EP1456186A1 - Novel inhibitor compounds specific of secreted non-pancreatic human a sb 2 /sb phospholipase of group ii - Google Patents

Abstract

The invention concerns compounds of general formula (I), wherein: D represents the group Z-HET, -HET being a heterocycle with 5 members such as oxadiazolone (II) or thiazolidine (III) corresponding to formulae (II) and (III) and Z being selected in the group consisting of -(CR1R2)n- and -(CR1=CR2)n; or D represents the group Z=HET, -Z- representing, with the heterocycle, a -Z=HET group of formula (IV) or (V), wherein: -Z= represents -CR1. Said compounds are useful as inhibitors specific of secreted non-pancreatic human phospholipase A2 (PLA2-snph) of group II, in particular for treating inflammatory pathologies .

Description

PIPERAZINE DERIVATIVES AND THEIR USE AS PHOSPHOLIPASE INHIBITORS

The present invention relates to new specific inhibitor compounds for secreted non-pancreatic phospholipase A ₂ (PLA ₂ -snph) group II, their preparation process, compositions containing them and their use in particular in therapy of inflammatory pathologies.

10 Acting upon the penetration into the body of pathogenic agents (viruses, bacteria, parasites or antigens) or even in response to inflammatory stimuli such as trauma, burns or irradiation, PLA ₂ play a pivotal role in the spread and amplification of inflammation. These enzymes catalyze the hydrolysis of phospholipids

15 in position sn-2 and release lysophospholipids and fatty acids such as arachidonic acid. These can be the precursors of a variety of lipid mediators including the platelet activating factor (PAF), leukotrienes and prostaglandins. They are endowed with multiple biological activities (proliferation and cell migration,

20 contraction, neurosecretion, hormonal release, etc.) and are involved in various inflammatory pathologies and certain cancers.

Among the phospholipases A ₂ referenced under No. EC 3.114 according to the International Classification, the phospholipases A ₂ of group II constitute a particular class of enzymes. Phospholipase A ₂

25 secreted non-pancreatic human (PLA ₂ -snph) group II plays a central role, probably acting in an autocrine / paracrine way by participating in the production of proinflammatory lipid mediators, stimulating migration and cell proliferation and through its properties antibacterial. In many situations

30 pathological, the level of circulating PLA ₂ -snph is closely correlated with the severity and outcome of the disease. This is the case in septic shock caused by either a gram negative infection, peritonitis, malaria or even aspirin poisoning. In this case, an excessive release of PLA ₂ -snp contributes to circulatory collapse,

35 hypotension, development of respiratory distress syndrome and mortality. In rheumatoid arthritis, PLA ₂ -snp accumulates in cartilage, joint and extra-articular matrix, chondrocytes and synovial fluid and the level of circulating enzyme is in accordance with the size and number of inflamed joints. In the respiratory tract and the lung, PLA ₂ -snp is involved in asthma, allergic rhinitis, asbestosis. In the cardiovascular system, this enzyme is activated by ischemia (its expression is also increased after an ischemic brain shock) and plays an important role in the deposition of high density lipoproteins (it is indeed found to be strongly expressed in the atheroma plaque) suggesting a potential role in atherosclerosis and cardiovascular morbidity. In the gastrointestinal tract, high concentrations of this enzyme are measured in Crohn's disease, ulcerative colitis and intestinal inflammation, as well as in cirrhosis and acute pancreatitis. In psoriasis, an increase in its activity is demonstrated in lesions of the skin. At the brain level, it is implicated in cellular and tissue damage from cerebral ischemia and schizophrenia. Finally, it is attributed a role in multiple sclerosis and in various cancers, in particular of the breast and the gastrointestinal tract. PLA ₂ -snph group II is not the only secreted PLA ₂ present in the human body where the PLA ₂ of groups I (pancreas), N (heart, lung) and X (spleen, leukocytes, lung) play also an important role. If the last two, groups V and X recently discovered and whose functions are still poorly defined, are also, like that of group II, involved in inflammation, that of group I, secreted pancreatic PLA ₂ has a role physiological primordial since its catalytic activity is responsible for the digestion of lipids of food origin. It is therefore important not to influence this function, this being complicated by the fact that all of these enzymes have a great similarity in their size (13 to 14 kDa), their three-dimensional structure (three oc helices more or less connected by 6 with 8 disulfide bridges) and their dependence on calcium necessary for catalytic activity at concentrations of the order of mM. They also have the same action mechanism based on a relay system of protons which involves several residues of the active site: histidine 48, glycine 30, aspartates 49 and 99 and tyrosines 52 and 73.

According to the French patent application filed under the number FR 99 06366, it has been shown that compounds having a heterocycle of the oxadiazolone type were capable of inducing a very good selectivity for PLA ₂ -snp of group II compared to PLA2 pancreatic group I. Compounds of this series, which are very active in vitro, have revealed in vivo an activity analogous to indomethacin (reference anti-inflammatory) on carrageenan edema on the rat's paw when administered intraperitoneally. On the other hand, the compounds described in French patent application No. FR 99 06366 had a low bioavailability by the oral route.

There is provided according to the invention a new family of group II PLA2 selective inhibitor compounds having an inhibitory activity much higher than that shown for previously known compounds, in particular the compounds described in French patent application No. FR 99 06366. supra. The new compounds according to the invention are characterized in particular by the presence of a substituted or unsubstituted piperazinyl ring on carbon atoms. The compounds according to the invention not only have the selective inhibitory activity on group II PLA ₂ while remaining completely inactive on group I pancreatic PLA _2, but also have an in vivo activity greater than that of indometacin. In addition, the compounds according to the invention have excellent bioavailability when administered orally.

The subject of the present invention is therefore a compound having the following general formula (I):

in which:

- D signifies the Z-HET group or the Z≈HET group and

(i) when D signifies the Z-HET group:

-HET is a 5-membered heterocycle such as oxadiazolone (II) or thiazolidine dione (III) of the following formulas: .4.

-Z- is chosen from the group consisting of - (CRιR2) _n - and - (CRi≈ CR ₂ ) n

- with n being an integer having a value ranging from 1 to 6, Ri and

R ₂ , identical or different, independently of one another represent a hydrogen atom or a linear or branched alkyl group having from 1 to 6 carbon atoms.

(ii) when D signifies the Z≈HET group:

-Z- represents, with the heterocycle, a group -Z≈HÉT of formula (IV) or (V) below:

in which -Z = represents -CRi≈, in which Ri represents a hydrogen atom or a linear or branched alkyl group of 1 to 6 carbon atoms; - p is an integer having the value 0 or 1;

-Y- is chosen from the group consisting of C≈O, SO2 and - (CR3R) m- with m being an integer having a value ranging from 1 to 6, R ₃ and R, identical or different, independently represent the one of the other a hydrogen atom or a linear or branched alkyl group having from 1 to 6 carbon atoms.

- A and B on the piperazine cycle, identical or different, independently of one another represent a carbon atom linked to hydrogens, a carbon atom linked both to a hydrogen and to a linear or branched alkyl group having 1 to 3 carbon atoms, or a -C≈O group.

- q is an integer having the value 0 or 1;

0 o

-W- is chosen from the group consisting of - c -, - 0 - c - and

0

__ _N _J__

H ^C ; -R is chosen from the group consisting of linear or branched alkyl groups having from 1 to 22 carbon atoms, the polyaryl groups and the aryl-alkyl, alkyl-Q-alkyl, alkyl-Q-aryl, aryl-Q-aryl and aryl-Q-alkyl groups for which:

"aryl" represents an aryl group having from 5 to 10 members, it is understood any aryl compound, substituted or unsubstituted, known to those skilled in the art, in particular the phenyl, naphthyl, phenylphenyl (or biphenyl) group or else a heterocyclic aryl such as the indolyl group. The aryl group is preferably substituted by one or more halogen atoms, such as F, Cl or Br, or also by groups chosen from CF ₃ , OH, MeO and N0 ₂ . "alkyl" represents a linear or branched alkyl group having from 1 to 12 carbon atoms, and

"Q" is selected from the group consisting of -O-, -S-, -NH-, -NRs-, -

O 0

0 0 M II

-NOT-

NH-CO-NH-, - o - c -, - c - o -, H and H, with R5 representing a linear or branched alkyl group having from 1 to 6 carbon atoms.

Overall, the compounds of formula (I) defined above have a selective inhibitory activity on PLA2 from group II, expressed as the concentration of compound (I) capable of inhibiting 50% of the enzymatic activity (IC50), which is in general less than 1 μM, often less than 0.5 μM and for some of these compounds is approximately 0.1 μM, while the most active of the compounds described in application FR 99 06366 has an inhibitory activity, expressed as an IC50 , 3 μM.

A family of preferred compounds according to the invention having a very high inhibitory activity selectively on human PLA ₂ of Group II is the family of compounds for which p is equal to 1 and Y represents the group C≈O, groups D, A , B, q, W and R having the meanings defined above.

According to a particularly advantageous embodiment of the present invention, the compounds of formula (I) above are chosen from the group consisting of: a) l- [4 ′ - (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-ylmethyl) benzyl] - 4-tetradecyl piperazine b) l- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo- oxadiazol-3-ylmethyl) benzoyl] -4-octa decylpiperazine c) l- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-ylmethyl) benzoyl] -2,5-di methyl-4-dodecylpiperazine d) l- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-ylmethyl) phenyl] - 4-octa decylpiperazine e) la [4- (4'- octadecylpiperazin-l'-ylcarbonyl) benzylidene] -1,3-thiazolidine-2,4-dione f) l- [4 '- (2,4-dioxo-1,3-thiazolidin-5-ylmethyl) benzoyl] - 4-octadecyl piperazine g) l- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-ylmethyl) benzyl] -4- tetradecylpiperazin-2-one h ) l- [4 '- (4,5-dihydro-l ₅ 2,4 (4H) -5-oxo-oxadiazol-3-ylethyl) benzoyl] -4- tetradecylpiperazine i) la 1 - [4' - ( 4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-ylpropyl) benzoyl] -4- tetradecylpiperazine j) l- [4 '- (4,5-dihydro-l, 2 , 4 (4H) -5-oxo-oxadiazol-3-yl-methylbenzoyl] -4- (N- octadecylaminocarbonyl) piperazine

The present invention also relates to a process for the preparation of the compounds of formula (I) described above. In general, as illustrated by the examples below, the preparation process can be chosen from processes I and II below:

- Method I: reacting hydroxylamine hydrochloride with a derivative of formula (VI):

where R, q, W, A, B, p, and Y are defined as above and:

- Z- is chosen from the group consisting of - (CRiR∑Jn- and - (CRi≈ CR2) n - with n being an integer having a value ranging from 1 to 6, Ri and

R2, identical or different, independently of one another represent a hydrogen atom or a linear or branched alkyl group having from 1 to 6 carbon atoms, or to form the corresponding intermediate oxime, then to subject this oxime cyclization by reaction with a chlorocarbonate (or chloroformate) followed by heating to a temperature sufficient to achieve substantially complete cyclization; a) - Method II: reacting the thiazolidine-2,4-dione on the aldehyde function of the derivative of formula (VII):

in which R, q, W, A, B, p and Y are defined as above and, -r is an integer having the value 0 or 1, -U- is chosen from the group consisting of - (CR ₆ R7) s - and - (CRβ≈ CR7) _S - with s being an integer having a value ranging from 1 to 6 and with R ₆ and R ₇ , identical or different, representing a hydrogen atom or a linear or branched alkyl group having from 1 to 6 carbon atoms. This reaction is carried out in refluxing toluene in the presence of pyridinium benzoate to form the ethylenic derivative (V) as defined above, and followed by a possible reduction of the Z≈C double bond by catalytic hydrogenation (Parr apparatus ) in the presence of 100% black palladium and hydrogen under pressure in absolute ethanol at 50 ° C. The starting materials for the above process steps can be prepared according to methods well known to those skilled in the art (see in particular Examples 1 to 6 below).

The invention also relates to the use of a compound of formula (I) as defined above for selectively inhibiting a group II PLA ₂ , preferably in an in vitro test.

The present invention further relates to a pharmaceutical composition characterized in that it comprises at least one compound of general formula (I) described above in association with at least one excipient chosen from the group consisting of pharmaceutically acceptable excipients.

To formulate a pharmaceutical composition according to the invention, a person skilled in the art may advantageously refer to the last edition of the European Pharmacopoeia or the Pharmacopoeia of the United States of America (USP).

The skilled person may especially advantageously refer to the ^4th edition "2002" of the European Pharmacopoeia, or the edition USP 25-NF20 the USP (US Pharmacopeia).

Advantageously, a pharmaceutical composition as defined is suitable for daily oral or parenteral administration of an amount of a compound of formula (I) of between 1 μg and 10 mg and preferably between 1 μg and 1 mg per kg of weight of the patient.

Advantageously, a pharmaceutical composition as defined above is suitable for local, preferably topical, daily administration of an amount of a compound of formula (I) of between Iμg and 100 mg and preferably between 100 μg and 10 mg per kg of patient weight.

When the composition according to the invention comprises at least one pharmaceutically acceptable excipient, it is in particular an excipient suitable for administration of the composition by topical route, an excipient suitable for administration of the composition by oral route and / or an excipient suitable for administration of the composition by the parenteral route.

Finally, with particular reference to the study of biological activity below, the present invention also relates to a compound of general formula (I) as described above, for its use as a therapeutically active principle in a drug.

