, using the appropriate carbyxylic acid chloride and 4-azido-3-hydroxytetrahydrofuran to produce the desired stereochemistry consistent with the final product, the following compound was prepared.

¹ H NMR δ (d ₆ DMSO, 400 MHz) 8.62 (d, 1H, J = 3.8 Hz), 8.52 (s, 1H), 8.50 (d, 1H, J = 3.8 Hz), 8.04 (d, 1H, J =

3.8 Hz), 8.02-7.95 (m, 3H), 7.62-7.58 (m, 2H), 4.63-4.60 (m, 1H), 4.32 (apparent) t, 1H, J = 8.8 Hz), 4.23 (ddd, 1H, J = 8.8 and 16.0 Hz), 4.04 (apparent d, 1H, J = 16.4 Hz), 3 86 (apparent d, 1H, J = 16.4 Hz), 3.81 (apparent t, 1H, J = 8.0 Hz), 1.75-1.64 (m, 2H), 1.62- 1.55 (m, 1H), 0.92 (d, 3H, J = 6.0 Hz), 0.88 (d, 3H, J = 6.0 Hz).

MS calculated for (C ₂₁ H ₂₄ N ₂ O ₄ - H) ⁺ : 367, found: 367.

Example 98 • · · ·

Preparation of 4-S-amino-N - [(quinoline-2-carbonyl) -S-leucine] tetrahydrofuran-3-one

Following the procedure of Example 94, using the appropriate carbyxylic acid chloride and 4-azido-3-hydroxytetrahydrofuran to produce the desired stereochemistry consistent with the final product, the following compound was prepared.

1 H NMR δ (d ₆ DMSO, 400 MHz) 8.80 (d, 1H, J = 9.0 Hz), 8.70 (d, 1H, J = 6.9 Hz), 8.63 (d, 1H) J = 8.5 Hz), 8.39 (d, 1H, J = 2.5 Hz), 8.27 (d, 1H, J = 2.5 Hz), 8.21 (d, 1H, J = 7.9 Hz), 7.95 (apparent t, 1H, J = 8.2 Hz), 7.77 (apparent t, 1H, J = 7.1 Hz), 4.75-4.66 (m (1H), 4.41-4.26 (m, 2H), 4.16-3.79 (m, 3H), 1.82-1.63 (m, 3H), 0.97-0.88 (m, 6 H).

MS calculated for (C ₂₀ H ₂₃ N ₃ O ₄ + H) ⁺ : 370, found: 370.

Example 99

Preparation of 4-R-amino-N - [(quinoline-2-carbonyl-5-leucine) tetrahydrofuran-3-one

Following the procedure of Example 94, using the appropriate carbyxylic acid chloride and 4-azido-3-hydroxytetrahydrofuran to produce the desired stereochemistry consistent with the final product, the following compound was prepared.

1 H NMR δ (d ₆ DMSO, 400 MHz) 8.88 (d, 1H, J = 9.0 Hz), 8.82 (d, 1H, J = 6.7 Hz), 8.70 (d, 1H) Δ = 8.5 Hz), 8.30

9 9 9

· 9

- 82 (d, 1H, J = 2.5 Hz), 8.27 (d, 1H, J = 2.5 Hz), 8.21 (d, 1H, J = 7.9 Hz), 8.02 (apparent t, 1H, J = 7.0 Hz), 7.87 (apparent t, 1H, J = 7.0 Hz), 4.82-4.73 (m, 1H), 4.48-3, 88 (m, 5H) 1.88-1.70 (m, 3H), 1.05-1.02 (m, 6H).

MS calculated for (C ₂₀ H ₂₃ N ₃ O ₄ + H) ⁺ : 370, found 370.

Example 100

Preparation of 4-S-amino-N - [(5-methoxybenzofuran-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one

Following the procedure of Example 94, using the appropriate carbyxylic acid chloride and 4-azido-3-hydroxytetrahydrofuran to produce the desired stereochemistry consistent with the final product, the following compound was prepared.

Ή NMR delta (d ₆ DMSO, 400 MHz) 8.73 (d, 1H, J = 8.4 Hz), 8.52 ( d, 1H, J = 7.0 Hz), 7,65-7,62 (m, 2H) , 7, 33 (d, 1 H, J) = 2.6 Hz), 7 , 12 (dd, 1H, J = 2.6 and 9.2 Hz) , 4, 64-4.55 (m,

1H), 4.42-4.30 (m, 2H), 4.18-3.87 (m, 3H), 3.86 (s, 3H), 1.84-1.57 (m, 3H) 0.90-0.87 (m, 6H).

MS calcd for (C ₂₀ H ₂₄ N ₂ O ₆ + H) ⁺ : 38 9, found: 38 9 ·

Examples 101 to 108

Procedures analogous to those described in Example 15 using the appropriate amino acid and acid and acid chloride as reagents consistent with

99

9 9 9 ·· 999

83 NMR spectra and / or mass spectra were consistent with the structures in Table 3.

Table 3

102 5-hydroxybenzo [b] thiophen-2-yl SPS

103 4- (3-Hydroxymethyl) phenyl) phenyl SPS

104 3-phenylphenyl SPS

105 4- (3-Oxophenyl) phenyl SPS

Examples 109 to 126

According to the procedures of Example 94, using the chloride using the appropriate chloride, respectively. Examples of carboxylic acids and 4-azido-3-hydroxytetrahydrofuran having the desired stereochemistry to be consistent with the end products were prepared by Examples 109-126. ² H NMR and / or mass spectra were consistent with the structures in Table 4.

