4-VINYL-2-CYCLOPENTEN-1-ONE DERIVATIVES, THE PRODUCTION THEREOF, AND THE USE OF SAME AS AN ANTIBIOTIC AGENT

Abstract

The invention relates to a compound of general formula (I) wherein: R1 is a hydrogen atom or a C1 to C4 alkyl radical; R2 is a C1 to C4 hydroxyl or alkoxyl radical; and R3 and R4 are independently a hydrogen atom or a C1 C4 alkyl radical; and the enantiomers and mixtures of enantiomers thereof, especially in a racemic form.

Claims

1. Compound of the following general formula (I): ##STR00010## wherein: R.sub.1 is a hydrogen atom or a C.sub.1 to C.sub.4 alkyl radical; R.sub.2 is a C.sub.1 to C.sub.4 hydroxyl or alkoxyl radical, and R.sub.3 and R.sub.4 are independently a hydrogen atom or a C.sub.1 to C.sub.4 alkyl radical; the enantiomers and mixtures of enantiomers thereof, particularly in a racemic form.

2. A compound according to claim 1 of the following formula (Ia): ##STR00011##

3. A method for producing a compound of formula (I) or (Ia) according to claim 1 or claim 2, characterized in that it involves implementing the following operational steps: i. culturing the filamentous fungus Trichoderma, in particular Trichoderma atroviride, in a medium and under suitable conditions; ii. during culture, in situ capture by solid-phase extraction of a compound of formula (I) or (Ia); iii. isolating and purifying the compound of formula (I) or (Ia).

4. The method according to claim 3, characterized in that the in situ capture step is carried out by solid-phase extraction after introducing into the culture medium at least a nonpolar neutral resin.

5. The method according to claim 3, characterized in that the in situ capture step is carried out by solid-phase extraction after introducing into the culture medium at least a nonpolar neutral resin, wherein said resin is a polystyrene-divinylbenzene copolymer.

6. A method for producing a compound of formula (I) or (Ia), according to claim 1 or claim 2, characterized in that it involves implementing the steps illustrated in the reaction scheme below, the initial reagents being optionally substituted in a suitable manner in order to obtain the compounds of formula (I) with the meanings of R.sub.1 to R.sub.4 being as defined above: ##STR00012##

7. A compound of formula (I) or (Ia) according to claim 1 or claim 2, for use as an antibiotic agent.

8. A compound according to claim 7, for use as a broad-spectrum antibiotic agent.

9. A compound according to claim 7, for use as an antibiotic agent against multiresistant Gram-positive and Gram-negative pathogenic bacteria.

10. A pharmaceutical composition characterized in that it contains as an active ingredient at least a compound of formula (I) or (Ia) according to claim 1 or claim 2 associated with a pharmaceutically acceptable excipient.

Description

EXAMPLE 1

[0037] 1.1. General Procedure

[0038] Unless otherwise specified, all the reactions were carried out under an inert atmosphere of argon, in glassware first dried in a 120 C. oven.

[0039] 1.1.1. Solvents

[0040] Tetrahydrofuran (THF) was distilled under argon over sodium/benzophenone. Dichloromethane (CH.sub.2Cl.sub.2) was distilled under argon over calcium hydride (CaH.sub.2). Unless otherwise specified, all the reagents employed are commercial reagents of reagent-grade quality.

[0041] 1.1.2. TLC Analysis

[0042] The crude reaction mixtures obtained, the fractions and the purified products were analyzed by thing layer chromatography (TLC) on silica gel plates (Merck 60 F.sub.254 0.2-mm thickness on aluminum sheets). The plates are observed under a UV lamp (at 254 and 312 nm) before being developed by spraying with ammonium molybdate solution (NH.sub.4).sub.2Mo.sub.2O.sub.7 (100 g/L in 10% sulfuric acid) and heating.

[0043] Retention factor is defined as the ratio between the distance covered by the compound on the plate and the solvent front.