More particularly, the subject of the invention is the use of at least one compound of general formula (I) as described above for the preparation of a medicinal composition intended to inhibit the activity of non-secreted PLA ₂ human pancreatic group IL

The present invention also relates to the use of at least one compound of general formula (I) as described above, for the preparation of a medicament intended for the prevention or treatment of inflammation, in particular of chronic inflammation and acute inflammation, that is to say, in particular inflammatory pathologies in which the non-pancreatic secreted PLA2 is involved.

It is therefore very particularly the inflammatory pathologies mentioned above, namely rheumatoid arthritis, septic shock, whether the cause is an infection, peritonitis, malaria or even aspirin poisoning, respiratory collapse , hypotension, respiratory distress syndrome, asthma, allergic rhinitis, acute lung injury, asbestosis, ischemia, atherosclerosis, cardiovascular morbidity, Crohn's disease, ulcerative colitis, intestinal inflammation, cirrhosis, acute pancreatitis, psoriasis, cell and tissue damage from cerebral ischemia and schizophrenia and other conditions such as multiple sclerosis and certain cancers of the breast and gastrointestinal tract .

The present invention also relates to the use of at least one compound of general formula (I) as described above, for the preparation of a medicament intended for the treatment of rheumatic disorders. The invention also relates to a method for treating an inflammation state in a patient, preferably a chronic inflammation state or an acute inflammation state, said method comprising a step during which the patient is administered a therapeutically effective amount of a compound of formula (I) or of a pharmaceutical composition containing a compound of formula (I).

The subject of the invention is also a method for preventing a state of inflammation in a patient, said method comprising a step during which the patient is administered a therapeutically effective amount of a compound of formula (I) or of a pharmaceutical composition containing a compound of formula (I).

The compound of formula (I) or the pharmaceutical composition containing the compound of formula (I) can be administered orally, parenterally or even be applied topically, locally to the skin of a patient. The following examples are intended to illustrate the present invention and should in no way be interpreted as being capable of limiting its scope.

Example 1: Preparation of l-r4 4,5-dihydro-1,2,4 (4110-5-oxo-oxadiazol-3-ylmethyl) benzyl1-4-tetradecylpiperazine

(compound of formula (I) in which D = Z-HET with HET ≈ oxadiazolone (II), Z = -CH ₂ -, or n ≈ 1, Y = -CH ₂ -, A = B = -CH ₂ -, R = -

1.1- Preparation of 1-tetradecylpiperazine

13 g (0.151 mole) of piperazine dissolved in 100 ml of a THF / CH ₂ Cl2 mixture, 3: 1, v / v, are stirred in a 250 ml enmeyer flask. 4.24 g (15 mmol) of 1-bromotetradecane are added to the mixture and stirring is continued at room temperature for one hour. The solvents are then evaporated, the residue obtained is taken up in dichloromethane and then washed twice with water. The organic phase is dried over MgSO ₄ , filtered and evaporated. Crystallization from an acetone / ether mixture at -18 ° C leads to 3.4 g of white crystals which melt at room temperature. Yield: 80%. Rf: 0.40 (CH ₂ Cl2 / MeOH / NH ₄ OH, 80: 20: 2, v / v / v).

IR (KBr): 3440 (NH) cm- ⁱ

N NMR (200 MHz, CDCls, HMDS) δ ppm: 6-8 (solid, 1H, NH), 2.85 and 2.33 (2t, 8H, / = 4.88 and 4.50 Hz, H of the piperazine), 2.21 (t, 2H, / ≈ 7.56 Hz, CH ₂ -N), 1.40 (m, 2H, CH ₂ -CN), 1.20 (si, 22H, CH2), 0 , 80 (t, 3H, / ≈ 6.62 Hz, CH ₃ ).

1.2- Preparation of 1- (4'-cyanomethylbenzyl) -4- tetradecylpiperazine a) Preparation of 4-bromomethylphenyl acetonitrile

In a liter liter erlenmeyer flask, 25 g (0.19 mole) of

4-methylphenyl acetonitrile in 300 ml of carbon tetrachloride. 41 g (0.23 mol) of N-bromosuccinimide (NBS), previously recrystallized from acetic acid, are added, as well as 0.5 g of 2.2'- azobis (2-methyl-piOpionitrile) (AIBN). The solution is heated at reflux for 3 hours. At the end of the reaction, the mixture is cooled and then washed three times with water. The organic phase is then dried over MgSO ₄ , filtered and evaporated in vacuo. A distillation under reduced pressure (1 mm Hg) of the residue obtained makes it possible to collect, successively, three fractions at 95, 110 and 140 ° C. The latter corresponds to the expected 4-bromomethylphenyl acetonitrile which, by crystallization from ether at -18 ° C, leads to 14 g of white crystals. Yield: 35%. Melting point: 63 ° C. Rf: 0.19 (ether / petroleum ether, 30:70, v / v).

IR (KBr): 2224 (CNN), 1594 (C≈Car) cm- ¹

N NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.34 and 7.24 (2d, 4H, / = 8.30 and

8.27 Hz, aromatic H), 4.41 (s, 2H CH ₂ -Br), 3.68 (s, 2H, CH ₂ -G≡N).

b) Preparation of 1- (4'-cyanomethylbenzyl) -4-tetradecylpiperazine

7 g (24 mmol) of 1- tetradecylpiperazine, 6 g (28 mmol) of 4-bromomethylphenyl acetonitrile, 9 7 g (24 mmol) are mixed in a 250 ml erlen eyer, topped with a condenser and a calcium chloride guard. , 93 g (71 mmol) of potassium carbonate as well as 0.5 g of potassium iodide in 200 ml of acetonitrile. The reaction mixture is heated at reflux for 6 hours. At the end of the reaction, the suspension is filtered and the K ₂ C0 ₃ rinsed several times with dichloromethane. The solvents are then evaporated under vacuum and the residue obtained is taken up in 150 ml of dichloromethane then washed with water until neutral pH. The organic phase is then dried over MgSO ₄ , filtered and concentrated under reduced pressure. The residue is purified by chromatography on a column of silica gel using the MeOH / CTfcC mixture, 1:99, v / v as eluent, to give 8.2 g of the title nitrile in the form of a brownish oil. Yield: 83%. Rf: 0.33 (CH ₂ Cl ₂ / MeOH, 95: 5, v / v).

IR (KBr): 2248 (C≡N), 1607 (C≈Car) cm- ⁱ

NMR m (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.26 and 7.18 (2d, 4H, / = 8.09 and 9.52 Hz, aromatic H), 3.64 (s, 2H, CH -C≡N), 3.42 (s, 2H, Ph-CH ₂ -N), 2.40 (m, 8H, H of piperazine), 2.25 (t, 2H, / = 7.64 Hz , CH2-N), 1.39 (m, 2H, CH2-CN), 1.18 (si, 22H, CH ₂ ), 0.80 (t, 3H, / = 6.13 Hz, CH3). 1.3- Preparation of l- [4 '- (N- hydroxyamidinomethyl) benzyl] -4-tetra decyl piperazine

13.08 g (94 mmol) of potassium carbonate 5.48 g (78 mmol) of hydroxylamine hydrochloride in 150 ml are suspended in a 250 ml Erlenmeyer flask topped with an addition funnel and a condenser of absolute ethanol and the mixture is heated to reflux. 6.5 g (15 mmol) of 1- (4'-cyanomethyl benzyl) -4- tetradecylpiperazine dissolved in 50 ml of absolute ethanol are then added dropwise to this suspension. The reaction mixture is kept under stirring and at reflux for 24 hours. At the end of the reaction, the salt is filtered hot and washed several times with dichloromethane. The filtrate is concentrated under reduced pressure and then taken up in dichloromethane. The organic phase obtained is washed with water until neutral, dried over MgSO ₄ and then filtered. After evaporation of the solvent, the product crystallizes from acetone and leads to 4.62 g of white crystals of title amidoxime. Yield: 65%. Melting point: 74 ° C. Rf: 0.28 (CH ₂ Cl ₂ / MeOH, 90:10, v / v).

IR (KBr): 3490 (OH), 3374 (NH ₂ ), 1655 (C = N), 1607 (C≈Car) cm- ¹

NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.20 and 7.14 (2d, 4H, / = 7.39 and 8.10 Hz, aromatic H), 4.41 (s, 2H, NH ₂ ), 3.41 (s, 2H, CH ₂ -C = N), 3.35 (s, 2H, Ph-CH ₂ -N), 2.41 (m, 8H, piperazine H), 2, 25 (t, 2H, / = 7.64 Hz, CH ₂ -N), 1.36 (m, 2H, CH2-CN), 1.18 (si, 22H, CH ₂ ), 0.80 (t, 3H, / = 6.13 Hz, CH ₃ ).

1.4- Preparation of l- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-yl methyl) benzyl] -4-tetradecylpiperazine

This synthesis is carried out in two stages. 1.8 g (4 mmol) of amidoxime and 0.66 ml (4 mmol) of triethylamine in 40 ml of anhydrous dichloromethane are dissolved in a 100 ml flask. The solution is stirred at 0 ° C for one hour and then 0.60 ml (5 mmol) of phenyl chloroformate is added dropwise to the reaction mixture. After one hour of stirring at 0 ° C, the solution is washed successively with an alkaline solution (saturated Na ₂ Cθ ₃ ) and three times with water, dried over MgS0 ₄ , filtered and then concentrated in vacuo. The carbonate intermediate obtained is then taken up in 40 ml of dry toluene and heated for 12 hours at reflux. The toluene is then evaporated under reduced pressure and the residue obtained is purified by chromatography on a column of silica gel eluted with a mixture of EfeCfe / MeOH, 98: 2, v / v. The product crystallizes from an acetone / ether mixture and leads to 500 mg of white crystals of the terminal compound. Yield: 26%. Melting point: 98 ° C. Rf: 0.33 (CH ₂ Cl ₂ / MeOH, 90:10, v / v).

IR (KBr): 1732 (NC≈O), 1688 (C = N), 1599 (C≈Car) cm- ¹

N NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 8-12 (solid, IH, NH), 7.27 and 7.18 (2d, 4H, / = 8.07 and 9.12 Hz, aromatic H ), 3.67 (s, 2H, CH ₂ -C = N), 3.35 (s, 2H, PI1-CH2-N), 2.56 and 2.34 (2m, 8H, H of piperazine) , 2.46 (t, 2H, / = 7.64 Hz, CH ₂ -N), 1.47 (m, 2H, CH ₂ -CN), 1.18 (si, 22H, CH ₂ ), 0, 80

Example 2: Preparation of l-f4 '- (4,5-dihydro-1,2,4 (4jH) -5-oxo-oxadiazol-3-yl methyl) benzoyl -4-octadecylpiperazine (compound of formula (I) in which D = Z-HET with HET = oxadiazolone (II), Z = -CH ₂ -, i.e. n = 1, Y = C≈O, A = B = -CH ₂ -, R = -

2.1- Preparation of 1-octadecylpiperazine Following the same protocol as that described for step 1.1 of Example 1, but starting with 13 g (0.151 mole) of piperazine and 5 g (15 mmol) of 1-bromooctadecane 4.5 g of white crystals are obtained after crystallization from acetone. Yield: 89%. Melting point: 61.5 ° C. Rf: 0.40 (CH ₂ Cl ₂ / MeOH / NH ₄ OH, 80: 20: 2, v / v / v).

IR (KBr): 3440 (NH) cm- ¹

N NMR (200 MHz, CDCI3, HMDS) δ ppm: 6-8 (si, IH, NH), 2.85 and 2.33 (2t, 8H, J = 4.88 and 4.50 Hz, H of the piperazine), 2.21 (t, 2H, / = 7.56 Hz, CH ₂ -N), 1.40 (m, 2H, CH ₂ -CN), 1.20 (si, 30H, CH ₂ ), 0.80 (t, 3H, / = 6.62 Hz, CH ₃ ). 048139

14 -

2.2- Preparation of 4-bromomethylbenzoyl chloride

8.4 g (54 mmol) of 4-methylbenzoyl chloride and 9.66 g (54 mmol) of N-brornosuccinimide are dissolved in a 250 ml flask surmounted by a condenser and a calcium chloride guard. (NBS) previously recrystallized from acetic acid and 0.5 g of 2,2'-azobis (2-methylproρionitrile) (AIBN) in 150 ml of carbon tetrachloride. The solution is heated at reflux for 3 hours. At the end of the reaction, the salt is filtered, the solution is cooled and then washed three times with water. The organic phase is dried over MgSO ₄ , filtered and concentrated in vacuo. The residue is then crystallized from pentane and 9 g of white crystals are obtained. Yield: 72%. Melting point: 86.7 ° C.

NMR * H (200 MHz, CDC1 ₃ , HMDS) δ ppm: 8.02 and 7.46 (2d, 4H, / = 8.38 and 8.29 Hz, aromatic H), 4.43 (s, 2H, CH ₂ -Br).

2.3- Preparation of l- (4'-bromomethylbenzoyl) -4- octadecylpiperazine

4.4 g (13 mmol) of octadecylpiperazine and 2.7 ml (19 mmol) of triethylamine in 100 ml are dissolved in a 250 ml Erlenmeyer flask topped with an addition vial and a calcium chloride guard. dry benzene. The mixture is stirred at 0 ° C. and then 3.04 g (13 mmol) of 4-bromomethylbenzoyl chloride are added dropwise. After two hours of stirring at room temperature, the solvent is evaporated, the residue obtained is taken up in dichloromethane, washed with an alkaline solution and then several times with water until neutral. The organic phase is then dried over

MgS0 ₄ , filtered and concentrated in vacuo. The product is purified by chiOmatography on a column of silica gel with dichloromethane as eluent. 5 g of pure product are thus obtained in the form of an oil.