Table 4

Example l ³ R stereochemistry at position 4 109 / with 4- (pyrid-3-yl) phenyl WITH 110 F 4- (pyrid-2-yl) phenyl WITH 111 4-acetylphenyl WITH 112 benzyloxy WITH 113 3,4-dimethoxyphenyl WITH 114 benzofuran-2-yl WITH 115 4- (6-methylpyrid-3-yl) phenyl WITH 116 5-chlorobenzo [b] thiophen-2-yl with 117 4- (pyrid-4-yl) phenyl with

- 85 Table 4 - continued

Example R ³ R stereochemistry at position 4 118 II 2-chlorophenyl WITH 119 II 4-bromophenyl WITH 120 If 4-chlorobenzo [b] thiophen-2-yl WITH 121 II 4-Benzyl-piperidin-1-yl WITH 122 If 3,4-dichlorophenyl WITH 123 ff 3,4-dimethoxyphenyl R 124 ff benzofuran-2-yl R 125 If 3-chlorophenyl WITH 126 II 5-chlorobenzo [b] thiophen-2-yl R

Examples 127 to 129

Using the SPS procedure of Example 15, except that 3,3-dimethoxy-4-aminotetrahydropyran was used in place of 3,3-dimethoxy-4-aminotetrahydrofuran and using the appropriate amino acid and carboxylic acid as reagents consistent with the end product, prepares examples 127 to 129. 1 H NMR and / or mass spectra were consistent with the structures in Table 5.

Table 5

«* T t t t t t * * * * * * t t t t t t t t t t t t

Example R ^{* 3} R synthesis procedure

127 14-Phenoxyphenyl SPS 128 rr quinolin-2-yl SPS 129 M 3,4-dimethoxyphenyl SPS

Example 130

Preparation of 4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrothiophen-3-one (a) N-Benzo [b] thiophen-2-ylcarbonyl methyl ester -L-leucine

Benzo [b] thiophene-2-carbonyl chloride (4.9 g, 25 mmol) was added to a stirred solution of L-leucine methyl ester (4.5 g, 25 mmol) and diisopropylethylamine (9 mL, 51 mmol) in DCM (200 mL). ). After stirring at room temperature for 2 hours, the mixture was poured into water and washed with brine and dried (MgSO ₄ ). Evaporation under reduced pressure gave the title compound as a white solid, 7.6 g, 100% yield.

³ H NMR δ (CDCl ₃ , 250 MHz) 7.88-7.78 (m, 3H), 7.44-7.38 (m, 2H), 6.53 (d, 1H, J = 7.6Hz) 4.91-4.82 (m, 1H), 3.78 (s, 3H), 1.82-1.60 (m, 3H), 1.00 (apparent t, 6H, J = 5, 9 Hz).

· E · e e e e · · ·

- 87 • <9 (b) N-Benzo [b] thiophen-2-ylcarbonyl-L-leucine

Lithium hydroxide (1.41 g, 59 mmol) was added in one portion to a stirred solution of N-benzo [b] thiophen-2-ylcarbonyl-L-leucine methyl ester (8.99 g, 29.4 mmol) in THF / H ₂₀ (1: 1, 300 mL). After stirring at room temperature for 12 hours, the mixture was poured into water and acidified to pH 1 with concentrated HCl and extracted with Et ₂ O (2x). Evaporation under reduced pressure gave the title compound as a yellow solid, 6.1 g, 71% yield.

1 H NMR δ (d ₆ DMSO, 250 MHz) 8.78 (d, 1H, J = 9.1 Hz), 8.12 (s, 1H), 7.92-7.83 (m, 2H), 7 36-7.30 (m, 2H), 4.37-4.28 (m, 1H), 1.88-1.47 (m, 3H), 0.82 (d, 3H, J = 6, 0.7 Hz (d, 3H, J = 6.0 Hz).

(c) N-Benzo [b] thiophen-2-ylcarbonyl-L-leucine-S- (methoxycarbonylmethyl) -L, D-cysteine methyl ester

Isopropyl chloroformate (5.9 mL, 1.0 M in toluene) was added to a stirred solution of N-benzo [b] thiophen-2-ylcarbonyl-L-leucine (1.65 g, 5.7 mmol) and triethylamine (1.6 ml,

11.8 mmol) in DCM (20 mL). After stirring at room temperature for 1 h, L-cysteine ethyl ester was added followed by triethylamine (1.6 mL, 11.8 mmol) and the mixture was allowed to stir for an additional 6 h. Methyl bromoacetate (0.6 mL, 6.3 mmol) was added and before being poured into water, extracted with EtOAc (3x), and allowed to stand for an additional 0.5 h. The combined organic layers were washed with brine, dried (MgSO 4 and evaporated under reduced pressure) to give the title compound by flash column chromatography (40% hexane-Et, O). φ 9 9 9 · 9 · 9 • · ··· · · Λ

88 as a white solid, 1.6 g, 70% yield.

