EP3140280A1 - 4-vinyl-2-cyclopenten-1-one, the production thereof, and the use of same as an antibiotic agent - Google Patents

4-vinyl-2-cyclopenten-1-one, the production thereof, and the use of same as an antibiotic agent

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Publication number
EP3140280A1
EP3140280A1 EP15721221.8A EP15721221A EP3140280A1 EP 3140280 A1 EP3140280 A1 EP 3140280A1 EP 15721221 A EP15721221 A EP 15721221A EP 3140280 A1 EP3140280 A1 EP 3140280A1
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EP
European Patent Office
Prior art keywords
compound
formula
antibiotic agent
ppm
hydrogen atom
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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Application number
EP15721221.8A
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German (de)
French (fr)
Inventor
Jamal Ouazzani
Emilie ADELIN
Géraldine LE GOFF
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Centre National de la Recherche Scientifique CNRS
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Centre National de la Recherche Scientifique CNRS
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Publication of EP3140280A1 publication Critical patent/EP3140280A1/en
Withdrawn legal-status Critical Current

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C69/00—Esters of carboxylic acids; Esters of carbonic or haloformic acids
    • C07C69/66—Esters of carboxylic acids having esterified carboxylic groups bound to acyclic carbon atoms and having any of the groups OH, O—metal, —CHO, keto, ether, acyloxy, groups, groups, or in the acid moiety
    • C07C69/73—Esters of carboxylic acids having esterified carboxylic groups bound to acyclic carbon atoms and having any of the groups OH, O—metal, —CHO, keto, ether, acyloxy, groups, groups, or in the acid moiety of unsaturated acids
    • C07C69/738—Esters of keto-carboxylic acids or aldehydo-carboxylic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04—Antibacterial agents
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C45/00—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds
    • C07C45/56—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds from heterocyclic compounds
    • C07C45/57—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds from heterocyclic compounds with oxygen as the only heteroatom
    • C07C45/59—Preparation of compounds having >C = O groups bound only to carbon or hydrogen atoms; Preparation of chelates of such compounds from heterocyclic compounds with oxygen as the only heteroatom in five-membered rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C67/00—Preparation of carboxylic acid esters
    • C07C67/475—Preparation of carboxylic acid esters by splitting of carbon-to-carbon bonds and redistribution, e.g. disproportionation or migration of groups between different molecules
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P5/00—Preparation of hydrocarbons or halogenated hydrocarbons
    • C12P5/007—Preparation of hydrocarbons or halogenated hydrocarbons containing one or more isoprene units, i.e. terpenes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/24—Preparation of oxygen-containing organic compounds containing a carbonyl group
    • C12P7/26—Ketones
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P7/00—Preparation of oxygen-containing organic compounds
    • C12P7/62—Carboxylic acid esters
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07C—ACYCLIC OR CARBOCYCLIC COMPOUNDS
    • C07C2601/00—Systems containing only non-condensed rings
    • C07C2601/06—Systems containing only non-condensed rings with a five-membered ring
    • C07C2601/10—Systems containing only non-condensed rings with a five-membered ring the ring being unsaturated

Definitions

  • the present invention relates to novel 4-vinyl-2-cyclopentene-1-one derivatives, their preparation and their use as an antibiotic agent.
  • Infectious diseases are the leading cause of morbidity and mortality worldwide. Infections involving germs that are resistant to current treatments pose a major public health problem and lead to an exorbitant excess cost of treatment estimated at 62 billion euros. In Europe, 6.2% of patients who spend more than two days in intensive care contract lung infections and 3.0% of blood infections. 70% of FDA (Food and Drug Administration) -approved antibiotics and more than 80% of known antibiotics are of natural origin. These antibiotics have mainly been isolated from microorganisms (bacteria and filamentous fungi), inexhaustible resources for the discovery of antibiotics and preferentially broad-spectrum, effective against multiple community and nosocomial infections.
  • the present application relates to a new family of antibiotic molecules of general formula (I)
  • R 1 represents a hydrogen atom or a C 1 -C 4 alkyl radical
  • R2 represents a hydroxyl or C 1 -C 4 alkoxyl radical
  • R3 and R independently of one another a hydrogen atom or an alkyl radical -C 4; their enantiomers and enantiomeric mixtures, in particular in racemic form.
  • the compound of formula (Ia) was isolated from a filamentous Trichoderma fungus, particularly the atrovirid species.
  • the Trichoderma LMA strain used has been deposited with the CNCM. This molecule has been isolated and tested under special conditions including:
  • Trichoderma in particular Trichoderma atroviride
  • In-situ capture during culture of natural molecules produced by Trichoderma Natural products, secondary metabolites. This capture of the molecules is carried out thanks to a known protocol but little used in the field and called “solid phase extraction” or SPE,
  • Trichoderma consists of a petri dish on which the microorganism has developed, spores and fragments of mycelium are recovered without distinction by scraping the surface of the agar plate with a sterile liquid, preferably the liquid culture medium.
  • the spores and mycelium are then spread or introduced into a solid or liquid microorganism culture medium, preferably liquid and preferably PDB (Potatoes Dextrose Broth) medium consisting of potato and sugar extracts, the composition of which is for example the following: potato starch 4 g / L, dextrose 20 g / L.
  • PDB Pantatoes Dextrose Broth
  • the fungal strain is cultured at a temperature of between 0 ° C. and 50 ° C., preferably between 20 ° C. and 37 ° C., and preferably at 30 ° C. with rotary stirring or with a turbine driven by a motor between 25 and 25 ° C. 300 rotations per minute, preferably at a speed of 150 rotations per minute.
  • the culture is constantly aerated either by spontaneous aeration by the oxygen contained in the ambient air, or by forced aeration by oxygen injection, for example that included in the compressed air.
  • the culture thus described can be carried out in any solid or liquid microbiological culture device known to those skilled in the art, preferably in Erlenmeyer flasks or fermenters regardless of their size or size.
  • the duration of the culture must correspond to the maximum production of the molecule, it can vary from 1 to 15 days, preferably from 3 to 7 days and preferentially another 5 days.
  • In situ capture during culture, of the natural molecules produced by Trichoderma is carried out thanks to a protocol little used in the field and called "Extraction in situ and in solid phase" or in situ SPE.
  • This protocol consists in introducing an inert solid element during the culture of the microorganism capable of trapping the molecules of interest released by the latter throughout the culture.
  • this protocol involves resins of various natures and preferentially neutral resins such as those described in the following table and preferably apolar resins composed of polystyrene-divinylbenzene copolymers (PS-DVB) of XAD type and preferentially XAD-16 resin.
  • PS-DVB polystyrene-divinylbenzene copolymers
  • XAD polystyrene-divinylbenzene copolymers
  • the resin is then introduced into the culture medium in a sterile manner, either before or after the sterilization thereof.
  • the resin is introduced into the culture medium before sterilization and then sterilized at the same time as the medium.
  • the amounts of resin vary from 1 g / l to 100 g / l, preferably from 10 g / l to 50 g / l and preferably to 30 g / l. Cut
  • the resin is recovered alone or mixed with the biomass by any separation technique between a solid matrix and a liquid container, when the Trichoderma culture lasted the time necessary for the optimal production of the compound of formula (Ia).
  • separation techniques include, without limitation, all the means and techniques of filtration, centrifugation, decantation or drying.
  • the resin alone or mixed with the biomass is recovered by spontaneous or forced filtration under vacuum, preferably on a porous filter regardless of its nature and preferably on an inert fabric.
  • the resin is subsequently treated either after separation from the biomass or mixed with the biomass.
  • the molecules captured by the resin are recovered by bringing it into contact with a solvent whatever its nature, preferably an organic solvent, and preferably dichloromethane or methanol, and several times until the recovery of all the molecules of interest.
  • the solvent having dissolved the molecules of interest is then recovered by filtration and then evaporated by various techniques known to those skilled in the art. This results in a residue that contains among others the molecule of interest of formula (Ia) mixed with other molecules produced by the microorganism, or initially present in the culture medium.
  • the presence of the molecule of interest in this mixture is confirmed by chromatographic or spectrometric analyzes known to those skilled in the art, among which the various chromatography techniques, nuclear magnetic resonance or mass spectrometry.
  • chromatographic or spectrometric analyzes known to those skilled in the art, among which the various chromatography techniques, nuclear magnetic resonance or mass spectrometry.
  • the presence of the compound of formula (Ia) is evidenced by HPLC high performance liquid chromatography coupled with detectors of PDA, DEDL and mass spectrometry type.
  • the target molecule of formula (Ia) is subsequently purified from the total residue by chromatographic techniques known to those skilled in the art, including, but not limited to, chromatography on a silica column or on a reversed phase column. atmospheric pressure or under pressure.
  • the molecule of the compound of formula (Ia) is pre-purified by chromatography on a normal silica column and then purified by preparative HPLC under the following conditions: chromatography on silica in normal phase with a mixture of heptane / ethyl acetate solvent ( 9/1) for 20 min at a flow rate of 30 mL / min, followed by preparative HPLC on a C18 reverse phase column using a linear gradient of water and acetonitrile ranging from 0% to 100% acetonitrile in 10 minutes. minutes followed by a 100% acetonitrile step for 5 minutes at a flow rate of 4 mL / min. After lyophilization, 12 mg of the compound of formula (Ia) were obtained from 8L of culture.
  • the NMR structural analyzes (FIGS. 1 to 5) correlated with data from the HR-ESI-MS (high resolution mass spectrometry) made it possible to determine the following empirical formula CgHi 0 O 4 of molecular mass 182.0574.
  • 13 C NMR ( Figure 2) shows the presence of 9 signals (49.4, 51.7, 77.7, 120.2, 133.9, 149.2, 163.4, 166.4, 205.7 ppm), the proton spectrum ( Figure 1) shows the presence of six groups. protons (2.60, 3.74, 6.17, 6.20, 6.91, 7.33 ppm).