Yield: 72%. Rf: 0.50 (CH ₂ Cl ₂ / MeOH, 95: 5, v / v).

IR (KBr): 1624 (NC = 0), 1607 (C≈Car) cm- ¹

NMR (200 MHz, CDCI3, HMDS) δ ppm: 7.34 (s, 4H, aromatic H), 4.52 (s, 2H, CH ₂ -Br), 3.72 and 3.38 (2m, 4H, CH ₂ -NC≈Q of piperazine), 48139

15

2.43 (m, 4H, H of piperazine), 2.33 (t, 2H, / = 7.65 Hz, CH ₂ -N), 1.41 (m, 2H, CH ₂ -CN), 1 , 18 (if, 30H, CH ₂ ), 0.80 (t, 3H, / = 6.13 Hz, CH3).

2.4- Preparation of 1- (4'-cyanomethylbenzoyl) -4- octadecylpiperazine

5.35 g (10 mmol) of the bromine derivative prepared in step 2.3 above are dissolved in 70 ml of dimethyl sulfoxide in a 250 ml erlenmeyer flask surmounted by a condenser and a calcium chloride guard. The solution is stirred at 0 ° C. and then 1.96 g (40 mmol) of sodium cyanide are added little by little to the reaction mixture. The whole is then brought to room temperature and then heated at 80 ° C for 1 hour. The reaction mixture is diluted with dichloromethane and with water. The organic phase is washed with water several times, dried over MgSO 4, filtered and then concentrated in vacuo. The residue is purified by chromatography on a column of silica gel with dichloromethane as eluent. 3 g of pure title nitrile are then obtained in the form of a dark honey-colored oil. Yield: 61%. Rf: 0.5 (CH ₂ Cl ₂ / MeOH, 97: 3, v / v).

IR (KBr): 2251 (C≡N), 1620 (NC≈O), 1607 (C≈Car) cm- ¹

NMR m (200 MHz, CDCI3, HMDS) δ ppm: 7.36 and 7.30 (2d, 4H, / = 8.51 and 8.48 Hz, aromatic H), 3.71 (s, 2H, CH ₂ -C≡N), 3.72 and 3.38 (2m, 4H, CH ₂ -NC = 0 of piperazine), 2.43 (m, 4H, H of piperazine), 2.33 (t, 2H , / = 7.65 Hz, CH ₂ -N), 1.41 (m, 2H, CH ₂ -CN), 1.18 (si, 30H, CH ₂ ), 0.81 (t, 3H, / = 6.13 Hz, CH ₃ ).

2.5- Preparation of l- [4 '- (N- hydroxyamidinomethyl) benzoyl] -4 ~ octadecyl piperazine

Following the same protocol as that described for step 1.3 of Example 1, but starting with 6 g (12 mmol) of 1- (4'- cyanomethylbenzoyl) -4-octadecylpiperazine, 10.26 g (74 mmol ) potassium carbonate and 4.30 g (61 mmol) hydroxylamine hydrochloride. The crude product is crystallized from acetone and leads to 4 g of white crystals of the title oxime. Yield: 67%. Melting point: 105.2 ° C. Rf: 0.39 (CH ₂ Cl ₂ / MeOH, 90:10, v / v). 048139

16 -

IR (KBr): 3486 (OH), 3373 (NH ₂ ), 1657 (NC≈O), 1625 (C = N), 1582 (C≈Car) cm--

NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.30 and 7.24 (2d, 4H, J = 8.28 and 8.11 Hz, aromatic H), 4.43 (s, 2H, NH ₂ ), 3.42 (s, 2H, CH ₂ -C = N), 3.73 and 3.40 (2m, 4H, CH ₂ -NC = 0 of piperazine), 2.55 (m, 4H, H piperazine), 2.29 (t, 2H,} = 6.64 Hz, CH ₂ -N), 1.39 (m, 2H, CH2-CN), 1.18 (si, 30H, CH ₂ ) , 0.81 (t, 3H, J = 6.03 Hz, CH ₃ ).

2.6- Preparation of l- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-yl methyl) benzoyI] -4-octadecylpiperazine

Following the same protocol as that described for step 1.4 of Example 1, but starting with 1.3 g (2.53 mmol) of amidoxime prepared in the previous step 2.5, 0.45 ml (3 , 28 mmol) of triethylamine as well as 0.4 ml (3.03 mmol) of phenyl chloroformate. The product crystallizes from acetone and leads to 500 mg of white crystals of terminal compound. Yield: 37%. Melting point: 121 ° C. Rf: 0.38 (CH ₂ Cl ₂ / MeOH, 90:10, v / v).

IR (KBr): 1780 (OC≈O), 1734 (C≈N), 1640 (NC≈O), 1607 (C≈Car) cm- ¹

¹ H NMR (200 MHz, CDCI3, HMDS) δ ppm: 8-12 (solid, IH, NH), 7.16 (s, 4H, aromatic H), 3.79 (s, 2H, CH ₂ -C = N), 3.77 and 3.36 (2m, 4H, CH ₂ -N- C≈O of piperazine), 2.52 (m, 4H, H of piperazine), 2.35 (t, 2H, } = 5.86 Hz, CH ₂ -N), 1.43 (m, 2H, CH2-CN), 1.18 (si, 30H, CH ₂ ), 0.81 (t, 3H, J

Example 3: Preparation of l-r4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-yl methyl) benzovn-2,5-dimethyl-4-dodecylpiperazine

(compound of formula (I) in which D = Z-HET with HET = oxadiazolone (II), Z = -CH ₂ -, or n = 1, Y = C≈O, A = B = CH-CH3, R =

3.1- Preparation of 2,5-dimethyl-1-dodecylpiperazine

Following the same protocol as that described for step 1.1 of Example 1, but starting with 3.27 g (13 mmol) of bromide 048139

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dodecane and 12 g (0.105 mol) of trans-2,5-dimethylpiperazine in 170 ml of THF, 2.8 g of the title substituted piperazine are recovered in the form of an oil. Yield: 76%. Rf: 0.3 (CH ₂ Cl ₂ / MeOH / NH4θH, 80: 20: 2, v / v / v).

IR (KBr): 3440 (NH) cm " ¹

N NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 6-8 (solid, IH, NH), 2.29 (m, 8H, CH2-N and H of piperazine), 1.36 (m, 5H , CH3 on piperazine and CH2-CN), 1.19 (if, 18H, CH ₂ ), 0.98 (if, 3H, CH3 on piperazine), 0.81 (t,

3.2- Preparation of l- (4'-chloromethylbenzoyl) -2,5- dimethyl-4-dodé cylpiperazine

In a 250 ml Erlenmeyer flask are dissolved 6 g (21 mmol) of the substituted piperazine prepared in step 3.1 above and 3.18 g (31 mmol) of triethylamine in 150 ml of benzene. The mixture is stirred at 0 ° C. and then 4.82 g (25 mmol) of 4-chloromethylbenzoyl chloride (commercial or prepared according to the same protocol as that described in step 2.2 of Example 2 but using N- chlorosuccinimide) are added dropwise. After 3 hours of stirring at room temperature, the benzene is evaporated, the residue taken up in dichloromethane is washed with a saturated solution of Na ₂ Cθ3 and then twice with water. The organic phase is dried over MgSO ₄ , filtered and then concentrated in vacuo. The residue is then purified by chromatography on a column of silica gel with dichloromethane as eluent. 4.33 g of the pure title chlorinated derivative are thus obtained in the form of an oil. Yield: 48%. Rf: 0.36 (CH ₂ Cl ₂ / MeOH, 95: 5, v / v).

IR (KBr): 1624 (NC≈O), 1595 (C≈Car) cm- ¹

¹ H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.34 and 7.27 (2d, 4H, J = 8.40 and 7.96 Hz, aromatic H), 4.52 (s, 2H, CH2-CI), 3.36 and 2.62 (2d, 4H, J = 7.40 and 7.67 Hz, CH ₂ of piperazine), 2.88 and 2.28 (2m, 2H, CH of the piperazine), 2.24 (t, 2H, J = 5.52 Hz, CH ₂ -N), 1.40 (m, 2H, CH ₂ -CN), 1.35 048139

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and 0.94 (2d, 6H, CH ₃ on piperazine), 1.18 (if, 18H, CH ₂ ), 0.81 (t, 3H,} = 6.74 Hz, CH ₃ ).

3.3- Preparation of l- (4'-cyanomethylbenzoyI) -2,5-dimethyl-4-dodecyl piperazine

4.33 g (9.96 mmol) of the chlorine derivative prepared in step 3.2 above are dissolved in 50 ml of DMSO. To this solution stirred at 0 ° C. are gradually added 1.49 g (29 mmol) of sodium cyanide. After total addition, the solution is heated at 80 ° C for 1 hour. At the end of the reaction, an extraction is carried out by adding to the mixture, dichloromethane and water. The organic phase is washed twice with water, dried over MgSθ4, filtered and then concentrated in vacuo. The residue obtained is purified by chromatography on a column of silica gel with dichloromethane as eluent. 4 g of the pure title nitrile are then recovered in the form of an oil. Yield: 94%. Rf: 0.5 (CH ₂ Cl ₂ / MeQH, 95: 5, v / v).

IR (KBr): 2245 (G≡N), 1611 (NC≈O), 1607 (C≈Car) cm- ¹ NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.30 (s, 4H, H aromatic), 3.71 (s, 2H, CH ₂ - C≡N), 3.36 and 2.62 (2d, 4H,} ≈ 7.40 and 7.67 Hz, piperine CH ₂ ), 2 , 88 and 2.28 (2m, 2H, piperazine CH), 2.24 (t, 2H, J = 5.52 Hz, CH -N), 1.40 (m, 2H, CH2-CN), 1.28 and 0.84 (2d, 6H, CH ₃ on piperazine), 1.18 (if, 18H, CH ₂ ), 0.81 (t, 3H, J = 6.74 Hz, CH3).

3.4- Preparation of 1- [4 '- (N-hydroxyamidinomethy) benzoyl] -2,5-di methyl-4-dodecylpiperazine

Following the same protocol as that described for step 1.3 of Example 1, but starting with 4 g (9.41 mmol) of the nitrile prepared in the previous step 3.3, 3.26 g (47 mmol) of hydroxylamine hydrochloride and 7.79 g (56 mmol) of potassium bicarbonate.

After purification, 1.6 g of the title amidoxime are recovered in the form of an oil. Yield: 37%. Rf: 0.46 (CH ₂ Cl ₂ / MeOH, 90:10, v / v). 048139

19

IR (KBr): 3369 (OH), 3328 (NH.), 1661 (NC≈O), 1612 (C≈N), 1595 (C ^≈ Car) cm- ¹

N NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.30 (s, 4H, aromatic H), 3.36 and 2.62 (2d, 4H, J = 7.40 and 7.67 Hz, CH ₂ of piperazine), 5.70 (si, IH, OH), 4.44 (si, 2H, NH ₂ ), 3.39 (s, 2H, CH ₂ - C≈N), 2.88 and 2 , 28 (2m, 2H, piperazine CH), 2.24 (t, 2H,} = 5.52 Hz, CH ₂ -N), 1.40 (m, 2H, CH2-CN), 1.28 and 0.84 (2d, 6H, CHs on piperazine), 1.18 (if, 18H, CH ₂ ), 0.81 (t, 3H, 7 = 6.74 Hz, CH ₃ ).

3.5- Preparation of l- [4 '- (4,5 ~ dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-yl methyl) benzoyl] -2,5-dimethyl -4- do décylpipérazine

Following the same protocol as that described for step 1.4 of Example 1, but starting with 1.6 g (3.49 mmol) of the amidoxime prepared in the previous step 3.4, 0.58 ml ( 4.19 mmol) of triethylamine and 0.48 ml (3.83 mmol) of phenyl chloroformate. The crude residue is purified by chromatography on a column of silica gel with dichloromethane as eluent. 600 mg of pure terminal compound are thus obtained in the form of a foam. Yield: 35%. Rf: 0.4 (CH ₂ Cl ₂ / MeOH, 90:10, v / v).

IR (KBr): 1670 (NOC≈O), 1634 (C≈N), 1595 (C = C _ar ) cm- ¹ NMR (200 MHz, CDCI3, HMDS) δ ppm: 7.14 (s, 4H, H aromatic), 6.11 (si, 1H, NH), 3.74 (s, 2H, CH ₂ -C = N), 3.36 and 2.62 (2d, 4H,] = 7.40 and 7, 67 Hz, piperazine CH ₂ ), 2.88 and 2.28 (2m, 2H, piperazine CH), 2.24 (t, 2H, 7 = 5.52 Hz, CH2-N), 1, 40 (m, 2H, CH2-CN), 1.28 and 0.84 (2d, 6H, CH ₃ on piperazine), 1.18 (si, 18H, CH ₂ ), 0.81 (t, 3H, J = 6.74 Hz, CH ₃ ).

Example 4 Preparation of l-r4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-yl methyl) phenyπ-4-oeta decylpiperazine

(compound of formula (I) in which D = Z-HET with HET = oxadiazolone (II), Z = -CH ₂ -, or n = 1, p is 0, A = B = -CH ₂ -, R = - 48139

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4.1- Preparation of N-octadecyldiethanoamine

10 g (95 mmol) of diethanolamine, 37.96 g (0.114 mole) of octadecane bromide, 39.33 g (0.285 mole) of potassium bicarbonate and 0.5 g are mixed in a 500 ml erlenmeyer flask potassium iodide in 200 ml of acetonitrile. The reaction mixture is stirred and heated to reflux for 3 hours. At the end of the reaction, the salts are filtered, the solvent is evaporated and the residue is taken up in dichloromethane. The organic phase is washed twice with water, dried over MgSO ₄ , filtered and then concentrated in vacuo and the residue is allowed to crystallize from acetone. 33 g of white crystals are thus obtained. Yield: quantitative. Melting point: 49 ° C. Rf: 0.20 (CH ₂ Cl ₂ / MeOH, 90:10, v / v).

