WO2012062996A1

WO2012062996A1 - Derivatives of 6-amino-6-deoxyglucosamine and use of same as antibacterial agents

Info

Publication number: WO2012062996A1
Application number: PCT/FR2011/052604
Authority: WO
Inventors: Jean-Luc Decout; Isabelle Baussanne; Jérôme DESIRE; Indrajit Das; Marie-Paule Mingeot-Leclercq
Original assignee: Universite Joseph Fourier
Priority date: 2010-11-09
Filing date: 2011-11-08
Publication date: 2012-05-18
Also published as: FR2967068A1

Abstract

The invention relates to compounds of formula (I) where: R₁ ≠ 2-deoxystreptamine and the derivatives thereof substituted in positions 5 and/or 6; R₂ and R₆ are the same or different and represent a primary, secondary or tertiary amine function: NH₂, NHR, NRR' where R and R' = alkyl, aryl, cyclic or otherwise, substituted or otherwise; OR₃ and OR₄ form alcohol, ether, ester, carbonate, carbamate, sulfonate or sulfamate functions; and if R₃ = R₄, then R₃and R₄ ≠ H; if R₃ = H, then R₄ ≠ H and R₁ ≠ OH; and if R₄ = H, then R₃ ≠ H and R₁ ≠ OH; if R₁ = OH, then R₃ and R₄ ≠ H, and if R₁ = OR₅, then R₃ and/or R₄ and/or R₅ are not a sugar or a polysaccharide. The invention further relates to the use of said compounds as drugs and, specifically, as antibiotics.

Description

6-AMINO-6-SOXYGLUC O-AMINE DERIVATIVES AND THEIR USE AS ANTIBACTERIAL AGENTS

TECHNICAL AREA

The present invention relates to the field of antibacterial agents.

More specifically, new derivatives of 6-amino-6-deoxyglucosamine have been synthesized and appear to be very active.

PRIOR STATE OF THE TECHNIQUE

Aminoglycosides are natural polyamino pseudo-polysaccharides synthesized by Gram-positive bacteria to control other bacteria. The first compounds of this family were discovered in the 1940s and identified as powerful antibiotic agents quickly used in the hospital. Streptomycin, neomycin, kanamycin and gentamicin were the first molecules in the aminoglycoside family to be isolated.

The majority of aminoglycosides have as a common structural element the streptamine or 2-deoxystreptamine ring linked, by glycosidic linkage, to one or two mono- or disaccharide units.

Among them are shown below paromamine (1) and neamine (2), consisting of ring I (2-deoxystreptamine) linked on position 4 to ring II (glucosamine if R = OH or 6-amino-6-deoxy glucosamine if R = NH ₂ ) by glycosidic linkage. These two compounds constitute the basic units of two major families of aminoglycosides:

the family of 5-substituted derivatives to which paromomycin (3) and neomycin (4) belong;

the family of 6-substituted derivatives (5), to which kanamycins and tobramycin belong.

1: R = OH, Paromamine

2: R = NH ₂ , Namine

Among the current antibiotic agents, aminoglycosides represent a very important class of molecules that are mainly used in hospitals. Restricted use in this medium minimizes the risk of resistant bacteria developing, which provides access to active molecules in an emergency.

Most aminoglycosides have an antibiotic effect against both Gram-positive and Gram-negative bacteria. They are often administered in combination with β-lactams or fluoroquinolones to broaden their spectra. In contrast, all aminoglycosides are inactive on anaerobic bacteria and mycoplasmas.

Aminoglycosides have several modes of action, the main one of which is the disruption of protein synthesis due to a strong interaction with the A site of the 16S bacterial ribosomal AR. In 1987, Mozaed and Noller showed that some aminoglycosides interact specifically with ribosomal RNA. Since this discovery, other RNAs for which aminoglycosides are good ligands have been identified. They are, for example, able to bind to TAR RNA, RRE and HIV-1 DIS.

Their antibiotic effects are also due to changes in the properties of the bacterial membrane they cause. Unfortunately, aminoglycosides have certain disadvantages when used as drugs. Indeed, their toxicity limits the therapeutic use, and as for many other agents, phenomena of resistance appear. Thus, resistant bacterial strains are responsible for nosocomial diseases that constitute a major health problem.

These resistances involve enzymatic modifications of the aminoglycoside structure rendering them incapable of binding to bacterial ribosomal RNA, the reduction of their intracellular efflux concentration, the alteration of the 30S subunit structure of the target AR and the methylation of the binding site of aminoglycosides in RNA.

With regard to the resistance caused by the modification enzymes synthesized by certain bacteria, we find three classes of resistance enzymes:

aminoglycosides O-phosphotransferases (APH) and aminoglycoside nucleotidyltransferases (ANT) which phosphorylate aminoglycosides on hydroxyl functions using ATP;

aminoglycoside N-acetyltransferases (AACs) which catalyze the acylation of amino functions of aminoglycoside using coenzyme A.

The modifying enzymes are specific for different positions of the aminoglycoside. An enzyme that phosphorylates the 3 'hydroxyl function on ring II will bear the name APH3'. Kanamycin B is, for example, modified by the following enzymes:

Note that a bifunctional enzyme AAC (6 ') - APH (2 ") was discovered in 1997.

With regard to the mechanisms that contribute to the decrease of the cellular concentration in active molecules, the efflux pumps present in the cell membrane of the prokaryotic cells constitute a system of rejection of the xenobiotic agents which are liable to poison the cell. These efflux pumps can be overactivated because of a mutation. Some cells have mutations in the regulatory genes of these pumps so that they will be overexpressed and the rejection of xenobiotics will be less selective allowing the efflux of aminoglycosides ("Multi-Drug Resistant" or MDR systems). The major antibiotic effects of aminoglycosides and resistance problems have therefore led to a renewed interest in the chemistry of these molecules in order to find new antibiotic and / or antiviral agents. In particular, it involves identifying new active agents on currently available aminoglycoside-resistant bacteria and therefore current treatments.

Thus, the document WO2009 / 095588 reports antimicrobial activities of interest for molecules derived from neamine, more specifically derivatives of disubstituted neamine or paromamine (3 '+6 or 3' + 4 'or 4' + 6), or trisubstituted (3 '+ 4' + 6) at positions 3 ', 4' and / or 6 or involving position 5 (3 '+ 6 + 5 or 3' + 4 '+ 5 or 4' + 6 + 5) . However, these molecules, having at least two cycles, one of which is a sugar, are obtained from expensive starting materials (paromomycin and neomycin).

There is therefore a real need to develop new molecules presenting the desired antibacterial activities. It is in this approach that the present invention falls. Indeed, the Applicant has demonstrated that molecules derived from 6-amino-6-deoxyglucosamine have such properties.

As a reminder, 6-amino-6-deoxyglucosamine has the following formula:

6-amino-6-désoxyglucosamine

This atom numbering will be adopted in the rest of the description.

SUMMARY OF THE INVENTION

Thus and according to a first aspect, the invention relates to compounds having the following formula:

in which :

Ri ≠ 2-deoxystreptamine and its derivatives substituted at the 5 and / or 6 positions;

R ₂ _O and R, identical or different, represent a primary amine function, secondary or tertiary: NH _2, NHR, NRR 'with R and R' = alkyl, aryl, cyclic or noncyclic, substituted or unsubstituted;

OR ₃ and OR ₄ form alcohol, ether, ester, carbonate, carbamate, sulfonate or sulfamate functions; and

. if R ₃ = R ₄ , then R ₃ and R ₄ ≠ H;

. if R ₃ = H, then R 4 ≠ H and R 1 ≠ OH; and

. if R ₄ = H, then R ₃ ≠ H and R 1 ≠ OH;

. if Ri = OH, then R ₃ and R 4 ≠ H.

