FR2817867A1 - ISOCOUMARINS INHIBITING PRODUCTION OF AMYLOID PEPTIDE, PREPARATION, COMPOSITIONS CONTAINING THEM AND USES - Google Patents

Abstract

The invention concerns novel compounds inhibiting production of amyloid peptide, their preparation and their uses. The invention also concerns methods for identifying or characterising compounds inhibiting production of amyloid peptide (and in particular non-toxic for embryonic development). The invention further concerns methods and uses of said compounds for treating nervous system disorders, in particular neurodegenerative pathologies such as Alzheimer's disease.

Description

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The present invention relates to new compounds which inhibit the production of amyloid peptide ss, their preparation and their uses. It also relates to methods for the identification or the characterization of compounds which inhibit the production of amyloid peptide ss, in particular selective inhibitors of the production of amyloid peptide ss (and in particular non-toxic for embryonic development). The invention also relates to methods and uses of the compounds for the treatment of disorders of the nervous system, in particular of neurodegenerative pathologies such as Alzheimer's disease.

 The search for protease inhibitors acting on the gamma cleavage of the APP protein and usable in Alzheimer's disease represents a major therapeutic challenge.

One of the constant signs of Alzheimer's disease is the existence in patients of amyloid deposits (senile plaques) whose main constituent is the amyloid peptide P (peptide Ap, in particular Ass42, which is mainly found in the formation of senile plaque).

l'APP. Les isoformes de l'APP sont des sialoglycoprotéines transmembranaires qui sont codées par un simple gène sur le chromosome 21 qui contient 19 exons. Les 3 isoformes de l'APP sont APP APP7i et l'APP Le clivage proteolytique de l'APP fait intervenir 3 protéases, appelées secretases a, ss et y. Le peptide Ass résulte du clivage ss et y (Murphy et al (1999)"y-secretase, evidence for multiple proteolytic activities and influence of membrane positioning of substrate on generation of amyloid ss peptides of varying length" J. Biol. Chem. 274 (17) 11914-11923). The Ass peptide is derived from a proteolytic process of one or more isoforms of

APP. APP isoforms are transmembrane sialoglycoproteins that are encoded by a single gene on chromosome 21 which contains 19 exons. The 3 APP isoforms are APP APP7i and APP The proteolytic cleavage of APP involves 3 proteases, called secretases a, ss and y. The Ass peptide results from the ss and y cleavage (Murphy et al (1999) "y-secretase, evidence for multiple proteolytic activities and influence of membrane positioning of substrate on generation of amyloid ss peptides of varying length" J. Biol. Chem. 274 (17) 11914-11923).

 The Ap peptide (Ass42 or Ass40 form) is toxic to neurons as well as other fragments of cleavage by y secretase, released in the cytoplasm. It would therefore be particularly advantageous to have compounds capable of inhibiting the production of these peptides, which are not toxic to organs or processes which involve the same or similar enzymes. This objective is not trivial because the evidence has accumulated tending to approximate or identify γ-secretetase and preselinin-1, which suggests that γ-secretase inhibitors will also be inhibitors of the pathways.

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 metabolic pathways in which presenilin-1 is involved, in particular the Notch Delta pathway. This path is found in embryonic development and in hematopoiesis. In this regard, the y secretase inhibitors described so far are also toxic for the NotchDelta pathway and are generally not very specific, inhibiting both chymotrypsin, elastase and calpains.

présente demande décrit une nouvelle sous famille d'iso-coumarines ayant des propriétés améliorées inhibitrices de la production de peptide amyloïde ss. Il s'agit plus spécifiquement des composés formule générale (I) ci-après :

dans laquelle : - RI et R2, indépendamment l'un de l'autre, sont choisis parmi un atome d'hydrogène, un groupe (CI-C24) alkyle, (C2-C24) alkényle, (C3-CI5) cycloalkyle, (C5-C 12) aryle, (C5-Cl2) aryle (CI-C24) alkyle, (C5-CI2) aryle (C2-C24) alkényle, (C3-C15) alkoxy, (C3-CI5) thioalkoxy, COR', COOR'ou CONHR', R'étant un atome d'hydrogène ou un groupe (CI-C24) alkyle, (C2-C24) alkényle, (C3-