IR (KBr): 3310 (OH) cm " ¹ NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 3.53 (t, 4H,} = 5.43 Hz, CH ₂ - O), 3.27 ( if, 2H, OH), 2.57 (t, 4H, J = 5.43 Hz, N-CH ₂ -C-0), 2.44 (t, 2H,} = 7.06 Hz, CH ₂ - N), 1.34 (m, 2H, CH ₂ -CN), 1.18 (si, 30H, CH2), 0.80 (t, 3H,} = 5.85 Hz, CH3).

4.2- Preparation of N, N'-di (chloroethyl) octadecylamine

13 g (36 mmol) of N-octadecyldiethanolamine in 100 ml of chloroform are dissolved and cooled to 0 ° C. in a 250 ml Erlenmeyer flask. 7.95 ml (0.109 mole) of thionyl chloride are then added dropwise. At the end of the addition, the reaction mixture is heated at reflux of chloroform for 3 hours. The solvent and excess thionyl chloride are evaporated and the residue taken up in dichloromethane is washed with a saturated solution of Na2Cθ3 and then several times with water until neutral. The organic phase is dried over MgSO ₄ , filtered then concentrated in vacuo and the residue purified by chromatography on a column of silica gel with an ether / petroleum ether mixture, 5:95, v / v as eluent. 10 g of pure chlorinated amine are thus recovered in the form of an oil. Yield: 70%. Rf: 0.43 (ether / petroleum ether, 5:95, v / v). 048139

• 21

^I H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 3.38 (t, 4H,} = 5.43 Hz, CH ₂ - Cl), 2.75 (t, 4H, 7 = 7.30 Hz , N-CH2-C-CI), 2.43 (t, 2H,} = 6.67 Hz, CH2-N), 1.36 (m, 2H, CH2-CN), 1.16 (si, 30H , CH ₂ ), 0.80 (t, 3H,} = 5.85 Hz, CH3).

4.3- Preparation of 1- (4'-cyanomethylphenyl) -4- octadecylpiperazine

In a 250 ml flask are mixed 3 g (7.6 mmol) of N, N'-di (chloroethyl) octadecylamine, 2 g (15 mmol) of 4-aminophenylacetonitrile and 0.2 g of potassium iodide in 100 ml of acetonitrile. The suspension is stirred at reflux for 16 hours. At the end of the reaction, the solvent is evaporated, the residue taken up in dichloromethane is washed with a basic solution and then with water several times. The organic phase is dried over MgSθ4, filtered and then concentrated in vacuo. The residue obtained crystallizes from acetone and leads to 2.66 g of white crystals of the title disubstituted piperazine. Yield: 77%. Melting point: 94 ° C. Rf: 0.33 (CH ₂ Cl ₂ / MeOH, 98: 2, v / v).

IR (KBr): 2252 (C≡N), 1607 (C≈Car) cm- ¹

^I H NMR (200 MHz, CDCI3, HMDS) δ ppm: 7.17 and 6.84 (2d, 4H, J = 8.60 and 8.63 Hz, aromatic H), 3.62 (s, 2H, CH ₂ -C≡N), 3.61 and 3.22 (2sl, 8H, piperazine H), 2.92 (t, 2H, J = 8.32 Hz, CH2-N), 1.85 (m , 2H, CH ₂ -CN), 1.19 (si, 30H, CH ₂ ), 0.81 (t, 3H, J = 5.90 Hz, CH3).

4.4- Preparation of l- [4 '- (N- hydroxyamidinomethyl) phenyl] -4-octa decylpiperazine

Following the same protocol as that described for step 1.3 of Example 1, but starting from 1.52 g (21 mmol) of hydroxylamine hydrochloride, 3.64 g (26 mmol) of potassium carbonate and 2 g (4 mmol) of the nitrile obtained in step 4.3 above. The crude product is purified by chromatography on a column of silica gel with dichloromethane as eluent and gives an oil which crystallizes from acetone. 600 mg of white crystals of the title amidoxime are thus recovered. Yield: 28%. Melting point: 110.1 ° C. Rf: 0.40 (CH ₂ Cl ₂ / MeOH, 95: 5, v / v). 48139

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IR (KBr): 3489 (OH), 3375 (NH ₂ ), 1655 (C≈N), 1607 (C≈Car) cm- ¹ NMR Η (200 MHz, CDCI3, HMDS) δ ppm: 7.08 and 6 , 80 (2d, 4H,} = 8.60 and 8.62 Hz, aromatic H), 4.36 (s, 2H, NH ₂ ), 3.44 (s, 2H, CH ₂ -C = N), 3.14 and 2.56 (2sl, 8H, piperazine H), 2.34 (t, 2H,} = 7.34 Hz, CH ₂ -N), 1.47 (m, 2H, CH2-CN ), 1.19 (if, 30H, CH ₂ ), 0.81 (t, 3H, J = 6.05 Hz, CH3).

4.5- Preparation of l- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-yl methyl) ρhenyl] -4-octadecyl piperazine

Following the same protocol as that described for step 1.4 of Example 1, but starting from 600 mg (1.2 mmol) of the amidoxime prepared in the previous step 4.4, 0.22 ml (1, 6 mmol) of triethylamine and 0.2 ml (1.6 mmol) of phenyl chloroformate. The product crystallizes from acetone and leads to 210 mg of white crystals of terminal compound. Yield: 33%. Melting point: 147.3 ° C. Rf = 0.35 (CH ₂ Cl ₂ / MeOH, 95: 5, v / v).

IR (KBr): 1740 (OC≈O), 1716 (C≈N), 1607 (C≈Car) cm- ¹ NMR m (200 MHz, CDCI3, HMDS) δ ppm: 7.33 (s, IH, NH ), 7.12 and 6.74 (2d, 4H, 7 = 8.62 and 8.60 Hz, aromatic H), 3.71 (s, 2H, CH ₂ -C = N), 3.14 and 2 , 56 (2sl, 8H, piperazine H), 2.34 (t, 2H,} = 7.34 Hz, CH2-N), 1.47 (m, 2H, CH2-CN), 1.19 ( if, 30H, CH ₂ ), 0.81 (t, 3H, J = 6.05 Hz, CH3).

Example 5 Preparation of f4- (4'-octadecylpiperazin-1 ^/ -ylcarbonyl) benzylidene1-1,3-thiazolidine-2,4-dione

(compound of formula (I) in which D≈Z≈HET with HET≈ the compound of formula (V), Z = CH≈ Y = C≈O, A ≈ B = -CH ₂ -, R = - (CH ₂ ) ι ₇ -CH ₃ )

5.1- Preparation of 4-formylbenzoyl chloride

In a 250 ml Erlenmeyer flask, 5 g (33 mmol) of acid are dissolved

4-formyl benzoic in 100 ml of chloroform. The mixture is stirred at

0 ° C then 3.63 ml (49 mmol) of thionyl chloride dissolved in 50 ml of chloroform are added dropwise. Once the addition is complete, the reaction mixture is heated at 40 ° C for 3 hours. In 048139

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end of reaction, the mixture is evaporated and the residue obtained taken up in dichloromethane, is washed with a saturated solution of Na ₂ Cθ3 and then twice with water. The organic phase is dried quickly over MgSO ₄ , filtered and then concentrated in vacuo. 4 g of title acid chloride are thus recovered in the form of a colorless oil used in the next step without purification. Yield: 71%.

IR (KBr): 1779 (aldehyde C≈O), 1756 (CIC≈O) cm- ¹ NMR Ή (200 MHz, CDC1 ₃ , HMDS) δ ppm: 10.10 (s, IH, H aldehyde), 8, 18 and 7.96 (2d, 4H, J = 8.42 and 8.22 Hz, aromatic H).

5.2- Preparation of l- (4'-formylbenzoyl) -4- octadecylpiperazine

In a 250 ml Erlenmeyer flask surmounted by an addition funnel and a calcium chloride guard, 4 g (11 mmol) of octadecylpiperazine (dissolved according to the protocol of step 2.1 of Example 2) are dissolved and 2.46 ml (17 mmol) of triethylamine in 150 ml of dry benzene. The mixture is stirred at 0 ° C. and then 2.99 g (17 mmol) of the acid chloride prepared in step 5.1 above are added dropwise. The whole is stirred for 2 hours at room temperature.

At the end of the reaction, the solvent is evaporated, the residue is taken up in dichloromethane, washed with an alkaline solution and then twice with water.

The organic phase is then dried over MgSO ₄ , filtered and then concentrated in vacuo. The crude product obtained is purified by chromatography on a column of silica gel with dichloromethane as eluent. 5.5 g of the title aldehyde are thus recovered in the form of an oil. Yield:

98%. Rf: 0.41 (CH ₂ Cl ₂ / MeOH, 97: 3, v / v).

IR (KBr): 1705 (aldehyde C≈O), 1642 (NC≈O), 1609 (C≈Car) cm- ¹ NMR (200 MHz, CDCI3, HMDS) δ ppm: 10.02 (s, IH, H aldehyde), 7.87 and 7.50 (2d, 4H, J = 7.72 and 8.08 Hz, aromatic H), 3.86 and 3.48 (2sl, 4H, CH ₂ -NC = 0 of the piperazine), 2.67 (m, 4H, piperazine H), 2.49 (t, 2H, 7 = 7.66 Hz, CH ₂ -N), 1.51 (m, 2H, CH2-CN) , 1.18 (if, 30H, CH ₂ ), 0.81 (t, 3H, 7 = 6.16 Hz, CH3). 48139

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5.3- Preparation of [5- (4'-octadecylρiperazin-l'-ylcarbonyl) benzylidene] -l, 3-thiazolidine-2,4-dione

In a 100 ml flask surmounted by a condenser and a Dean and Starck apparatus, 4.2 g (8.9 mmol) of the aldehyde prepared in the previous step 5.2, 1.04 g ( 8.8 mmol) of 2,4-thiazolidinedione and 0.5 g of pyridinium benzoate in 50 ml of toluene. The mixture is stirred at reflux for 3 hours, the water being removed progressively. At the end of the reaction, the toluene is evaporated, the residue obtained is taken up in hot ethanol and a yellow precipitate appears while cooling. The crystals are filtered and 2.34 g of the pure terminal compound are thus recovered. Yield: 46%. Melting point: 86.3 ° C. Rf: 0.3 (CH ₂ Cl ₂ / MeOH, 95: 5, v / v).

IR (KBr): 1736 (NHC≈O), 1700 (NC≈O), 1604 (C≈Car) cm- ¹ NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.69 (s, IH, CH ≈), 7.45 (s, 4H,

Aromatic H), 4.73 (s, 1H, NH), 3.79 and 3.46 (2sl, 4H, CH ₂ -NC = 0 of piperazine), 2.48 (m, 4H, H of piperazine ), 2.39 (t, 2H, J = 7.33 Hz, CH ₂ -

N), 1.45 (m, 2H, CH2-CN), 1.17 (si, 30H, CH ₂ ), 0.80 (t, 3H, J = 5.89 Hz,

CH ₃ ).

Example 6 Preparation of l-r4 '- (2,4-dioxo-1,3,3-thiazolidin-5-ylmethyl) benzoyπ-4-octadecylpiperazine

Compound of formula (I) in which D = Z-HET with HET = thiazolidine dione (III), Z = -CH ₂ -, Y = C≈O, A = B = -CH ₂ -, R = - (CH ₂ ) ι ₇ - CH ₃ )

A suspension of 210 mg (3.69 10 ⁻⁴ mole) of the compound of Example 5, dissolved in 50 ml of absolute ethanol is subjected to catalytic hydrogenation in a Parr apparatus, in the presence of 100% black palladium and of hydrogen under pressure (40 to 50 psi) and stirred for more than 5 hours at 60 ° C. At the end of the reaction, the palladium is filtered, the ethanol evaporated and the residue obtained is purified by chromatography on a gel column. silica with a dichloromethane / methanol mixture, 99: 1, v / v as eluent. The product is then crystallized from acetonitrile and 126 is recovered. 48139

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mg of slightly yellowish crystals of the terminal compound. Yield: 60%. Melting point: 106.7 ° C. Rf: 0.55 (CH ₂ Cl ₂ / MeOH, 95: 5, v / v). IR (KBr): 1736 (NHC≈O), 1700 (NC≈O), 1604 (C≈Car) cm- ¹ NMR Η (200 MHz, CDCls, HMDS) δ ppm: 7.5 (si, IH, NH ), 7.30 and 7.20 (2d, 4H, 7 = 8.15 and 8.18 Hz, aromatic H), 4.44 (si, IH, CH-C≈O), 3.70 and 3, 40 (2sl, 4H, CH ₂ -NC = 0 of piperazine), 3.43 (dd, 2H, J = 3.90 Hz, Ph- CH ₂ ), 2.48 (m, 4H, H of piperazine ), 2.39 (t, 2H, J = 7.33 Hz, CH2-N), 1.45 (m, 2H, CH ₂ -CN), 1.17 (si, 30H, CH ₂ ), 0, 80 (t, 3H, J = 5.89 Hz, CH ₃ ).