Moreover, and in a preferred manner, if R 1 = OR ₅ , then R ₃ and / or R 4 and / or R ₅ is not a sugar (or monosaccharide or monosaccharide) or a polysaccharide (or polyose or oligosaccharide). In the context of the invention, the term "sugar" means a ring formed from 5 or 6 atoms, including an intracyclic oxygen atom bonded to an equally intracyclic carbon atom, itself linked to a second exocyclic oxygen atom. (sugar in hemiacetal or acetal form). In the case of a polysaccharide, such a ring is bonded, via an oxygen atom, to one or more other "sugar" type cycles as defined above.

In practice, the compounds which are excluded are disubstituted or trisubstituted derivatives in which: R ₃ 0U R 4 or R ₅ is a sugar or a polysaccharide; R ₃ and R ₄ , R ₃ and R ₅ or R ₄ and R ₅ are sugars or polysaccharides; or R ₃ and R ₄ and R ₅ are sugars or polysaccharides.

It should be noted that Ri can be in both alpha and beta configuration (alpha anomer or beta anomer).

Moreover, the various diastereoisomers of these compounds are also covered in the context of the present application.

In addition, the compounds according to the invention may be in the form of salts resulting from the reaction of the amine functional groups with an acid, for example in the form of hydrochlorides or fluoroacetates. This new class of antibiotic agents has certain and unexpected advantages:

While all the natural aminoglycosides of interest have at least two monosaccharide units (rings), it has been demonstrated in the context of the present invention that only the glucosamine-type ring II is essential for the desired activity. ;

The starting material for obtaining the active derivatives is inexpensive, especially N-acetylglucosamine which is the major constituent of the polysaccharide forming the shell of crustaceans such as crab, namely chitosan.

In other words, the present invention relates in particular to the di- (3 + 1 or 4 + 1 or 3 + 4) and trisubstituted derivatives (3 + 4 + 1) of the following known substances:

6-amino-6-deoxyglucosamine;

6-amino-6-désoxyglucosamine

the isomeric derivatives of the preceding 6-amino-6-deoxyglucosamine derivatives, such as those of 6-amino-6-deoxymannosamine or 6-amino-6-deoxygalactosamine, prepared from mannosamine and galactosamine more expensive, respectively:

mannosamine 6-amino-6-deoxymannosamine galactosamine 6-amino-6-deoxygalactosamine

As already stated, in these molecules, the primary amine functions in position 2 and 6 can be replaced by secondary or tertiary amine functions included in a ring or not, by substitution with alkyl or aryl groups of one or more atoms of hydrogen of the primary amine function.

The present invention is based on the demonstration by the Applicant that the antibacterial activity of the compounds derived from neamine or paromamine disubstituted on ring II (6-amino-6-deoxyglucosamine), especially at the 3 'positions. and 4 'of neamine or paromamine, retained activity even in the absence of cycle I, namely 2-deoxystreptamine. It is therefore possible to introduce any other substituent in position 1 of this ring, embodied in the context of the present invention by the residue Ri.

As a reminder, the various molecules mentioned have the following formula:

e

In order to differentiate from the prior art, the residue R 1 is advantageously different from 2-deoxystreptamine and its derivatives substituted at the 4 and / or 5 and / or 6 positions, as described in the document WO2009 / 095588.

In other words, Ri does not present the following formula:

with R ' ₅ , R' ₆ defined as R ₃ and R ₄ in the context of the present application.

In the context of the invention, the residue R 1 may be of any kind, in particular it may represent a substituted or unsubstituted carbon chain which may contain hetero atoms and which may be cyclic or non-cyclic.

For example, the residue R 1 corresponds to CR ₅ , OR ₅ or SR ₅ , R ₅ being defined as R ₃ and R ₄ above. According to this embodiment, the present invention relates to compounds of formula:

However, in a preferred manner, the present invention relates to compounds of formula:

in which :

OR ₃ , OR ₄ and OR ₅ form alcohol, ether, ester, carbonate, carbamate, sulfonate or sulfamate functions; and

. if R3 = R4 = R5, then R3, R4 and R ₅ ≠ H;

. if R ₃ = H, then R ₄ and R ₅ ≠ H;

. if R ₄ = H, then R ₃ and R ₅ ≠ H;

. if R ₅ = H, then R ₃ and R ₄ ≠ H.

By the way and as already said, if

then R ₃ and / or R ₄ and / or R ₅ is not a sugar or a polysaccharide.

According to a particular embodiment, the targeted derivatives are substituted at the same time at the positions 1, 3 and 4, that is to say that R ₃ ≠ H, R ₄ ≠ H and R ₅ ≠ H. However, the disubstituted derivatives can be preferred because of lower potential toxicity.

Advantageously, the residues R ₃ and R ₄ (and optionally R ₅ ) are identical.

Thus, according to the invention, one or both alcohol functions in position 3 and 4 present in the known molecules are replaced by ether, ester, carbonate, carbamate, sulfonate or sulfamate functions.

In the alcohol function, oxygen has a simple bond with a hydrogen atom (OH).

In the ether function, the oxygen has a simple bond with a carbon atom, itself engaged in a group of the alkyl (OCRR'R ") or aryl (OAr) type.

In the ester function, oxygen has a simple bond with a carbon atom, itself engaged in a double bond with another oxygen atom (OCOR). In the carbonate function, the oxygen has a single bond with a carbon atom, itself engaged in a double bond with a second oxygen atom and in another single bond with a third oxygen atom (OC (O )GOLD).

In the carbamate function, oxygen has a single bond with a carbon atom, itself engaged in a double bond with another oxygen atom and in a single bond with a nitrogen atom (OC (O) NHR or OC (O) NRR ').

In the sulphonate function, oxygen has a simple bond with a sulfur atom, itself involved in two double bonds, each formed with an oxygen atom, and in a single bond with a carbon atom (OS0 ₂ R ).

In the sulphamate function, oxygen has a simple bond with a sulfur atom, itself involved in two double bonds, each formed with an oxygen atom, and in a single bond with a nitrogen atom (OSO ₂ NHR or OSO ₂ NRR ').

The nature of these functions derives in particular from the process implemented for the synthesis of these derivatives.

It is clear that in the compounds targeted by the present invention, the OR ₃ and OR _{4 functions} can independently be selected from the group consisting of alcohol, ether, ester, carbonate, carbamate, sulfonate or sulfamate functions. All combinations are therefore envisaged.

Without wishing to be bound by any theory, the modification and / or congestion of these strategic positions by substituents could prevent their degradation by the bacterial resistance enzymes involved in the modifications of the aminoglycoside structure. This could explain the remarkable antibacterial activity observed for these compounds.

According to a preferred embodiment of the invention, the residues R 3, R 4 and optionally R _5, identical or different, represent:

♦ H with at least two of them different from H;

An alkyl, haloalkyl or heteroalkyl group containing between 1 and 30 carbon atoms in a linear or branched chain;

One or more alkenyl or alkynyl groups containing between 2 and 30 carbon atoms in a linear or branched chain; One or more substituted or unsubstituted cycloalkyl, cycloalkenyl or cycloalkynyl groups containing between 3 and 30 carbon atoms in a linear or branched chain;

One or more aryl or heteroaryl groups containing between 3 and 10 carbon atoms per cycle;

An alkaryl or aralkyl group containing between 1 and 30 carbon atoms, the terms aryl and alkyl having the above definitions;

One or more alkoxy, thioalkyl, sulphonylalkyl or aminoalkyl groups containing between 1 and 30 carbon atoms in a linear or branched chain;

One or more alkoxyalkyl, alkylthioalkyl, alkylsulfonylalkyl, alkylaminoalkyl, alkylketoalkyl or alkyl or aryl alkyl ester containing between 1 and 30 carbon atoms in a linear or branched chain;

One or more heterocyclic groups containing between 5 and 10 carbon atoms per ring;

An alkylcarbonyl or arylcarbonyl group (-C (O) R), R being defined as R ₃ , R 4 and

R ₅ ;

An alkyloxycarbonyl or aryloxycarbonyl (-C (O) OR) group, R being defined as R ₃ , R 4 and R 5;

An alkylcarbamoyl or arylcarbamoyl group (-C (O) NHR or (-C (O) NRR '), R and R' identical or different being defined as R ₃ , R 4 and R ₅ ;

A sulfonyl group (-SO ₂ R), R being defined as R ₃ , R 4 and R ₅ ;

An aminosulfonyl group (-SO ₂ NHR or -SO ₂ NHRR '), R and R' identical or different being defined as R ₃ , R 4 and R 5;

said groups being all optionally substituted by one or more nitro, cyano, hydroxy, carboxy, carbonyl or amino groups, by one or more halogens or by one or more functions nitrile, cyanohydrin, aldehyde.