C15) cycloalkyle, (C5-C12) aryle, (C5-CI2) aryle (CI-C24) alkyle, (C5-CI2) aryle (C2- C24) alkényle, un acide gras, saturé ou insaturé, ou un acide aminé, protégé ou non, ou, RI et R2 pris ensemble forment un système monocyclique ou bicyclique ayant de 5 à 15 chaînons dont éventuellement 1 à 4 hétéroatomes choisis parmi N, 0 et S, There is therefore a real need for compounds having improved pharmacological properties, ensuring better selectivity and lower toxicity during therapeutic uses. Surprisingly, the Applicant has now found, using an original method, inhibitor compounds having improved biological properties, from the point of view of efficacy, selectivity and / or toxicity. More specifically, the

The present application describes a new sub-family of iso-coumarins having improved properties which inhibit the production of amyloid peptide ss. More specifically, these are the compounds of general formula (I) below:

in which: - RI and R2, independently of each other, are chosen from a hydrogen atom, a (CI-C24) alkyl group ((C2-C24) alkenyl, (C3-CI5) cycloalkyl, ( C5-C 12) aryl, (C5-Cl2) aryl (CI-C24) alkyl, (C5-CI2) aryl (C2-C24) alkenyl, (C3-C15) alkoxy, (C3-CI5) thioalkoxy, COR ', COOR'or CONHR ', R' being a hydrogen atom or a (CI-C24) alkyl, (C2-C24) alkenyl, (C3-

C15) cycloalkyl, (C5-C12) aryl, (C5-CI2) aryl (CI-C24) alkyl, (C5-CI2) aryl (C2-C24) alkenyl, a fatty acid, saturated or unsaturated, or an amino acid, protected or not, or, RI and R2 taken together form a monocyclic or bicyclic system having 5 to 15 links, of which possibly 1 to 4 heteroatoms chosen from N, 0 and S,

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- R3 représente un groupe (CI-C24) alkyle, (C2-C24) alkényle, (C3-CI5) cycloalkyle (C5-C12) aryle, (C5-C12) aryle (CI-C24) alkyle, (C5-C12) aryle (C2-C24) alkényle ou un hétérocyle en (C3-Cl5) ayant de 1 à 3 hétéroatomes identiques ou différents choisis parmi N, 0 et S, - A représente un atome d'azote, de phosphore ou un groupe CX dans lequel X est un atome d'halogène (de préférence un atome de chlore) ou un groupe OR", R"étant un atome d'hydrogène ou un groupement (C5-CI2) aryle, (Cl-C6) alkyle ou tosyle, et - Y représente un atome d'oxygène, de soufre ou d'azote, les groupes alkyle, alkényle, cycloalkyle, aryle, alkoxy, thioalkoxy et le système cyclique formé éventuellement par RI et R2 étant non substitués ou substitués par un ou plusieurs

substituants choisis de préférence parmi un atome d'halogène, un groupe (Cl-C6) alkyle, (C3-C15) cycloalkyle, (C5-C12) aryle, (C3-C15) alkoxy, (C3-C15) thioalkoxy, amino, amino (Cl-C15) (cyclo) alkyle, aminodi (C 1-C 15) (cyclo) alkyle, COOH, (Cl-C15) acyle, (ClC15) acyloxy, nitro et trifluorométhyle, ainsi que leurs sels et, le cas échéant, énantiomères.

- R3 represents a (CI-C24) alkyl group ((C2-C24) alkenyl, (C3-CI5) cycloalkyl (C5-C12) aryl, (C5-C12) aryl (CI-C24) alkyl, (C5-C12) (C2-C24) alkenyl or a (C3-Cl5) heterocyle having from 1 to 3 identical or different heteroatoms chosen from N, 0 and S, - A represents a nitrogen, phosphorus or CX group in which X is a halogen atom (preferably a chlorine atom) or a group OR ", R" being a hydrogen atom or a (C5-CI2) aryl, (Cl-C6) alkyl or tosyl group, and - Y represents an oxygen, sulfur or nitrogen atom, the alkyl, alkenyl, cycloalkyl, aryl, alkoxy, thioalkoxy groups and the ring system optionally formed by RI and R2 being unsubstituted or substituted by one or more