Example 7 Preparation of 1- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-ylmethyl) benzyl] -4-tetradecylpiperazin-2-one

(compound of formula (I) in which D = Z-ΗET with ΗET = oxadiazolone (II), Z = -CΗ ₂ -, or n = 1, Y = A = -CH ₂ -, B = CO, R ≈ - (CH ₂ ) ₁₃ -CH ₃

7.1- Synthesis of N-benzylaminoacetaldehyde diethyl acetal A mixture consisting of 42 ml (0.2 mol) of aminoacetaldehyde diethyl acetal, 29.3 ml (0.2 mol) of benzaldehyde, 48 g of magnesium sulfate, and 300 ml toluene is heated at reflux for six hours. The solution is filtered and the solvent evaporated. The residue obtained is used without purification. Resumed in methanol, 12 g (0.3 mol) of sodium borohydride are added slowly and allowed to stir for 30 minutes. After hydrolysis and evaporation of the solvents, the residue obtained is dissolved in dichloromethane and then washed with water. The organic phase is dried (MgSO ₄ ), filtered and evaporated under reduced pressure. Flash chromatography with dichloromethane as eluent leads to 52 g of a viscous oil. Yield: 81%. Rf: 0.34 (CH ₂ Cl ₂ / MeOH, 95: 5, v / v).

IR (film): 3300 (NH), 1594 (C≈Car) cm- ¹ NMR 1H (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.23 (m, 5H, H ^, 4.56 (t, IH, J≈ 5.58 Hz, CH), 3.75 (s, 2H, PhCH ₂ ), 3.54 (m, 4H, OCH ₂ ), 2.68 (d, 2H, J≈ 5.58 Hz , CH ₂ ), 1.71 (s, 1H, NH), 1.14 (t, 6H, J≈ 7.04 Hz, CH ₃ ). 048139

• 26 -

7.2- Synthesis of (JV-tetradecyl-iV-benzyl) aminoacetaldehyde diethyl acetal

64 g (0.23 mol) of tetradecyl bromide is added to a mixture of 51.6 g (0.23 mol) of N-benzylaminoacetaldehyde diethyl acetal, 63.9 g (0.46 mol) of potassium carbonate and a catalytic amount of potassium iodide (1 g) in 700 ml of acetonitrile then the mixture is heated overnight at reflux. The solution is filtered, evaporated and the residue obtained after evaporation is taken up with dichloromethane and washed with water. The organic phase is dried over MgSO ₄ , filtered and evaporated under reduced pressure. The crude product is purified by flash chromatography with dichloromethane as eluent and gives 88 g of the expected product in the form of a yellow oil. Yield: 90% Rf: 0.65 (CH ₂ Cl ₂ / MeOH, 95: 5, v / v).

IR (film): 1594 (C≈Car) cm- ¹

NMR * H (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.21 (m, 5H, H _* ), 4.47 (t, IH, 7 = 5.16 Hz, CH), 3.56 (s , 2H, PhCH ₂ ), 3.50 (m, 4H, OCH2), 2.55 (d, 2H, J = 5.17 Hz, CH ₂ N), 2.41 (t, 2H,} = 7, 23 Hz, CH ₂ ), 1.39 (t, 2H, J = 6.87 Hz, CH) 1.13 (m, 28H, CH ₂ , CH ₃ ), 0.81 (t, 3H, J = 6 , 37 Hz, CH3).

7.3- Preparation of iV-tetradecylaminoacetaldehyde diethyl acetal

A mixture of 88 g (0.2 mol) of (N-tetradecyl-N-benzyl) aminoacetaldehyde diethyl acetal, dissolved in 300 ml of ethanol and supplemented with 20 mg of Pd-C at 10% is subjected to hydrogenation under pressure at 40 ° C for 48 h. The catalyst is filtered and the solvent evaporated under reduced pressure leads to a residue which is purified by flash chromatography with dichloromethane as eluent and leads to 59 g of a yellow oil. Yield: 90%. Rf: 0.29 (CH ₂ Cl ₂ / MeOH, 95: 5, v / v).

IR (film): 3300 (ΝH) cm ^' 48139

• 27 -

1 H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 4.59 (t, IH, CH), 3.57 (m, 4H, CH ₂ O), 2.63 (d, 2H, J≈ 5, 57 Hz, CHCH ₂ ). 2.58 (t, 2H, J≈ 7.25 Hz, NHCH ₂ ), 2.32 (IH, NH), 1.45 (t, 2H, J≈ 7.03 Hz, CH ₂ ), 1.15 (m, 28H, CH ₃ , CH ₂ ), 0.81 (t, 3H, J≈ 6.39 Hz, CH ₃ ).

7.4- Preparation of ethyl N-benzyloxycarbonylaminoacetate. a) A mixture of 138 g (1 mol) of potassium carbonate, 70 g (0.5 mol) of ethyl ester of glycine in 300 ml of tetrahydrofuran is stirred for 10 min, cooled to 0 ° C and then added slowly 71 ml (0.5 mol) of benzyl chloroformate. The solution is stirred for 30 min then filtered and the solvent evaporated. The residue obtained is dissolved in dichloromethane and washed with water. The organic phase is dried over MgSO ₄ , filtered and the solvent evaporated under reduced pressure. The residue obtained is used without purification, for the next step.

IR (film): 1735 (-O-CO-), 1750 (-O-CO-N) cm ^"1

NMR ^! H (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.21 (m, 5H, H ^), 5.81 (si, IH, NH), 5.01 (s, 2H, PhCH ₂ ), 4, 06 (q, 2H, J≈ 7.14 Hz, CH ₂ CH ₃ ), 3.80 (d, 2H, J≈ 14.27 Hz, NHCH ₂ ), 1.14 (t, 3H, J≈ 7, 13 Hz, CH ₃ ).

b) The residue obtained previously, dissolved in 250 ml of ethanol is treated with 250 ml of a 10% potassium hydroxide solution and then heated at reflux overnight. The ethanol is evaporated and the aqueous phase is acidified (pH = 1) with concentrated HCl. The precipitate obtained filtered and dried gives 105 g of a white solid. Yield: 90%. Melting point: 110 ° C. Rf: 0.25 (CH ₂ Cl ₂ / MeOH, 90:10, v / v).

IR (KBr): 1678 (C≈C), 1727 (OC≈O) cm ^-1 1H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.24 (m, 5H, H _ar ), 5.23 (if IH, NH), 5.06 (s, 2H, CH ₂ , PhCH ₂ ), 4.63 (s, IH, OH), 3.94 (d, 2H, J = 5.50 Hz, CTLCOOH) . 048139

- 28 -

7.5- Preparation of N-benzyloxycarbonyïamino-N'-2,2-diethoxyethyl-N'-tetradecylacetamide

To a mixture of 18.7 g (5.6 mmol) of tetadadecylaminoacetaldehyde dietylacetal, 12.6 g (56 mmol) of ethyl N-benzyloxycarbonylaminoacetate, 15 ml (0.112 mol) of triethylamine, 9 g (67 mmol) of 1-hydroxybenzotriazole in 120 ml of dichloromethane is added 24.7 g (0.12 mole) of Ν, Ν'- dicyclohexylcarbodiimide. After heating at reflux for two hours, the solution is filtered and washed with water. The organic phase is dried over MgSO ₄ , filtered and evaporated. The residue obtained is purified on a column of silica gel using a CH Cl ₂ / MeOH mixture, 99: 1, v / v) and leads to 25 g of a colorless oil. Yield: 96%. Rf: 0.42 (CH ₂ Cl ₂ / MeOH, 98: 2, v / v).

IR (film): 1649 (C≈O), 1720 (OC≈O) cm ^"1 NMR 1H (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.17 (m, 5H, H _ar ), 5.85 (if, IH,

NH), 5.01 (s, 2H, PhCH ₂ ), 4.54 (t, 1H, J = 5.26 Hz, CH), 3.92-4.02 (m, 2H,

NHCH ₂ CO), 3.48-3.65 (m, 2H, NCH ₂ CH), 3.2 (m, 6H, CH ₂ , OCH ₂ ), 1.44 (if,

2H, CH ₂ ), 1.17 (si, 22H, CH ₂ ), 1.09 (t, 6H, J≈ 6.98 Hz, CH ₃ ), 0.79 (t, 3H, J≈

6.2 Hz, CH ₃ ).

7.6-Preparation of 4-benzyloxycarbonyl-1-tetradecylpiperazin-2-one

23 g (47 mmol) of the amide prepared previously dissolved in 250 ml of toluene is stirred and added with a catalytic amount of paratoluene sulfonic acid (780 mg, 4.1 mmol) at room temperature. Then the solution is stirred at 75 ° C for 3 h. After cooling, the mixture is washed with water and the organic phase is dried over MgSO ₄ , filtered and concentrated. The residue obtained is chromatographed on a silica gel column with dichloromethane as eluent to give 14 g of a yellow oil. Yield: 72%. Rf: 0.61 (CH ₂ Cl ₂ / MeOH, 98: 2, v / v).

IR (film): 1669 (C≈O, C≈C), 1700 (OC≈O) cm ^-1 48139

- 29 -

1H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.25 (m, 5H, H _ar ), 6.29 (dd, IH, / = 21.58 and 5.98 Hz, CH≈CH), 5.42 (dd, IH, / = 18.78 and 6.01 Hz, CH≈CH), 5.12 (s, 2H, PhCH ₂ O), 4.59 (s, 2H, COCH ₂ N), 3.39 (t, 2H, / = 7.24 Hz, CH ₂ ), 1.33 (if, 2H, CH ₂ ), 1.18 (if, 22H, CH ₂ ), 0.81 (t, 3H , J≈ 6.32 Hz, CH ₃ ).

7.7- Preparation of l-tetradecylpiperazin-2-one hydrochloride

To a solution of 10.5 g (24 mmol) of 4-benzyloxycarbonyl-1-tetradecylpiperazin-2-one dissolved in 100 ml of ethanol are added 5 ml of concentrated HCl. The mixture is subjected to catalytic hydrogenation under hydrogen pressure in the presence of 1 g of 10% Pd-C at 40 ° C for 24 h. After filtration, the solvent is evaporated and the residue is crystallized from ether to yield 7 g of a yellow solid. Yield: 86%. Melting point: 161.6 ° C (dec). Rf: 0.25 (CH ₂ Cl ₂ / MeOH, 90:10, v / v).

IR (KBr): 1655 (C≈O), 3451 (NH ₂ ) cm ^-1

1 H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 9.87 (si, IH, NH ₂ ), 7.7-8.9 (si, IH, NH ₂ ), 4.18 (si, 2H, NCH ₂ ), 3.93 (if, 2H, NCH ₂ ), 3.63 (if, 2H, NCH ₂ ), 3.30 (s, 2H, CH ₂ ), 1.46 (if, 2H, CH ₂ ), 1.19 (si, 22H, CH ₂ ), 0.81 (s, 3H, CH ₃ ).

7.8- Preparation of 4- (4-cyanomethylbenzyl) -l- tetradecylpiperazin-2-one

2.2 g (10 mmol) of 4-bromomethylphenylacetonitrile is added to a mixture consisting of 2.95 g (8.8 mmol) of 1-tetradecylpiperazin-2-one hydrochloride, 2.6 g (17.6 mmol) of potassium carbonate and 0.5 g of potassium iodide in 100 ml of acetonitrile. The solution is stirred at reflux for 4 h then filtered and evaporated. The residue obtained is dissolved in dichloromethane and washed with a saturated solution of sodium carbonate. The organic phase is dried over MgSO ₄ , filtered and evaporated. The crude compound obtained is purified by flash chromatography with a mixture of CH ₂ C1 ₂ MeOH, 99: 1, v / v and gives 3.6 g of a yellow oil. Yield: 95%. Rf: 0.48 (CH ₂ Cl ₂ / MeOH, 95: 5, v / v) 48139

- 30 -

IR (film): 1647 (C≈O), 2249 (CN) cm ⁴

^X NMR H (200 MHz, CDC1 _3, HMDS) δ ppm: 7.19 (m, 4H, H _ar), 3.67 (s, 2H, PhCH ₂ CN), 3.47 (s, 2H, NCH ₂ Ph), 3.26 (m, 4H, NCH ₂ ), 3.07 (s, 2H, COCH ₂ N), 2.58 (t, 2H, J≈ 5.39 Hz, CH ₂ ), 1.47 (if, 2H, CH ₂ ), 1.18 (if, 22H, CH ₂ ), 0.81 (t, 3H, J≈ 6.40 Hz, CH ₃ ).

7.9- Preparation of 4- [4- (-hydroxyamidino methyl) benzyl] -1-tetradecylpiperazin-2-one To a mixture of 7 g (50 mmol) of potassium carbonate, 2.9 g (41 mmol) of hydrochloride of hydroxylamine in 80 ml of ethanol heated at reflux for 15 min is added dropwise 3.6 g (8.4 mmol) of 4- (4-cyanomethylbenzyl) -l-tetradecylpiperazin-2-one dissolved in 20 ml of ethanol. At the end of the addition, the mixture is heated at reflux for 12 h. After filtration and evaporation of the solvents, the residue obtained is taken up in dichloromethane and washed with water. The organic phase is dried over MgSO _4d, filtered and evaporated. The residue is then purified on a column of silica gel with a mixture of CH ₂ Cl / MeOH, 98: 2, v / v and leads to 2.2 g of a yellow oil. Yield: 56%. Rf: 0.43 (CH ₂ Cl ₂ / MeOH, 90:10, v / v).