By "amine" is meant a group comprising a nitrogen atom. It may be a primary amine NH ₂ , secondary amines NH or tertiary amines NR'R "R and R", which may be identical or different, are defined as R ₃ , R 4 and R ₅ .

According to the invention, the term "alkyl" denotes a linear or branched hydrocarbon radical of 1 to 30 carbon atoms, such as, by way of indication, methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, isobutyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, pentadecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl or icosyl. The alkyl group defined above may comprise one or more halogen atoms (fluorine, chlorine, bromine or iodine). In this case, we speak of "haloalkyl" group. The alkyl group may further comprise heteroatoms selected from P, O, N, S and Se. In this case, we speak of "heteroalkyl" group. By "alkenyl" is meant a linear or branched hydrocarbon chain of 2 to 30 carbon atoms comprising one or more double bonds. Examples of alkenyl groups are alkenyl groups carrying a single double bond such as -CH-CH = CH-CH ₂ , H ₂ C = CH- (vinyl) or H ₂ C = CH-CH ₂ - (allyl).

By "alkynyl" is meant a linear or branched hydrocarbon chain of 2 to 30 carbon atoms comprising one or more triple bonds. Examples of alkynyl groups are alkynyl groups carrying a single triple bond such as -CH ₂ -C≡CH.

The term "cycloalkyl" refers to saturated hydrocarbon groups which may be mono- or polycyclic and comprise from 3 to 10 carbon atoms. These are, for example, monocyclic cycloalkyl groups such as cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, cyclononyl, cyclodecyl, cycloundecyl and cyclododecyl.

By "cycloalkenyl" is meant according to the invention a group derived from a cycloalkyl group as defined above, having one or more double bonds, for example two double bonds. It is for example the cyclohexene group (a double bond) or cyclopenta-1,3-diene (two double bonds).

By "cycloalkynyl" is meant according to the invention a group derived from a cycloalkyl group as defined above, having one or more triple bonds, for example a triple bond.

The term "aryl" represents an aromatic monocyclic or polycyclic hydrocarbon group comprising 3 to 10 carbon atoms per ring, such as phenyl or naphthyl.

The term "heteroaryl" refers to a monocyclic or polycyclic aromatic group comprising between 3 and 10 carbon atoms per ring and comprising 1, 2 or 3 endocyclic heteroatoms per ring selected from P, O, N, S and Se. Examples are furyl, thienyl, pyrrolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, imidazolyl, pyrazolyl, oxadiazolyl, triazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrazinyl and triazinyl groups.

By "alkaryl" is meant an alkyl group, substituted by an aryl group, these two groups being defined above.

By "aralkyl" is meant an aryl group substituted with an alkyl group, both of which groups are defined above. By "alkoxy" is meant an O-alkyl group having 1 to 30 carbon atoms, especially methoxy, ethoxy, propoxy and butoxy. By "alkoxyalkyl" is meant an alkyl-O-alkyl group having 1 to 30 carbon atoms. "Thioalkyl" or "alkylthioalkyl", "sulfonylalkyl" or "alkylsulfonylalkyl" and "aminoalkyl" or "alkylaminoalkyl" groups furthermore contain one or more sulfur atoms, one or more sulphonyl groups and one or more amine functions, respectively.

The term "heterocyclic group" refers to saturated or unsaturated, monocyclic or polycyclic carbon rings having 1, 2 or 3 endocyclic heteroatoms selected from P, O, N, S and Se. These are generally derivatives of the heteroaryl groups described above. Examples of unsaturated heterocycles are dihydrofuryl, dihydrothienyl, dihydropyrrolyl, pyrrolinyl, oxazolinyl, thiazolinyl, imidazolinyl, pyrazolinyl, isoxazolinyl, isothiazolinyl, oxadiazolinyl, pyranyl and the monounsaturated derivatives of piperidine, dioxane, piperazine, trithiane, morpholine of dithiane, thiomorpholine, as well as tetrahydropyridazinyl, tetrahydropyrimidinyl, and tetrahydrotriazinyl.

According to a preferred embodiment, R ₃ and / or R 4, (and optionally R ₅ ) are chosen from the following group: alkyl, advantageously nonyl (C ₉ H ₁₉ ) or hexyl, 1-naphthyl alkyl, advantageously naphthyl-1 - methylene, 1-naphthylpropyl or 1-naphthyl-butyl, 2-naphthylalkyl, advantageously naphthyl-2-methylene, naphthyl-2-propyl or naphthyl-2-butyl.

As already stated, Ri (OR5) does not correspond to 2-deoxystreptamine and its derivatives substituted at the 5 and / or 6 positions and fixed by the 4-position.

According to a preferred embodiment, R ₅ is advantageously chosen from the following group: methyl (CH ₃ ), allyl or CH ₂ -CHOH-CH ₂ OH, CH ₂ -CHOH-CH ₂ R (R = OH, NH ₂ , NH- (CH ₂ ) ₂ -OH, NH- (CH ₂ ) _n -NH ₂ ) or CH ₂ -CHOH-CHOH-CH ₂ -NH ₂ . More generally, it is therefore an alkyl group substituted or not with at least one hydroxyl and / or amine function.

In the particular case where the residue R ₅ is an allyl group, various subsequent modifications are possible, as illustrated below: Oxidation of the double bond of the allyl group

Coupling by metathesis reaction:

In general, R, R 'and R "are defined as a substituted or unsubstituted carbon chain which may contain hetero atoms, cyclic or otherwise.

All of these reactions are well known and can easily be implemented by those skilled in the art.

Thus, preferred compounds according to the invention are the following: onyl

In particular, compounds according to the invention are for example the following compounds:

Remarkably, it has been shown in the context of the present invention that these compounds may be of interest in the context of a therapeutic application. Therefore and in another aspect, the present invention is directed to the use of a compound as described above as a medicament and a pharmaceutical composition comprising it.

In such a composition, the compound according to the invention is advantageously in the form of pharmaceutically acceptable salt, especially hydrochloride or methylsulfonate.

Of course, such a composition may also contain any excipient or pharmaceutically inert carrier, and optionally one or more other active ingredients. In the context of antibiotic cocktails, a preferred active ingredient is an antibiotic of another class, preferably a β-lactam or a fluoroquinolone.

As already stated, the compounds according to the invention are advantageously used for the preparation of medicaments intended for the prophylaxis or treatment of microbial infections, advantageously bacterial. These bacterial infections can be caused by both Gram-positive bacteria, such as Staphylococcus aureus, and Gram-negative bacteria such as Escherichia coli. Remarkably, the compounds according to the invention can be active both with respect to susceptible strains, and with respect to strains considered to be particularly resistant to conventional aminoglycosides such as neamine or neomycin. EXAMPLES OF REALIZATION

The invention and the advantages thereof will become more apparent from the following exemplary embodiments. These are however in no way limiting.