  • the COZY spectrum (FIG. 4) shows correlations between the protons of alkenes showing the existence on the molecule of two ethylenic type unsaturations.
  • the combination of NMR data and information obtained by mass spectrometry confirms that it is a cyclopentenone type molecule hydroxylated in position 4 substituted by a linear chain of methyl acrylate type.
  • the infrared spectrum (FIG. 6) shows absorption bands that reinforce the presence of a ketone, an alcohol and double bonds.
  • MW MicroWave (microwaves)
  • Red-AI® sodium bis (2-methoxyethoxy) aluminumhybrid Reaction scheme for preparing the compound of formula (la)
  • the various compounds of formula (I) may be obtained according to the above reaction scheme by choosing the starting reaction compounds substituted correspondingly to the compounds of formula (I) to be prepared.
  • the groups R 3 and R 4 can be introduced in the first step by choosing the appropriate substituent on the chain of furfuryl alcohol (Microwave-gold Microreactor-Assisted Conversion of Furfuryl Alcohols into 4-Hydroxy-2-cyclopentenones, K. Ulbrich et al., Synlett 2010, No. 13, pp. 2037-2040) or later in the synthetic sequence, in particular to convert the hydroxy group to the corresponding alkoxy OR2.
  • the molecule of the compound of formula (Ia) is redissolved in appropriate solvents and at an appropriate concentration, preferably in dimethyl sulfoxide DMSO at a concentration of 10 mg / ml. This initial solution is then diluted according to the different biological tests to be performed. Two series of tests were carried out.
  • FIG. 7 shows that the antibiotic activity of the compound of formula (Ia) is dependent dose.
  • Figure 7 illustrates the antibiotic activity of the compound of formula (Ia) on various target pathogenic microorganisms in amounts of 1 to 100 g in comparison with chloramphenicol and gentamycin, respectively at 30 g and 10 g. It shows, for example in the case of Escherichia coli ATCC 25922, that the compound (Ia) has a much higher activity than that of chloramphenicol. In fact, the 1 g inhibition of the compound of formula (la) is equivalent to that obtained with 30 g of chloramphenicol.
  • FIG. 7 shows that, in addition to the broad spectrum of sensitive pathogenic bacteria, the molecule of the compound of formula (Ia) is effective at very low doses.
  • TLC analysis The reaction crude obtained, the fractions and the purified products were analyzed by thin layer chromatography (TLC) on gel plates. Silica 60 F 2 54 with a thickness of 0.2 mm on an aluminum support (Merck). The plates are observed under a UV lamp (at 254 and 312 nm) before being revealed by spraying a solution of ammonium molybdate ( ⁇ ) 2 ⁇ 2 ⁇ 7 (100 g / l in 10% sulfuric acid) and heating.
  • the frontal ratio is defined as the ratio between the distance traveled by the compound on the plate and the solvent front.
  • the high resolution mass spectra were performed on a mass spectrometer equipped with an atmospheric pressure ionization source (ESI) and a TOF time-of-flight type mass analyzer (LCT®, Waters).
  • ESI atmospheric pressure ionization source
  • LCDT® TOF time-of-flight type mass analyzer
  • the nuclear magnetic resonance experiments were carried out on Bruker Avance 300 and 500 units using deuterated chloroform CDCl3.
  • the chemical shifts are expressed in ppm (parts per million) and calibrated relative to the reference solvent.
  • the coupling constants are expressed in Hertz (Hz).
  • the multiplicity of signals is expressed by the following abbreviations: s (singlet), bs (wide singlet), d (doublet), dd (doublet split), t (triplet), m (multiplet), q (quadruplet).
  • the rotatory powers of the compounds were measured using a Jasco TM P1010 polarimeter equipped with Spectro Manager software.
  • Source monochromatic luminous is the D-line of sodium.
  • the experiments were carried out with a 100 mm quartz tank of 350 ⁇ , and the products were solubilized in methanol.
  • the infrared (IR) absorption spectra of the described compounds were measured on the Perkin-Elmer Spectrum 100 FT-IR spectrometer. The device is equipped with Spectrum software (version 6.3.5) from Perkin-Elmer. The compounds were prepared in solution in methanol and then dried with compressed air. The absorption bands are given in cm -1 .
  • 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 maintained at -78 ° C. After 30 minutes, the methyl propiolate (277 ⁇ l, 1.0 eq) is added to the previously obtained LDA solution. The reaction is stirred for 1 h at -78 ° C. Compound 3 (660 mg, 3.1 mmol) is solubilized in 5 ml of THF at -78 ° C. and the solution is slowly added to the previously generated lithium acetylide solution. The reaction is stirred at -78 ° C for 1 hour.
  • Compound 7 was analyzed by chiral HPLC equipped with a UV detector in order to highlight the two enantiomers and to be able to separate them. (Alliance Waters 2695 HPLC chain coupled to a Waters 996 PDA detector, chiral IC Daicel Chiral 4.6 x 250 mm (5 ⁇ ) column, eluent n-Heptane / Isopropanol 80:20).
  • the present invention relates to the compounds of formula (I) or (la) for their use as an antibiotic agent in particular as a broad-spectrum antibiotic agent and in particular against gram-positive and gram-negative pathogenic bacteria that are multidrug-resistant.
  • the present invention relates to a pharmaceutical composition containing as active principle at least one compound of formula (I) or (la) associated with a pharmaceutically acceptable excipient, which can be determined easily according to the general knowledge of the skilled person according to the chosen route of administration.

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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 à C4 alkyl radical; and the enantiomers and mixtures of enantiomers thereof, especially in a racemic form.

Description

DERIVES DE 4-VINYL-2CYCLOPENTENE-1 -ONE LEUR PREPARATION ET LEUR UTILISATION EN TANT QU'AGENT ANTIBIOTIQUE  4-VINYL-2CYCLOPENTENE-1 -ONE DERIVATIVES FOR THEIR PREPARATION AND THEIR USE AS ANTIBIOTIC AGENT
La présente invention concerne de nouveaux dérivés de 4-vinyl- 2cyclopentène-1 -one, leur préparation et leur utilisation en tant qu'agent antibiotique. The present invention relates to novel 4-vinyl-2-cyclopentene-1-one derivatives, their preparation and their use as an antibiotic agent.
Les maladies infectieuses représentent la principale cause de morbidité et de mortalité dans le monde. Les infections impliquant des germes résistants aux traitements actuels posent un grand problème de santé publique et engendre un surcoût de traitement exorbitant estimé à 62 milliards d'euros. En Europe, 6,2 % des patients qui passent plus de deux jours en soins intensifs contractent des infections pulmonaires et 3,0 % des infections sanguines. 70 % des antibiotiques approuvés par la FDA (Food and Drug Administration, USA) et plus de 80 % des antibiotiques connus sont d'origine naturelle. Ces antibiotiques ont principalement été isolés à partir de microorganismes (bactéries et champignons filamenteux), ressources inépuisables pour la découverte d'antibiotiques et préférentiellement à large spectre, efficaces contre les infections multiples communautaires et nosocomiales.  Infectious diseases are the leading cause of morbidity and mortality worldwide. Infections involving germs that are resistant to current treatments pose a major public health problem and lead to an exorbitant excess cost of treatment estimated at 62 billion euros. In Europe, 6.2% of patients who spend more than two days in intensive care contract lung infections and 3.0% of blood infections. 70% of FDA (Food and Drug Administration) -approved antibiotics and more than 80% of known antibiotics are of natural origin. These antibiotics have mainly been isolated from microorganisms (bacteria and filamentous fungi), inexhaustible resources for the discovery of antibiotics and preferentially broad-spectrum, effective against multiple community and nosocomial infections.
La présente demande concerne une nouvelle famille de molécules antibiotiques de formule générale (I)  The present application relates to a new family of antibiotic molecules of general formula (I)
dans laquelle :  in which :
Ri représente un atome d'hydrogène ou un radical alkyle en Ci à C4 ; R2 représente un radical hydroxyle ou alkoxyle en Ci à C4, et R 1 represents a hydrogen atom or a C 1 -C 4 alkyl radical; R2 represents a hydroxyl or C 1 -C 4 alkoxyl radical, and
R3 et R représentent indépendamment l'un de l'autre un atome d'hydrogène ou un radical alkyle en Ci à C4 ; leurs énantiomères et mélanges d'énantiomères en particulier sous forme racémique. R3 and R independently of one another a hydrogen atom or an alkyl radical -C 4; their enantiomers and enantiomeric mixtures, in particular in racemic form.
Plus particulièrement la présente demande concerne une famille de molécules antibiotiques de formule générale (la)  More particularly the present application relates to a family of antibiotic molecules of general formula (la)
(la)  (the)
Le composé de formule (la) a été isolé à partir d'un champignon filamenteux Trichoderma, en particulier de l'espèce atroviride. La souche Trichoderma LMA utilisée a fait l'objet d'un dépôt auprès de la CNCM. Cette molécule a été isolée et testée dans des conditions particulières incluant :  The compound of formula (Ia) was isolated from a filamentous Trichoderma fungus, particularly the atrovirid species. The Trichoderma LMA strain used has been deposited with the CNCM. This molecule has been isolated and tested under special conditions including:
1 . La mise en culture de Trichoderma, en particulier de Trichoderma atroviride, 2. La capture in-situ, en cours de culture, des molécules naturelles produites par Trichoderma (produits naturels, métabolites secondaires). Cette capture des molécules est réalisée grâce à un protocole connu mais peu utilisé dans le domaine et appelé « Extraction en phase solide » ou SPE, 1. Cultivation of Trichoderma, in particular Trichoderma atroviride, 2. In-situ capture during culture of natural molecules produced by Trichoderma (natural products, secondary metabolites). This capture of the molecules is carried out thanks to a known protocol but little used in the field and called "solid phase extraction" or SPE,
3. L'isolement et la purification du composé de formule (la) et l'identification de sa structure chimique, 3. The isolation and purification of the compound of formula (Ia) and the identification of its chemical structure,
4. La réalisation des tests biologiques pour mettre en évidence des activités antibiotiques. 4. Performing biological tests to highlight antibiotic activities.