[0044] 1.1.3. Purification

[0045] Purification of the reaction mixtures was carried out by flash chromatography on silica gel (Merck 60 (35-70 m)).

[0046] 1.1.4. Spectroscopic Analyses

[0047] 1.1.4.1 High-Resolution Mass Spectrometry

[0048] High-resolution mass spectra were obtained on a mass spectrometer equipped with an atmospheric pressure ionization source (ESI) and with a time of flight (TOF)-type mass analyzer (LCT, Waters).

[0049] 1.1.4.2 Nuclear Magnetic Resonance (NMR)

[0050] Nuclear magnetic resonance experiments were carried out on Bruker Avance 300 and 500 devices using deuterated chloroform (CDCl.sub.3). Chemical shifts are expressed in parts per million (ppm) and calibrated relative to the reference solvent. Coupling constants are expressed in hertz (Hz). Signal multiplicity is expressed by the following abbreviations: s (singlet), bs (broad singlet), d (doublet), dd (doublet of doublets), t (triplet), m (multiplet), q (quadruplet). Attribution of the proton and carbon signals was carried out from one-dimensional (1D) .sup.1H and .sup.13C, and two-dimensional (2D).sup.1H-.sup.1H COSY, .sup.1H-.sup.13C HSQC or HMQC, .sup.1H-.sup.13C HMBC, .sup.1H-.sup.1H NOESY experiments. NMR spectra are processed using the dedicated TopSpin software (Brucker).

[0051] 1.1.4.3 Optical Rotation

[0052] The optical rotations of the compounds were measured using a Jasco P1010 polarimeter equipped with the Spectro Manager software. The monochromatic light source is the sodium D-line. The experiments were conducted with a 100 mm, 350 L quartz cuvette and the products were dissolved in methanol.

[0053] 1.1.4.4 Infrared Spectroscopy

[0054] Infrared (IR) absorption spectra of the described compounds were measured on the Perkin-Elmer Spectrum 100 FT-IR spectrometer. The device is equipped with the Spectrum software (version 6.3.5) from Perkin-Elmer. The compounds were produced in solution in methanol then dried with compressed air. Absorption bands are given in cm.sup.1.

[0055] 1.2. Synthesis of Compounds 2 to 7

Compound 2: 4-hydroxycyclopent-2-enone

[0056] ##STR00004##

[0057] A solution of furfuryl alcohol (1) (3.0 ml, 34.7 mmol) in distilled water (5 ml) is placed in a microwave tube (CEM Discover) and heated in a closed vessel at 300 W for 10 minutes. After cooling and decanting, the aqueous phase is collected and the residue washed with 25 ml of distilled water. The aqueous phases are combined and washed by liquid-liquid extraction with ethyl acetate (250 ml). The aqueous phase is then concentrated to give 668 mg (20%) of compound 2 in the form of an orange-red oil. The product obtained does not require purification before being employed in the following reaction (K. Ulbrich, P. Kreitmeier, O. Reiser, Synlett. 2010, 13, 2037-2040)

[0058] Rf=0.32 (Heptane/AcOEt, 2:8)

[0059] .sup.1H NMR (300 MHz, CDCl.sub.3): .sub.H (ppm) 7.57 (dd, 1H, J=2.3 Hz, J=5.7 Hz), 6.22 (dd, 1H, J=1.1 Hz, J=5.7 Hz), 5.06-5.02 (m, 1H), 2.77 (dd, 1H, J=6.1 Hz, J=18.5 Hz), 2.27 (dd, 1H, J=2.1 Hz, J=18.5 Hz).

[0060] .sup.13C NMR (75 MHz, CDCl.sub.3): .sub.C (ppm) 207.1, 163.7, 135.0, 70.3, 44.2.