¹ H NMR δ (CDCl ₃ , 250 MHz) 7.84-7.77 (m, 3H), 7.66 (d, 0.5H, J = 8.4 Hz), 7.52 (d, 0) 5 H, J = 8.4 Hz), 7.40-7.35 (m, 2H), 7.24 (d, 0.5 H, J = 8.4 Hz), 7.16 (d, 0.5 H, J =

8.4 Hz), 4.88-4.82 (m, 2H), 4.23-4.15 (m, 2H), 3.70 (s, 1.5H), 3.69 (s, 1, H) 3.36-3.22 (m, 2H), 3.15-3.08 (m, 2H), 1.82-1.75 (m, 3H), 1.28 (t, 1.5 H) J = 4.5 Hz), 1.23 (t,

1.5 H, J = 4.5 Hz), 0.98-0.95 (m, 6H).

(d) 4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrothiophen-3-one

A freshly prepared solution of sodium methoxide (from 75 mg of sodium and 2 ml of methanol) was added to N-benzo [b] thiophen-2-ylcarbonyl-L-leucine-S- (carbomethoxymethyl) -L, D-cysteine ethyl ester (1,5 g, 3.2 mmol) in methanol (2 mL) at room temperature. After 1 h, Et ₂ O (80 mL) was added, then the solution was cooled to 0 ° C and the resulting white solid was filtered off. The solid was dissolved in 15 mL glacial acetic acid, 10 mL concentrated HCl and 15 mL H ₂ O. The mixture was stirred under reflux for 0.5 h, then cooled to room temperature and extracted with CHCl ₃ (5x). The combined organic layers were washed with NaHCO ₃ , brine, dried (MgSO ₄ ) and evaporated under reduced pressure. Bleskouvou column chromatography (40% hexane-Et ₂ O) gave upon the title compound as a white solid, 120 mg, 10% yield.

1 H NMR δ (CDCl ₃ , 400 MHz) 7.88-7.78 (m, 3H), 7.58 (d, 0.5H, J = 6.4 Hz), 7.54 (d, 0.5) H, J = 6.4 Hz), 7.42-7.35 (m, 3H), 4.85-4.80 (m, 1H), 4.43-4.34 (m, 1H), 3 , 34-3.23 (m, 3H), 3.02-2.91 (m, 1H), 1.76-1.74 (m, 3H), 0.98-0.85

A ·

89 (m, 6H).

MS calcd for (C ₁₉ H ₂₂ N ₂ O ₃ S + Η) ^4: 391, found: 391st

Example 131

Preparation of 4-amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] -3,3-dimethoxytetrahydrofuran diastereomers 1

a) 4- (R, S) -Amino-N - [(benzyloxycarbonyl) -S-leucine] -3,3-dimethoxytetrahydrofuran

4- (R, S) -amino-3,3-dimethoxytetrahydrofuran (2.2 g, 15 mmol), N-benzyloxycarbonyl-L-leucine (3.98 g, 15 mmol), EDC (3.17 g, 16 , 5 mmol) and hydroxyaminobenzotriazole (0.45 g, 3.3 mmol) were stirred together in a mixture of dichloromethane and tetrahydrofuran (1: 1, 100 mL) at room temperature for 12 hours. The mixture was added to ethyl acetate (300 mL), washed with water (2 x 100 mL), dried (MgSO ₄ ) and evaporated to dryness. Chromatography of the resulting oil on silica gel (ethyl acetate / hexane, gradient 1: 3 to 1: 1) gave the title compound as an oil, 5.27 g, 89% yield.

¹ H NMR δ (CDCl ₃ ) 7.34 (s, 5H), 6.55 (d, 1H), 5.11 (m,

3H), 4.34 (q, 1H, J = 6.7 Hz), 4.20 (m, 2H), 3, , 77 (s, 2H) 3.51 (m, 1 H), 3.26 (d, 3H, J = 2.3 Hz ), 3.20 (d, 3H, J = 4.7 Hz), 1.54 (m, 3H), 0.95 (s, , 3H) , 0.93 (s, 3 H).

b) Separation of 4-amino-N- (S-leucine) -3,3-dimethoxytetrahydrofuran diastereomers

4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] -3,3-dimethoxytetrahydrofuran (5.27g, 13.4 mmol) is subjected to hydrogenation at a pressure of 10 psig in methanol containing 10 % palladium on coal. After 3.5 hours, the mixture was filtered through celite and the solvent was removed under reduced pressure. Chromatography of the resulting oil (3.5 g) on silica gel (MeOH containing CH ₂ Cl ₂ , gradient 0-4%) separates the title compound into two pure diastereomers alone:

diastereoisomer 1 (faster moving) (0.63 g) ^X H NMR delta (CDC1 ₃₎ 7.76 (d, 1H, J = 7.3Hz), 4.36 (dd, 1H,

J = 6.3, 7.2 Hz), 4.21 (dd, 1H, J = 6.4, 9.0 Hz), 3.80 (s, 2H), 3.57 (dd, 1H, J = 5.3, 9.1 Hz), 3.41 (dd, 1H, J = 3.8, 10.1 Hz), 3.30 (s, 3H), 3.24 (s, 3H), 1 73 (m, 2H), 1.35 (m, 1H), 0.96 (d, 3H, J = 6.7 Hz), 0.94 (d, 3H, J = 6.6 Hz).

diastereomer 2 (moving slower) (0.91 g)

1 H NMR δ (CDCl ₃ ) 7.77 (d, 1H, J = 7.1 Hz), 4.36 (dd, 1H, J = 6.4, 12.9 Hz), 4.21 (dd, 1H) J = 6.5, 9.1 Hz), 3.80 (s, 2H), 3.55 (dd, 1H, J = 5.3, 9.0 Hz), 3.38 (dd, 1H, J = 4.0, 10.0 Hz), 3.30 (s, 3H), 3.25 (s, 3H), 1.72 (m, 2H), 1.38 (m, 1H), O, 96 (d, 3H, J = 6.7 Hz), 0.94 (d, 3H, J = 6.8 Hz).

c) Diastereomer 1 of 4-amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] -3,3-dimethoxytetrahydrofuran