The present invention will be further illustrated with the aid of the following compounds 10, 17a, 17b, 31, 32, and 33:

From an intermediate in the synthesis of compound 10, a reference compound 12 was synthesized to highlight the importance of R3 and R4 substituents in the observed antimicrobial properties:

EXPERIMENTAL PART

/ - Synthesis of the compounds:

The experimental part below describes the synthesis protocols followed for obtaining two glucosamine derivatives, as well as the characterizations of the new products. The summary scheme is detailed below:

12: R = H

Reagents and conditions: (a) Allyl alcohol, CH ₃ COCl, reflux, 20 h, quantitative (b) TsCl, pyridine, room temperature (ta), 13 h, 63% (c) NaN ₃ , DMF, 80 C, 3 h, 91% (d) 2 NMBr, NaH, DMF, Ta, 48 h, 28% (e) KOH, EtOH, reflux or Ba (OH) ₂ , H ₂ O, reflux, 8 h, 78% (f) TrCl, DMF, and ₃ N, Ta, 8 h, 82% (g) 2NMBr, NaH, DMF, Ta, 10h, 79% (h) Ph ₃ P, THF / H ₂ 0 (19/1), 80 ° C, 6 h, (i) TFA / anisole (1/1), 0 ° C, 3 h, 0 ^' ) Dowex resin (Cl ^" ), 62% for 10, 76% for 12 (3 steps).

Derivatives 2, 3, 4 and 7 are already described in the literature:

Wong, C.-H; Hendrix, M .; Manning, D.D .; Rosenbohm, C; Greenberg, W.A. A library approach to the discovery of small molecules that recognize RNA: use of 1,3-hydroxyamine motif as core. J. Am. Chem. Soc. 1998, 120, 8319-8327;

Wu, B .; Yang, j .; Robinson, D .; Hofstadler, S .; Griffey, R .; Swayze, EE; He, Y. Synthesis of linked carbohydrates and evaluation of their binding for 16S RNA by mass spectrometry. Bioorg. Med. Chem. Lett. 2003, 13, 3915-3918. 1-Allyl-6-azido-2,6-dideoxy-2-tritylamino-α-D-glucopyranoside 8:

To a solution of the derivative 7 (0.30 g, 1.23 mmol) in DMF (8 mL) and triethylamine (0.5 mL) is added a solution of TrCl (1.03 g, 3.68 mmol, 3 equiv) in DMF (3 mL) and triethylamine (0.5 mL), then the mixture is stirred at room temperature for 8 h under an argon atmosphere. A saturated solution of NH ₄ Cl is added and the mixture is then extracted with ethyl acetate (3 x 10 mL). The combined organic phases are dried over MgSO ₄ , filtered and concentrated. The residue obtained is purified by silica gel chromatography column (eluent: 1/3 ethyl acetate / cyclohexane, containing 0.1% triethylamine) to give compound 8 (0.49 g, 82%) under form of a white solid.

Melting point: 128-130 ° C.

1 H NMR (400 MHz, CDCl ₃ ) δ 7.54-7.56 (m, 5H, arom), 7.23-7.32 (m, 10H, arom), 5.81 (m, 1H, allyl CH), 5.21 (dd, 1H, J = 4.0, 16.0 Hz, allyl CH ₂ ), 5.13 (dd, 1H, J = 4.0, 12.0 Hz, allyl CH ₂ ), 3.75-3.80 (m, 2H, H-3, OCH ₂ ), 3.60 (m, 1H, H-5), 3.43-3.48 (m, 2H, H-4). , H-6), 3.27-3.38 (m, 2H, H-6 ', OCH ₂ ), 3.07 (d, 1H, J = 4.0 Hz, H 1), 2.96 (dd , 1H, J = 4.0, 12.0 Hz, H-2).

¹³ C NMR (100 MHz, CDCl ₃ ) δ 146.7, 134.0 (allyl CH), 129.0, 128.2, 126.9, 117.1 (allyl CH ₂ ), 97.5 (Cl) , 73.9 (C-3), 71.6 (C-4), 70.4 (C-5, CPh ₃ ), 68.7 (OCH ₂ ), 57.6 (C-2), 51, 7 (C-6).

HRMS (ESL) m / z: [M + K] ⁺ calcd. 525.1941, found 525.1931, [M + Na] ⁺ calcd. 509.2165, found 509.2164. 1-Allyl-6-azido-2,6-dideoxy-3,4-di-O - [(2-naphthyl) methyl] -2-tritylamino-α-D-glucopyranoside 9:

To a solution of the derivative 8 (1.22 g, 2.51 mmol) in DMF (10 mL) are added NaH (0.4 g, 10.04 mmol, 4 equiv) and then 2-bromomethylnaphthalene (2.22 g, 10.04 mmol, 4 equiv). The reaction mixture is stirred at room temperature for 10 h under an argon atmosphere. A saturated solution of NH ₄ Cl is added and the mixture is then extracted with ethyl acetate (3 x 30 mL). The combined organic phases are dried over MgSO ₄ , filtered and concentrated. The residue obtained is purified by column of silica gel chromatography (eluent: 1/45 ethyl acetate / cyclohexane, containing 0.1% triethylamine) to give compound 9 (1.52 g, 79%) as a white solid.

Fusion: 118-120 ° C.

1 H NMR (400 MHz, CDCl ₃ ) δ 7.17-8.01 (m, 29H, arom), 5.80 (m, 1H, CH allyl), 5.70 (m, 1H, CH ₂ naphthyl) , 5.18-5.23 (m, 2H, CH ₂ allyl, CH ₂ naphthyl), 5,11-5,14 (m, 2H, CH ₂ allyl, CH ₂ naphthyl), 4.76 (m, 1H , CH ₂ naphthyl), 3.94 (t, 1H, J = 8.0 Hz, H-3), 3.65-3.72 (m, 2H, H-5, OCH ₂ allyl), 3.50 (dd, 1H, J = 8.0, 12.0 Hz, H-4), 3.42 (m, 1H, H-6), 3.14- 3.32 (m, 3H, H-2, H-6, OCH ₂ allyl), 2.71 (d, 1H, J = 4.0 Hz, H1).

¹³ C NMR (100 MHz, CDCl ₃ ) δ 147.4, 136.6, 135.7, 134.2 (CH allyl), 133.6, 133.4, 133.1, 133.0, 129.2 , 128.4, 128.3, 128.1, 127.9, 127.8, 126.7, 126.6, 126.3, 126.2, 126.1, 126.0, 125.9, 117 , 0 (CH ₂ allyl), 97.9 (Cl), 83.4 (C-3), 79.7 (C-4), 77.2 (CH ₂ naphthyl), 75.5 (CH ₂ naphthyl) , 70.8 (CPh ₃ ), 70.0 (C-5), 68.8 (OCH ₂ allyl), 58.3 (C-2), 51.6 (C-6).

HRMS (ESI ⁺ ) m / z: [M + Na] ⁺ calcd. 789.3417, found 789.3434, [M-N2 + Na] ⁺ calculated 761.3353, found 761.3339. l-Allyl-2,6-diamino-2,6-dideoxy-3,4-di-0 - [(2-naphthyl) methyl] -ad-glucopyranoside

10:

To a solution of 9 (0.41 g, 0.53 mmol) in THF / H ₂ O (19/1) is added triphenylphosphine (0.21 g, 0.80 mmol, 1.5 equiv). ), then the mixture is heated at 80 ° C for 8 h. After concentration, the residue is purified by silica gel chromatography column (eluent: 1/4 ethyl acetate / cyclohexane, containing 0.1%) of triethylamine) to give the reduced compound at the 6-position.

This is treated with a TFA / anisole mixture (1/1) for 3 h at 0 ° C under an argon atmosphere, and then the mixture is co-evaporated 3 times with toluene. The resulting residue is washed with dry diethyl ether (3 x 5 mL) to give a yellow compound. This is purified by chromatography on an ion exchange resin column (Dowex Cl ^- ) in methanol, to give compound 10 (0.19 g, 62%> for the 3 steps) as a yellowish solid. Melting point: 183-185 ° C.

