PHOSPHINIC PEPTIDE DERIVATIVES FOR USE AS MMP-12 INHIBITORS

Abstract

This invention relates to compounds that are selective inhibitors of Matrix Metalloprotease-12 (MMP-12), to cosmetic and pharmaceutical compositions containing them, and to their use in the prevention and/or treatment of ailments associated with MMP-12. Formula (I).

##STR00001##

Claims

1. A compound of formula (I) ##STR00044## wherein n is 0, 1 or 2, R.sup.1 is H, or an amino acid side chain which, if a functional group is present, may optionally be substituted by a C.sub.1-C.sub.6alkyl group, a C.sub.1-C.sub.6alkylcarbonyl group or a C.sub.1-C.sub.6alkoxycarbonyl group, R.sup.2 is H or Methyl, and R.sup.3 is selected from the group consisting of H, a C.sub.1-C.sub.6alkyl group and an arylC.sub.1-C.sub.6alkanyl group or a cosmetically acceptable salt thereof.

2. The compound according to claim 1, wherein the amino acid side chain is a nonpolar or an uncharged polar side chains of an amino acid.

3. A compound according to claim 1, wherein the amino acid side chain is selected from the side chains of glutamine, phenylalanine, methionine, valine, glycine, 2,4-diaminobutyric acid, 2-aminobutyric acid, alanine, leucine, isoleucine, norleucine, tryptophan, thiotryptophan, cyclohexylglycine, -amino-2-naphthalenepropionic acid, serine, threonine, tyrosine, proline and cysteine, preferably of phenylalanine, methionine, valine, thiotryptophan, isoleucine, tryptophan, 2,4-diaminobutyric acid, leucine, -amino-2-naphthalenepropionic acid, norleucine, asparagine, alanine, 2-amino butyric acid, and cyclohexylglycine, most preferably of thiotryptophan, isoleucine, tryptophan, 2,4-diaminobutyric acid, leucine, -amino-2-naphthalenepropionic acid, norleucine, asparagine, alanine, 2-amino butyric acid, and cyclohexylglycine

4. A compound according to claim 1, wherein the amino acid side chain has a functional group which is substituted with a C.sub.1-C.sub.6alkoxycarbonyl group, preferably with a tert-Butyloxycarbonyl (Boc) group.

5. A compound according to claim 1, wherein n is 0, 1 or 2, R.sup.1 is H or an amino acid side chain of thiotryptophan, isoleucine, tryptophan, 2,4-diaminobutyric acid, leucine, a-amino-2-naphthalenepropionic acid, norleucine, asparagine, alanine, 2-amino butyric acid, cyclohexylglycine, 2-amino-4-(tert-butoxycarbonylamino)butyric acid and 1-[(1,1-dimethylethoxy)carbonyl]-tryptophan R.sup.2 is H or methyl, preferably H, and R.sup.3 is selected from the group consisting of H, a methyl group, a tert.-butyl group and a benzyl group, or a cosmetically acceptable salt thereof.

6. A compound according to claim 1, wherein n is 0 or 2, R.sup.1 is H or an amino acid side chain of thiotryptophan, isoleucine, tryptophan, 2,4-diaminobutyric acid, leucine, -amino-2-naphthalenepropionic acid, norleucine, asparagine, alanine, 2-amino butyric acid, cyclohexylglycine, 2-amino-4-(tert-butoxycarbonylamino)butyric acid and 1-[(1,1-dimethylethoxy)carbonyl]-tryptophan R.sup.2 is H and R.sup.3 is selected from the group consisting of H, a methyl group, a tert.-butyl group and a benzyl group, or a cosmetically acceptable salt thereof.