substituents preferably chosen from a halogen atom, a (Cl-C6) alkyl, (C3-C15) cycloalkyl, (C5-C12) aryl, (C3-C15) alkoxy, (C3-C15) thioalkoxy, amino group, amino (Cl-C15) (cyclo) alkyl, aminodi (C 1-C 15) (cyclo) alkyl, COOH, (Cl-C15) acyl, (ClC15) acyloxy, nitro and trifluoromethyl, and their salts and, if applicable if necessary, enantiomers.

The invention therefore relates to the above compounds as inhibitors of the production of amyloid peptide Ap. The invention further relates to specific new compounds corresponding to the general formula (I) above, having advantageous properties.

According to the invention, the term "alkyl" denotes a linear or branched hydrocarbon radical having from 1 to 24 carbon atoms. It may be a lower alkyl having from 1 to 6 carbon atoms, such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert-butyl, pentyl, neopentyl, n-hexyl. C; -C groups are preferred. It may also be an alkyl with an aliphatic chain, linear or branched, comprising for example from 12 to 24 carbon atoms, preferably from 12 to 20. Mention may in particular be made of the linoleyl, linolenyl or oleyl groups.

According to the invention, the term "alkylene" denotes an unsaturated, linear or

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 branched, having 1 to 24 carbon atoms. It can have one or more double bonds. It can be a lower alkylene having from 2 to 6 carbon atoms, such as methylene, ethylene, propylene, butylene, etc. It may also be an alkylene with an unsaturated, linear or branched aliphatic chain, comprising for example from 12 to 24 carbon atoms, preferably from 12 to 20. Mention may in particular be made of palmityl, arachidonyl groups, etc.

The term cycloalkyl designates any cyclic hydrocarbon system, which can typically comprise from 3 to 15 carbon atoms and be mono- or poly-cyclic. A preferred cycloalkyl is the adamantyl group.

Aryl groups are mono-, bi- or tricyclic aromatic hydrocarbon systems having from 2 to 18 carbon atoms and optionally one or more heteroatoms, preferably from 5 to 12 carbon atoms.

The term arylalkyl or arylalkylene designates any alkyl or alkylene group as defined above, in which one or more aryl groups as defined above is incorporated.

The "acyl" groups are CO- (cyclo) alkyl or CO-aryl groups, (cyclo) alkyl and aryl being as defined above.

The alkoxy groups correspond to the (cyclo) alkyl groups defined above linked to the nucleus via an O-bond (ether).

The "thioalkoxy" groups correspond to the (cyclo) alkyl groups defined above linked to the nucleus via an S-bond (thio).

The term “heterocycle” means any carbon mono- or multicyclic system advantageously containing from 3 to 15 links, including optionally 1 to 4 heteroatoms chosen from 0, N and S. Mention may in particular be made of the pyridinyl, pyrimidyl, imidazolyl, benzymidazolyl, piperazinyl groups, etc.

By halogen is meant a fluorine, chlorine, bromine or iodine atom.

By amino acid is meant any natural amino acid, in particular alanine, valine, proline, tryptophan, glutamine, etc.

By heteroatom is meant an atom chosen from 0, N and S.

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As illustrated in the examples, the preferred compounds according to the invention have the required properties of inhibiting the production of Ap peptide, but respect the embryonic development. In this regard, the present invention indeed shows in an advantageous and unexpected manner that it is possible to discriminate between the action on the production of Ap peptide and on other phenomena in which presenilinel also intervenes (for example the Notch Delta pathway). , embryonic development or hematopoiesis), and to produce or select compounds with a particularly advantageous pharmacological profile. The invention therefore also relates to a method of selection, identification, characterization or qualification of selective protease inhibitor compounds, consisting of a double test of the compound on the one hand on the production of Ap peptide (from APP) and on the other hand on a toxic activity dependent on protease.