La mise en culture de Trichoderma consiste à partir d'une boîte de pétri sur laquelle le microorganisme s'est développé, les spores et fragments de mycélium sont récupérés sans distinction en grattant la surface de la boîte de gélose avec un liquide stérile, préférentiellement du milieu de culture liquide.  The cultivation of Trichoderma consists of a petri dish on which the microorganism has developed, spores and fragments of mycelium are recovered without distinction by scraping the surface of the agar plate with a sterile liquid, preferably the liquid culture medium.
Les spores et mycélium sont ensuite étalés ou introduits dans un milieu de culture de microorganismes solide ou liquide, préférentiellement liquide et préférentiellement le milieu PDB (Potatoes Dextrose Broth) composé d'extraits de pomme de terre et de sucre, dont la composition est par exemple la suivante : fécule de pomme de terre 4 g/L, dextrose 20 g/L. La souche fongique est cultivée à une température comprise entre 0°C et 50°C, préférentiellement entre 20°C et 37°C, et préférentiellement à 30°C sous agitation soit rotative soit grâce à une turbine entraînée par un moteur entre 25 et 300 rotations par minute, préférentiellement à une vitesse de 150 rotations par minute. La culture est constamment aérée soit par aération spontanée par l'oxygène contenu dans l'air ambiant, soit par aération forcée par injection d'oxygène par exemple celui compris dans l'air comprimé. La culture ainsi décrite peut être réalisée dans n'importe quel dispositif de culture microbiologique solide ou liquide connu de l'homme du métier, préférentiellement dans des Erlenmeyers ou dans des fermenteurs quelles que soient leurs dimensions ou leur tailles. La durée de la culture doit correspondre au maximum de production de la molécule, elle peut varier de 1 à 15 jours, préférentiellement de 3 à 7 jours et préférentiellement encore 5 jours. The spores and mycelium are then spread or introduced into a solid or liquid microorganism culture medium, preferably liquid and preferably PDB (Potatoes Dextrose Broth) medium consisting of potato and sugar extracts, the composition of which is for example the following: potato starch 4 g / L, dextrose 20 g / L. The fungal strain is cultured at a temperature of between 0 ° C. and 50 ° C., preferably between 20 ° C. and 37 ° C., and preferably at 30 ° C. with rotary stirring or with a turbine driven by a motor between 25 and 25 ° C. 300 rotations per minute, preferably at a speed of 150 rotations per minute. The culture is constantly aerated either by spontaneous aeration by the oxygen contained in the ambient air, or by forced aeration by oxygen injection, for example that included in the compressed air. The culture thus described can be carried out in any solid or liquid microbiological culture device known to those skilled in the art, preferably in Erlenmeyer flasks or fermenters regardless of their size or size. The duration of the culture must correspond to the maximum production of the molecule, it can vary from 1 to 15 days, preferably from 3 to 7 days and preferentially another 5 days.
La capture in situ, en cours de culture, des molécules naturelles produites par Trichoderma est réalisée grâce à un protocole peu utilisé dans le domaine et appelé « Extraction in situ et en phase solide » ou in situ SPE. Ce protocole consiste à introduire un élément solide inerte durant la culture du microorganisme capable de piéger les molécules d'intérêt libérées par ce dernier tout au long de la culture. Idéalement, ce protocole implique des résines de natures variées et préférentiellement des résines neutres comme celles décrites dans le tableau suivant et préférentiellement des résines apolaires composées de copolymères polystyrène-divinylbenzène (PS-DVB) de type XAD et préférentiellement la résine XAD-16. La résine est lavée avant utilisation avec une solution aqueuse, et récupérée soit par décantation soit par filtration. La résine est ensuite introduite dans le milieu de culture de manière stérile, soit avant, soit après la stérilisation de celui-ci. Préférentiellement, la résine est introduite dans le milieu de culture avant stérilisation puis stérilisée en même temps que le milieu. Les quantités de résine varient de 1 g/L à 100 g/L, préférentiellement entre 10 g/L et 50 g/L et préférentiellement à 30 g/L. Taille In situ capture, during culture, of the natural molecules produced by Trichoderma is carried out thanks to a protocol little used in the field and called "Extraction in situ and in solid phase" or in situ SPE. This protocol consists in introducing an inert solid element during the culture of the microorganism capable of trapping the molecules of interest released by the latter throughout the culture. Ideally, this protocol involves resins of various natures and preferentially neutral resins such as those described in the following table and preferably apolar resins composed of polystyrene-divinylbenzene copolymers (PS-DVB) of XAD type and preferentially XAD-16 resin. The resin is washed before use with an aqueous solution, and recovered either by decantation or by filtration. The resin is then introduced into the culture medium in a sterile manner, either before or after the sterilization thereof. Preferably, the resin is introduced into the culture medium before sterilization and then sterilized at the same time as the medium. The amounts of resin vary from 1 g / l to 100 g / l, preferably from 10 g / l to 50 g / l and preferably to 30 g / l. Cut
Exemples de Surface Porosité  Examples of Surface Porosity
Matrice particule  Particle matrix
résines (m2/g) (ml/ml) resins (m 2 / g) (ml / ml)
(mm)  (Mm)
Ester  Ester
XAD 7-HP 450 0.43 - 0.69 1 .4  XAD 7-HP 450 0.43 - 0.69 1 .4
acrylique  acrylic
Polystyrène  polystyrene
XAD 4 > 725 0.49 - 0.69 > 0.5  XAD 4> 725 0.49 - 0.69> 0.5
DVB  DVB
Polystyrène  polystyrene
Diaion HP-20 600 > 0.25 1 .3  Diaion HP-20 600> 0.25 1 .3
DVB  DVB
Polystyrène  polystyrene
XAD 16 > 800 0.56 - 0.71 > 0.55  XAD 16> 800 0.56 - 0.71> 0.55
DVB  DVB
Polystyrène  polystyrene
Sepabeads 1000 > 0.25 1 .2  Sepabeads 1000> 0.25 1 .2
DVB  DVB
En ce qui concerne l'isolement et la purification du composé de formule (la) et l'identification de sa structure chimique, la résine est récupérée seule ou mélangée à la biomasse par toute technique de séparation entre une matrice solide et un contenant liquide, lorsque la culture de Trichoderma a duré le temps nécessaire pour la production optimale du composé de formule (la). Ces méthodes connues de l'homme du métier incluent de façon non limitative tous les moyens et techniques de filtration, centrifugation, décantation ou séchage. Préférentiellement, la résine seule ou mélangée à la biomasse est récupérée par filtration spontanée ou forcée sous vide, préférentiellement sur un filtre poreux quelle que soit sa nature et préférentiellement sur un tissu inerte. With regard to the isolation and purification of the compound of formula (Ia) and the identification of its chemical structure, the resin is recovered alone or mixed with the biomass by any separation technique between a solid matrix and a liquid container, when the Trichoderma culture lasted the time necessary for the optimal production of the compound of formula (Ia). These methods known to those skilled in the art include, without limitation, all the means and techniques of filtration, centrifugation, decantation or drying. Preferably, the resin alone or mixed with the biomass is recovered by spontaneous or forced filtration under vacuum, preferably on a porous filter regardless of its nature and preferably on an inert fabric.
La résine est traitée par la suite soit après sa séparation de la biomasse, soit en mélange avec la biomasse. Les molécules captées par la résine sont récupérées par mise en contact de celle-ci avec un solvant quelle que soit sa nature, de préférence un solvant organique, et de préférence le dichlorométhane ou le méthanol, et ce plusieurs fois jusqu'à la récupération de la totalité des molécules d'intérêt. Le solvant ayant dissous les molécules d'intérêt, est ensuite récupéré par filtration puis évaporé par différentes techniques connues de l'homme du métier. Il en résulte un résidu qui contient entre autres la molécule d'intérêt de formule (la) mélangée à d'autres molécules produites par le microorganisme, ou présentes initialement dans le milieu de culture. La présence de la molécule d'intérêt dans ce mélange est confirmée par des analyses chromatographiques ou spectrométriques connues de l'homme de l'art, parmi lesquelles les différentes techniques de chromatographie, la résonnance magnétique nucléaire ou la spectrométrie de masse. De préférence, la présence du composé de formule (la) est mise en évidence grâce à la chromatographie liquide haute performance HPLC couplée à des détecteurs de type PDA, DEDL et spectrométrie de masse. The resin is subsequently treated either after separation from the biomass or mixed with the biomass. The molecules captured by the resin are recovered by bringing it into contact with a solvent whatever its nature, preferably an organic solvent, and preferably dichloromethane or methanol, and several times until the recovery of all the molecules of interest. The solvent having dissolved the molecules of interest, is then recovered by filtration and then evaporated by various techniques known to those skilled in the art. This results in a residue that contains among others the molecule of interest of formula (Ia) mixed with other molecules produced by the microorganism, or initially present in the culture medium. The presence of the molecule of interest in this mixture is confirmed by chromatographic or spectrometric analyzes known to those skilled in the art, among which the various chromatography techniques, nuclear magnetic resonance or mass spectrometry. Preferably, the presence of the compound of formula (Ia) is evidenced by HPLC high performance liquid chromatography coupled with detectors of PDA, DEDL and mass spectrometry type.