Saturated sodium bicarbonate solution (5 mL) was added to a solution of diastereomer 1 4-amino-N- (S-leucine) 91

-3,3-dimethoxytetrahydrofuran (0.13 g, 0.5 mmol) in 1,4-dioxane (5 mL) then benzo [b] thiophen-2-ylcarbonyl chloride (0.39 g, 2 mmol) was added. After 45 minutes the mixture was added to ethyl acetate, washed with water, sodium bicarbonate solution and water. The organic solution was then dried (MgSO ₄ ) and evaporated to give a white solid (0.47 g). Silica gel chromatography (MeOH containing CH ₂ Cl ₂ , gradient 0-5%) gave the title compound as a white solid 0.18g, 89% yield.

di NMR delta ( CDC1 ₃ ) 7.88 (s, 1H), . 7.74 (m, 3H), 7.34 (m, 2H), 7 , 18 (d, 1H, J = 7.1 Hz ), 4, 82 (m, 1H) [Α] 4.38 (dd, 1H, J = 6, 4, 12.3 Hz), 4.21 (dd, 1H, 1 = 6.5, 9.2 Hz), 3.78 (ABq, 2H, J = 3,8, 9.8 Hz), 3.62 (dd, 1 H, J = 5,2,9,1 Hz), 1 78 (m 3H), 0.99 (s, 3H), 0.96 (s, 3H).

Examples 132-142

Procedures analogous to those described in Example 131 using the appropriate diastereomers of 4-amino-N- (S-leucine) -3,3-dimethoxytetrahydrofuran and a carboxylic acid halide as reagents consistent with the end product also prepared the compounds of Table 6. ³ H NMR and / or mass spectra were consistent with the structures in Table 6.

Table 6

- 92 .a

STEED

Example R ³ R R ^a R ^a ' diastereomer 132 / Ί benzo [b] - thiophen-2-yl Me Me 2 133 TI indol-5yl IV IV 1 134 Tl II TV IV 2 135 1 » quinolin-2-yl IV vv 1 136 II II IV vv 2 137 II 3-bromophenyl II IV 1 138 II II II vv 2 139 1 » 4-Phenoxyphenyl II vv 1 140 II IV IV vv 2 141 II indol-6-yl IV vv 2 142 IV benzimidazole-5 -yi 1

Example 143

Preparation of diastereomer 1 4-Amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] -3,3-dimethoxytetrahydropyran

a) 4- (R, S) -Amino-N - [(benzyloxycarbonyl) -S-leucine] -3,3-dimethoxytetrahydropyran

4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] •

93-tetrahydropyran-3-one (Example 67c, 3.1 g, 8.6 mmol) was refluxed with trimethylorthoformate (2.8 mL) and p-toluenesulfonic acid (0.080 g) in methanol (50 mL) for 12 h. ). After evaporation of the solvent under reduced pressure, the resulting oil was chromatographed on silica gel (ethyl acetate / hexane, gradient) to give the title compound as a white solid 2.89 g, 80% yield.

¹ H NMR δ (CDCl ₃ ) 7.29 (s, 5H), 6.99 (m, 1H), 6.15 (m,

1H), 5.07 (s, 2H), 4.28 (m, 1H), 4.18 (m, 1H), 3.52 (m 4H), 3.19 (s, 3H), 3.14 (s, 3H), 1.85 (m, 1H), 1.63 (m, 4H),

0.93 (s, 6H).

b) Separation of 4-amino-N- (S-leucine) -3,3-dimethoxytetrahydropyran diastereomers

4- (R, S) -Amino-N - [(benzyloxycarbonyl) -S-leucine] -3,3-dimethoxytetrahydropyran was hydrogenated as in Example 131b to afford pure diastereomers of the title compound after chromatography:

diastereomer 1: ^X H NMR .delta (CDCl ₃ ) 7.75 (d, 1H) , J = 8.1 Hz), 4.17 (m, 1 H), 3.58 (m, 3H), 3.41 (dd, 1 H, J = 3.5, 9.7 Hz), 3.28 (s, 3H) 3.24 (with, 3H), 1.92 (m, 1 H), 1.71 (m, 4H), 1.30 (m, 1 H), 0.97 (d, 3H, J = 6.4 Hz), 0 94 (d, 3H, J) = 6.3 Hz). diastereomer 2: ^X H NMR .delta (CDCl ₃ ) 7.8 (s, 1H) 4.16 (m, 1H), 3.59 (m, 3H) 3.49 (m, 1H), 3.28 (s, 3H), 3.24 (s, 3H), 1.92 (m, 1 H), 1, 72 (m, 4H), 1.41 (m, 1 H), 0.97 (d, 3H, J = 6.1 Hz) 0.95 (d, 3H, J = = 6.0 Hz).

c) Diasteromer 1 of 4-amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] -3,3-dimethoxytetrahydropyran

Diastereomer 1 of 4-amino-N- (S-leucine) -3,3-dimethoxytetrahydropyran was treated with benzo [b] thiophen-2-ylcarbonyl chloride as in Example 131c to give the title compound.

^X H NMR .delta (CDC1 ₃ ) 7 , 98 (d, IH, J = 8.3 Hz), 7 , 93 (with, ÍH), 7.78 (d, 1H, J = 7.7 Hz), 7 71 (d, 1 H, J = 8.3 Hz), 7.33 (m, 3H) 4.83 (m, ÍH) , 4.19 (m, 1H), 3.69 (m, ÍH), 3.57 (m, 2H) 3.40 (d, OH, J = 13 Hz), 3.23 (s, 3H), 3.13

(s, 3H), 1.85 (m, 5H), 0.98 (d, 3H, J = 4.8 Hz), 0.96 (d, 3H, J = 5.2 Hz).