[0061] HRMS (ESI): m/z calculated for C.sub.5H.sub.7O.sub.2 [M+H.sup.+] 99.0446 experimental value 99.0449

Compound 3: 4-((tert-butyldimethylsilyl)oxy)cyclopent-2-enone

[0062] ##STR00005##

[0063] Compound 2 (334 mg, 3.41 mmol) is dissolved in dichloromethane (6 ml). The reaction mixture is then held at 0 C. Triethylamine (700 l, 4.78 mmol), DMAP (33 mg, 0.27 mmol) and tert-butyldimethylsilyl chloride (1.0 M in THF, 3.75 ml) are then added. The reaction is held at 0 C. with stirring for 12 hours. After adding distilled water (10 ml), the medium is extracted with dichloromethane (320 ml). The organic phase is dried over anhydrous sodium sulfate, then evaporated to dryness. The crude reaction mixture obtained is purified by flash chromatography on silica gel (eluent: heptane/AcOEt, 80:20) to give 664 mg (92%) of compound 3 in the form of a transparent oil (white crystals at 4 C.) (K. Ulbrich, P. Kreitmeier, O. Reiser, Synlett. 2010, 13, 2037-2040).

[0064] Rf=0.53 (Heptane/AcOEt, 3:7)

[0065] .sup.1H NMR (300 MHz, CDCl.sub.3): .sub.H (ppm) 7.46 (dd, 1H, J=2.3 Hz, J=5.7 Hz), 6.19 (dd, 1H, J=1.1 Hz, J=5.7 Hz), 5.01-4.98 (m, 1H), 2.71 (dd, 1H, J=6.1 Hz, J=18.5 Hz), 2.25 (dd, 1H, J=2.2 Hz, J=18.5 Hz), 0.91 (s, 9H), 0.14 (s, 3H), 0.13 (s, 3H).

[0066] .sup.13C NMR (75 MHz, CDCl.sub.3): .sub.C (ppm) 206.5, 163.8, 134.4, 70.9, 45.0, 25.7, 18.1, 4.8

[0067] HRMS (ESI): m/z calculated for C.sub.22H.sub.41O.sub.4Si.sub.2 [2M+H.sup.+] 425.2583 experimental value 425.2601

Compound 4: methyl 3-(4-((tert-butyldimethylsilyl)oxy)-1-hydroxycyclopent-2-en-1-yl)propiolate

[0068] ##STR00006##

[0069] n-Butyl lithium (1.6 M in hexane, 2.14 ml, 1.1 eq) is added to a solution of diisopropylamine (529 l, 1.2 eq) in 10 ml of THF and held at 78 C. At the end of 30 minutes, methyl propiolate (277 l, 1.0 eq) is added to the LDA solution obtained beforehand. The reaction is held at 78 C. with stirring for 1 hour. Compound 3 (660 mg, 3.11 mmol) is dissolved in 5 ml of THF at 78 C. then the solution is slowly added to the lithium acetylide solution generated beforehand. The reaction is held at 78 C. with stirring for 1 hour. After adding 20 ml of saturated NH.sub.4Cl solution, the mixture is extracted with 350 ml of ethyl acetate. The organic phase is dried over anhydrous sodium sulfate, then evaporated to dryness. The crude reaction mixture obtained is purified by flash chromatography on silica gel (eluent: heptane/AcOEt, 80:20) to give 718 mg (78%) of compound 4 in the form of a yellow translucent oil.

[0070] Rf=0.44 (Heptane/AcOEt, 6:4)

[0071] .sup.1H NMR (300 MHz, CDCl.sub.3): .sub.H (ppm) 5.99 (dd, 1H, J=1.7 Hz, J=5.4 Hz), 5.92 (d, 1H, J=5.5 Hz), 4.83 (t, 1H, J=5.4 Hz), 3.78 (s, 3H), 2.87 (dd, 1H, J=6.7 Hz, J=13.6 Hz), 2.02 (dd, 1H, J=4.3 Hz, J=13.7 Hz), 0.90 (s, 9H), 0.10 (s, 6H).