La molécule cible de formule (la) est par la suite purifiée à partir du résidu total par des techniques chromatographiques connues de l'homme du métier, incluant de façon non limitative, la chromatographie sur colonne de silice ou sur colonne en phase inverse, à pression atmosphérique ou sous pression. Préférentiellement la molécule du composé de formule (la) est pré- purifiée par chromatographie sur une colonne de silice normale puis purifiée par HPLC préparative dans les conditions suivantes : chromatographie sur silice en phase normale par un mélange de solvant heptane / acétate d'éthyle (9 / 1 ) pendant 20 min à un débit de 30 mL/min, puis HPLC préparative sur une colonne en phase inverse C18 en utilisant un gradient linéaire d'eau et d'acétonitrile allant de 0 % à 100 % d'acétonitrile en 10 minutes suivi d'un palier à 100 % d'acétonitrile pendant 5 minutes à un débit de 4 mL/min. Après lyophilisation, 12 mg du composé de formule (la) ont été obtenus à partir de 8L de culture.  The target molecule of formula (Ia) is subsequently purified from the total residue by chromatographic techniques known to those skilled in the art, including, but not limited to, chromatography on a silica column or on a reversed phase column. atmospheric pressure or under pressure. Preferably, the molecule of the compound of formula (Ia) is pre-purified by chromatography on a normal silica column and then purified by preparative HPLC under the following conditions: chromatography on silica in normal phase with a mixture of heptane / ethyl acetate solvent ( 9/1) for 20 min at a flow rate of 30 mL / min, followed by preparative HPLC on a C18 reverse phase column using a linear gradient of water and acetonitrile ranging from 0% to 100% acetonitrile in 10 minutes. minutes followed by a 100% acetonitrile step for 5 minutes at a flow rate of 4 mL / min. After lyophilization, 12 mg of the compound of formula (Ia) were obtained from 8L of culture.
La structure de la molécule du composé de formule (la) a été réalisée par des méthodes spectroscopiques. Les spectres et autres données analytiques sont rapportés dans les figures annexées.  The structure of the molecule of the compound of formula (Ia) was carried out by spectroscopic methods. Spectra and other analytical data are reported in the accompanying figures.
Les analyses structurales RMN (figures 1 à 5) corrélées aux données issues de l'HR-ESI-MS (spectrométrie de masse haute résolution) ont permis de déterminer la formule brute suivante CgHi0O4 de masse moléculaire 182.0574. La RMN 13C (figure 2) montre la présence de 9 signaux (49.4 ; 51 .7 ; 77.7 ; 120.2 ; 133.9 ; 149.2 ; 163.4 ; 166.4 ; 205.7 ppm), le spectre proton (figure 1 ) montre la présence de six groupes de protons (2.60 ; 3.74 ; 6.17 ; 6.20 ; 6.91 ; 7.33 ppm). The NMR structural analyzes (FIGS. 1 to 5) correlated with data from the HR-ESI-MS (high resolution mass spectrometry) made it possible to determine the following empirical formula CgHi 0 O 4 of molecular mass 182.0574. 13 C NMR (Figure 2) shows the presence of 9 signals (49.4, 51.7, 77.7, 120.2, 133.9, 149.2, 163.4, 166.4, 205.7 ppm), the proton spectrum (Figure 1) shows the presence of six groups. protons (2.60, 3.74, 6.17, 6.20, 6.91, 7.33 ppm).
L'analyse du spectre HSQC (figure 3) corrélé au spectre 1 D 1 H et 13C montre la présence d'un groupement méthoxy à 51 .7 ppm, un groupe méthylène à 49.4 ppm et quatre groupements alcènes à 120.2 ; 133.9 ; 149.2 et 163.4 ppm. L'absence de tâches de corrélations de trois carbones permet de conclure à la présence de trois carbones quaternaires : un de type cétone à 205.4 ppm ; un de type ester ou lactone à 166.4 ppm et un de type alcool tertiaire à 77.7 ppm. Analysis of the HSQC spectrum (FIG. 3) correlated with the 1 D 1 H and 13 C spectrum shows the presence of a methoxy group at 51.7 ppm, a methylene group at 49.4 ppm and four alkene groups at 120.2; 133.9; 149.2 and 163.4 ppm. The absence of correlation tasks of three carbons makes it possible to conclude to the presence of three quaternary carbons: a ketone type at 205.4 ppm; one of the ester or lactone type at 166.4 ppm and one of the tertiary alcohol type at 77.7 ppm.
Le spectre COSY (figure 4) montre des corrélations entre les protons d'alcènes montrant l'existence sur la molécule de deux insaturations de type éthyléniques.  The COZY spectrum (FIG. 4) shows correlations between the protons of alkenes showing the existence on the molecule of two ethylenic type unsaturations.
La structure de la molécule du composé de formule (la) est confirmée par les corrélations du spectre HMBC (figure 5) et ses interactions scalaires longues distances résumées dans le tableau ci-dessous.  The structure of the molecule of the compound of formula (Ia) is confirmed by the correlations of the HMBC spectrum (FIG. 5) and its long-range scalar interactions summarized in the table below.
La combinaison des données RMN et des informations obtenues par spectrométrie de masse confirme qu'il s'agit d'une molécule de type cyclopentènone hydroxylée en position 4 substituée par une chaîne linéaire de type acrylate de méthyle. Le spectre d'absorption UV montre un maximum à λ max = 257 nm. Le spectre infrarouge (figure 6) montre des bandes d'absorption qui confortent la présence d'une cétone, d'un alcool et des doubles liaisons. Le signe du pouvoir rotatoire mesuré pour la molécule du composé de formule (la) [a]D 20 = +42,6 (20 mg/10mL CH2CI2) a été comparé à celui de molécules ressemblantes comme la trichodénone A naturelle [OI] D20 = +56 qui possède un pouvoir rotatoire positif et une stéréochimie R confirmée par synthèse totale énantioséléctive (Tetrahedron asymetry 2000, 1 1 (371 1 -3725). The combination of NMR data and information obtained by mass spectrometry confirms that it is a cyclopentenone type molecule hydroxylated in position 4 substituted by a linear chain of methyl acrylate type. The UV absorption spectrum shows a maximum at λ max = 257 nm. The infrared spectrum (FIG. 6) shows absorption bands that reinforce the presence of a ketone, an alcohol and double bonds. The sign of the rotatory power measured for the molecule of the compound of formula (la) [a] D 20 = +42.6 (20 mg / 10 mL CH 2 CI 2 ) was compared with that of similar molecules such as natural trichodenone A [ OI] D 20 = +56 which possesses a positive rotatory power and a stereochemistry R confirmed by total enantioselective synthesis (Tetrahedron asymetry 2000, 1 1 (371 1 -3725).
Les composés de formules (I) et (la) peuvent également être synthétisés conformément au schéma réactionnel mentionné ci-après.  The compounds of formulas (I) and (Ia) can also be synthesized according to the reaction scheme mentioned below.
MW : MicroWave (microondes) MW: MicroWave (microwaves)
LDA : Lithium Diisopropylamide LDA: Lithium Diisopropylamide
n-BuLi : n-ButylLithium n-BuLi: n-ButylLithium
THF : Tetrahydrofurane THF: Tetrahydrofuran
RT : Room Température (température ambiante)  RT: Room Temperature (room temperature)
PCC : Pyridinium ChloroChromate  PCC: Pyridinium ChloroChromate
Red-AI® : sodium bis(2-methoxyethoxy)aluminumhybride Schéma réactionnel de préparation du composé de formule (la) Red-AI®: sodium bis (2-methoxyethoxy) aluminumhybrid Reaction scheme for preparing the compound of formula (la)
Les différents composés de formule (I) pourront être obtenus conformément au schéma réactionnel ci-dessus en choisissant les composés réactionnels de départ substitués de façon correspondante aux composés de formule (I) à préparer. En particulier, les groupements R3 et R4 peuvent être introduits à la première étape en choisissant le substituant approprié sur la chaîne du furfuryl alcool (Microwave- or Microreactor- Assisted Conversion of furfuryl Alcohols into 4-Hydroxy-2-cyclopentenones, K. Ulbrich et al., Synlett 2010, N° 13, pp 2037-2040) ou ultérieurement dans la séquence synthétique, en particulier pour transformer le radical hydroxy en alcoxy correspondant OR2. The various compounds of formula (I) may be obtained according to the above reaction scheme by choosing the starting reaction compounds substituted correspondingly to the compounds of formula (I) to be prepared. In particular, the groups R 3 and R 4 can be introduced in the first step by choosing the appropriate substituent on the chain of furfuryl alcohol (Microwave-gold Microreactor-Assisted Conversion of Furfuryl Alcohols into 4-Hydroxy-2-cyclopentenones, K. Ulbrich et al., Synlett 2010, No. 13, pp. 2037-2040) or later in the synthetic sequence, in particular to convert the hydroxy group to the corresponding alkoxy OR2.
Une fois purifiée, la molécule du composé de formule (la) est redissoute dans des solvants appropriés et à une concentration appopriée, préférentiellement dans du diméthyl-sulfoxyde DMSO à la concentration de 10 mg/nnl. Cette solution initiale est ensuite diluée en fonction des différents tests biologiques à réaliser. Deux séries de tests ont été réalisées.  Once purified, the molecule of the compound of formula (Ia) is redissolved in appropriate solvents and at an appropriate concentration, preferably in dimethyl sulfoxide DMSO at a concentration of 10 mg / ml. This initial solution is then diluted according to the different biological tests to be performed. Two series of tests were carried out.
Des tests de cytoxicité sur une lignée de cellules cancéreuses KB (carcinome) et une lignée de cellules équivalentes non-cancéreuses MRC5. Le tableau qui suit rapporte le pourcentage d'inhibition de la croissance par la molécule du composé de formule (la) aux concentrations de 1 et 10 micromolaires. Il se dégage de ce résultat qu'aux concentrations utilisées aucune cytotoxicité n'est observée, ni vis-à-vis de cellules tumorales, ni vis-à- vis de cellules saines.  Cytoxicity tests on a KB cancer cell line (carcinoma) and a non-cancerous equivalent cell line MRC5. The following table reports the percent inhibition of growth by the molecule of the compound of formula (Ia) at concentrations of 1 and 10 micromolar. It emerges from this result that at the concentrations used, no cytotoxicity is observed, either with respect to tumor cells or towards healthy cells.