1 H NMR (400 MHz, CD ₃ OD, hydrochloride) δ 7.34-7.81 (m, 14H, arom), 6.03 (m, 1H, CH allyl), 5.43 (m, 1H, CH ₂ allyl), 5.32 (m, 1H, CH ₂ allyl), 5.21 (d, 1H, J = 4.0 Hz, H 1), 5.07 (dd, 2H, CH ₂ naphthyl), 4, 91 (dd, 2H, naphthyl CH ₂ ), 4.37 (dd, 1H, J = 8.0, 12.0 Hz, OCH ₂ allyl), 4.20 (dd, 1H, J = 8.0, 12; , 0 Hz, OCH ₂ allyl), 4.06-4.15 (m, 2H, H-3, H-5), 3.70 (t, 1H, J = 8.0 Hz, H-4), 3.57 (dd, 1H, J = 4.0, 12.0 Hz, H-2), 3.35-3.40 (m, 1H, H-6), 3.08-3.11 (m , 1H, H-6).

¹³ C NMR (100 MHz, D ₂ O) δ 136.7, 136.3, 134.8, 134.7, 134.6 (CH allyl), 134.5, 129.4, 129.2, 129, 1, 128.8, 127.7, 127.4, 127.3, 127.2, 126.8, 126.7, 119.5 (CH ₂ allyl), 96.4 (Cl), 81.5 ( C-4), 79.5 (C-3), 76.5 (OCH ₂ naphthyl), 76.4 (OCH ₂ naphthyl), 70.8 (OCH ₂ allyl), 69.8 (C-5), 54.9 (C-2), 41.8 (C-6).

HRMS (ESI ⁺ ) m / z: [M + Na] ⁺ calcd. 521.2416, found 521.2404, [M + H] ⁺ calcd. 499.2597, found 499.2594.

Elemental analysis for C ₃ H ₃₆ N ₂ O ₄ + 1.5H ₂ O: calculated C 62.20, H 6.57, N 4.68; found C 62.44, H 6.30, N 4.88. 1-Allyl-6-amino-2,6-dideoxy-2-tritylamino-α-D-glucopyranoside 11:

To a solution of compound 8 (0.22 g, 0.45 mmol) in THF / H ₂ O (19/1) is added triphenylphosphine (0.15 g, 0.59 mmol, 1.3 equiv. ), then the mixture is heated at 80 ° C for 6 h. After concentration, the residue is purified by chromatography column on silica gel (eluent: methanol / ethyl acetate 1/9 to 1/1, containing 0.1% triethylamine) to give compound 11 (0.19 g , 91%>) in the form of a white solid.

Fusion: 139-14FC.

1 H NMR (400 MHz, CD ₃ OD) δ 7.66-7.69 (m, 5H, arom), 7.15-7.27 (m, 10H, arom), 5.85 (m, 1H). , CH allyl), 5.25 (dd, 1H, J = 4.0, 16.0 Hz, CH ₂ allyl), 5.12 (m, 1H, CH ₂ allyl), 3.84 (dd, 1H, J = 4.0, 12.0 Hz, OCH ₂ ), 3.74 (t, 1H, J = 8.0, 12.0 Hz, H-3), 3.58 (m, 1H, H-5), 3.24 (m, 2H, H-6, OCH ₂ ), 3.15 (m, 1H, H-4), 2.90 (dd, 1H, J = 4.0, 12.0 Hz, H-6), 2.87 (d, 1H, J = 4.0 Hz, H1), 2.75 (dd, 1H, J = 4.0, 12.0 Hz, H-2 ).

¹³ C NMR (100 MHz, CD ₃ OD) δ 148.7, 135.5 (CH allyl), 130.3, 129.0, 127.6, 17.0 (CH ₂ allyl), 99.4 ( Cl), 74.5 (C-3), 73.8 (C-4), 71.7 (CPh ₃ ), 69.8 (OCH ₂ ), 69.3 (C-5), 59.2 ( C-2), 42.4 (C-6).

HRMS (ESI ⁺ ) m / z: [M + Na] ⁺ calc. 483.2260, found 483.2260, [M + H] ⁺ calc. 461, 2440, found 461, 2442. 1-Allyl-2,6-diamino-2,6-dideoxy-α-D-glucopyranoside 12

A solution of compound 11 (0.1 g, 0.24 mmol) in a TFA / anisole mixture (1/1) is stirred for 6 h at 0 ° C under argon atmosphere. The mixture is co-evaporated 3 times with toluene, and the residue is washed with dry diethyl ether (3 x 5 mL). The oil obtained is purified by chromatography on an ion exchange resin column (Dowex Cl ^" ) in water, to give compound 12 (0.06 g, 84% for both steps) in the form of a yellow oil.

1 H NMR (400 MHz, D ₂ 0) δ 5.96 (m, 1H, CH allyl), 5.36 (dd, 1H, J = 4.0, 16.0 Hz, CH ₂ allyl), 5.28 (dd, 1H, J = 4.0, 12.0 Hz, CH ₂ allyl), 5.21 (d, 1H, J = 4.0 Hz, H1), 4.26 (m, 1H, OCH ₂ ) , 4.10 (m, 1H, OCH ₂ ), 3.85-3.93 (m, 2H, H-4, H-5), 3.36-3.46 (m, 3H, H-2, H-3, H-6), 3, 16 (dd, 1H, J = 4.0, 12.0 Hz, H-6).

¹³ C NMR (100 MHz, D ₂ 0) δ 133.0 (CH allyl), 1 18.7 (CH ₂ allyl), 94.3 (Cl), 71.1 (C-3), 69.4 ( C-4), 69.0 (OCH ₂ allyl), 68.0 (C-5), 53.6 (C-2), 40.2 (C-6).

HRMS (ESI ⁺⁾ m / z: [M + H] ⁺ calcd for C ₉ Hi ₈ N ₂ 0 ₄ 219.1345, found 219, 1353.

Reference compound 12 in the antimicrobial tests was synthesized from Intermediate 7 in the previous synthesis:

Synthesis of compounds 17a and 17b

Reagents and conditions: (a): C ₉ Hi ₉ Br, toluene, TBAF, _aq 50%; b: NaH, DMF, 2NPBr, 50 ° C, 20 h, quantitative; (b) (tBuOCO) ₂ 0, DMAP, THF, 45 ° C, 6 h; (c) (CH ₃ ^0- , Na ⁺ ) / CH ₃ OH, ta, 6 h; (d) PPh ₃ , THF / H ₂ O, rt, 4 h; (f) TFA, DCM, t. a, 4 h.

Step a: obtaining compounds 13a and 13b

Compound 13a

A solution of the compound 4 (1.0 g) in toluene (20 mL) and a 50% aqueous solution of NaOH (10 mL) are stirred at 45 ° C in the presence of 1.65 g of TBAF (1.5 mL). eq) and 14 mL of 1-bromononane (2 eq). After 16 h, ethyl acetate (20 mL) is added to the cooled solution. After extraction, the organic phase is recovered and the aqueous phase is taken up with ethyl acetate (3 × 20 mL). The combined organic phases are dried over MgSO 4, filtered and concentrated. The residue obtained is purified by chromatography on silica gel (eluent: ethyl acetate / cyclohexane 1/5 to 2/3) to give the compound 13a (1.08 g, 57%) in the form of a solid White.

Compound 13b

A solution of compound 4 (1.0 g) in DMF (40 mL) is stirred at room temperature in the presence of NaH (0.42 g at 60% in oil) and 2- (3-bromopropyl) naphthalene (1.73 g, 2 eq). After 16 h, toluene and methanol are added and the solution is evaporated. The residue obtained is purified by chromatography on silica gel (eluent ethyl acetate / cyclohexane 1/5 to 2/3) to give compound 13b (0.64 g, 28%) as a white solid.