 The subject of the invention is also compositions, in particular pharmaceutical compositions comprising the compounds as defined above, or obtained by the method described above. The invention further relates to the use of these compounds or compositions for the treatment of neurodegenerative pathologies, in particular for the inhibition of the cleavage of APP in subjects, in particular for treating or inhibiting or preventing the appearance or development Alzheimer's disease.

 Inhibiting compounds which are particularly preferred within the meaning of the invention are compounds of general formula (I) above, in which: - Y represents 0 or N, more preferably 0, and / or - A represents a nitrogen atom or a CX group in which X is a halogen atom (preferably a chlorine atom) or an OR group ", R" being a hydrogen atom or a (Cl-C3) alkyl or tosyl group, and / or - RI is a hydrogen atom or an alkyl or alkylene group comprising from 1 to 6 carbon atoms, preferably a hydrogen atom, and / or - R2 is a hydrogen atom or a COOR group in which R ' is a (C 1 -C 24) alkyl, (C2-C24) alkenyl, (C3-C15) cycloalkyl, (C5-C12) aryl, (C5-C12) aryl (Cl-C24) alkyl or (C5-CI2) group aryl (C2-C24) alkenyl, even more

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préférentiellement un groupe (C3-C15) cycloalkyle, notamment adamantyle, et/ou - R3 est un groupe alkyle substitué, de préférence un groupe alkylalkoxy.

preferably a (C3-C15) cycloalkyl group, in particular adamantyl, and / or - R3 is a substituted alkyl group, preferably an alkylalkoxy group.

In another particular embodiment, the compounds correspond to the general formula (I) in which R2 is a CONHR 'group, R' being a hydrogen atom or a (Cl-C24) alkyl, (C2-C24) alkenyl group, (C3-CI5) cyc1oalkyle, (C5-CI2) aryl, (C5-CI2) aryl (ClC24) alkyl or (C5-C12) aryl (C2-C24) alkenyl.

 In another particular mode, the compounds correspond to the general formula (I) in which R2 is a group COR ', COOR'or CONHR', R'étant, a fatty acid, saturated or unsaturated, or an amino acid, protected or no.

 In a particular embodiment, R3 represents an alkyl, alkenyl or aryl group omega-functionalized by an alcohol, acid or amine terminal function.

 In another more preferred embodiment of the compounds according to general formula (I), A represents a group CX in which X is a halogen atom, preferably a chlorine atom. A particularly advantageous family of compounds is in fact made up of halogenated isocoumarins.

 A particularly preferred family is that in which A represents a CX group in which X is a halogen atom, preferably a chlorine atom, and Y represents 0 or N, more preferentially O. Include more preferentially, it is d a family in which RI further represents a hydrogen atom or an alkyl or alkylene group comprising from 1 to 6 carbon atoms, preferably a hydrogen atom.

 Preferred compounds are characterized by a chloro-isocoumarin ring 4 substituted in position 7 by an amine which can be engaged in a carbamate or amide bond. It is particularly preferred that this hydrophobic structure consists of a hydrophobic alcohol carbamate, such as the adamantyl substituent.

 A particularly preferred family consists of compounds of formula

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générale (I) telle que définie ci-avant, dans laquelle A est un groupe C-Cl, RI est un atome d'hydrogène, R2 est un atome d'hydrogène ou un groupe COOR'dans lequel R'est un groupe (C3-CI5) cycloalkyle, Y est un atome d'oxygène, et R3 est un groupe alkyle substitué, de préférence un groupe alkylealkoxy. De tels composés constituent en outre un objet particulier de la présente invention.

general (I) as defined above, in which A is a C-Cl group, RI is a hydrogen atom, R2 is a hydrogen atom or a COOR group in which R is a group (C3 -CI5) cycloalkyl, Y is an oxygen atom, and R3 is a substituted alkyl group, preferably an alkylalkoxy group. Such compounds further constitute a particular object of the present invention.