7. A compound according to claim 1, which is ##STR00045## ##STR00046## ##STR00047## ##STR00048##

8. A compound according to claim 1, which is TABLE-US-00007 Name ((S)-2-([1,1-biphenyl]-4-ylmethyl)-3-(((S)-3-(benzo[b]thiophen-3-yl)-1-methoxy- (I)-A 1-oxopropan-2-yl)amino)-3-oxopropyl)(phenethyl)phosphinic acid ((2S)-3-([1,1-biphenyl]-4-yl)-2- (I)-B ((hydroxy(phenethyl)phosphoryl)methyl)propanoyl)-L-alloisoleucine ((2S)-3-([1,1-biphenyl]-4-yl)-2- (I)-C ((hydroxy(phenethyl)phosphoryl)methyl)propanoyl)-L-tryptophan salt with 2,2,2- trifluoroacetic acid (1:1) ((R)-2-([1,1-biphenyl]-4-ylmethyl)-3-(((S)-4-((tert-butoxycarbonyl)amino)-1- (I)-D methoxy-1-oxobutan-2-yl)amino)-3-oxopropyl)(phenethyl)phosphinic acid ((S)-2-([1,1-biphenyl]-4-ylmethyl)-3-(((S)-1-(tert-butoxy)-4-methyl-1-oxopentan- (I)-F 2-yl)amino)-3-oxopropyl)(phenethyl)phosphinic acid ((R)-2-([1,1-biphenyl]-4-ylmethyl)-3-(((S)-1-(benzyloxy)-3-(naphthalen-2-yl)-1- (I)-G oxopropan-2-yl)amino)-3-oxopropyl)(phenethyl)phosphinic acid ((S)-2-([1,1-biphenyl]-4-ylmethyl)-3-(((S)-1-methoxy-1-oxohexan-2-yl)amino)-3- (I)-H oxopropyl)(phenethyl)phosphinic acid ((2R)-3-([1,1-biphenyl]-4-yl)-2- (I)-I ((hydroxy(phenethyl)phosphoryl)methyl)propanoyl)-L-asparagine ((2S)-3-([1,1-biphenyl]-4-yl)-2- (I)-J ((hydroxy(phenethyl)phosphoryl)methyl)propanoyl)-L-alanine ((2R)-3-([1,1-biphenyl]-4-yl)-2- (I)-K ((hydroxy(phenethyl)phosphoryl)methyl)propanoyl)-L-tryptophan compound with 2,2,2-trifluoroacetic acid (1:1) 4-(3-([1,1-biphenyl]-4-yl)-2- (I)-L ((hydroxy(phenethyl)phosphoryl)methyl)propanamido)butyric acid (2S)-2-((2R)-3-([1,1-biphenyl]-4-yl)-2- (I)-M ((hydroxy(phenethyl)phosphoryl)methyl)propanamido)-3-(benzo[b]thiophen-3- yl)propanoic acid (2S)-2-((2R)-3-([1,1-biphenyl]-4-yl)-2- (I)-N ((hydroxy(phenethyl)phosphoryl)methyl)propanamido)hexanoic acid ((2S)-3-([1,1-biphenyl]-4-yl)-2- (I)-O ((hydroxy(phenethyl)phosphoryl)methyl)propanoyl)-L-asparagine ((S)-2-([1,1-biphenyl]-4-ylmethyl)-3-(((S)-1-(tert-butoxy)-1-oxobutan-2- (I)-P yl)amino)-3-oxopropyl)(phenethyl)phosphinic acid ((S)-2-([1,1-biphenyl]-4-ylmethyl)-3-(((S)-1-(benzyloxy)-3-(naphthalen-2-yl)-1- (I)-Q oxopropan-2-yl)amino)-3-oxopropyl)(phenethyl)phosphinic acid (2-([1,1-biphenyl]-4-ylmethyl)-3-(((S)-1-(tert-butoxy)-3-(1-(tert-butoxycarbonyl)- (I)-R 1H-indol-3-yl)-1-oxopropan-2-yl)amino)-3-oxopropyl)(phenethyl)phosphinic acid ((S)-2-([1,1-biphenyl]-4-ylmethyl)-3-(((S)-1-cyclohexyl-2-methoxy-2- (I)-S oxoethyl)amino)-3-oxopropyl)(phenethyl)phosphinic acid ((R)-2-([1,1-biphenyl]-4-ylmethyl)-3-(((2S,3R)-1-(tert-butoxy)-3-methyl-1- (I)-T oxopentan-2-yl)amino)-3-oxopropyl)(phenethyl)phosphinic acid ((S)-2-([1,1-biphenyl]-4-ylmethyl)-3-(((2S,3R)-1-(tert-butoxy)-3-methyl-1- (I)-U oxopentan-2-yl)amino)-3-oxopropyl)(phenethyl)phosphinic acid (2-([1,1-biphenyl]-4-ylmethyl)-3-((1-methoxy-2-methyl-1-oxopropan-2- (I)-V yl)amino)-3-oxopropyl)(phenethyl)phosphinic acid

9. A cosmetic composition comprising a compound of formula (I) according to claim 1 and a cosmetically acceptable carrier.