Step b, obtaining compounds 14a and 14b

compound 14a compound 14b

A solution of compound 13a (0.15 g) or 13b (0.14 g), DMAP (0.033 g) and t-butyloxycarbonyl anhydride (0.067 g) in anhydrous THF (6 mL) is heated with stirring for 2 h at 45 ° C. Two fractions of DMAP (0.033 g) and anhydride (0.067 g) are successively added at intervals of 2 h. After heating the mixture at 45 ° C for 6 hours in total, the mixture is then diluted with dichloromethane (20 mL) and washed several times with water until the aqueous phase is neutral (4 x 30 mL). The organic phase is dried over MgSO ₄ , filtered and concentrated. The residues obtained are purified by chromatography on silica gel (eluent: ethyl acetate / cyclohexane 1/19 to 1/9) to give as white solids the compounds 14a (0.134 g, 77%) and 14b (0. 14 g, 86%).

Step

Compounds 14a (0.122g) and 14b (0.138g) are respectively dissolved in a solution of sodium methanolate (0.03g) in anhydrous methanol (5mL). After stirring at room temperature for 2 h, two further methanolate fractions (0.03 g) are added at 2 h intervals. After stirring for a total of 6 h, the mixture is concentrated and diluted with dichloromethane (20 mL). The solution is then washed several times with water (3 x 30 mL). The organic phase is dried over MgSO ₄ , filtered and then evaporated to dryness to give compounds 15a (0.095 g) and 15b (0.125 g). Step of obtaining compounds 16a and 16b

compound 16a compound 16b

A solution of the compound 16a (0.093 g) or 16b (0.1 g) and triphenylphosphine (3 eq for each azide function to be reduced) in a THF / water mixture (3: 1, 4 ml) is stirred for 4 h at room temperature. room. The solution is then evaporated to dryness. The residue obtained is then chromatographed on silica gel (eluent: 1/9 ethyl acetate / cyclohexane) to obtain compound 16a (0.085 g, yield for steps c and d: 81%) and compound 16b (0.083 g yield for steps c and d: 83%) as yellow residues.

Step e, obtaining compounds 17a and 17b

compound 17a compound 17b

The compounds 16a (0.074 g) and 16b (0.073 g) are respectively stirred in a dichloromethane / trifluoroacetic acid (TFA) mixture (2/3, 5 mL) for 4 h at room temperature. The solutions are then evaporated to dryness and the residues are dissolved in water (20 ml). The aqueous phase is then washed with ethyl ether (3 x 20 ml). The organic phases are combined and extracted once with water (20 mL). The aqueous phases are combined and evaporated to dryness to give the compounds 17a (0.09 g, 97%) and

17b (0.085 g, 98%) as highly hygroscopic transparent gums.

1H NMR

2 '2'

compound 17a compound 17b 17a: δ, 04-5.94 (m, 1H, H-2 '), 6.04-5.94 (m, 1Η, H-2'), 5.38 (dd, 1Η, J = 1); , 45 Hz, J ₂ = 17.3 Hz, H-3 '), 5.27 (dd, 1Η, J = 1.0 Hz, J ₂ = 10.8 Hz, H-3'), 5.13 (d, 1Η, J = 3.3Hz, H-1), 5.56 (dd, 1H, J = 5.6Hz, J ₂ = 12.5Hz, H-1 '), 4.09 ( dd, 1Η, J = 6.5 Hz, J ₂ = 12.5 Hz, H-1 '), 3.89-3.76 (m, 3Η, H-1 "H-5), 3.72- 3.65 (m, 2Η, H-1 "H-3), 3.59 (td, 1Η, J = 6.9 Hz, J ₂ = 8.8 Hz, H-1"), 3.35- 3.27 (m, 2Η, H-6H-2), 3.22 (t, 1Η, J = 9.3, H-4), 3.11 (dd, 1Η, J = 9.6 Hz, J ₂ = 12.9 Hz, H-6), 1.69-1.56 (m, 4Η, H-2 "), 1.33-1.28 (m, 24Η, H-3"), 0.91-0.88 (m, 6H, H-4 ").

17b: δ 7.76-7.18 (m, 14Η, H-naphth), 6.05-5.95 (m, 1Η, H-2 '), 5.39 (dd, 1Η, J = 1); , 5 Hz, J ₂ = 17.3 Hz, H-3 '), 5.28 (dd, 1Η, J = 1.0 Hz, J ₂ = 10.8 Hz, H-3'), 5.13 (d, 1Η, J = 3.1 Hz, H-1), 4.31 (dd, 1Η, J = 5.5 Hz, J ₂ = 12.6 Hz, H-1 '), 4.10 (dd, 1Η, J = 6.3 Hz, J ₂ = 12.6 Hz, H-1 '), 3.88-3.78 (m, 2Η, H-1 "H-5), 3.72-3, 65 (m, 3Η, H-1 "H-3), 3.54 (td, 1Η, J = 6.7 Hz, J ₂ = 9.0 Hz, H-1"), 3.33-3, 29 (m, 2Η, H-6H-2), 3.23 (t, 1Η, J = 9.47, H-4), 3.09 (dd, 1Η, J = 9.5 Hz, J ₂ = 12.9 Hz, H-6), 2.71 (t, 2Η, J = 7.5 Hz, H-3 "), 2.63 (t, 2Η, J = 7.6 Hz, H-3 "), 2.00-1.93 (m, 2Η, H-2"), 1.81-1.73 (m, 2Η, H-2 ").

Reagents and conditions: (a) mCPBA, DCM, TA, 24 h; (b) NaN ₃ , DMF, 60 ° C, 24 h or ethanolamine, 45 ° C, 16 h or 1,3-diaminopropane, 16 h, 45 ° C; (c) (tBuOCO) ₂ 0, DMAP, THF, 45 ° C, 16 h; (d) (CH ₃ ^0- , Na ⁺ ) / CH ₃ OH, ta, 8 h; (e) PPh ₃ , THF / H ₂ O, rt, 6 h; (f) TFA, DCM, Ta, 4h.

Step a, compound 18

To a solution of compound 13a (0.9 g) in dry dichloromethane (25 mL) is added 3 times every 8 h of mCPBA (0.38 g, 3 x 1.3 eq). After stirring for 24 h at room temperature, dichloromethane is added (15 mL) and the mixture is washed with aqueous 5% NaOH solution (30 mL) and then with water (2 x 30 mL). The organic phase is dried over MgSO 4, filtered and then evaporated to dryness to give the mixture 18 of two diastereoisomers a and b (in a proportion of 2: 1) in the form of an opaque solid.

(0.880 g).

1H NMR (400 MHz, CDC1 _3), 18a + b:

δ 5.70 (d, 0.66H, J = 9.6Hz, NHaAc), 5.66 (d, 0.33H, J = 9.6Hz, NH 4Ac), 4.80-4.78 (m , 1Η, H1), 4.01 (dd, 0.33H, J _; = 2.5 Hz J ₂ = 1 1.4 Hz, H1 'b), 3.84-3.69 (m, 3.7H) , 5.6 (dd, 0.66H, J _; = 6.0 Hz J ₂ = 11.9 Hz, H (a)), 3.56-3.46 (m, 3Η, H-6H-1 "), 3.44-3.23 (m, 3.33H, H-3H-4H-6Hl 'b), 3.21-3.15 ( m, 1Η, H-2 '), 2.87-2.81 (m, 1Η, H-3'), 2.66 (dd, 0.66H, J _; = 2.7 Hz, J ₂ = 4, 9 Hz, H-3 'a), 2.60 (dd, 0.33H, J _; = 2.7 Hz J ₂ = 4.8 Hz, H-3'b), 2.02 (s, 3Η, CH ₃ ), 1.57-1.48 (m, 4Η, H-2 "), 1.35-1.21 (m, 24Η, H-3"), 0.90-0.86 (m, 6H, H-4 ").