IR (film): 1636 (C≈O, C≈N) on ¹

1H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.17 (m, 4H, H *), 4.51 (s, 2H, NH ₂ ), 3.44 (s, 2H, PhCH ₂ CN) , 3.35 (s, 2H, NCH ₂ Ph), 3.24 (m, 4H, NCH ₂ ), 3.05 (s, 2H, COCH ₂ N), 2.58 (t, 2H, J≈ 5 , 21 Hz, CH ₂ ), 1.45 (if, 2H, CH ₂ ), 1.18 (if, 22H, CH ₂ ), 0.80 (t, 3H, J≈ 6.38 Hz, CH ₃ ) .

7.10- Preparation of 1- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-yl methyl) benzyl] -4-tetradecylpiperazin-2- one A une 1.1 g (2.4 mmol) solution of 4- [4- (N-hydroxyamidinomethyl) benzyl] -1-tetradecylpiperazin-2-one and 0.4 ml (2.9 mmol) 048139

of triethylamine in 130 ml of dichloromethane cooled at 0 ° C for 15 min is added dropwise 0.36 ml (2.8 mmol) of phenyl chlorocarbonate. The mixture is stirred for 2 h then treated with a saturated solution of Na ₂ CO _3> then washed with water. The organic phase is dried over MgSO _4d filtered and concentrated to give a residue which is dissolved in 50 ml of toluene and heated at reflux for 6 h. The toluene is evaporated off and the residue obtained is chromatographed on a column of silica gel with CHCl ₂ / MeOH, 98: 1, v / v as eluent, then crystallized from ether to give 300 mg of a yellow solid. Yield: 30%. Rf: 0.52 (CH ₂ Cl ₂ / MeOH, 90:10, v / v).

IR (KBr): 1636 (CON), 1775 (-O-CO-N) cm ^-1

N NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.16 (m, 4H, Har), 5.23 (s, IH, NH), 3.76 (s, 2H, PhCH ₂ CN), 3 , 38 (s, 2H, NCH ₂ Ph), 3.25 (m, 4H, CH ₂ N), 2.86 (s, 2H, COCH2N), 2.57 (t, 2H, J = 5.23 Hz , CH ₂ ), 1.45 (m, 2H, CH ₂ ), 1.18 (if, 22H, CH ₂ ), 0.81 (t, 3H, J = 6.42 Hz, CH3).

Example 8. Preparation of 1- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-ylethyl) benzoyl] -4-tetradecylpiperazine

(compound of formula (I) in which D = Z-HET with HET = oxadiazolone (II),

Z = - (CH ₂ ) ₂ -, i.e. n = 2, A = B = -CH ₂ -, Y = CO, R ≈ - (CH ₂ ) ₁₃ -CH ₃

8.1- Preparation of 4- (2-chloro-2-cyanoethyl) benzoic acid In a 250 ml ground Erlenmeyer flask, 10 g (72 mmol) of para-aminobenzoic acid are dissolved in 70 ml of acetic acid to which is added 6 ml of 12N hydrochloric acid. The mixture is cooled to 0 ° C. and 2.5 g (36.2 mmol) of NaNO ₂ are added in small portions. After 30 min of stirring, the viscous liquid obtained is added dropwise to a mixture of 6.5 ml (94.8 mmol) of acrylonitrile and a few milligrams of copper oxide (CuO) suspended in 20 ml anhydrous acetone. The reaction mixture is stirred at room temperature for two hours, the solid obtained is filtered under vacuum and then washed several times with water. The product is purified by recrystallization in water and leads to a white solid. Yield: 65%. Melting point: 157 ° C. Rf: 0.29 (CH Cl ₂ / MeOH, 50:50, v / v).

IR (KBr): 1690 (C ^≈ O), 2240 (CN) cm ^-1 1H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.9 (d, 2H, J = 8.22 Hz, H _ar ), 7.4 (d, 2H, J = 8.18 Hz, H _ar ), 5.5 (t, IH, J = 7 Hz, CNCHC1), 3.43 (d, 2H, J = 6.96 Hz, CH ₂ CHCN).

8.2- Preparation of 4- (2-cyanoethyl) benzoic acid To 10 g (47 mmol) of 4- (2-chloro-2-cyanoethyl) benzoic acid dissolved in 250 ml of glacial acetic acid is added in small batches portions 1.56 g (23 mmol) of zinc powder. The mixture is heated at reflux for two hours.

The salt formed (ZnCl) is filtered under vacuum and washed several times with water. A precipitate is deposited cold in the filtrate, it is filtered and washed several times with water and then dried. A white solid is recovered. Yield: 68%. Fusion point :

165 ° C. Rf: 0.25 (CH ₂ Cl ₂ / MeOH, 50:50, v / v).

IR (KBr): 1700 (C≈O), 2252 (CN) cm ^-1

1H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.9 (d, 2H, J = 8.17 Hz, H _ar ), 7.25 (d, 2H, J = 8, 13 Hz, H _ar ), 2.95 (t, 2H, J = 7.22 Hz, CH ₂ CN), 2.6 (t, 2H, J = 7.36 Hz, CH ₂ CH ₂ CN).

All the condensation steps were carried out by following the same experimental protocol of Example 2 described above with substantially the same yields and lead to the final product with 1 Oxadiazolone named above.

Example 9. Preparation of l-f4 ^, - (4,5-dihydro-l., 2,4 (4H ^r ) -5-oxo-oxadiazol-3-ylpropyDbenzoyπ-4-tetradecy-Ipiperazine

(compound of formula (I) in which D = Z-HET with HET = oxadiazolone (II),

Z = - (CH ₂ ) ₃ -, i.e. n = 3, A = B = -CH ₂ -, Y = CO, R = - (CH ₂ ) ₁₃ -CH ₃

9.1- Preparation of l-bromo-3-phenylpropane 048139

- 33 -

To 10 g (73 mmol) of 3-phenylpropan-1-ol dissolved in 150 ml of anhydrous dichloromethane are added in small portions 50 ml of a 1M solution of PBr ₃ (36 mmol) in dichloromethane. The reaction mixture is stirred at room temperature for one hour. After washing the latter several times with water, the organic phases are dried and then evaporated. The residue obtained is purified by chromatography on silica gel with an ether / petroleum ether mixture, 5:95, v / v as eluent, to give the bromine derivative of viscous liquid appearance. Yield: 80%. Rf: 0.25 (ether / petroleum ether, 5:95, v / v).

IR (film): 1605 (C = C _ar ) cm ^-1

N NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.25-7.06 (m, 5H, H _a r), 3.29 (t, 2H, J = 6.59 Hz, CH ₂ Br) , 2.68 (t, 2H, J = 7.34 Hz, PhCH ₂ CH CH ₂ Br), 2.14-1.99 (m, 2H, CH ₂ CH ₂ CH ₂ Br).

9.2- Preparation of 4- (3-bromopropyl) acetophenone A a mixture of 17.5 g (88 mmol) of aluminum trichloride and 50 ml of acetyl chloride dissolved in 100 ml of CS are added to 0 ° C dropwise, 25 g (125 mmol) of the bromine derivative 1-bromo-3-phenylpropane dissolved in 20 ml of acetyl chloride. The mixture is stirred at room temperature for two hours. The excess acetyl chloride and the CS ₂ are removed by evaporation under reduced pressure. The residue obtained is taken up in dichloromethane, washed several times with water, dried over MgSO ₄ and then concentrated under vacuum. Distillation under reduced pressure gives the acetophenone substituted in the form of a yellowish liquid with a yield of 79%. Boiling point: 140-145 ° C / 3 mmHg. Rf: 0.25 (ether / petroleum ether, 50:50, v / v).

IR (film): 1670 (C≈O), 1605 (C = C _ar ) cm ^-1

1H NMR (200 MHz, CDC1 _3, HMDS) δ ppm: 7.79 (if, 2H, H ^), 7.23 (if, 2H, H ^), 3.26 (t, 2H, J = 6, 49 Hz, CH ₂ Br), 2.7 (t, 2H, J ≈ 6.26 Hz, PhCH ₂ CH ₂ CH ₂ Br), 2.48 (s, 3H, CH ₃ ), 2.08-1, 9 (m, 2H, CH ₂ CH ₂ CH ₂ Br). 048139

• 34 -

9.3 - Preparation of 4- (3-bromopropyl) benzoic acid

To 33 g of NaOH dissolved in 200 ml of water are successively added 50 ml of Br ₂ dropwise and 100 ml of dioxane. The mixture is cooled to 0 ° C. and 22 g of 4- (3-bromopropyl) acetophenone are added dropwise. Stirring is continued at room temperature until the brown color of the bromine has disappeared (one hour). The mixture is carefully acidified using an aqueous 12N HCl solution (20 ml). The precipitate formed, filtered under vacuum, rinsed several times with water and then dried leads to a yellow solid. Yield: 85%. Melting point: 120 ° C. Rf: 0.25 (MeOH / CH ₂ Cl ₂ , 20:80, v / v).

IR (KBr): 3340 (OH), 1700 (C≈O) cm ⁴

1H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.9 (d, 2H, J = 8.16 Hz, Har), 7.2 (d, 2H, J = 8.3 H _ar ), 3 , 3 (t, 2H, J = 6.47 Hz, CH ₂ Br), 2.78 (t, 2H, J = 7.7 Hz, PhCH ₂ CH ₂ CH ₂ Br), 2.18-2.04 (m, 2H, CH ₂ CH ₂ CH ₂ Br).

9.4- Preparation of 4- (3-cyanopropyl) benzoic acid

The bromine derivative prepared above is transformed into nitrile according to the same protocol described in Example 1 above with a yield of 75% and of viscous appearance. Rf: 0.29 (MeOH / CH ₂ Cl ₂ , 15:85, v / v).

IR (film): 3345 (OH), 1700 (C≈O), 2253 (CN) cm ^"1

1H NMR (200 MHz, CDC1 _3, HMDS) δ ppm: 7.9 (d, 2H, J = 8.09 Hz, H _ar ), 7.2 (d, 2H, J = 8.09 Hz, Ha _r ), 2.8 (t, 2H, J = 7.44 Hz, CH ₂ CN), 2.28 (t, 2H, J = 6.99 Hz, PhCH ₂ CH ₂ CH ₂ CN), 2.02- 1.91 (m, 2H, CH ₂ CH ₂ CH ₂ CN).

9.5- Continuation of steps

The condensation of tetradecylpiperazine on 3-chloromethylbenzoyl chloride in basic medium as described previously in Example 2 leads to the corresponding chloride. The latter is transformed into nitrile then into amidoxy and finally cyclized into oxadiazolone: l- [4 ′ - (4,5-dihydro- 1,2,4 (4H) -5-oxo-oxadiazol-3-ylpropyl) benzoyl] -4-tétradécylpipérazin-2-one, / 048139

^• 35 -

according to the same protocols already described. The characteristics of all the intermediaries as well as of the final product are given below:

1 - (3-chloromethylbenzoyl) -4-tetradecylpiperazine Appearance: viscous oil. Yield: 67%.

Eluent (MeOH / CH ₂ Cl ₂ , 5:95, v / v).

Rf: 0.25 (MeOH / CH ₂ Cl ₂ , 10:90, v / v).

IR (film): 1660 (NCO), 1610 (C≈C _ar ) cm ^-1 1H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.38 (si, 4H, Ha _r ), 4.55 ( s, 2H, CH ₂ C1), 3.72 (m, 2H, NCH ₂ ), 3.38 (m, 2H, NCH ₂ ), 2.52 (m, 4H, NCH ₂ ), 2.36 (t , 2H, NCH ₂ ), 1.41 (m, 2H, NCH ₂ CH ₂ ), 1.25-1.15 (si, 22H, CH ₂ ), 0.81 (t, 3H, J≈ 6.09 Hz, CH ₃ ).

1 - (3-cyanomethylbenzoyl) -4-tetradecylpiperazine

Appearance: viscous oil. Yield: 67% Eluent (MeOH / CH ₂ Cl ₂ , 5:95, v / v). Rf: 0.25 (MeOH / CH ₂ Cl ₂ , 10:90, v / v).

IR (film): 1665 (NCO), 2253 (CN), 1605 (C≈C _ar ) cm ^-1

1H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.36-7.23 (si, 4H, H _ar ), 3.71 (s, 2H, CH ₂ CN), 3.74-3.71 (m, 2H, NCH ₂ ), 3.43 (m, 2H, NH ₂ ), 2.54-2.25 (m, 4H, NCH ₂ ), 2.29 (t, 2H, J≈ 7.45 Hz, NCH ₂ ), 1.5-1.35 (m, 2H, NCH ₂ CH ₂ ), 1.3-1.1 (m, 22H, CH ₂ ), 0.81 (t, 3H, J≈ 6.36 Hz, CH ₃ ).

l- [3- (N-hydroxyamidino) methylbenzoyl] -4-tetradecylpiperazine Appearance: viscous oil. Yield: 58%.

Eluent (MeOH / CH ₂ Cl ₂ , 5:95, v / v).

Rf: 0.25 (MeOH / CH ₂ Cl ₂ , 10:90, v / v).

IR (film): 3430 (ΝH), 1660 (ΝCO), 1605 (C = C _ar ) cm ^"1 048139

- 36 -

NMR ^! H (200 MHz, CDC1 _3> HMDS) δ ppm: 7.24-7.07 (m, 4H, H ^), 3.65-3.53 (m, 2H, NCH ₂ ), 3.33-3 , 28 (m, 2H, NCH ₂ ), 2.41-2.11 (m, 6H, NCH ₂ ), 2.09 (s, 2H, CH ₂ CN), 1.5-1.3 (m, 2H, NCH ₂ CH ₂ ), 1.25-1.15 (m, 22H, CH ₂ ), 0.81 (t, 3H, J≈ 6.41 Hz, CH ₃ ).