10. The cosmetic composition according to claim 9, wherein the compounds of formula (I) is present in an amount selected in the range of 0.00001 to 0.5 wt.-% based on the total weight of the composition.

11. A compound according to claim 1 for the use as MMP-12 inhibitor.

12. A compound according to claim 11 for the use to inhibit the degradation of elastin and/or collagen in human skin.

13. A compound according to claim 1, for the use in a method to protect the extracellular matrix proteins, preferably elastin and/or collagen in human skin, said method comprising the step of applying a cosmetic composition to the affected area.

14. A compound according to claim 1, for the use in a method to prevent and/or treat aged or senescent skin, said method comprising the step of applying a cosmetic composition to the affected area.

15. A compound according to claim 14 to smoothen wrinkles and fine lines, to decrease their volume and depth, to treat skin sagging and/or to improve skin firmness.

Description

EXAMPLE 1

Preparation of ((2S)-3-([1,1-biphenyl]-4-yl)-2-((hydroxy(phenethyl)phosphoryl)methyl)-propanoyl)-L-alanine (I)-J and ((2R)-3-([1,1-biphenyl]-4-yl)-2-((hydroxyl-(phenethyl)-phosphoryl)methyl)propanoyl)-L-alanine (I)-J

[0088] 350 mg (1.47 mmol) of 2-biphenyl-4-yl-acrylic acid was dissolved in 4 ml of DMF and chilled to 5 C. Subsequently 495 mg TBTU and 512 l DIPEA were added, followed by a solution of 320 mg of H-Ala-OtBu.HCl (1.76 mmol) in 3 ml of DMF along with 299 l of DIPEA. After stirring for 30 min at 5 C. and overnight at RT, the crude product was isolated by evaporation of solvent. The residue was dissolved in 30 ml of EtOAc and washed consecutively with 10% citric acid, 10% NaHCO.sub.3 and saturated NaCl-solution. After drying over Na.sub.2SO.sub.4 and evaporation of solvent, 500 mg of an oil was obtained. HPLC (220 nm)95%, [M+H].sup.+, found 366 (theo. 366). The obtained intermediate (475 mg, 1.3 mmol) was dissolved in 5 ml of BSA along with 265 mg (1.56 mmol) of (2-phenethyl)-phosphinic acid under a stream of argon, followed by heating to ca. 70 C. for 62 h (94% conversion). The reaction mixture was diluted with 100 ml of EtOAc and BSA quenched by addition of 10 ml of water. After washing thoroughly with fresh water and saturated NaCl-solution, drying over Na.sub.2SO.sub.4, filtering and evaporation of solvent, 630 mg of a yellowish oil was obtained. The crude product was chromatographed using preparative HPLC to separate the two formed diastereomers yielding after lyophilisation:

[0089] 295 mg of (2-([1,1-biphenyl]-4-ylmethyl)-3-(-1-(tert-butoxy)-1-oxopropan-2-yl)amino)-3-oxopropyl)(phenethyl)phosphinic acid consisting mainly of the (S/S)-isomer (80%) and 146 mg consisting of mainly the (R/S)-isomer (82%)).

[0090] ESI-MS [M+H].sup.+: found 536.4 for both peaks (theo: 536.3).

[0091] Each fraction was hydrolyzed to the corresponding acid by dissolving the respective compound in 2 ml of DCM and adding 2 ml of 95% TFA. Upon completion of hydrolysis of the ester and evaporation of the solvent, the crude products were chromatographed with preparative HPLC yielding

[0092] 171 mg of (I)-J as colorless powder, purity: 98% (HPLC), ESI-MS [M+H].sup.+: 480.3 (theo: 480.2) and

[0093] 62 mg of (I)-J as off-white powder, purity: 97% (HPLC), ESI-MS [M+H].sup.+: found 480.3 (theo: 480.2).