Step b

Compound 19

A solution of 18 (0.1 g) NaN ₃ (0.18 g, 15 eq) in anhydrous DMF (10 mL) was stirred at 60 for 24 h. After addition of dichloromethane (20 mL), the mixture is washed with water (2 x 30 mL). The organic phase is dried over MgSO 4, filtered and concentrated to give compound 19 as a yellow residue (0.094 g).

Compound 20

A solution of compound 18 (0.1 g) in ethanolamine (20 mL) is stirred for 16 h at 45 ° C. After addition of water (20 mL), the solution is extracted 4 times with ethyl acetate (4 x 20 mL). The organic phases are combined and washed with water (2 x 20 mL). The aqueous phases are then washed with ethyl acetate (2 x 20 mL). The organic phases are then combined and evaporated to dryness to give compound 20 as a yellow residue (0.1 g).

Compound 21

A solution of compound 18 (0.1 g, 0.18 mmol) in 1,3-diaminopropane (10 mL) is stirred for 16 h at 45 ° C and then evaporated. After addition of ethyl acetate (20 mL), the solution is washed with water (3 x 20 mL). The aqueous phases are combined and washed with ethyl acetate (2 x 20 mL). The organic phases are then combined and evaporated to dryness to give compound 21 as a yellow residue (0.1 12 g).

Step c, compounds 22-24

compound 22 compound 23 compound 24

A solution of compound 19 (0.094 g) or (0.11 g) or 21 (0.1 g), DMAP (0.030 g) and t-butyloxycarbonyl anhydride (0.067 g) in anhydrous THF (6 mL) ) is heated with stirring for 2 h at 45 ° C. Two fractions of DMAP (0.033 g) and anhydride (0.067 g) are successively added at intervals of 2 h. After heating the mixture at 45 ° C for 6 hours in total, the mixture is then diluted with dichloromethane (20 mL) and washed several times with water until the aqueous phase is neutral (4 x 30 mL). The organic phase is dried over MgSO ₄ , filtered and concentrated. The residues obtained are purified by chromatography on silica gel (eluent: ethyl acetate / cyclohexane 1/19 to 1/9) to give compounds 22, 23 and 24 as yellow residues (0.078 g, 0.030, respectively). g, 0.060 g, yield for steps a, b and c: 60, 23 and 39%). Step d, compounds 25-27

compound 26 compound 26 27

Compounds 22 (0.078 g), 23 (0.030 g) and 24 (0.060 g) are respectively dissolved in a solution of sodium methanolate (0.03 g) in anhydrous methanol (5 mL). After stirring at room temperature for 2 h, two further methanolate fractions (0.03 g) are added at 2 h intervals. After stirring for a total of 6 h, the mixture is concentrated and diluted with dichloromethane (20 mL). The solution is then washed several times with water (3 x 30 mL). The organic phase is dried over MgSO 4, filtered and then evaporated to dryness to give the compounds (0.059 g), 26 (0.021 g) and 27 (0.34 g).

compound 28 compound 29 compound 30

A solution of the compound 25 (0.059 g), 26 (0.021 g) or 27 (0.034 g) and triphenylphosphine (3 eq. Per azide function to be reduced) in a THF / water mixture (3: 1, 4 mL) is stirred for 4 h at room temperature. The solution is then evaporated to dryness. The residue obtained is chromatographed on silica gel (eluent: 1/9 ethyl acetate / cyclohexane) to obtain the compounds 28, 29 and 30 as yellow residues (0.055 g, 0.017 g, 0.032 g, yield, respectively). steps d and e: 79, 62 and 58%).

Step f, obtaining compounds 31-33

compound 31 compound 32 compound 33 Compounds 28 (0.055 g), 29 (0.017 g) and 30 (0.032 g) are respectively stirred in a dichloromethane / trifluoroacetic acid (TFA) mixture (2/3, 5 mL) for 4 h at room temperature. The solutions are then evaporated to dryness and the residues are dissolved in water (20 ml). The aqueous phase is then washed with ethyl ether (3 x 20 ml). The organic phases are combined and extracted once with water (20 mL). The aqueous phases are combined and evaporated to dryness to give compounds 31 (0.073 g, 92%), 32 (0.020 g, 96%) and 33 (0.039 g, 99%) in the form of highly hygroscopic gums in an R / S mixture. .

compound 31 compound 32 compound 33

31: δ 5.13 (d, 1H, J = 2.5 Hz), 3.95-3.53 (m, 9H, H-3H-5H1H-H1-), 3.35-3 , 29 (m, 2Η, H-2H-6), 3.25-3.18 (m, 2Η, H-4H-3 '), 3.11 (dd, 1Η, J = 9.5 Hz J ₂ = 13.0 Hz, H-6), 3.01 (dd, 1H, Ji = 10.0 Hz J ₂ = 11.8 Hz, H-3 '), 1.70-1.56 (m , 4Η, H-2 "), 1.33-1.28 (m, 24Η, H-3"), 0.92-0.88 (m, 24Η, H-4 ").

32: δ 5.13 (d, 1H, J = 3.3 Hz, H1), 3.96-3.52 (m, 1 1Η, H-3 H-5 HH, H-2 'H1 "), 3.36- 3.03 (m, 8Η, H-2H-4H-6H-3'H-4 '), 1.70-1.56 (m, 4Η, H-2 "), 1 , 33-1.28 (m, 24Η, H-3 "), 0.92-0.88 (m, 6H, H-4").

33: δ 5.13 (d, 1Η, J = 3.4 Hz, H1), 3.95-3.50 (m, 9Η, H-3H-5Hl, H-2'Hl "), 3 36- 3.29 (m, 2Η, H-2H-6), 3.27-2.95 (m, 8Η, H-4H-6H-3'H-4'H-6 ') , 3.13 (q, 2Η, J = 7.1Hz, H-5 '), 1.70-1.56 (m, 4Η, H-2 "), 1.33-1.28 (m, 24Η, H-3 "), 0.92-0.88 (m, 6Η, H-4").

77 - Antibacterial activity 1 / Molecules tested:

The molecules are dissolved in water or DMSO at a concentration of 5 or 10 mg / ml, and then the solutions are diluted if necessary. For each of them a volume of 200 μΐ is arranged in a 96-well plate. 2 / Strains tested:

The following strains are used to test the antimicrobial activity:

- Staphylococcus aureus ATCC25923 = gram (+), sensitive clinical strain;

- Staphylococcus aureus 1199B (Pump NorA) = gram (+), a mutant clinical strain overexpressing a NorA efflux pump of the MFS family, resistant to fluoroquinolones, including ciprofloxacin;

Staphylococcus aureus RN4220 / pUL5054 (Pump MsrA) = gram (+), strain overexpressing the transporter ABC MrsA, containing the multicopy plasmid pUL5054 carrying the msr (A) gene: EryR.

- Staphylococcus aureus APH2 "-AAC6 '= gram (+), resistant strain, capable of enzymatic modification of aminoglycosides at the 2" and 6' position: enzymatic resistance by action of aminoglycoside-6'-N-acetyltransferase / 2 "- O-phosphoryltransferase overexpressed;

- Staphylococcus aureus APH3 '= gram (+), capable of enzymatic modification of aminoglycosides: enzymatic resistance by the action of overexpressed raminoglycoside-3'-O-phosphoryltransferase;

Staphylococcus aureus ANT4 '= gram +, resistant strain, capable of enzymatic modification of aminoglycosides: enzymatic resistance by the action of overexpressed raminoglycoside-4'-O-phosphoryltransferase;

S. aureus ATCC33592 HA-MRSA: gram (+) resistant to methicillin;

S. aureus VRSA-VRS-2: gram (+) resistant to vancomycin

A. Iwoffi ATCC 17925: = gram (-), susceptible strain;

E. coli 25922: = gram (-), susceptible strain;

E. coli 25922 AAC6'-IB: = gram (-), a strain capable of enzymatic modification of aminoglycosides: enzymatic resistance by the action of overexpressed aminoglycoside-6'-O-acetyltransferase;

E. coli arm: = gram (-), a clinical strain capable of enzymatic resistance to aminoglycosides by the action of the enzyme methyltransferase on its own ribosomal RNA;

Pseudomonas. aeruginosa ATCC 27853, Psa.F03 = Gram (-)

Pseudomonas aeruginosa PsaF03 AAC6'-IIA and Pseudomonas aeruginosa PA22 overexMexXY = gram (-) overexpressing respectively an aminoglycoside-resistant enzyme and efflux pumps. 3 / Microbiological tests:

A- Validation of strains

Before being used, each strain is tested individually for its antibiotic resistance pattern. This means that for each strain, the MIC (minimum inhibitory concentration) of one or more reference antibiotics is tested to ensure the phenotypic stability of the bacteria.