Example 144

Preparation of 4-amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] -3,3-dimethoxytetrahydropyran diastereomer 2

Using the 4-amino-N- (S-leucine) -3,3-dimethoxytetrahydropyran diastereomer 2 in the procedure of Example 143c, the title compound was prepared.

The above description and examples fully demonstrate how to make and use the compounds of the present invention. However, the present invention is not limited to the particular embodiments described above, but includes all modifications thereof within the scope of the following claims. The various references to magazines, patents, and other publications cited herein include the prior art and are incorporated herein by reference as if fully set forth.

Claims (34)

PATENT CLAIMS 1. A compound of formula (I): in which R ¹ R ² R ⁴ (I) is R, R 'C (O), RC (S), RSO ₂ , ROC (O), RR'NC (O) or ROC (O) NR'CH (R ⁶ ) C (O) is H, C ^ alkyl, c _{2. 6} alkenyl, Ar-C ₀ _ _g alkyl or Het-C ₀ _ ₆ alkyl, is H, C ^ alkyl, C ₂ _ ₆ alkenyl, c _{2. 6} alkynyl, C ₃ _ _g cycloalkyl-C _{0th 6} alkyl, Ar-C _{0th g} alkyl or Het-C ₀ alkyl, is H, C _{1st 6} alkyl, c _{2. 6} alkenyl, Ar-C ₀ _ _g alkyl or Het-C ₀ _ ₆ alkyl, each R ⁵ is independently H, C ^ alkyl, c _{2. 6} alkenyl, Ar-C ₀ _ _g alkyl or Het-C ₀ _ ₆ alkyl, R ⁶ is H, C 1-6 alkyl, C ₂ . ₆ alkenyl, C ₃ _ ₆ cycloalkyl-C _0-6 alkyl, Ar-C ₀ _ _g alkyl or Het-C ₀ _ _g alkyl, R 1 is H, C 1-4 alkyl, C ₂ . ₆ alkenyl, Ar-C ₀ _ _g alkyl or Het-C ₀ _ _g alkyl, • 9 - R 96 is C ^ alkyl, Ar-C ₀ _ _g alkyl, Het-C ₀ _ _g alkyl, 'Ar-C ₂ _ ₆ alkenyl phenyl, or Het-C ₂ _ _g alkenyl, X is O or S, and n is 1, 2 or 3, or a pharmaceutically acceptable salt thereof. The compound of claim 1, wherein R ² and R ⁴ are each H. 3. A compound according to claim 1, wherein R ³ is C ^ alkyl or C ₂ _ ₆ alkenyl. The compound of claim ³ , wherein R ³ is isobutyl. A compound according to claim 1, wherein R ¹ is ROC (O), RSO ₂ or RC (O) wherein R is Ar-C ₀ - _G alkyl or Het-C ₀ _ _g alkyl. The compound of claim 5, wherein R is wherein B ₂ is OH, CN, OCF ₃ , OC 1-6 alkyl or 0— YOU ^! 0 ^ U ¹ ' OAr, SO ₂ C ₁ . ₆ alkyl, C alkyl φ φ φ φ - 97 or halogen. The compound of claim 1, wherein n is 1 or 2 The compound of claim 7, wherein n is 1. The compound of claim 1, wherein X is O. The compound of claim 1, wherein each R ⁵ is H Compound according to claim 1, of the general formula (IIa): (Ha). A compound according to claim 1, having the general formula (IIb): (Hb). A compound according to claim 1, having the general formula (IIc): The compound of claim 1 which is: (HC). 9 99 9 99 99 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 · 9 9 • 9 • 9 9 9 9 9 999 9 9 9 9 4- (R, S) -amino-N - [(3,4-methylenedioxybenzoyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(3,4-dichlorobenzoyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(2-quinolinecarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(8-quinolinecarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] -2,2-dibenzyltetrahydrofuran-3-one, 4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(3,4-dimethoxybenzoyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(indol-6-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(benzofuran-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(5-aminobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, • 9999 99 9 99 99 99 · 9 · 99 9 9 · 9 · 9 · 9 · 9 9 9 9 · 9 · 9 · 9 · 999 · 999 99 99 99 99 4- (R, S) -amino-N - [(5-chlorobenzofuran-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(5-methoxybenzofuran-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(3-bromobenzoyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(4-bromobenzoyl) -S-leucine] -tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(5-chlorobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(4-fluorobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(4-phenoxybenzoyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(4-phenylbenzoyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(6-trifluoromethylbenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(4-ethylbenzoyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [{4- (tert-butyl) benzoyl) -S-leucine] tetrahydrofuran-3-one, • • • 9 9 9 9 9 9 9 9 