EXAMPLE 2

((S)-2-([1,1-biphenyl]-4-ylmethyl)-3-(((S)-1-methoxy-1-oxohexan-2-yl)amino)-3-oxopropyl)(phenethyl)phosphinic acid (I)-H and (2S)-2-((2R)-3-([1,1-biphenyl]-4-yl)-2-((hydroxy(phenethyl)phosphoryl)methyl)propanamido)hexanoic acid (I)-N

[0094] 350 mg (1.47 mmol) of 2-biphenyl-4-yl-acrylic acid was dissolved in 4 ml of DMF and chilled to 5 C. Subsequently 495 mg TBTU and 512 l DIPEA were added, followed by a solution of 320 mg of HNIe-OMe.HCl (1.76 mmol) in 3 ml of DMF along with 299 l of DIPEA. After stirring for 30 min at 5 C. and overnight at RT, the crude product was isolated by evaporation of solvent. The residue was dissolved in 30 ml of EtOAc and washed consecutively with 10% citric acid, 10% NaHCO.sub.3 and saturated NaCl-solution. After drying over Na.sub.2SO.sub.4, filtration and evaporation of solvent, 523 mg of an oil was obtained. HPLC (220 nm)83%, LC-MS: [M+H].sup.+, found 366 (theo. 366). The intermediate (523 mg, 1.33 mmol) was dissolved in 5 ml of BSA along with 285 mg (1.68 mmol) of (2-phenethyl)-phosphinic acid under a stream of argon, followed by heating to ca. 70 C. After 15 h, HPLC monitoring indicated approx. 70% conversion. Fresh BSA along with 200 mg of (2-phenethyl)-phosphinic acid was added and stirring continued under argon for a total of 4 days. The reaction mixture was diluted with 100 ml of EtOAc and BSA quenched by addition of 10 ml water. After washing thoroughly with fresh water and saturated NaCl-solution, drying over Na.sub.2SO.sub.4, filtration and evaporation of solvent, 710 mg of a yellowish oil was obtained. The crude product was chromatographed using preparative HPLC to separate the two formed diastereomers yielding after lyophilization

[0095] 36 mg of ((S)-2-([1,1-biphenyl]-4-ylmethyl)-3-(((S)-1-methoxy-1-oxohexan-2-yl)amino)-3-oxopropyl)(phenethyl)phosphinic acid (I)-H as off-white powder and

[0096] 69 mg of ((R)-2-([1,1-biphenyl]-4-ylmethyl)-3-(((S)-1-methoxy-1-oxohexan-2-yl)amino)-3-oxopropyl)(phenethyl)phosphinic acid (I)-H as off-white powder and

[0097] 257 mg of a mixture of (I)-H (51%) and (I)-H (47%)

[0098] ESI-MS [M+H].sup.+: found 536.4 for both peaks (theo: 536.3).

[0099] Each fraction was mildly saponified to the corresponding acid by dissolving in THF and adding 1.1 eq of LiOH. Upon completion of hydrolysis of the ester, each reaction mixture was acidified and extracted with EtOAc and the crude product chromatographed with preparative HPLC yielding

[0100] 88 mg (2S)-2-((2S)-3-([1,1-biphenyl]-4-yl)-2-((hydroxy(phenethyl)phosphoryl)methyl)-propanamido)hexanoic acid (I)-N as off-white powder. Purity: 99% (HPLC); ESI-MS [M+H].sup.+: found 522.2 (theo: 522.2) and

[0101] 80 mg (2S)-2-((2R)-3-([1,1-biphenyl]-4-yl)-2-((hydroxy(phenethyl)phosphoryl)methyl)-propanamido)hexanoic acid (I)-N as off-white powder. Purity 90% (HPLC); ESI-MS [M+H].sup.+: found 522.2 (theo: 522.2)

EXAMPLE 3: INHIBITION OF MMP-12

[0102] The MMP-12 inhibitor activity was quantified by fluorometric analysis using the MMP-12 Fluorometric Drug Discovery Kit (Enzo Life Sciences Inc., Farmingdale, N.Y., USA) according to the manufacturers' instructions.