La molécule du composé de formule (la) a été engagée dans des tests antibiotiques selon la technique des zones d'inhibition, sur différentes bactéries Gram positives et Gram négatives, à la quantité de 100 g. En parallèle, des tests similaires sont réalisés avec des antibiotiques témoins avec des quantités variables selon l'antibiotique soit 10 g, soit 30 g. Zone d'inhibition Zone d'inhibition The molecule of the compound of formula (Ia) was engaged in antibiotic tests according to the inhibition zone technique, on different Gram-positive and Gram-negative bacteria, in the amount of 100 g. In parallel, similar tests are carried out with control antibiotics with varying amounts depending on the antibiotic either 10 g or 30 g. Inhibition zone Inhibition zone
Antibiotique de  Antibiotic
Microorganisme pour 100 g du par les référence et  Microorganism for 100 g of by reference and
cible composé de la antibiotiques de quantité déposée target compound of antibiotics deposited amount
formule (la) référence (cm) formula (the) reference (cm)
Escherichia coli Chloramphénicol Escherichia coli Chloramphenicol
2,7 1 ,0  2.7 1, 0
(ATCC 25922) (30Mg) (ATCC 25922) (30 M g)
Klebsiella  Klebsiella
Chloramphénicol  chloramphenicol
pneumoniae 2,9 2,5 pneumoniae 2.9 2.5
(30Mg) (30 M g)
(ATCC 1001) (ATCC 1001)
Enterobacter  Enterobacter
Chloramphénicol  chloramphenicol
cloacae 1 ,2 1 ,5 cloacae 1, 2 1, 5
(30Mg) (30 M g)
(ATCC 13047) (ATCC 13047)
Micrococcus  Micrococcus
Chloramphénicol  chloramphenicol
luteus 3,2 3,2 Luteus 3.2 3.2
(30Mg) (30 M g)
(ATCC 10240)  (ATCC 10240)
Bacillus subtillis Gentamycine  Bacillus subtillis Gentamycin
2,7 1 ,2  2.7 1, 2
(ATCC 6633) (10Mg) (ATCC 6633) (10 g M)
Streptococcus  Streptococcus
pneumoniae 2,0 Pénicilline (30 g) 3,5 pneumoniae 2.0 Penicillin (30 g) 3.5
(ATCC 49619) (ATCC 49619)
Acinetobacter  Acinetobacter
Gentamycine  gentamicin
baumanii 3,2 0,9 Baumanii 3,2 0.9
(10Mg) (10 M g)
(ATCC 19606) (ATCC 19606)
Pseudomonas  Pseudomonas
Gentamycine  gentamicin
aeruginosa - 1 ,3 aeruginosa - 1, 3
(10Mg) (10 M g)
(ATCC 27853) (ATCC 27853)
Staphylococcus  Staphylococcus
Gentamycine  gentamicin
aureus 3,0 1 ,2 aureus 3.0 1, 2
(10Mg) (10 M g)
(ATCC 25923) La figure 7 annexée montre que l'activité antibiotique du composé de formule (la) est dose dépendante. La Figure 7 illustre l'activité antibiotique du composé de formule (la) sur différents microorganismes pathogènes cibles à des quantités de 1 à 100 g en comparaison avec le chloramphénicol et la gentamycine, respectivement à 30 g et à 10 g. Elle montre par exemple dans le cas à'Escherichia coli ATCC 25922, que le composé (la) présente une activité bien supérieure à celle du chloramphénicol. En effet, l'inhibition par 1 g du composé de formule (la) équivaut à celle obtenue avec 30 g de chloramphénicol. Les mêmes observations et conclusions peuvent être faites pour toutes les souches sensibles au composé de formule (la) comme le montre la figure 7. Ceci montre qu'en plus du large spectre de bactéries pathogènes sensibles, la molécule du composé de formule (la) est efficace à très faibles doses. (ATCC 25923) The attached FIG. 7 shows that the antibiotic activity of the compound of formula (Ia) is dependent dose. Figure 7 illustrates the antibiotic activity of the compound of formula (Ia) on various target pathogenic microorganisms in amounts of 1 to 100 g in comparison with chloramphenicol and gentamycin, respectively at 30 g and 10 g. It shows, for example in the case of Escherichia coli ATCC 25922, that the compound (Ia) has a much higher activity than that of chloramphenicol. In fact, the 1 g inhibition of the compound of formula (la) is equivalent to that obtained with 30 g of chloramphenicol. The same observations and conclusions can be made for all the strains sensitive to the compound of formula (Ia) as shown in FIG. 7. This shows that, in addition to the broad spectrum of sensitive pathogenic bacteria, the molecule of the compound of formula (Ia) is effective at very low doses.
On mentionne ci-après à titre d'illustration un exemple de préparation du composé de formule la.  An illustration of an example of preparation of the compound of formula la is given below by way of illustration.
Exemple 1 : 1 .1 . Procédure générale Example 1: 1 .1. General procedure
Sauf mention contraire, toutes les réactions ont été réalisées sous une atmosphère inerte d'argon, dans de la verrerie préalablement séchée à l'étuve à 120 °C. Unless otherwise stated, all the reactions were carried out under an inert atmosphere of argon, in glassware previously dried in an oven at 120 ° C.
1 .1 .1 . Solvants Le tétrahydrofurane (THF) a été distillé sous argon sur sodium/benzophénone. Le dichlorométhane (CH2CI2) a été distillé sous argon sur hydride de calcium (CaH2). Sauf indication contraire, tous les réactifs engagés sont des réactifs commerciaux de qualité « reagent-grade ». 1 .1 .1. Solvents Tetrahydrofuran (THF) was distilled under argon on sodium / benzophenone. The dichloromethane (CH 2 Cl 2 ) was distilled under argon on calcium hydride (CaH 2 ). Unless otherwise stated, all reagents involved are reagent-grade commercial reagents.
1 .1 .2. Analyse par CCM Les bruts réactionnels obtenus, les fractions et les produits purifiés ont été analysés par chromatographie sur couche mince (CCM) sur plaques de gel de silice 60 F254 de 0.2 mm d'épaisseur sur support d'aluminium (Merck). Les plaques sont observées sous lampe UV (à 254 et 312 nm) avant d'être révélées par pulvérisation d'une solution d'ammonium molybdate (ΝΗ )2Μθ2θ7 (100g/L dans 10% acide sulfurique) et chauffage. Le rapport frontal est défini comme étant le rapport entre la distance parcourue par le composé sur la plaque et le front de solvant. 1 .1 .2. TLC analysis The reaction crude obtained, the fractions and the purified products were analyzed by thin layer chromatography (TLC) on gel plates. Silica 60 F 2 54 with a thickness of 0.2 mm on an aluminum support (Merck). The plates are observed under a UV lamp (at 254 and 312 nm) before being revealed by spraying a solution of ammonium molybdate (ΝΗ) 2 Μθ2θ 7 (100 g / l in 10% sulfuric acid) and heating. The frontal ratio is defined as the ratio between the distance traveled by the compound on the plate and the solvent front.
1 .1 .3. Purification  1 .1 .3. Purification
La purification des mélanges réactionnels a été réalisée par chromatographie flash sur gel de silice (Merck 60 (35-70 μιτι)). 1 .1 .4. Analyses spectroscopiques The purification of the reaction mixtures was carried out by flash chromatography on silica gel (Merck 60 (35-70 μιτι)). 1 .1 .4. Spectroscopic analyzes
1 .1 .4.1 Spectrométrie de Masse Haute Résolution 1 .1 .4.1 High Resolution Mass Spectrometry
Les spectres de masse haute résolution ont été réalisés sur un spectromètre de masse équipé d'une source d'ionisation à pression atmosphérique (ESI) et d'un analyseur de masse de type temps de vol TOF (LCT®, Waters). The high resolution mass spectra were performed on a mass spectrometer equipped with an atmospheric pressure ionization source (ESI) and a TOF time-of-flight type mass analyzer (LCT®, Waters).
1 .1 .4.2 Résonance magnétique nucléaire (RMN) 1 .1 .4.2 Nuclear Magnetic Resonance (NMR)
Les expériences de résonance magnétique nucléaire ont été réalisées sur des appareils Bruker Avance 300 et 500 en utilisant le chloroforme deutérié CDCI3, Les déplacements chimiques sont exprimés en ppm (partie par million) et calibrés par rapport au solvant de référence. Les constantes de couplage sont exprimées en Hertz (Hz). La multiplicité des signaux est exprimée par les abréviations suivantes : s (singulet), bs (singulet large), d (doublet), dd (doublet dédoublé), t (triplet), m (multiplet), q (quadruplet). L'attribution des signaux des protons et des carbones a été effectuée à partir des expériences monodimensionnelles 1 D 1 H et 13C, et bidimensionnelles 2D 1 H-1 H COSY, 1 H- 13C HSQC ou HMQC, 1 H-13C HMBC, 1 H-1 H NOESY. Les spectres RMN sont traités grâce aux logiciels dédiés TopSpin (Brucker). The nuclear magnetic resonance experiments were carried out on Bruker Avance 300 and 500 units using deuterated chloroform CDCl3. The chemical shifts are expressed in ppm (parts per million) and calibrated relative to the reference solvent. The coupling constants are expressed in Hertz (Hz). The multiplicity of signals is expressed by the following abbreviations: s (singlet), bs (wide singlet), d (doublet), dd (doublet split), t (triplet), m (multiplet), q (quadruplet). The assignment of the protons and carbons signals was performed from the one-dimensional experiments 1 D 1 H and 13 C, and two-dimensional 2D 1 H- 1 H COZY, 1 H- 13 C HSQC or HMQC, 1 H- 13 C HMBC, 1 H- 1 H NOESY. NMR spectra are processed using TopSpin dedicated software (Brucker).
1 .1 .4.3 Pouvoir rotatoire 1 .1 .4.3 Rotatory power
Les pouvoirs rotatoires des composés ont été mesurés à l'aide d'un polarimètre Jasco™ P1010, équipé du logiciel Spectro Manager. La source lumineuse monochromatique est la raie D de sodium. Les expériences ont été menées avec une cuve en quartz 100 mm de 350 μί, et les produits ont été solubilisés dans le méthanol. The rotatory powers of the compounds were measured using a Jasco ™ P1010 polarimeter equipped with Spectro Manager software. Source monochromatic luminous is the D-line of sodium. The experiments were carried out with a 100 mm quartz tank of 350 μί, and the products were solubilized in methanol.