B- Determination of the MIC

In the case of S. aureus strains, the solution of the selected molecule is diluted by a factor of 2 to 2 in 96-well plates so as to cover a wide concentration range. To each of these wells is added an inoculum volume consisting of the preculture of the diluted bacterial strain to l / ^100th. Each well is doubled. Again, evidence of non-contamination, growth, and resistance / sensitivity to a reference antibiotic is incorporated into the experiment. The plate is incubated at 37 ° C, and bacterial proliferation kinetics are monitored over 24 hours, with measurements of OD 600 nm at 0, 1, 4, 7, and 24 hours. From these values, the minimum inhibitory concentration (MIC) is determined: the MIC is the lowest concentration found to inhibit bacterial growth after 24 hours of incubation. Having no idea a priori of this value, it is possible that it is not in the range of dilution chosen. It is then necessary to repeat the operation by adjusting this range.

In the case of the A. Iwoffi and E. coli strains, the MIC was determined by geometric microdilutions in accordance with CLSI guidelines (Clinical and Laboratory Standards Institute, Methods for Antimicrobial Dilution. Approved standards M7-A7 and 17 ^th informational supplement M100-S17, CLSI: Wayne, PA, 2007, vol 27, No. 1).

RESULTS

The antibiotic effects were therefore measured in terms of minimal bacterial growth inhibitory concentrations (MICs) on different Gram-positive S. aureus bacteria, resistant or not.

The MIC results, expressed in micrograms / mL, are shown in Tables I to IV below for the representative compounds 10, 17a, 17b, 31, 32 and 33 of the invention. and the reference compound 12 selected in the following table and are to be compared with those of neomycin B and neamine.

Enzyme tested molecule

ATCC Pump Pump Enzyme Enzyme

APH2 ''

25923 NorA MsrA APH3 'AN 4'

-AAC6 '

Neomycin B 2 1 2 1> 128 32

Neamine 32 32 16 16> 128> 128

Compound according to g 4 4 g 4 4

the invention 10

Composed of

> 128> 128> 128> 128> 128> 128 reference 12

Table I: Minimal inhibitory concentrations (MIC) in micrograms / mL against different strains of wild Staphylococcus aureus or overexpressing efflux pumps or aminoglycoside resistance enzymes

Table II: Minimum inhibitory concentrations (MIC) in micrograms / mL against different strains of wild or methicillin-resistant Staphylococcus aureus or vancomycin (nd: not determined) Molecule tested A. Iwoffi E. coli E. E. coli coli ATCC 17925 25922 AAC6 '-IB arm

Neomycin B 2 4 1

Composed according

64 16 64 the invention 10

Composed of> 12i

> 12i> 12E> 12i reference 12

Table III: Minimum Inhibitory Concentrations (MIC) in micrograms / mL different strains of Gram (-) bacteria

Table IV: Minimum inhibitory concentrations (MIC) in micrograms / mL different strains of Gram (-) bacteria

The compounds according to the invention, in particular compounds 10 and 33, prove to be good antibacterial agents against Gram (+) or even Gram (-) bacteria. These results further show the importance of the presence of the substituents carried in positions 3 and 4 to obtain an antimicrobial activity. REFERENCES

Moazed, D .; NoUer, H. F. Interaction of antibiotics with functional sites in 16S ribosomal A. Nature 1987, 327, 389-394.

Baussanne, I .; Bussière, A.; Halder, S .; Ganem-Elbaz, C; Ouberai, M .; Riou, M.; Paris, J.-M .; Ennifar, E.; Mingeot-Leclercq, M. -P. ; Decout, J.-L. Synthesis and antimicrobial evaluation of amphiphilic neamine derivatives. J. Med. Chem. 2010, 59, 119-127.

Claims

1 - Compound of formula:

in which :

. if R ₃ = R ₄ , then R ₃ and R ₄ ≠ H;

. if R ₃ = H, then R ₄ ≠ H and R 1 ≠ OH; and

. if R ₄ = H, then R ₃ ≠ H and R 1 ≠ OH;

. if R 1 = OH, then R ₃ and R ₄ ≠ H; and

if R 1 = OR ₅ , then R ₃ and / or R ₄ and / or R ₅ is not a sugar or a polysaccharide.

2 - Compound according to claim 1 characterized in that it has the following formula:

wherein OR ₃ , OR ₄ and OR ₅ form alcohol, ether, ester, carbonate, carbamate, sulfonate or sulfamate functions; and

. if R3 = R4 = R5, then R3, R4 and R ₅ ≠ H;

if R ₃ = H, then R ₄ and R ₅ ≠ H;

if R ₄ = H, then R ₃ and R ₅ ≠ H;

if R ₅ = H, then R ₃ and R ₄ ≠ H.

3 - Compound according to claim 1 or 2 characterized in that R1 is in alpha or beta configuration. 4 - Compound according to one of the preceding claims characterized in that R ₃ , R 4 and optionally R 5, represent:

^♦ H;

One or more alkenyl or alkynyl groups containing between 2 and 30 carbon atoms in a linear or branched chain;

One or more substituted or unsubstituted cycloalkyl, cycloalkenyl or cycloalkynyl groups containing between 3 and 30 carbon atoms in a linear or branched chain;

R ₅ ;

A sulfonyl group (-SO ₂ R), R being defined as R ₃ , R 4 and R ₅ ;

5 - Compound according to one of the preceding claims characterized in that R ₃ and R4, and optionally R _5i are identical. 6 - Compound according to one of the preceding claims, characterized in that R ₃ and / or R 4 are chosen from the following group: alkyl, advantageously nonyl, 1-naphthyl alkyl, advantageously naphthyl-1-methylene or naphthyl-1-propyl, 2-naphthyl alkyl, advantageously naphthyl-2-methylene or naphthyl-2-propyl.

7 - Compound according to one of the preceding claims characterized in that R ₅ is an alkyl group substituted or not by at least one hydroxyl function and / or amine, preferably selected from the following group: methyl, allyl, CH ₂ -CHOH- CH ₂ R with R = OH, NH ₂ , NH- (CH ₂ ) ₂ -OH, NH- (CH ₂ ) _n -NH ₂ , or CH ₂ -CHOH-CHOH-CH ₂ -NH ₂ .

8- Compound according to one of the preceding claims characterized in that it has a formula selected from the following group:

9 - Compound according to one of the preceding claims characterized in that it has one of the following formulas:

Pharmaceutical composition comprising a compound according to one of claims 1 to 9. 11 - pharmaceutical composition according to claim 10 characterized in that it further comprises another antibiotic such as a β-lactam or a fluoroquinolone.

12 - Compound as defined in one of claims 1 to 9 for use as a medicament.

13 - Compound as defined in one of claims 1 to 9 for its use as an antimicrobial agent, preferably antibacterial.

14 - Use of a compound as defined in one of Claims 1 to 9 for the preparation of a medicinal product directed against Gram-positive bacteria such as Staphylococcus aureus or Gram-negative, such as Escherichia coli, in particular against resistant strains to aminoglycosides.