TABLE-US-00003 TABLE 3-1 Results of the assay % Inhibition after 20 min # Compound 10 M 1 M 0.5 M 1 [00023] 92 82 75 2 [00024] 92 94 94 3 [00025] 92 94 93 4 [00026] 91 90 85 5 [00027] 91 82 73 6 [00028] 91 85 78 7 [00029] 91 89 83 8 [00030] 90 89 84 9 [00031] 90 94 93 10 [00032] 90 79 69 11 [00033] 90 82 75 12 [00034] 89 64 52 13 [00035] 89 n.a. n.a. 14 [00036] 88 n.a. n.a. 15 [00037] 88 n.a. n.a. 16 [00038] 83 n.a. n.a. 17 [00039] 82 n.a. n.a. 18 [00040] 82 n.a. n.a. 19 [00041] 75 n.a. n.a. 20 [00042] 72 n.a. n.a. 21 [00043] 61 n.a. n.a.

EXAMPLE 4-1: INHIBITION OF THE DEGRADATION OF THE ELASTIN FIBERS PRODUCED BY HUMAN DERMAL FIBROBLASTS

[0103] Human dermal fibroblasts were pre-cultured in DMEM, 10% FCS, 1% P/S for 48 hrs before starving them in DMEM, 0.2% FCS, 1% P/S for 24 hrs. To induce elastin fiber production TGFb1 (10 ng/ml) was added and incubated for 5 days. After incubation with TGFb1, medium was carefully discarded and cells were washed with assay buffer (HBSS, 25 M HEPES). In the meantime MMP-12 (200 U/ml) and compounds were mixed in assay buffer and allowed to interact for 30 min. This mixture was then added to the cells and incubated for 2 hrs at 37 C.

[0104] Cells were then fixed with ice-cold methanol for 5 min and rehydrated by PBS. Unspecific sites were blocked with 1% BSA in PBS for 1 h at room temperature. Elastin fibers were detected using a primary antibody against elastin and the first antibody was detected with AlexaFluor 488 conjugated secondary antibody. Finally, nuclei were counterstained with DAPI. Stained cells were analyzed with the Thermo Scientific ArrayScan XTI Live High Content Platform. 49 pictures per well were acquired with a 20 objective and as readout we used the area covered by the elastin fibers. % of inhibition of elastin fiber degradation was calculated the following way:

% protection of elastin fiber network degradation=100[(max areamin area)/(max areainhibitor area)*100] in which

max area=area covered by elastin fibers of untreated control cells
min area=area covered by elastin fibers of MMP12 treated cells
inhibitor area=area covered by elastin fibers of MMP12 and inhibitor treated cells.

[0105] The results are outlined below outlined in table 4-1.

TABLE-US-00004 TABLE 4-1 % protection of elastin fiber network degradation Compound 20 M 1 M 0.05 M (I)-C.sup. 92.03 84.78 44.07 (I)-c* 87.95 71.96 24.86 (I)-J.sup. 91.72 68.42 26.46 (I)-m* 80.76 64.88 8.95 (I)-B.sup. 90.12 81.75 17.38 (I)-e* 83.73 68.17 12.44 (I)-k* 98.72 84.79 25.93 (I)-O.sup. 89.07 45.61 13.75 (I)-l* 105.48 54.65 18.25 (I)-n* 76.78 30.36 41.01 .sup.Pure stereoisomer *Mixture of stereoisomers

EXAMPLE 4-2: TOTAL DERMAL ELASTIN AFTER UV-IRRADIATION

[0106] Human skin from abdominal plastic surgery classified as Intermediate (ITA angle=53) has been used. The skin samples were cut in pieces of approx. 83 mm (thickness) and cultured up to day 6 in an air-liquid interface in a perforated ring of stainless steel in contact with a culture medium (modified Williams'E medium), the culture medium was renewed every three days. Eight skin specimens were used for each treatment. Each test sample (4 l) was topically applied twice a day, immediately after UV irradiation and renewed 4 hours later top of each piece after gentle cleaning of the surface with a cotton pad followed by covering with a 6 mm delivery membrane, which procedure was repeated daily. The samples were irradiated daily with 800% of the Biological Effective Dose of daylight (BED; Del Bino et al, Pigment Cell research, 19, 2006) equal to a dose of 45 J/cm.sup.2 UVA using an adopted BIO-SUN sytem (Vilber Lourmat). At day 6 twelve skin sections were immunostained with mouse monoclonal anti-Elastin antibody (Sigma cat#E4013). The papillary dermis is selected for the analysis. The evaluation has been performed by estimating both color intensity and distribution with IMAGE J (NIH) analysis software Two slides of each skin sample have been processed by image acquisition and related analysis (i.e. 12 images for each treatment). The score of the dermal Elastin of the non-UV treated sample was set to 100%.