1 .1 .4.4 Spectroscopie infrarouge 1 .1 .4.4 Infrared spectroscopy
Les spectres d'absorption infrarouge (IR) des composés décrits ont été mesurés sur le spectromètre Perkin-Elmer Spectrum 100 FT-IR. L'appareil est équipé du logiciel Spectrum (version 6.3.5) de Perkin-Elmer. Les composés ont été préparés en solution dans le méthanol puis séchés à l'air comprimé. Les bandes d'absorption sont données en cm"1. The infrared (IR) absorption spectra of the described compounds were measured on the Perkin-Elmer Spectrum 100 FT-IR spectrometer. The device is equipped with Spectrum software (version 6.3.5) from Perkin-Elmer. The compounds were prepared in solution in methanol and then dried with compressed air. The absorption bands are given in cm -1 .
1 .2. Synthèse des composés 2 à 7 1 .2. Synthesis of compounds 2 to 7
Composé 2 : 4-hydroxycyclopent-2-enone  Compound 2: 4-hydroxycyclopent-2-enone
C5H6O2 C 5 H 6 O 2
98,1 g.mol"1 98.1 g.mol "1
Une solution d'alcool furfurylique (1 ) (3.0 ml, 34.7 mmol) dans de l'eau distillée (5 ml) est placée dans un tube pour micro-onde (CEM Discover) et chauffé en vase clos à 300 W pendant 10 minutes. Après refroidissement et décantation, la phase aqueuse est récupérée et le résidu lavé par 2 X 5 ml d'eau distillée. Les phases aqueuses sont rassemblées et lavées par extraction liquide-liquide à l'acétate d'éthyle (2 X 50 ml). La phase aqueuse est ensuite concentrée pour donner 668 mg (20%) du composé 2 sous forme d'une huile rouge orangée. Le produit obtenu ne nécessite pas de purification avant d'être engagé dans la réaction suivante (K. Ulbrich, P.Kreitmeier, O. Reeiser, Synlett. 2010, 13, 2037-2040) Rf = 0.32 (Heptane/AcOEt, 2:8) 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 vacuum at 300 W for 10 minutes. . After cooling and decantation, the aqueous phase is recovered and the residue washed with 2 × 5 ml of distilled water. The aqueous phases are combined and washed by liquid-liquid extraction with ethyl acetate (2 × 50 ml). The aqueous phase is then concentrated to give 668 mg (20%) of compound 2 as an orange-red oil. The product obtained does not require purification before being engaged in the next reaction (K. Ulbrich, P. Kreitmeier, O. Reeiser, Synlett, 2010, 13, 2037-2040) Rf = 0.32 (Heptane / AcOEt, 2: 8)
RMN 1H (300 MHz, CDCI3) : δΗ (ppm) 7.57 (dd, 1 H, J = 2.3 Hz, J = 5.7 Hz), 6.22 (dd, 1 H, J = 1 .1 Hz, J = 5.7 Hz), 5.06-5.02 (m, 1 H), 2.77 (dd, 1 H, J = 6.1 Hz, J = 18.5 Hz), 2.27 (dd, 1 H, J = 2.1 Hz, J = 18.5 Hz). 1 H NMR (300 MHz, CDCl 3 ): δ Η (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 = 6.1Hz, J = 18.5Hz), 2.27 (dd, 1H, J = 2.1Hz, J = 18.5Hz).
RMN 13C (75 MHz, CDCI3) : 5C (ppm) 207.1 , 163.7, 135.0, 70.3, 44.2. 13 C NMR (75 MHz, CDCI 3) 5 C (ppm) 207.1, 163.7, 135.0, 70.3, 44.2.
HRMS (ESI) : m/z calculée pour C5H7O2 [M+H+] 99.0446 HRMS (ESI): m / z calcd for C 5 H 7 O 2 [M + H + ] 99.0446
valeur expérimentale 99.0449  experimental value 99.0449
Composé 3 : 4-((tert-butyldimethylsilyl)oxy)cyclopent-2-enone Compound 3: 4 - ((tert-butyldimethylsilyl) oxy) cyclopent-2-enone
212.1 g. mol"1 212.1 g. mol "1
Le composé 2 (334 mg, 3.41 mmol) est solubilisé dans le dichlorométhane (6 ml). Le milieu réactionnel est ensuite maintenu à 0°C. La triéthylamine (700 μΙ, 4.78 mmol), le DMAP (33 mg, 0.27 mmol) et le chlorure de te/Î-butyldiméthylsilyl (1 .0M dans le THF, 3.75 ml) sont ensuite additionnés. La réaction est maintenue à 0°C sous agitation pendant 12 heures. Après addition d'eau distillée (10 ml), le milieu est extrait au dichlorométhane (3 X 20 ml). La phase organique est séchée sur du sulfate de sodium anhydre, puis évaporée à sec. Le brut réactionnel obtenu est purifié par chromatographie flash sur gel de silice (éluant Heptane/AcOEt, 80:20) pour donner 664 mg (92%) du composé 3 sous forme d'une huile transparente (cristaux blanc à 4°C) (K. Ulbrich, P.Kreitmeier, O. Reeiser, Synlett. 2010, 13, 2037-2040). Rf = 0.53 (Heptane/AcOEt, 3:7) Compound 2 (334 mg, 3.41 mmol) is solubilized in dichloromethane (6 ml). The reaction medium is then maintained at 0 ° C. Triethylamine (700 μl, 4.78 mmol), DMAP (33 mg, 0.27 mmol) and tert-butyldimethylsilyl chloride (1 M in THF, 3.75 ml) are then added. The reaction is maintained at 0 ° C with stirring for 12 hours. After addition of distilled water (10 ml), the medium is extracted with dichloromethane (3 × 20 ml). The organic phase is dried over anhydrous sodium sulfate and then evaporated to dryness. The crude reaction product obtained is purified by flash chromatography on silica gel (Heptane / AcOEt eluent, 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. Reeiser, Synlett, 2010, 13, 2037-2040). Rf = 0.53 (Heptane / AcOEt, 3: 7)
RMN 1H (300 MHz, CDCI3) : δΗ (ppm) 7.46 (dd, 1 H, J = 2.3 Hz, J = 5.7 Hz), 6.19 (dd, 1 H, J = 1 .1 Hz, J = 5.7 Hz), 5.01 -4.98 (m, 1 H), 2.71 (dd, 1 H, J = 6.1 Hz, J = 18.5 Hz), 2.25 (dd, 1 H, J = 2.2 Hz, J = 18.5 Hz), 0.91 (s, 9H), 0.14 (s, 3H), 0.13 (s, 3H). 1 H NMR (300 MHz, CDCl 3 ): δ Η (ppm) 7.46 (dd, 1H, J = 2.3Hz, J = 5.7Hz), 6.19 (dd, 1H, J = 1.1H, J = 5.7 Hz), 5.01 -4.98 (m, 1 H), 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).
RMN 13C (75 MHz, CDCI3) : 5C (ppm) 206.5, 163.8, 134.4, 70.9, 45.0, 25.7, 18.1 , -4.8 13 C NMR (75 MHz, CDCl 3 ): δ C (ppm) 206.5, 163.8, 134.4, 70.9, 45.0, 25.7, 18.1, -4.8
HRMS (ESI) : m/z calculée pour C22H4iO4Si2 [2M+H+] 425.2583 HRMS (ESI): m / z calcd for C 22 H 4 iO 4 Si 2 [2M + H + ] 425.2583
valeur expérimentale 425.2601  experimental value 425.2601
Composé 4 : methyl 3-(4-((tert-butyldimethylsilyl)oxy)-1 - hydroxycyclopent-2-en-1 -yl)propiolate Compound 4: methyl 3- (4 - ((tert-butyldimethylsilyl) oxy) -1-hydroxycyclopent-2-en-1-yl) propiolate
TBDMS  TBDMS
296.1 g. mol" 296.1 g. mol "
Le n-butyl lithium (1 ,6 M dans l'hexane, 2.14 ml, 1 .1 éq) est ajouté à une solution de diisopropylamine (529 μΙ, 1 .2 éq) dans 10 ml de THF et maintenue à -78°C.A bout de 30 minutes, le methyl propiolate (277 μΙ, 1 .0 éq) est ajouté à la solution de LDA préalablement obtenue. La réaction est maintenue sous agitation pendant 1 h à -78°C. Le composé 3 (660 mg, 3.1 1 mmol) est solubilisé dans 5 ml de THF à -78°C puis la solution est lentement additionnée à la solution d'acétylide de lithium préalablement générée. La réaction est maintenue sous agitation à -78°C pendant 1 heure. Après addition de 20 ml d'une solution saturée de NH CI, le mélange est extrait par 3 x 50 ml d'acétate d'éthyle. La phase organique est séchée sur du sulfate de sodium anhydre, puis évaporée à sec. Le brut réactionnel obtenu est purifié par chromatographie flash sur gel de silice (éluant Heptane/AcOEt, 80 :20) pour donner 718 mg (78%) du composé 4 sous forme d'une huile translucide jaune. N-Butyl lithium (1.6 M in hexane, 2.14 ml, 1.1 eq) is added to a solution of diisopropylamine (529 μl, 1.2 eq) in 10 ml of THF and maintained at -78 ° C. After 30 minutes, the methyl propiolate (277 μl, 1.0 eq) is added to the previously obtained LDA solution. The reaction is stirred for 1 h at -78 ° C. Compound 3 (660 mg, 3.1 mmol) is solubilized in 5 ml of THF at -78 ° C. and the solution is slowly added to the previously generated lithium acetylide solution. The reaction is stirred at -78 ° C for 1 hour. After addition of 20 ml of a saturated solution of NH Cl, the mixture is extracted with 3 x 50 ml of ethyl acetate. The organic phase is dried over anhydrous sodium sulfate and then evaporated to dryness. The crude reaction product obtained is purified by flash chromatography on silica gel (Heptane / AcOEt eluent, 80:20) to give 718 mg (78%) of compound 4 in the form of a yellow translucent oil.