[0072] .sup.13C NMR (75 MHz, CDCl.sub.3): .sub.C (ppm) 153.8, 138.5, 134.7, 88.2, 75.6, 74.9, 52.8, 51.0, 25.8, 18.0, 4.7.

[0073] HRMS (ESI): m/z calculated for C.sub.15H.sub.23O.sub.3Si [MH.sub.2O+H.sup.+] 279.1416 experimental value 279.1414

Compound 5: (E)-methyl 3-(4-((tert-butyldimethylsilyl)oxy)-1-hydroxycyclopent-2-en-1-yl)acrylate

[0074] ##STR00007##

[0075] Compound 4 (700 mg, 2.36 mmol) is dissolved in 20 ml of distilled THF and the solution is cooled to 78 C. Commercial Red-Al solution (3.21 M in toluene, 1.54 ml, 2.0 eq) is added dropwise and the reaction is held at 78 C. with stirring for 30 minutes. After adding 20 ml of distilled water, the crude reaction mixture is extracted with 3100 ml of ethyl acetate. The organic phase is washed with 10 ml of saturated NaHCO.sub.3 solution, then dried over anhydrous sodium sulfate before being filtered and evaporated to dryness. The residue obtained is purified by flash chromatography on silica gel (eluent: heptane/AcOEt, 80:20) to give 668 mg (95%) of compound 5 in the form of a light yellow translucent oil.

[0076] Rf=0.63 (Heptane/AcOEt, 6:4)

[0077] .sup.1H NMR (300 MHz, CDCl.sub.3): .sub.H (ppm) 6.84 (d, 1H, J=15.4 Hz), 6.09 (d, 1H, J=15.6 Hz), 5.97 (dd, 1H, J=2.0 Hz, J=5.4 Hz), 5.77 (d, 1H, J=5.5 Hz), 4.80-4.77 (m, 1H), 3.75 (s, 3H), 2.52 (dd, 1H, J=6.5 Hz, J=13.8 Hz), 1.87 (dd, 1H, J=3.9 Hz, J=13.8 Hz), 0.90 (s, 9H), 0.10 (s, 6H).

[0078] .sup.13C NMR (75 MHz, CDCl.sub.3): .sub.C (ppm) 167.4, 150.5, 137.4, 136.7, 118.5, 82.9, 75.3, 51.6, 49.8, 25.8, 18.1, 4.7.

[0079] HRMS (ESI): m/z calculated for C.sub.15H.sub.25O.sub.3Si [MH.sub.2O+H.sup.+] 281.1573 experimental value 281.1562

Compound 6: (E)-methyl 3-(1,4-dihydroxycyclopent-2-en-1-yl)acrylate

[0080] ##STR00008##

[0081] Compound 5 (650 mg, 2.18 mmol) is dissolved in 20 ml of distilled THF at room temperature. Commercial TBAF solution (1 M in THF, 4.37 ml, 2.0 eq) is added dropwise and the reaction is held at room temperature with stirring for 2 hours. After adding 10 ml of saturated NaCl solution, the reaction mixture is extracted with 3100 ml of ethyl acetate. The combined organic phases are dried over anhydrous sodium sulfate before being filtered and evaporated to dryness. The residue obtained is purified by flash chromatography on silica gel (eluent: heptane/AcOEt, 80:20) to give 357 mg (89%) of compound 6 in the form of a translucent oil.

[0082] Rf=0.41 (Heptane/AcOEt, 3:7)

[0083] .sup.1H NMR (300 MHz, CDCl.sub.3): .sub.H (ppm) 6.87 (d, 1H, J=15.7 Hz), 6.09 (d, 1H, J=15.6 Hz), 6.09 (dd, 1H, J=2.0 Hz, J=5.4 Hz), 5.84 (d, 1H, J=5.5 Hz), 4.85-4.81 (m, 1H), 3.76 (s, 3H), 2.60 (dd, 1H, J=6.7 Hz, J=14.3 Hz), 1.91 (dd, 1H, J=3.5 Hz, J=14.3 Hz).