1 - [3 - (4,5-dihydro-1,2,4 (4H) -oxadiazol-5-one-3 -yl) methylbenzoyl] -4- tetradecyl-piperazine Appearance: viscous oil. Yield: 67%. Eluent (MeOΗ / CΗ ₂ Cl ₂ , 5:95, v / v).

Rf: 0.25 (MeOH / CH ₂ Cl ₂ , 10:90, v / v).

IR (film): 3435 (NH), 1665 (NCO), 1610 (C = C _ar ) cm ^-1

NMR ^] H (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.3-7.2 (m, 4H, H _ar ), 6.57 (si, IH, NH), 3.68-3.35 ( m, 4H, NCH ₂ ), 2.52-2.29 (m, 6H, NCH ₂ ), 2.1 (s, 2H, CH ₂ CN), 1.4-1.35 (m, 2H, NCH ₂ CH ₂ ), 1.3-1.1 (m, 22H, CH ₂ ), 0.8 (t, 3H, J≈ 6 Hz, CH ₃ ).

Example 10. Preparation of 1- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-yl-methylbenzoyl] -4- (N-octadecylaminocarbonyl) piperazine ( compound of formula (I) in which D = Z-ΗET with ΗET = oxadiazolone (II), Z = - (CΗ ₂ ) -, or n = 1, A = B = -CH ₂ -, Y = CO, R = - (CH ₂ ) ₁₇ -CH ₃

10.1- Preparation of N-octadecylaminocarbonylpiperazine In a 250 ml ground Erlenmeyer flask are stirred 13 g (0.151 mol) of piperazine dissolved in 100 ml of dichloromethane. 4.42 g (15 mmol) of 1-octadecylisocyanate are added to the mixture and stirring is continued at room temperature for 1 h. At the end of the reaction, the solution is washed twice with water. The organic phase dried over MgSO _; filtered and evaporated gives 5.21 g of white crystals. Yield: 91%. Melting point: 72 ° C,

Rf: 0.46 (CH ₂ Cl ₂ / MeOH / NH ₄ θH, 80: 20: 2, v / v / v).

IR (KBr): 3364 (NH), 1620 (N-CO-N) cm ^"1 048139

- 37 -

1H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 4.68 (t, 1H, J≈ 5.01 Hz, NHCO), 3.26 (t, 4H, J≈ 5.22 Hz, CH ₂ NCO ), 3.14 (q, 2H, J≈ 7.14 Hz, CH NHCO), 2.77 (t, 4H, J≈ 5.21 Hz, CH ₂ NH), 1.73 (si, IH, NH ), 1.42 (m, 2H, CH ₂ CH ₂ NH), 1.19 (si, 3 OH, CH ₂ ), 0.81 (t, 3H, J≈ 6.20 Hz, CH ₃ ).

10.2- Preparation of 1- (4'-chloromethylbenzoyl) -4- (N- octadecylaminocarbonyl) piperazine

This intermediate was obtained under the same conditions as Example 2 above from 9.9 g (26 mmol) of N-octadecylcarbonylpiperazine, 5.4 ml (38 mmol) of triethylamine and 5 g (26 mmol) of chloride 4-chloromethylbenzoic acid. The purification carried out by chromatography on silica gel with dichloromethane as eluent leads to 12.2 g of product in the form of white crystals. Yield: 88%. Melting point: 68-70 ° C. Rf: 0.4 (CH ₂ Cl ₂ / MeOH, 95: 5, v / v).

IR (KBr): 2365 (CN), 1653 (COIN), 1730 (C = C _ar ) cm ¹

1H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.30-7.39 (si, 4H, H _ar ), 4.82 (t, IH, / = 5 Hz, NHCO), 4.48 ( s, 2H, PhCH ₂ Cl), 3.52 (if, 4H, CH ₂ NCONH), 3.34 (if, 4H, CH ₂ NCO), 3.15 (q, 2H, J = 7 Hz, CH ₂ NH), 1.43 (m, 2H, CH ₂ CH2NH), 1.19 (sm, 30H, CH ₂ ), 0.81 (t, 3H, / = 5.15 Hz, CH ₃ ).

10.3- Preparation of 1- (4'-cyanomethylbenzoyl) -4- (N- octadecylamino carbonyl) piperazine

This compound was obtained according to the same synthesis protocols described above in Example 2 starting from 5.33 g (10 mmol) of 1- (4'-chloromethylbenzoyl) -4- (octadecylaminocarbonyl) piperazine and 1.96 g (40 mmol) sodium cyanide. 3.84 g of white precipitate are obtained. Yield: 74%. Melting point: 90 ° C. Rf: 0.56 (CH ₂ Cl ₂ / MeOH, 93: 7, v / v).

IR (KBr): 2365 (CN), 1653 (CON), 1730 (C = C _ar ) cm ^-1

1H NMR (200 MHz, CDC1 ₃ , HMDS) δ ppm: 7.30-7.39 (si, 4H, H _ar ), 4.53 (t, IH, J = 5 Hz, NHCON), 3.73 ( s, 2H, PhCH ₂ CN), 3.52 (si, 4H, CH ₂ NCONH), 3.34 048139

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(sl, 4H, CH ₂ NCO), 3.15 (q, 2H, J≈ 7 Hz, CH ₂ NH), 1.43 (m, 2H, CH ₂ CH ₂ NH), 1.19 (si, 30H , CH ₂ ), 0.81 (t, 3H, J≈ 5.15 Hz, CH ₃ ).

10.4- Preparation of 1- [4 '- (N-hydroxyamidinomethyl) benzoyl] - 4- (N-octadecylaminocarbonyl) piperazine

This amidoxime is obtained under the same conditions as those described above from 5.8 g (84 mmol) of hydroxylamine hydrochloride, 14.07 (102 mmol) of potassium carbonate and 8.9 g (17 mmol) of 1 - (4'-cyanomethylbenzoyl) -4- (octadecylaminocarbonyl) piperazine. The residue obtained is purified by chromatography on silica gel with a mixture of CH ₂ Cl ₂ / MeOH, 98: 2, v / v as eluent. 2.36 g of white crystals are thus obtained. Yield: 38%. Melting point: 104-106 ° C. Rf: 0.43 (CH ₂ Cl ₂ / MeOH, 90:10, v / v).

IR (KBr): 3493 (OH), 3355 (NH ₂ ), 2200 (CN), 1615 (C = C _ar ) cm ^-1

1 H NMR (200 MHz, CD ₃ OD, HMDS) δ ppm: 7.35 and 7.30 (2d, 4H, J≈ 8.8 and 8.5 Hz, H _ar ), 4.7 (t, IH, J ≈ 6 Hz, NHCON), 4.8 (s, 2H, NH ₂ ), 3.61 (m, 4H, CH ₂ NCONH), 3.50 (s, 2H, PhCH ₂ ), 3.27 (m , 4H, CH ₂ NCO), 3.07 (t, 2H, J ≈ 6 Hz, CH ₂ NH), 1.39 (m, 2H, CH ₂ CH ₂ NH), 1.21 (si, 30H, CH ₂ ), 0.83 (t, 3H, J≈ 8 Hz, CH ₃ ).

10.5- Preparation of 1- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-yl-methylbenzoyl] -4- (N- octadecylaminocarbonyl) piperazine This synthesis is carried out in two stages, according to the same protocol already described in example 2 above, starting from 1.25 g (2.3 mmol) of the above amidoxine, 0.38 ml (2.75 mmol) of triethylamine and 0.34 ml (2.75 mmol) of phenyl chlorocarbonate The 0.8 g of white crystals are obtained after chromatography on silica gel with dichloromethane as eluent, yield: 59%, melting point: 150 -152 ° C. Rf: 0.43 (CΗ ₂ Cl ₂ / MeOΗ, 93: 7, v / v).

IR (KBr): 1780 (OCON), 1618 (C≈N), 1550 (C≈C _a r) cm ^"1 NMR ^] H (200 MHz, CD ₃ OD, HMDS) δ ppm: 7.36 (s, 4H, H _ar ), 4.94 (t, IH, J≈ 5.53 Hz, NHCON), 3.77 ( s, 2H, PhCH ₂ ), 3.56 (si, 2H, CH ₂ NCONH), 3.27 (si, 2H, CH ₂ NCO), 3.07 (q, 2H, J ≈ 7.37 Hz, CH ₂ NHCO), 1.41 (if, 2H, CH ₂ CH ₂ NHCO), 1.18 (if, 30H, CH ₂ ), 0.81 (t, 3H, J≈ 6.8 Hz, CH ₃ ).

Example 11 Study of the biological activity in vitro

Phospholipases A ₂ hydrolyze the ester bond in position sn-2 of the glycerol phospholipids and release a fatty acid and a lysophospholipid. The action of the compounds was in particular evaluated in vitro by the determination of fatty acid according to the fluorimetric method of Radvanyi et al. (Anal. Biochem. 1989, 177, 103-109) and by the determination of lysophospholipid according to the UN spectrophotometric method of Reynolds et al. (Anal. Biochem. 1992, 204, 190-197).

11.1.1- Materials and methods

11.1.1- Materials

- The enzymes used are two secreted group II enzymes, recombinant human PLA ₂ and the basic PLA ₂ subunit of Crotalus durissus terrificus and a secreted group I enzyme, pig pancreatic PLA ₂ .

- The substrates are: a fluorescent substrate, palmitoyl-2- (10-pyrenyl decanoyl) -sn-glycero-3-phosphatidylglycerol acid, for the fluorimetric method and a sulfur substrate, the lithium salt of 1,2-bis - (dihexanylthio) -dideoxy-rac-glycero-3-ρhosphorylglycerol for the spectrophotometric method UN.

- The fluorimetric assays are carried out on a Perkin Elmer LS50 device in single-use polystyrene tanks, 1 cm wide. The exact concentration of the fluorescent substrate is determined on a UN spectrometer. Unicam in quartz tanks.

- UN spectrophotometric assays. are carried out on an ELx 808 Ultra Micro Plate Reader spectro device (96-well plates).

11.1.2- Methods 048139

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a) Fluorimetric determination

PLA2 is an enzyme which hydrolyzes the ester bond in position sn-2 of phospholipids. In aggregated form, the fluorescent substrate has a maximum fluorescence emission at 490 nanometers (nm), and zero at 398 nm. After enzymatic hydrolysis, the fluorescence emitted by the fatty acid released (pyrenyl decanoic acid), complexed with bovine serum albumin (SAB) is exalted and a strong emission is then observed at 378 and 398 nm. The principle of the assay is based on the measurement of this difference in fluorescence at 398 nm which is used to study the appearance of the fatty acid released over time, in other words, the PLA2 activity.

The measurement of the enzymatic activity is carried out in tanks in which we sample: 960 μl of Tris buffer, 50 mM HCl at pH 7.5; 0.5 M NaCl, EGTA, ImM; 1 μM substrate. This mixture is stirred for one minute using a vortex, to allow the formation of substrate vesicles, then 10 μl of 10% BSA, 10 μl of solvent (efhanol or DMSO) or inhibitor solution, are added successively, with stirring. μl of PLA2 at a given concentration and finally, 10 μl of 1 M calcium chloride (CaCl ₂ ) which triggers the activity. Good conditions for measuring the enzyme activity require saturation of the enzyme, the initial concentrations used are therefore: (i) human recombinant PLA ₂ : 0.1 μg / ml; (ii) PL A ₂ pancreatic pork: 0.6 μg / ml; (iii) PLA ₂ Crotalus durissus terrificus (CB): 0.05 μg / ml. The mother solution containing the inhibitor is prepared at an initial concentration of 10 ′ ² M.

The enzymatic activity results in a curve whose slope at the origin makes it possible to calculate the initial speed of the reaction. Let So be the slope of the curve in the absence of calcium (control), S the slope in the presence of calcium, V the volume in μl of the substrate solution and F _ma χ the maximum fluorescence signal, obtained once the enzymatic reaction ended, then, the relation: 048139

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(S - S _Q ) XV

A = 2.1CT ⁴ x

F "max

calculates the enzymatic activity (A) in μmole of fatty acid released per minute. The residual activity in the presence of inhibitor is then evaluated by the ratio of the slopes obtained in the absence and in the presence of inhibitor.

_n , _" _. ._- -. , .. (S - Sn) in the presence of inhibitor _{^ ΛΛ} % Residual activity = - - - ^ - x 100

(S - S _Q ) in the absence of an inhibitor

The values obtained plotted as a function of the logarithms of the concentrations of inhibitor used, make it possible to determine the value of Clδo, that is to say the concentration of inhibitor for which the enzymatic activity is halved. The lower this CISQ value, the better the inhibitory activity of the test compound.

PLA ₂ have more affinity for organized substrates.

However, three reasons may explain the inhibition possibly observed.

- The inhibitor disorganizes the substrate micelles and makes them inaccessible to the enzyme. In this case, the inhibition is due to the unavailability of the substrate.

- Part of the inhibitor can be fixed on the vesicles of the substrate and thus, the IC50 can be underestimated.

- The inhibitor reacts either with a group of the active site or with another part of the enzyme, thus preventing hydrolysis of the substrate. In this case, the inhibition observed is real, it takes place at the active site, and may or may not have a reversible character.