Rf = 0.44 (Heptane/AcOEt, 6:4) Rf = 0.44 (Heptane / AcOEt, 6: 4)
RMN 1H (300 MHz, CDCI3) : δΗ (ppm) 5.99 (dd, 1 H, J = 1 .7 Hz, J = 5.4 Hz), 5.92 (d, 1 H, J = 5.5 Hz), 4.83 (t, 1 H, J = 5.4 Hz), 3.78 (s, 3H), 2.87 (dd, 1 H, J = 6.7 Hz, J = 13.6 Hz), 2.02 (dd, 1 H, J = 4.3 Hz, J = 13.7 Hz), 0.90 (s, 9H), 0.10 (s, 6H). 1 H NMR (300 MHz, CDCl 3 ): δ Η (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).
RMN 13C (75 MHz, CDCI3) : 5C (ppm) 153.8, 138.5, 134.7, 88.2, 75.6, 74.9, 52.8, 51 .0, 25.8, 18.0, -4.7. 13 C NMR (75 MHz, CDCl 3 ): δ C (ppm) 153.8, 138.5, 134.7, 88.2, 75.6, 74.9, 52.8, 51.0, 25.8, 18.0, -4.7.
HRMS (ESI) : m/z calculée pour C15H23O3S1 [M-H2O+H+] 279.1416 HRMS (ESI): m / z calcd for C15H23O3S1 [MH 2 O + H + ] 279.1416
valeur expérimentale 279.1414  experimental value 279.1414
Composé 5 : (E)-methyl 3-(4-((tert-butyldimethylsilyl)oxy)- 1 -hydroxycyclopent-2-en-1 -yl)acrylate Compound 5: (E) -methyl 3- (4 - ((tert-butyldimethylsilyl) oxy) -1-hydroxycyclopent-2-en-1-yl) acrylate
TBDMS  TBDMS
298.2 g.mol"1 Le composé 4 (700 mg, 2.36 mmol) est solubilisé dans 20 ml de THF distillé et la solution est refroidie à -78°C. Une solution commerciale de Red-AI (3.21 M dans le toluène, 1 .54 ml, 2.0 éq) est ajouté goutte à goutte et la réaction est maintenue sous agitation à -78°C pendant 30 minutes. Après addition de 20 ml d'eau distillée, le brut réactionnel est extrait par 3 x 100 ml d'acétate d'éthyle. La phase organique est lavée par 10 ml d'une solution saturée de NaHCO3, puis séchée sur du sulfate de sodium anhydre avant d'être filtrée et évaporée à sec. Le résidu obtenu est purifié par chromatographie flash sur gel de silice (éluant Heptane/AcOEt, 80 :20) pour donner 668 mg (95%) du composé 5 sous forme d'une huile translucide jaune pâle. Rf = 0.63 (Heptane/AcOEt, 6:4) 298.2 gmol "1 Compound 4 (700 mg, 2.36 mmol) is solubilized in 20 ml of distilled THF and the solution is cooled to -78 ° C. A commercial solution of Red-AI (3.21M in toluene, 1.54ml, 2.0eq) was added dropwise and the reaction was stirred at -78 ° C for 30 minutes. After addition of 20 ml of distilled water, the crude reaction product is extracted with 3 × 100 ml of ethyl acetate. The organic phase is washed with 10 ml of saturated NaHCO 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 (Heptane / AcOEt eluent, 80:20) to give 668 mg (95%) of compound 5 in the form of a pale yellow translucent oil. Rf = 0.63 (Heptane / AcOEt, 6: 4)
RMN 1H (300 MHz, CDCI3) : δΗ (ppm) 6.84 (d, 1 H, J = 15.4 Hz), 6.09 (d, 1 H, J = 15.6 Hz), 5.97 (dd, 1 H, J = 2.0 Hz, J = 5.4 Hz), 5.77 (d, 1 H, J = 5.5 Hz), 4.80- 4.77 (m, 1 H), 3.75 (s, 3H), 2.52 (dd, 1 H, J = 6.5 Hz, J = 13.8 Hz), 1 .87 (dd, 1 H, J = 3.9 Hz, J = 13.8 Hz), 0.90 (s, 9H), 0.10 (s, 6H). 1 H NMR (300 MHz, CDCl 3 ): δ Η (ppm) 6.84 (d, 1H, J = 15.4Hz), 6.09 (d, 1H, J = 15.6Hz), 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).
RMN 13C (75 MHz, CDCI3) : 5C (ppm) 167.4, 150.5, 137.4, 136.7, 1 18.5, 82.9, 75.3, 51 .6, 49.8, 25.8, 18.1 , -4.7. 13 C NMR (75 MHz, CDCI 3) 5 C (ppm) 167.4, 150.5, 137.4, 136.7, 1 18.5, 82.9, 75.3, 51 .6, 49.8, 25.8, 18.1, -4.7.
HRMS (ESI) : m/z calculée pour C15H25O3S1 [M-H2O+H+] 281 .1573 HRMS (ESI): m / z calcd for C15H25O3S1 [MH 2 O + H + ] 281 .1573
valeur expérimentale 281 .1562 Composé 6 : (E)-methyl 3-(1 ,4-dihydroxycyclopent-2-en-1 -yl)acrylate  Experimental value 281 .1562 Compound 6: (E) -methyl 3- (1,4-dihydroxycyclopent-2-en-1-yl) acrylate
H  H
-1  -1
184.2 g. mol Le composé 5 (650 mg, 2.18 mmol) est solubilisé dans 20 ml de THF distillé à température ambiante. Une solution commerciale de TBAF (1 M dans le THF, 4.37 ml, 2.0 éq) est ajouté goutte à goutte et la réaction est maintenue sous agitation à température ambiante pendant 2 heures. Après addition de10 ml d'une solution saturée de NaCI, le milieu réactionnel est extrait par 3 x 100 ml d'acétate d'éthyle. Les phases organiques rassemblées sont séchées sur du sulfate de sodium anhydre avant d'être filtrées et évaporées à sec. Le résidu obtenu est purifié par chromatographie flash sur gel de silice (éluant Heptane/AcOEt, 80 :20) pour donner 357 mg (89%) du composé 6 sous forme d'une huile translucide. Rf = 0.41 (Heptane/AcOEt, 3:7) 184.2 g. Molar 5 (650 mg, 2.18 mmol) is solubilized in 20 ml of distilled THF at room temperature. A commercial solution of TBAF (1 M in THF, 4.37 ml, 2.0 eq) is added dropwise and the reaction is stirred at room temperature for 2 hours. After addition of 10 ml of a saturated solution of NaCl, the reaction medium is extracted with 3 x 100 ml of ethyl acetate. The combined organic phases are dried over anhydrous sodium sulphate 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. Rf = 0.41 (Heptane / AcOEt, 3: 7)
RMN 1H (300 MHz, CDCI3) : δΗ (ppm) 6.87 (d, 1 H, J = 15.7 Hz), 6.09 (d, 1 H, J = 15.6 Hz), 6.09 (dd, 1 H, J = 2.0 Hz, J = 5.4 Hz), 5.84 (d, 1 H, J = 5.5 Hz), 4.85- 4.81 (m, 1 H), 3.76 (s, 3H), 2.60 (dd, 1 H, J = 6.7 Hz, J = 14.3 Hz), 1 .91 (dd, 1 H, J = 3.5 Hz, J = 14.3 Hz). 1 H NMR (300 MHz, CDCl 3 ): δ Η (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).
RMN 13C (75 MHz, CDCI3) : 5C (ppm) 167.1 , 150.8, 137.7, 136.9, 1 18.6, 83.0, 75.2, 51 .7, 48.9. 13 C NMR (75 MHz, CDCI 3) 5 C (ppm) 167.1, 150.8, 137.7, 136.9, 1 18.6, 83.0, 75.2, 51 .7, 48.9.
HRMS (ESI) : m/z calculée pour C9Hn03 [M-H2O+H+] 167.0708 HRMS (ESI): m / z calcd for C 9 HnO 3 [MH 2 O + H + ] 167.0708
valeur expérimentale 167.0700 Composé 7 : (E)-methyl 3-(1 -hydroxy-4-oxocyclopent-2-en-1 -yl)acrylate Experimental value 167.0700 Compound 7: (E) -methyl 3- (1-hydroxy-4-oxocyclopent-2-en-1-yl) acrylate
(±)-LMA-EA-2801 (±) -lma-EA-2801
CgHioO4 CgHioO 4
182.2 g.mol"1 Le composé (350 mg, 1 .92 mmol) est solubilisé dans 15 ml de CH2CI2 distillé à température ambiante, avant d'y ajouter l'acétate de sodium (313 mg, 1 .2 éq) et le chlorochromate de pyridinium (PCC, 497 mg, 1 .2 éq). La réaction est maintenue sous agitation à température ambiante pendant 1 heure. Le mélange réactionnel est filtré sur une cartouche de célite et le filtrat obtenu est évaporé à sec. Le résidu est ensuite purifié par chromatographie flash sur gel de silice (éluant Heptane/AcOEt, 60:40) pour donner 296 mg (84%) du composé 7 sous forme d'une huile translucide. Rf = 0.38 (Heptane/AcOEt, 3:7) 182.2 g.mol "1 The compound (350 mg, 1.92 mmol) is solubilized in 15 ml of CH 2 Cl 2 distilled at room temperature, before adding sodium acetate (313 mg, 1.2 eq) and pyridinium chlorochromate (PCC, 497 mg, 1.2 eq). The reaction is stirred at room temperature for 1 hour. The reaction mixture is filtered through 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 as a translucent oil. Rf = 0.38 (Heptane / AcOEt, 3: 7)
RMN 1H (300 MHz, CDCI3) : δΗ (ppm) 7.35 (d, 1 H, J = 5.3 Hz), 6.94 (d, 1 H, J = 15.8 Hz), 6.27 (d, 1 H, J = 5.4 Hz), 6.18 (d, 1 H, J = 15.8 Hz), 3.77 (s, 3H), 2.67 (d, 1 H, J = 12.0 Hz). 1 H NMR (300 MHz, CDCl 3 ): δ Η (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).