TABLE-US-00005 TABLE 4-2 Dermal Elastin at day 6 Concentration Dermal elastin Increase vs. Treatment [m] [%] untreated-UV NO UV 100 Untreated - UV 81 (IV-c) - UV 100 123 +52% (II-m) - UV 100 106 +30% (IV-k) - UV 100 125 +54% ((Dermal elastin sample Dermal Elastin - UV)/Dermal Elastin UV * 100%)

[0107] As can be retrieved from the results outlined in table 4-2, the compounds according to the present invention counteracted UV damage while restoring or even enhancing dermal elastin in the papillary dermis.

EXAMPLE 5: COSMETIC COMPOSITION

[0108] Table 5-1 outlines exemplary O/W emulsions, wherein one compound (eventually as mixture of stereoisomers) selected from the group of I (a-r) [Table 1] and (I)-(A-V) [Table 2], is incorporated in the indicated amount.

TABLE-US-00006 TABLE 5-1 Exemplary O/W emulsion O/W Emulsions 1 2 3 4 5 6 7 8 Glyceryl Stearate 2.5 2 1.2 1 1 1 PEG-40 Stearate 1 PEG-100 Stearate 2.5 1 Ceteareth-20 1 Glyceryl Stearate Citrate 0.5 Potassium Cetyl Phosphate 3 1.5 Stearic Acid 2.5 3 Cetearyl Alcohol 4 2 2 Stearyl Alcohol 2 1 Cetyl Alcohol 1 1 0.5 Acrylates/C.sub.10-30 Alkyl Acrylate 0.2 0.2 0.4 0.2 Crosspolymer Carbomer 0.1 0.2 Xanthan Gum 0.3 0.3 C.sub.12-15 Alkyl Benzoate 5 2 5 5 10 5 Petrolatum 5 3 Butylene Glycol Dicaprylate/Dicaprate 4 2 9 9 Hydrogenated Polydecene 3 2 2 Caprylic/Capric Triglyceride 1 3 5 5 5 Cyclomethicone 5 2 10 Methylpropanediol 2 3 3 Glycerine 4 7 3 4 3 5 3 Glyceryl Glucoside 3.5 3 1 1 2 2 Alcohol denat. 1 3 0.5 10 4 8 4 Butylene Glycol 3 Ascorbylglucoside 0.5 1.0 1.5 0.1 Ubiquinone (Coenzyme 10) 0.1 0.05 0.01 Hyaluronic acid 0.2 Bisabolol 0.5 0.2 Isotridecylsalicylate 1 3 5 2 3 5 Compound selected from the group of 0.001 0.25 0.0001 0.05 0.1 0.0003 0.03 0.002 I (a-r) and (I)-(A-V) Dibutyl Adipate 1.5 3 Diisopropyl sebacate 1 1 2 3 Ethylhexyl Benzoate 0.75 1.5 1 Titanium Dioxide (PARSOL TX) 0.5 2 Methylene Bis-Benztriazoyl 0.5 4 6 2 Tetramethylbutylphenol Ethylhexyl methoxycinnamate 2 Phenylbenzimidazole Sulfonic Acid 2 2 2 Butyl Methoxydibenzoylmethane 1 2 2 3 3 3 Methylbenzylidene Camphor 2 3 Octocrylene 5 2 10 Polysilicone-15 2 3 Ethylhexyl Salicylate 5 Homosalate 4 2 Bis-Ethylhexyloxyphenol 1.5 2 Methoxyphenyltriazine Silica 1 2.5 0.5 Silica & Methicone 4 1 2.5 Methyl Methacrylate Crosspolymer 1 2 Disodium EDTA 0.1 0.5 Fragrance, Preservatives q.s. Sodium Hydroxide q.s. Water Ad 100