 The compounds of the invention can be synthesized from commercial materials, such as homophthalic acid using conventional methods of organic chemistry, as illustrated in the examples. Thus, it is possible, from homophthalic acid, to synthesize the 7-amino-4-chloro-3-substituted alkyl isocoumarin nucleus according to the protocol described by Powers et al. (Powers et al. (1990) Biochemistry, 29, 3108-3118). From this key synthon, the "dressing" of the aromatic amine function can be carried out according to the acylation reaction already very long described using acid chloride, and the formation of urea by the use isocyanates. It is understood that any other conventional synthetic route, known to a person skilled in the art, can be used for the production of the above compounds.

- ELISA, détection fluorescence, selon la méthode proposée par G. Christie, & al. (1999), Alzheimer disease : correlation of the suppression b-amylO peptide secretion from cultured cells with inhibition of the chymotrypsin-like activity of the proteasome ; J. of Neurochemistry, 73,195-204 ; - Immunoprécipitation-spectro de masse, selon la technique de Ron Wang et al. The biological properties of the compounds of the invention can be verified and tested according to different models or tests. In this regard, the inhibitory activity of the production of Ap peptide can be determined for example by one of the following tests, described in the literature: - Immunoprecipitation-electrophoresis of radiolabelled products, according to the technique described by ME Figueiredo-Pereira & al. (1999), Distinct secretases, a cystein protease and a serine protease, generate the C-termini of amyloiä ss-proteins Ass 1-40 and Ass 1-42 respectively; J. Neurochem. 72, 1417-1422;

- ELISA, fluorescence detection, according to the method proposed by G. Christie, & al. (1999), Alzheimer disease: correlation of the suppression b-amylO peptide secretion from cultured cells with inhibition of the chymotrypsin-like activity of the proteasome; J. of Neurochemistry, 73, 195-204; - Mass spectro immunoprecipitation, according to the technique of Ron Wang et al.

(1996) The profile of soluble Amyloid? Protein in cultured cell media, detection and

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quantification of amyloid ss protein and variants by immunoprecipitation-mass spectrometry. J. Biol. Chem. 271, 31894-31902 ou de Michael P. Murphy & al. (1999), ySecretase, evidence for multiple proteolytic activities and influence of membrane positioning of substrate on generation of amyloiä ss peptides of varying length, J.

quantification of amyloid ss protein and variants by immunoprecipitation-mass spectrometry. J. Biol. Chem. 271, 31894-31902 or Michael P. Murphy & al. (1999), ySecretase, evidence for multiple proteolytic activities and influence of membrane positioning of substrate on generation of amyloiä ss peptides of varying length, J.

Biol. Chem. 274, 11914-11923.

 These various tests, alone or in combination (s), make it possible to evaluate the inhibitory activity of the compounds of the invention. It is understood that the inhibitory nature includes total or partial inhibition of the activity. However, it is preferred, within the framework of the invention, compounds capable of inhibiting at least 25%, more preferably at least 45% of the production of Ap peptide, in particular of the Ass42 form.

 As indicated above, the compounds according to the invention are preferably selective inhibitors of the production of Ap peptide. The selective term indicates that the preferred compounds according to the invention have an inhibiting action on the hydrolysis activity of the higher APP protein. their possible inhibitory action on other proteolytic processes (other substrates and / or other enzymes, such as elastase, calpain and / or chymotrypsin, for example). Preferred compounds within the meaning of the invention are compounds which are essentially inactive or have no effect on the Notch Delta function or the segmentation or expression of the lunatic fringe gene.

 The toxicity or non-toxicity on the enzyme-dependent pathways can be assessed by injection into embryos or pregnant females, followed by observation of disturbances in embryonic development. A preferred test according to the invention is the blocking of embryonic development observed on a recently embryonated egg, as described in R. Jelinek & al. (1985), Indian Journal of experimental biology, 23, 588-595.