RMN 13C (75 MHz, CDCI3) : 5C (ppm) 205.0, 166.4, 162.6, 148.5, 134.6, 120.5, 78.3, 51 .9, 49.3. 13 C NMR (75 MHz, CDCI 3) 5 C (ppm) 205.0, 166.4, 162.6, 148.5, 134.6, 120.5, 78.3, 51 .9, 49.3.
HRMS (ESI) : m/z calculée pour C9H9O3 [M-H2O+H+] 165.0552 HRMS (ESI): m / z calcd for C9H9O3 [MH 2 O + H + ] 165.0552
valeur expérimentale 165.0550  experimental value 165.0550
Séparation des énantiomères (+)-LMA-EA-2801 et (-)-LMA-EA-2801 Separation of enantiomers (+) - LMA-EA-2801 and (-) - LMA-EA-2801
Le composé 7 a été analysé par HPLC chirale équipée d'un détecteur UV afin de mettre en évidence les deux énantiomères et de pouvoir les séparer. (Chaîne HPLC Alliance Waters 2695 couplée à un détecteur PDA Waters 996, colonne chirale IC Daicel Chiral Technologie 4,6 x 250 mm, 5 μιτι), éluant n- Heptane/ Isopropanol 80 :20). Compound 7 was analyzed by chiral HPLC equipped with a UV detector in order to highlight the two enantiomers and to be able to separate them. (Alliance Waters 2695 HPLC chain coupled to a Waters 996 PDA detector, chiral IC Daicel Chiral 4.6 x 250 mm (5 μιτι) column, eluent n-Heptane / Isopropanol 80:20).
A partir des données analytiques, la séparation des énantiomères a ensuite été réalisée par SFC préparative.  From the analytical data, the separation of the enantiomers was then carried out by preparative SFC.
(Chaîne Investigator II Waters équipée d'un détecteur DEDL 2420 et d'un détecteur PDA 2996, colonne chirale IC Daicel Chiral Technologie 4,6 x 250 mm, 5 μιτι), éluant CO2 supercritique/ Isopropanol/Methanol, 80 :10 :10).  (Investigator II Waters range equipped with a DEDL 2420 detector and a PDA 2996 detector, chiral column IC Daicel Chiral Technology 4.6 x 250 mm, 5 μιτι), supercritical CO2 / Isopropanol / Methanol eluent, 80: 10: 10 ).
Les pouvoirs rotatoires des deux énantiomères ont été mesurés et comparé à celui obtenu pour le composé optiquement pur d'origine naturelle [a]20 D (-)-LMA-EA-2801 : - 45 ,7 The rotational powers of the two enantiomers were measured and compared to that obtained for the optically pure compound of natural origin [a] 20 D (-) - LMA-EA-2801 - 45, 7
[a]20 D (+)-LMA-EA-2801 : + 40,9 (valeur cohérente avec celle obtenue pour le composé de formule la optiquement pur d'origine naturelle) [a] 20 D (+) - LMA-EA-2801: + 40.9 (value consistent with that obtained for the compound of the naturally occurring optically pure form)
La présente invention concerne les composés de formule (I) ou (la) pour leur utilisation en tant qu'agent antibiotique en particulier en tant qu'agent antibiotique à large spectre et notamment à rencontre de bactéries pathogènes Gram positives et Gram négatives multirésistantes.  The present invention relates to the compounds of formula (I) or (la) for their use as an antibiotic agent in particular as a broad-spectrum antibiotic agent and in particular against gram-positive and gram-negative pathogenic bacteria that are multidrug-resistant.
La présente invention concerne une composition pharmaceutique contenant à titre de principe actif au moins un composé de formule (I) ou (la) associé à un excipient pharmaceutiquement acceptable, qui pourra être déterminée aisément selon les connaissances générales de l'homme du métier en fonction de la voie d'administration retenue.  The present invention relates to a pharmaceutical composition containing as active principle at least one compound of formula (I) or (la) associated with a pharmaceutically acceptable excipient, which can be determined easily according to the general knowledge of the skilled person according to the chosen route of administration.

Claims

REVENDICATIONS 1 . Composé répondant à la formule générale (I) suivante : CLAIMS 1. Compound corresponding to the following general formula (I):
dans laquelle :  in which :
Ri représente un atome d'hydrogène ou un radical alkyle en Ci à C4 ;R 1 represents a hydrogen atom or a C 1 -C 4 alkyl radical;
R2 représente un radical hydroxyle ou alkoxyle en Ci à C4, etR2 represents a hydroxyl or C 1 -C 4 alkoxyl radical, and
R3 et R représentent indépendamment l'un de l'autre un atome d'hydrogène ou un radical alkyle en Ci à C4 ; R3 and R are, independently of one another, a hydrogen atom or a C1-C4 alkyl radical ;
leurs énantiomères et mélanges d'énantiomères en particulier sous forme racémique. their enantiomers and enantiomeric mixtures, in particular in racemic form.
2. Composé selon la revendication 1 répondant à la formule (la) suivante 2. Compound according to claim 1 corresponding to the following formula (la)
(la)  (the)
3. Procédé de préparation d'un composé de formule (I) ou (la) selon l'une des revendications 1 ou 2, caractérisé en ce qu'il implique la mise en œuvre des étapes opérationnelles suivantes : 3. Process for the preparation of a compound of formula (I) or (la) according to one of claims 1 or 2, characterized in that it involves the implementation of the following operational steps:
i. mise en culture d'un champignon filamenteux Trichoderma en particulier de Trichoderma atroviride dans un milieu et des conditions appropriées ; ii. en cours de culture, capture in situ par extraction en phase solide d'un composé de formule (I) ou (la) ;  i. culturing a Trichoderma filamentous fungus, in particular Trichoderma atroviride, in a medium and under suitable conditions; ii. during culture, capture in situ by solid phase extraction of a compound of formula (I) or (la);
iii. isolement et purification du composé de formule (I) ou (la). iii. isolation and purification of the compound of formula (I) or (la).
4. Procédé selon la revendication 3, caractérisé en ce que l'étape de capture in situ est réalisée par extraction en phase solide après introduction dans le milieu de culture d'au moins une résine neutre apolaire. 4. Method according to claim 3, characterized in that the capture step in situ is carried out by solid phase extraction after introduction into the culture medium of at least one apolar neutral resin.
5. Procédé selon la revendication 4, caractérisé en ce que ladite résine est un copolymère polystyrène-divinylbenzène. 5. Method according to claim 4, characterized in that said resin is a polystyrene-divinylbenzene copolymer.
6. Procédé de préparation d'un composé de formule (I) ou (la), selon l'une des revendications 1 ou 2, caractérisé en ce qu'il implique la mise en œuvre des étapes illustrées dans le schéma réactionnel ci-après, les réactifs de départ étant éventuellement substitués de manière appropriée pour obtenir les composés de formule (I) avec les significations Ri à R4 telles que définies précédemment : 6. Process for the preparation of a compound of formula (I) or (la), according to one of claims 1 or 2, characterized in that it involves the implementation of the steps illustrated in the reaction scheme below the starting reagents being optionally appropriately substituted in order to obtain the compounds of formula (I) with the meanings R 1 to R 4 as defined above:
7. Composé de formule (I) ou (la) selon l'une des revendications 1 et 2, pour son utilisation en tant qu'agent antibiotique. 7. Compound of formula (I) or (la) according to one of claims 1 and 2 for its use as an antibiotic agent.
8. Composé selon la revendication 7, pour son utilisation en tant qu'agent antibiotique à large spectre. The compound of claim 7 for use as a broad spectrum antibiotic agent.
9. Composé selon la revendication 8, pour son utilisation en tant qu'agent antibiotique à encontre de bactéries pathogènes Gram positives et Gram négatives multirésistantes. The compound of claim 8 for use as an antibiotic agent against multidrug-resistant Gram-positive and Gram-negative pathogenic bacteria.
10. Composition pharmaceutique caractérisée en ce qu'elle contient à titre de principe actif au moins un composé de formule (I) ou (la) associé à un excipient pharmaceutiquement acceptable. 10. Pharmaceutical composition characterized in that it contains as active ingredient at least one compound of formula (I) or (la) associated with a pharmaceutically acceptable excipient.
EP15721221.8A 2014-05-06 2015-05-06 4-vinyl-2-cyclopenten-1-one, the production thereof, and the use of same as an antibiotic agent Withdrawn EP3140280A1 (en)

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FR1454090A FR3020809B1 (en) 2014-05-06 2014-05-06 4-VINYL-2CYCLOPENTENE-1-ONE DERIVATIVES FOR THE PREPARATION THEREOF AND THEIR USE AS ANTIBIOTIC AGENT
PCT/EP2015/059999 WO2015169876A1 (en) 2014-05-06 2015-05-06 4-vinyl-2-cyclopenten-1-one, the production thereof, and the use of same as an antibiotic agent

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CN106866345B (en) * 2015-12-12 2020-06-02 中国科学院大连化学物理研究所 Method for preparing JP-10 aviation fuel from furfuryl alcohol
CN106866364B (en) * 2015-12-12 2020-06-09 中国科学院大连化学物理研究所 Method for preparing 1, 3-cyclopentanediol from furfuryl alcohol
CN106866392B (en) * 2015-12-12 2020-06-05 中国科学院大连化学物理研究所 Method for preparing 4-hydroxycyclopenta-2-enone from furfuryl alcohol
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US9751826B2 (en) 2017-09-05
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