9 9 9 Φ Φ ΦΦ ΦΦΦΦΦΦ · · · · · · · - 100 4- (R, S) -amino-N - [(5-methoxybenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N- [N - (4-nitrobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(6-bromobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(5-bromobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - ((6-methoxybenzo (b) thiophen-2-ylcarbonyl) -S-leucine) tetrahydrofuran-3-one, 4-S-amino-N - [(benzo (b) thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4-R-amino-N - [(benzo (b) thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4-S-amino-N - [(2-naphthoyl) -S-leucine] tetrahydrofuran-3-one, 4-R-amino-N - [(2-naphthoyl) -S-leucine] tetrahydrofuran-3-one, 4-S-amino-N - [(quinoline-2-carbonyl) -S-leucine] tetrahydrofuran-3-one, 4-R-amino-N - [(quinoline-2-carbonyl) -S-leucine] tetrahydrofuran-3-one, 4-S-amino-N - [(S-methoxybenzofuran-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one; - 101 4-S-amino-N - [((4-pyrid-3-yl) benzoyl) -S-leucine) -tetrahydrofuran-3-one, 4-S-amino-N - [((4-pyrid-2-yl) benzoyl) -S-leucine] tetrahydrofuran-3-one, 4-S-amino-N - [(benzyloxycarbonyl) -S-leucine] tetrahydrofuran-3-one, 4-S-amino-N - [(3,4-dimethoxybenzoyl) -S-leucine] tetrahydrofuran-3-one, 4-S-amino-N - [(benzofuran-2-yicarbonyl) -S-leucine] tetrahydrofuran-3-one; 4-S-amino-N - ((4- [6-methylpyrid-3-yl) benzoyl) -S-leucine] tetrahydrofuran-3-one, 4-S-amino-N - [(5-chlorobenzo [b] thiophen-2-ylcarbonyl) -Sleucine] tetrahydrofuran-3-one, 4-S-amino-N - [((4-pyrid-4-yl) benzoyl) -S-leucine] tetrahydrofuran-3-one, 4-S-amino-N - [(2-chlorobenzoyl) -S-leucine] tetrahydrofuran-3-one, 4-S-amino-N - [(4-bromobenzoyl) -S-leucine] tetrahydrofuran-3-one, 4-S-amino-N - [(4-chlorobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one; - 102 4-S-amino-N - [(4-benzylpiperidin-1-ylcarbonyl) -S-leucine] tetrahydrofuran-3-one, 4-S-amino-N - [(3,4-dichlorobenzoyl) -S-leucine] tetrahydrofuran-3-one, 4-S-amino-N - [(3-chlorobenzoyl) -S-leucine] tetrahydrofuran-3-one, 4- (R, S) -amino-N - [(3,4-dimethoxybenzoyl) -S-leucine] tetrahydropyran-3-one, 4- (R, S) -amino-N - [(4-phenoxybenzoyl) -S-leucine] tetrahydropyran-3-one, 4- (R, S) -amino-N - [(quinolin-2-ylcarbonyl) -S-leucine] tetrahydropyran-3-one, 4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] tetrahydropyran-3-one, 4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydropyran-3-one or 4- (R, S) -amino-N - ((benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrothiophen-3-one or a pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising a compound according to any one of claims 1 to 1. And a pharmaceutically acceptable carrier. 16. A method of inhibiting a cysteine protease, comprising administering to a patient in need thereof an effective amount of a compound of claim 1. The method of claim 16, wherein the cysteine protease is cathepsin K. 18. A method of inhibiting bone loss, comprising administering to a patient in need thereof an effective amount of a compound of claim 1. 19. A method of treating osteoporosis comprising administering to a patient in need thereof an effective amount of a compound of claim 1. 20. A method of treating gingival or periodontal disease, comprising administering to a patient in need thereof an effective amount of a compound of claim 1. 21. A method of treating a disease characterized by excessive cartilage or matrix degradation, comprising administering to a patient in need thereof an effective amount of a compound of claim 1. 22. The method of claim 21, wherein said disease is osteoarthritis or rheumatoid arthritis. A compound according to any one of claims 1 to 14 for use as a medicament. • · · · Use of a compound of formula (I) according to claim 1 for the manufacture of a medicament for the treatment of diseases wherein inhibition of cysteine protease is a factor. 