The fluorimetric test is a very sensitive technique, but does not make it possible to differentiate between these three types of inhibition, the substrate being in micellar form. However, in the spectrophotometric test, described below, the monomeric state of the substrate 48139

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makes it possible to remove any ambiguity as to the reality of the inhibition, even if, in this test, the enzyme does not fully function under optimal conditions.

b) U.V spectrophotometric assay

The lysothiophospholipid (LTPL) released by the lipolytic action of PLA ₂ in the presence of calcium reacts with dithionitrobenzoic acid (DTNB) present in the medium to form an LTPL-TNB complex and a TNB- anion which induces a yellow coloration of the reaction medium. The measurement of the optical density at 412 nanometers (wavelength of absorption of the TNB- ion) reflects the appearance of the lysothiophospholipid and therefore a PLA ₂ activity. The enzymatic activity is measured in multi-well plates, each well containing 190 μl of buffer IX, 2 μl of DTNB at 10 mM, 2 μl of substrate at 20 mM, 2 μl of solvent or inhibitor solution, 2 μl of PLA ₂ at a given concentration. The plate is shaken and 2 μl of 1 M calcium chloride are added to trigger the enzymatic reaction. The substrate is used here at a concentration lower than the cmc, critical micellar concentration (it is of ImM), in monomeric form, and the substrate / enzyme ratio is respected. This justifies the use of the substrate at a concentration 5 times lower than the cmc (200 μM). Good measurement conditions also require enzyme saturation. The concentrations used are therefore: (i) pancreatic pig PLA ₂ : 1.5 mg / ml; (ii) PLA ₂ Crotalus durissus terrificus: 0.43 mg / ml. The mother solution containing the inhibitor is prepared at an initial concentration of 10 ' ² M. The IC50 is determined using software coupled to the UN spectrophotometer. It directly calculates the initial rate of the reaction. This speed is represented by the ratio: 048139

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For each well (3 wells per concentration) 15 readings are taken at 3 second intervals.

11.2- Results

The results are collated in Table 1 in which are detailed the respective meanings of R, W, A, B, Y and D (Z and HET) in the formula (I) of the molecule tested. The results presented in Table I show that all of the compounds of formula (I) tested are very selective with respect to PLA2 from the IL group.

Compounds n ° 6 to 9 and 13 to 15, for which p is 1 and Y = -CO- have the strongest inhibiting activity, compounds n ° 7, 8, 9, 12, 13 and 14 being the most active with an IC50 value less than or equal to 0.3 μM vis-à-vis the human PLA2 of the IL group

Example 12 Study of In Vivo Activity

12.1- Materials and methods

The search for in vivo activity was carried out by the well-known test for carrageenan edema in rats.

In the experimental protocol followed, the reference product, indomethacin, or compound No. 5 to be evaluated, were administered either intraperitoneally (ip) or orally (po) one hour before the injection of carrageenan in the hind paw of the rat. The volume of the edema was measured at times 0, 3 and 5 hours after the injection of carrageenan. The doses used were 5, 10 and 20 mg / kg for the two products tested. 048139

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12.2- Results

By i.p., the two products have equivalent activity. Thus, at a dose of 10 mg / kg, the inhibitions of edema by indomethacin and compound No. 5 are 79% and 73% respectively.

By the po route, compound No. 5 has a clearly superior activity to that of indomethacin since, after 5 hours after the injection of carrageenan, it inhibits edema by 65%, while the reference product has an inhibitory activity of 16%, the two products having been administered at a dose of 10 mg / kg po

EXAMPLE 13 Second Study of In Vivo Activity

Example 13 relates to a study of the anti-inflammatory activity in vivo of certain compounds of the invention, by means of the test of ear edema as an experimental model of acute inflammation.

A. MATERIALS AND METHODS

A. L. Material and reagent.

Six samples of the compounds PMS 1227, PMS 1237, PMS 1281, PMS 1289, PMS 1314 and PMS 1315 were prepared.

The list of the various compounds referenced above and their chemical identity is given in detail below:

PMS 1227 l- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-ylmethyl) ben2yl] -d- tetradecylpiperazine.

PMS 1281

C28H44N4O3 + 1/2 H ₂ 0 = 493 g / mole l- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-ylmethyl) benzyl] -4- tétradécylpipérazine.

PMS1289 1- (para ((1,2,4- (4H) -5-oxo) oxodiazol-3-ylmethyl) benzoyl) 4-dodecyl-2,5-dimethylpiperazine. PMS 1314 1- [4 '- (4,5-dihydiO-1,2,4 (4H) -5-oxo-oxadiazol-3-ylmethyl) benzoyl] - 4-dodecylpiperazine.

PMS 1315 C ₂₉ H43 3θ ₃ S + 1H ₂ 0 = 531g / mole l- [4 '- (2,4-dioxo-1,3-thiazolidine-5ylidene) benzoyl] -4- tetradecylpiperazine.

As reagents, special grade or first grade croton oil, indomethacin (Sigma co.), Acetone, chloroform (100%), chloroform (80%), carboxymethylcellulose

(CMC), ethanol, hexane, ether, polyethylene glycol (PEG) and a saline solution.

A.2. The animals used were male mice of the ICR strain having a weight of 25 g.

A3. As instruments, we used: a punch for taking skin samples, an ear thickness measuring device (Ozaki, Japan), a balance, an automatic pipette, tweezers, a Nortex shaker, a anesthesia chamber, hood, cage, Eppendorf ® tube, safety mat, tubes etc.

A.4. To evaluate the anti-inflammatory effect in vivo of the PMS compounds of the invention listed above, the ear edema test was used as an experimental model for acute inflammation. Measurement of the local anti-inflammatory effect:

After inducing edema on one of the ears of the mice by applying croton oil, samples of each of the above PMS compounds were dissolved in 80% chloroform, then the resulting solution was applied to the ear, at a rate of one milligram of each of the compounds per ear. On the other side of the ear, only the solvent was applied, that is, 80% chloroform.

5 hours after the start of the experiment, the ear tissue at the edema was removed by punching the skin and the weight 048139

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tissue thus removed was compared with a punch sample from the control portion of the ear to calculate the inhibition ratio. Systemic anti-inflammatory effect:

Samples of the PMS compounds listed above were suspended in CMC and administered orally at 80 mg of each of the compounds per mouse. One hour after the start of the experiment, the edema was induced by applying croton oil.

Five hours after the application of croton oil, the weight of the tissue developing an edema was taken by punching the skin and compared with a sample by punching the skin of a control part in order to calculate the inhibition ratio. .

AT 5. For the calculation of the statistical significance, the results obtained respectively in the control group and the tested group were evaluated using the Student's test (Student's t-test). B. RESULTS

B.l Local anti-inflammatory effect

The local anti-inflammatory effect of the croton oil-induced ear edema test in mice by topical administration of the PMS compounds listed in the Materials and Methods section are shown in Table 2 below.

TABLE 2

* The numbers in parentheses indicate the number of experiments performed.

According to a classification in decreasing order of magnitude, the anti-inflammatory activities in vivo of the PMS compounds are classified from the 048139

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as follows: PMS 1289> PMS 1227> PMS 1281> PMS1315> PMS1314.

B. 2 Systemic anti-inflammatory effect

The results obtained show that the oral administration of the PMS compounds listed in the Materials and Methods section does not induce a systemic anti-inflammatory effect in the experimental model of oral edema induced by croton oil.

48

Ό Table I (continued)

"measured on a fluorimeter with recombinant human PLA ₂ (group II) ^b measured on a fluorimeter, unless otherwise specified, with pancreatic pig PLA ₂ (group I) ^c measured as indicated with the PLA2 of Crotalus durissus terrifiais (group II)

Table 1 (Suite 2)

Claims

CLAIMS 1. Compound having the following general formula (I):

in which:

- D signifies the Z-HET group or the Z≈HET group and (i) when D signifies the Z-HET group:

-HET is a 5-membered heterocycle such as oxadiazolone (II) or thiazolidine dione (III) of the following formulas:

-Z- is chosen from the group consisting of ~ (CRιR ₂ ) n- and - (CRi≈ CR ₂ ) n - with n being an integer having a value ranging from 1 to 6, Ri and R ₂ , identical or different , independently of one another, represent a hydrogen atom or a linear or branched alkyl group having from 1 to 6 carbon atoms.

(ii) when D signifies the Z≈HET group:

-Z- represents, with the heterocycle, a group -Z≈HÉT of formula (IV) or (V) below:

in which -Z = represents -CRi≈, in which Ri represents a. hydrogen atom or a linear or branched alkyl group of 1 to 6 carbon atoms; - p is an integer having the value 0 or 1;

-Y- is chosen from the group consisting of C≈O, SO2 and - (CR3R4) m- with m being an integer having a value ranging from 1 to 6, R3 and R, identical or different, independently represent one on the other one hydrogen atom or a linear or branched alkyl group having from 1 to 6 carbon atoms.

- A and B on the piperazine cycle, identical or different, independently of one another represent a carbon atom linked to hydrogens, a carbon atom linked both to a hydrogen and to a linear or branched alkyl group having 1 to 3 carbon atoms, or a -C≈O group.

- q is an integer having the value 0 or 1;

O 0

-W- is chosen from the group consisting of - c -, - o - c - and

0

II - ^{N ~ c} -;

-R is selected from the group consisting of linear or branched alkyl groups having from 1 to 22 carbon atoms, polyaryl groups and aryl-alkyl, alkyl-Q-alkyl, alkyl-Q-aryl, aryl-Q groups -aryl and aryl-Q-alkyl for which "aryl" represents an aryl group having 5 to 10 substituted or unsubstituted members.

2. Compound of formula (I) according to claim 1 characterized in that when the group R represents aryl-alkyl, alkyl-Q-aryl, aryl-Q-aryl and aryl-Q-alkyl, the aryl group is chosen from phenyl, naphthyl, phenylphenyl (or biphenyl) group or alternatively a heterocyclic aryl such as the indolyl group, "alkyl" represents a linear or branched alkyl group having from 1 to 12 carbon atoms and "Q" is chosen from the

0 group consisting of -O-, -S-, -NH-, -NRs-, -NH-CO-NH-, - o— S -,

O 0

S Il _M NC il O, OC il N _,

- c - o -, H and H, with R ₅ representing a linear or branched alkyl group having from 1 to 6 carbon atoms.

3. Compound according to one of claims 1 or 2, characterized in that p is equal to 1 and Y represents the group C≈O, the groups D, A, B, q, W and R having the meanings defined in the claim 1.

4. Compound according to one of claims 1 or 2, characterized in that it is chosen from the following compounds: a) l- [4 '- (4,5-dihydro-1,2,4 (4H) - 5-oxo-oxadiazol-3-ylmethyl) benzyl] -4-tetradecyl piperazine; b) 1- [4 '- (4,5-dihydro-1,2 _/ 4 (4H) -5-oxo-oxadiazol-3-ylmethyl) benzoyl] -4- octa decylpiperazine; c) 1- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-ylmethyl) benzoyl] - 2,5-di methyl-4-dodecylpiperazine; d) 1- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-ylmethyl) ρhenyl] -4- octa decylpiperazine; e) [4- (4'-octadecylpiperazin-l'-ylcarbonyl) benzylidene] -1,3-thiazolidine-2,4-dione; f) 1- [4 '- (2,4-dioxo-1,3-thiazolidin-5-ylmethyl) benzoyl] -4-octadecyl piperazine. g) l- [4 '- (4,5-dihydro-1,2,4 (4H) -5-oxo-oxadiazol-3-ylmethyl) benzyl] -4- tetradecylpiperazin-2-one h) la l- [4 '- (4 ₅ 5-dihydro-2 ₅ 4 (4H) -5-oxo-oxadiazol-3-ylethyl) benzoyl] -4- tétradécylpipérazine i) l- [4' - (4,5- dihydro-2 ₅ 4 (4H) -5-oxo-oxadiazol-3-ylproρyl) benzoyl] -4- tétradécylpipérazine j) l- [4 '- _{(4,5-dihydro-5} 2.4 (4H ) -5-oxo-oxadiazol-3-yl-methylethylbenzoyl] -4- (N- octadecylaminocarbonyl) piperazine

5. Method for the preparation of a compound of formula (I) according to one of claims 1 to 4, characterized in that it comprises the following stages: a) reacting hydroxylamine hydrochloride with a derivative of

where R, q, W, A, B, p, and Y have the same meaning as in claims 1 and 2:

- Z- is chosen from the group consisting of - (CRιR2) _n - and - (CRι = CR2) n - with n being an integer having a value ranging from 1 to 6, Ri and R ₂ , identical or different, independently of one another represent a hydrogen atom or a linear or branched alkyl group having from 1 to 6 carbon atoms, or to form the corresponding intermediate oxime, then b) subject the oxime obtained in step a) to a cyclization by reaction with a chlorocarbonate (or chloroformate) followed by heating to a temperature sufficient to obtain an essentially complete cyclization.

6. Process for the preparation of a compound of formula (I) according to one of claims 1 to 6, characterized in that it comprises a step consisting in: a) reacting the thiazolidine-2,4-dione on the aldehyde function of

wherein R, q, W, A, B, p and Y have the same meaning as in claims 1 or 2; and

-r is an integer having the value 0 or 1, -U- is chosen from the group consisting of ~ (CReR7) s- and - (CR ₆ = CR7) s - with s being an integer having a value ranging from 1 to 6 and with Rβ and R7, identical or different, representing a hydrogen atom or a linear or branched alkyl group having from 1 to 6 carbon atoms, to form the ethylenic derivative of formula (V) as defined in claim 1.

7. Method according to claim 6, characterized in that it comprises an additional step b) consisting in reducing the Z≈C double bond by catalytic hydrogenation.

8. Pharmaceutical composition characterized in that it comprises a compound of formula (I) according to one of claims 1 to 4.

9. Compound of formula (I) according to one of claims 1 to 4, for its use as an active principle of a medicament.

10. Use of a compound of formula (I) according to one of claims 1 to 4 for the preparation of a medicament intended for the prevention or the treatment of inflarnmation.