[0084] .sup.13C NMR (75 MHz, CDCl.sub.3): .sub.C (ppm) 167.1, 150.8, 137.7, 136.9, 118.6, 83.0, 75.2, 51.7, 48.9.

[0085] HRMS (ESI): m/z calculated for C.sub.9H.sub.11O.sub.3 [MH.sub.2O+H.sup.+] 167.0708 experimental value 167.0700

Compound 7: (E)-methyl 3-(1-hydroxy-4-oxocyclopent-2-en-1-yl)acrylate ()-LMA-EA-2801

[0086] ##STR00009##

[0087] The compound (350 mg, 1.92 mmol) is dissolved in 15 ml of distilled CH.sub.2Cl.sub.2 at room temperature, before adding thereto sodium acetate (313 mg, 1.2 eq) and pyridinium chlorochromate (PCC, 497 mg, 1.2 eq). The reaction is held at room temperature with stirring for 1 hour. The reaction mixture is filtered on a Celite cartridge and the filtrate obtained is evaporated to dryness. The residue is then purified by flash chromatography on silica gel (eluent: heptane/AcOEt, 60:40) to give 296 mg (84%) of compound 7 in the form of a translucent oil.

[0088] Rf=0.38 (Heptane/AcOEt, 3:7)

[0089] .sup.1H NMR (300 MHz, CDCl.sub.3): .sub.H (ppm) 7.35 (d, 1H, J=5.3 Hz), 6.94 (d, 1H, J=15.8 Hz), 6.27 (d, 1H, J=5.4 Hz), 6.18 (d, 1H, J=15.8 Hz), 3.77 (s, 3H), 2.67 (d, 1H, J=12.0 Hz).

[0090] .sup.13C NMR (75 MHz, CDCl.sub.3): .sub.C (ppm) 205.0, 166.4, 162.6, 148.5, 134.6, 120.5, 78.3, 51.9, 49.3.

[0091] HRMS (ESI): m/z calculated for C.sub.9H.sub.9O.sub.3 [MH.sub.2O+H.sup.+] 165.0552 experimental value 165.0550

[0092] Separation of Enantiomers (+)-LMA-EA-2801 and ()-LMA-EA-2801

[0093] Compound 7 was analyzed by chiral HPLC equipped with a UV detector in order to detect the two enantiomers and to be able to separate same. (Waters Alliance 2695 HPLC system coupled to a Waters 996 PDA detector; Daicel Chiral Technologies IC chiral column (4.6250 mm, 5 m); eluent: n-heptane/isopropanol, 80:20).

[0094] Based on the analytical data, the separation of the enantiomers was then carried out by preparative SFC.

[0095] (Waters Chain Investigator II equipped with a 2420 ELS detector and a 2996 PDA detector; Daicel Chiral Technologies IC chiral column (4.6250 mm, 5 m); eluent: supercritical CO.sub.2/isopropanol/methanol, 80:10:10).

[0096] The optical rotations of the two enantiomers were measured and compare with that obtained for the naturally-occurring optically-pure compound

[].sup.20.sub.D ()-LMA-EA-2801: 45.7
[].sup.20.sub.D (+)-LMA-EA-2801: +40.9 (value consistent with that obtained for the naturally-occurring optically-pure compound of formula Ia)

[0097] The present invention relates to compounds of formula (I) or (Ia) for use as an antibiotic agent, particularly as a broad-spectrum antibiotic agent, and notably against multiresistant Gram-positive and Gram-negative pathogenic bacteria.

[0098] The present invention relates to a pharmaceutical composition containing as active ingredient at least a compound of formula (I) or (Ia) associated with a pharmaceutically acceptable excipient, which may be easily determined based on the general knowledge of persons skilled in the art according to the route of administration selected.