9 ·)))))))))))))))))) - 104 The use of a compound according to claim 24, wherein the cysteine protease is cathepsin K. Use of a compound of formula (I) according to claim 1 for the manufacture of a medicament for inhibiting bone loss. Use of a compound of formula (I) according to claim 1 for the manufacture of a medicament for the treatment of osteoporosis. Use of a compound of formula (I) according to claim 1 for the manufacture of a medicament for the treatment of gingival or periodontal disease. Use of a compound of formula (I) according to claim 1 for the manufacture of a medicament for the treatment of diseases characterized by excessive degradation of cartilage or matrix. The use of a compound according to claim 29, wherein the disease characterized by excessive cartilage or matrix degradation is osteoarthritis or rheumatoid arthritis. A process for preparing a compound of formula (I), according to claim 1, comprising (i) reacting a compound of formula (III): • 9 9 · 98 99 ♦ «9 9 9 9 9 9 9 9 9 R ¹ , R ² , R ³ , R ⁴ , R ⁵ and ⁿ are as defined in formula (I) of claim 1, wherein any reactive functional group is protected, with an oxidizing agent, or (ii) decarboxylating a compound of formula (I). IV): in which R ¹ , R ² , R ³ , R ⁴ , R ⁵ and ⁿ are as defined in formula (I) of claim 1, wherein any reactive functional group is protected, or (iii) reacting a compound of formula (V): (V) in which R ¹ , R ² , R ³ , R ⁴ , R ⁵ and ⁿ are as defined in formula (I) of claim 1, wherein any reactive functional 9 9 9 9 9 9 9 99 99 • 9 * 9 * 9 9 9 * 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 99 99 99 99 The 106 group is protected, with an acid followed by removal of any protecting group and optionally forming a pharmaceutically acceptable salt. 32. Compound of formula (III): (ΙΠ) in which R ¹ is R, R 'C (O), RC (S), RSO ₂ , ROC (O), RR'NC (O) or ROC (O) NR'CH (R ⁶ ) C (O), R ² is H, C 1-4 alkyl, C ₂ . ₆ alkenyl, Ar-C ₀ _ _g alkyl. or Het-C ₀ _ ₆ alkyl, R ³ is H, C 1-4 alkyl, C ₂ . _C6 alkenyl, c _{2. 6} alkynyl, C _{3rd g} cycloalkyl-C ₀ _ ₆ alkyl, Ar-C ₀ _ _g alkyl or Het-C ₀ _ _g alkyl, R ⁴ is H, C 1-4 alkyl, C ₂ . ₆ alkenyl, Ar-C ₀ _ _g alkyl or Het-C ₀ _ _g alkyl φφφφ Each R ⁵ is independently H, C 1-6 alkyl, C _2-10 . ₆ alkenyl, Ar-C ₀ _ ₆ alkyl or Het-C ₀ _ ₆ alkyl, R ⁶ is H, C ^ alkyl, C ₂ _ ₆ alkenyl, C ₃ -C _gCycloalkyl ₀ _ ₆ alkyl, Ar-C ₀ _ ₆ alkyl or Het-C ₀ _ _g alkyl, R 'is H, C ^ alkyl, C ₂ _ ₆ alkenyl, Ar-C ₀ _ ₆ alkyl or Het-C ₀ _ ₆ alkyl, R is C ^ alkyl, Ar-C _0-6 alkyl, Het-C ₀ _ ₆ alkyl, Ar-C _{2nd C6} alkenyl or Het-C ₂ _ ₆ alkenyl, and n is 1, 2 or 3, or a pharmaceutically acceptable salt thereof. R ¹ is R, RC (O), RC (S), RSO ₂ , ROC (O), RR'NC (O) or ROC (O) NR'CH (R ⁶ ) C (O), R ² is H, Ci.galkyl, C ₂ _ ₆ alkenyl, Ar-C ^ alkyl or Het-C ₀ . ₆ alkyl, R ³ is H, C ^ alkyl, C ₂ _ ₆ alkenyl, c _{2. 6} alkynyl, C _{3rd 6} cycloalkyl-C ₀ _galkyi, Ar-C _{0th 6} alkyl, or Het-C ₀ _ ₆ alkyl, - 108 R ⁴ is H, COalkyl, C ₂ _ ₆ alkenyl, Ar-C ₀ _ ₆ alkyl or Het-C ₀ _ ₆ alkyl, each R ⁵ is independently H, (alkyl, C ₂ _ ₆ alkenyl, Ar-C ₀ _ ₆ alkyl or Het-C ₀ _ _g alkyl, R ⁶ is H, COalkyl, C ₂ _ ₆ alkenyl, C ₃ _ ₆ cycloalkyl-Co_ ₆ -alkyl, Ar-C ₀ _ _g alkyl or Het-C ₀ _ ₆ alkyl, R 'is H, COalkyl, C ₂ _ _g alkenyl, Ar-C ₀ _ ₆ alkyl or Het-C ₀ _ ₆ alkyl, R is COalkyl, Ar-C ₀ _ ₆ alkyl, Het-C _{0th 6} alkyl, Ar-C ₂ _ ₆ alkenyl or Het-C _{2nd C6} alkenyl, and n is 1, 2 or 3, or a pharmaceutically acceptable salt thereof. in which R ¹ is R, RC (O), R C (S), RSO ₂ , ROC (O), RR'NC (O) or ROC (O) NR'CH (R ⁶ ) C (O), R ³ is H, Coalkyl, C ₂ . _C6 alkenyl, c _{2. 6} alkynyl, C ₃ _ ₆ cycloalkyl-C _{0th 6} alkyl, Ar-C ₀ _ ₆ alkyl or Het-C _{0th 6} alkyl, · alkyl alkyl alkyl alkyl alkyl alkyl alkyl alkyl alkyl ♦ - 109 R ⁴ is H, Ci.galkyl, C ₂ _ _g alkenyl, Ar-C ₀ _ _g alkyl or Het-C ₀ . _g alkyl, each R ⁵ is independently H, C ^ alkyl, C ₂ _ _g alkenyl, Ar-C ₀ _ _g alkyl or Het-C ₀ _ _g alkyl, R ⁶ is H, C ^ alkyl, C ₂ _ ₆ alkenyl, C ₃ -cycloalkyl _ _g _ _{0-C 6} -alkyl, Ar-C ₀ . _g alkyl or Het-C ₀ _ _g alkyl, R 'is H, C ^ alkyl, C ₂ _ ₆ alkenyl, Ar-C _{0th g} alkyl or Het-C ₀ _ _g alkyl, R is C 1-6 alkyl, Ar-C ₀ . _g alkyl, Het-C ₀ _ _g alkyl, Ar-C ₂ _ _g alkenyl or Het-C ₂ _ _g alkenyl, X is O or S, and n is 1, 2 or 3; or a pharmaceutically acceptable salt thereof. 