 This property can also be determined by testing the effect of the compounds on a function known to be important in embryogenesis, such as segmentation, as described below. Thus, a preferred toxicity test is based on gene expression profiles during the somitogenesis of the chicken embryo (MJ Mc Grew & al (1998) Curr. Biol., 8,979-982; Caroline Jouve, Thesis University of the Mediterranean, 28 Sept 2000). In this regard, the team of Dr. Pourquié, inventor of the present application, has highlighted

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l'expression, en vague de propagation, du gène lunatic fringe dans le mésoderme présomitique, qui précède la somitogenèse. Un test particulier au sens de l'invention réside plus particulièrement dans une mesure de l'expression (ou de la vague de propagation) du gène lunatic fringe, à partir d'embryons. La mesure de l'expression de ce gène peut être réalisée par toute technique connue (sondes marquées, amplification, électrophorèse, puces, hybridation in situ, etc. ). Pour cela, on sépare des embryons de poulet de 2 jours selon l'axe sagittal en leurs 2 moitiés. Une moitié est fixée immédiatement alors que la moitié contralatérale est mise en culture pendant 90 minutes. On observe la propagation de l'expression de Lunatic Fringe, vue par hybridation in situ, dans la moitié expérimentale par rapport à la moitié contrôle fixée. Selon l'invention on cultive les deux moitiés de l'embryon en parallèle pendant 2 heures, une moitié sert de témoin et l'autre est cultivée en présence d'un composé test sélectionné en b). L'analyse de l'image permet de quantifier l'effet du composé test. Lorsque l'on observe une modification de l'expression de lunatic fringe, il y a présomption de toxicité.

the expression, in a propagation wave, of the lunatic fringe gene in the presomitic mesoderm, which precedes somitogenesis. A particular test within the meaning of the invention lies more particularly in a measurement of the expression (or of the propagation wave) of the lunatic fringe gene, from embryos. The expression of this gene can be measured by any known technique (labeled probes, amplification, electrophoresis, chips, in situ hybridization, etc.). For this, we separate 2-day chicken embryos along the sagittal axis into their 2 halves. Half is fixed immediately while the contralateral half is cultured for 90 minutes. The propagation of the Lunatic Fringe expression, seen by in situ hybridization, is observed in the experimental half compared to the fixed control half. According to the invention, the two halves of the embryo are cultivated in parallel for 2 hours, one half serves as a control and the other is cultivated in the presence of a test compound selected in b). Analysis of the image makes it possible to quantify the effect of the test compound. When a change in the expression of lunatic fringe is observed, there is a presumption of toxicity.

 It is also possible to use a cell test of the Notch Delta function as described in Bart from Strooper & al. (1991) Nature 398,518-525.

inhibiteurs de la production de peptide Ap et essentiellement inactif sur le développement embryonnaire, notamment sur l'expression du gène lunatic fringe chez l'embryon ou sur la voie Notch. Comme indiqué précédemment, l'invention montre, de manière inattendue et avantageuse, qu'il est possible de discriminer entre l'action sur la production de peptide Ap et sur d'autres phénomènes où intervient aussi la présénilinel (par exemple la voie Notch Delta, le développement embryonnaire ou l'hématopoïèse), et de produire ou sélectionner des composés présentant un profil pharmacologique particulièrement avantageux. Another subject of the invention relates to methods for the selection, identification, characterization or qualification of selective protease inhibitor compounds, in particular the production of Ap peptide from APP. More particularly, these are methods for selecting, identifying, characterizing or qualifying compounds

inhibitors of Ap peptide production and essentially inactive on embryonic development, in particular on the expression of the lunatic fringe gene in the embryo or on the Notch pathway. As indicated above, the invention unexpectedly and advantageously shows that it is possible to discriminate between the action on the production of Ap peptide and on other phenomena in which presenilinel also intervenes (for example the Notch Delta pathway). , embryonic development or hematopoiesis), and to produce or select compounds with a particularly advantageous pharmacological profile.

 The methods of the invention more particularly comprise: a) a determination of the action of one or more test compounds on the production of Ap peptide (for example the form Ass40 or Ass42,

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notamment Ass42), en particulier par clivage de l'APP (sauvage ou d'un isoforme ou mutant), b) la sélection de composés tests capables d'inhiber l'activité de production en a), c) une détermination de l'action de composés sélectionnés en b) sur le développement embryonnaire ou l'expression de voies métaboliques associées au développement embryonnaire, et d) la sélection de composés ayant une activité d'inhibition de la production de peptide Ap en a) supérieure à l'activité d'inhibition sur le développement embryonnaire en c).

in particular Ass42), in particular by cleavage of APP (wild type or of an isoform or mutant), b) the selection of test compounds capable of inhibiting the production activity in a), c) a determination of the action of compounds selected in b) on embryonic development or expression of metabolic pathways associated with embryonic development, and d) selection of compounds having an activity of inhibiting the production of Ap peptide in a) greater than the activity inhibition on embryonic development in c).