35. A compound selected from the group consisting of: trans-4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] -3-hydroxytetrahydrofuran, trans-4- (R, S) -amino-N - [(tert-butoxycarbonyl) - S-leucine] -3-hydroxytetrahydrofuran, trans-4- (R, S) -amino-N- (S-leucine) -3-hydroxytetrahydrofuran, trans-4- (R, S) -amino-N - [(3 (4-methylenedioxybenzoyl) -S- • · · -110 -leucine] -3-hydroxytetrahydrofuran, trans-4- (R, S) -amino-N- [(3,4-dichlorobenzoyl) -S-leucine] -3-ydroxytetrahydrofuran, trans-4- (R, S) -amino-N - [(2-quinolinecarbonyl) -S-leucine] -3-hydroxytetrahydrofuran, trans-4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] -3-hydroxytetrahydropyran, trans-4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] -3-hydroxyletrahydropyran, trans-4- (R, S) -amino-N- [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] -3-hydroxytetrahydrofuran, trans-4- (R, S) -amino-N- [(indole-6-ylcarbonyl) -S-leucine] - 3-hydroxytetrahydrofuran, trans-4- (R, S) -amino-N - [(5-aminobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] -3-hydroxytetrahydrofuran, trans-4-amino-3- hydroxytetrahydrofuran, trans-3-hydroxy-4-benzyloxycarbonylaminotetrahydrofuran, 4-benzyloxycarbonylaminotetrahydrofuran-3-one, 3,3-dimethoxy-4-benzyloxycarbonylaminotetrahydrofuran, 3,3-dimethoxy-4-amino-tetrahydrofuran, 9 9 • · 9 9 99 9 9 9 9 9 9 9 9 9 9 9 9 • 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 9 111 trans-4-S-amino-3-R-hydroxytetrahydrofuran, trans-4-S-amino-N - [(benzyloxycarbonyl] -S-leucine] -3-R-hydroxytetrahydrofuran, trans-4-S-amino -N- (S-leucine) -3-R-hydroxytetrahydrofuran, trans-4-S-amino-N - [(3,4-dichlorobenzoyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans-4-S -amino-N- [(2-quinolinecarbonyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans -4-S-amino-N- [(benzo (b) thiophen-2-ylcarbonyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans-4-S-amino-N - [(benzofuran-2-ylcarbonyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans-4-S-amino-N - [(2 -naphthyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans-4-S-amino-N - [(5-methoxybenzofuran-2-ylcarbonyl) -S-leucine] -3-R-hydroxytecrahydrofuran, trans-4 -S-amino-N- [(5-chlorobenzo [b] thiophen-2-ylcarbonyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans-4-S-amino-N - [(4-chlorobenzo [b] thiophen-2-ylcarbonyl-5-leucine] -3-R-hydroxytetrahydrofuran, trans-4-S-amino-N - [(4-bromobenzoyl) -S-leucine] -3-R-hydroxytetrahydrofuran, - 112 trans -4-S-amino-N - [(4- (pyrid-2-yl) benzoyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans -4-S-amino-N - [(4 - (pyrid-3-yl) benzoyl) -S-leucine] -3-R-hydroxytetrahydrofuran, trans -4-S-amino-N- [(3,4-dimethoxybenzoyl) -S-leucine] -3-R- hydroxytetrahydrofuran, trans-4-amino-3-hydroxytetrahydropyran, trans-4- (R, S) -amino-N - [(benzyloxycarbonyl) -S-leucine] -3-hydroxytetrahydropyran, trans-4- (R, S) - amino-N- (S-leucine) -3-hydroxytetrahydropyran, trans-4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] -3-hydroxytetrahydropyran, N-benzo [b] thiophen-2-ylcarbonyl-L-leucine methyl ester, N-benzo [b] thiophen-2-ylcarbonyl-L-leucine, N-benzo [b] thiophen-2-ylcarbonyl-L-leucine-S- (methoxycarbonylmethyl) -L, D-cysteine ethyl ester or 2-methoxycarbonyl-4- (R, S) -amino-N - [(benzo [b] thiophen-2-ylcarbonyl) -S-leucine] tetrahydrothiophen-3-one, or salts thereof. 36. Compound of formula (VI): • · · · - 113 - in which R ¹ is R, R'C (O), RC (S), RSO ₂ , ROC (O), RR'NC (O) or ROC (O) NR'CH (R ⁶ ) C (O), R ² is H, C ^ alkyl, C ₂ _ ₆ alkenyl, Ar-C _{0th g} alkyl or Het-C ₀ _ ₆ alkyl, R ³ is H, C ^ alkyl, C ₂ _ ₆ alkenyl, C ₂ _ ₆ alkynyl, C _{3rd g} cycloalkyl-C ₀ _ ₆ alkyl, Ar-C ₀ _ _g alkyl or Het-C ₀ _ _g alkyl, R ⁴ is H, C ^ alkyl, C ₂ _ ₆ alkenyl, Ar-C ₀ _ _g alkyl or Het-C ₀ _ ₆ alkyl, each R ⁵ is independently H, C ^ alkyl, C ₂ _ ₆ alkenyl, Ar-C ₀ _ _g alkyl or het-C ₀ _ _s alkyl, R ⁶ is H, C ^ alkyl, C ₂ _ _g -alkenyl, C ₃ _ ₆ cycloalkyl-C ₀ _ ₆ alkyl, Ar-C ₀ _ _g alkyl or Het-C _{0th g} alkyl, R 'is H, C ^ alkyl, C ₂ _ ₆ alkenyl, Ar-C ₀ _ _g alkyl or Het-C ₀ _ ₆ alkyl, R is C ^ alkyl, Ar-C ₀ _ _g alkyl, Het-C _{0th g} alkyl, Ar-C ₂ _ _g alkenyl or Het-C ₂ _ ₆ alkenyl, X is O or S, and • ··· · • · ·· ·· · • • • · • • • • • · • • • • • • • • • · • • • • • • • ··· · • · · «· • · ·· - 114 n is 1, 2 or 3, R ^a and R ^a 'are independently H or C 1-4 alkyl, provided that when one of R ^a or R ^a ' is H, the other is C ₁ . ₂ alkyl or together are (CH _{2) 2} _ ₃ forming a 5- or 6-membered ring or a pharmaceutically acceptable salt thereof.