 More preferably, the inhibitory action on the production of Ap peptide in a) is much greater than the inhibitory activity on embryonic development in c), preferably by a factor of 2 or more. Even more preferably, the compounds selected in d) are essentially devoid of effect or action in test c), and therefore exhibit low toxicity and high selectivity for cleavage of APP.

 The determination of the inhibitory action of step a) can be carried out according to various methods, such as the techniques of immunoprecipitation-electrophoresis, ELISA, or immunoprecipitation-mass spectrometry mentioned above.

 Preferably, the inhibitory effect on the production of Ap peptide is determined using a mammalian cell, preferably of neuronal origin (for example a neuroblastoma), expressing and / or transfected with a nucleic acid coding for human APP, carrying or not a mutation favoring the cleavage (s) ss and / or y. Another preferred embodiment consists in the use of cortical neurons freed from glial cells. Indeed it is important to verify that the inhibition is effective on the neurons themselves, an inhibition limited to glial cells would not be sufficient, because even if it results in a significant reduction in amyloid deposits it would not prevent the neuron from dying cytoplasmic fragments of APP, the toxicity of which has recently been demonstrated (PCT / FROO / 02174).

 The products formed, after extraction and immunoprecitation are analyzed by

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spectrométrie de masse/électro-spray.

mass spectrometry / electro-spray.

production du peptide Ass42 ou Ass40, plus préférentiellement Ass42, qui est majoritairement présent dans les plaques séniles. In step b), a selection is made of the compounds capable of at least partially inhibiting the production of Ap peptide. Preferably, the compounds capable of inhibiting at least 25%, more preferably at least 45% of the production are selected of Ap peptide observed under the same conditions but in the absence of the test compound. Even more preferably, the compounds capable of inhibiting the

production of the Ass42 or Ass40 peptide, more preferably Ass42, which is mainly present in senile plaques.

 In step c), the action of the selected compounds on embryonic development or the expression of metabolic pathways associated with embryonic development can be determined according to the techniques mentioned above. A preferred test is based on determining the effect of the test compounds on a metabolic function or pathway involved in embryogenesis, in particular on the expression of the lunatic fringe gene.

 The method described above can be applied to different types of test compounds, of origin and of varied structure. They can be nucleic acids, polypeptides or peptides, lipids, sugars, fatty acids, biological extracts, environmental samples, synthetic or semi-synthetic molecules, organic or mineral compounds, etc. The tests can be carried out on several compounds in parallel (or as a mixture), for example in assay plates (multi-well) or any other device. The method according to the invention can be used to characterize or profile compounds known for their inhibitory activity on the production of Ap peptide, with the aim of selecting compounds having an appropriate selectivity / toxicity profile. It is understood that the order of the steps can be reversed, the test for inhibition of the production of Ap peptide being carried out in second.

On the other hand, the method may include additional steps for determining the activity of the compounds on other substances (eg, enzymes) or other substrates.

 In this regard, an object of the invention also lies in the use of compounds which are selective inhibitors of the production of Ap peptide (in particular inhibitors of the activity of APP hydrolysis by gamma secretase and which are essentially inactive or without effect on the

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fonction Notch Delta ou la segmentation ou l'expression du gène lunatic fringe) pour la préparation d'une composition pharmaceutique destinée au traitement de pathologies neurodégénératives, notamment de pathologies associées à l'hydrolyse de l'APP, en particulier de la maladie d'Alzheimer.

Notch Delta function or segmentation or expression of the lunatic fringe gene) for the preparation of a pharmaceutical composition intended for the treatment of neurodegenerative pathologies, in particular pathologies associated with APP hydrolysis, in particular Alzheimer.

 Another subject of the invention also resides in a composition, in particular a pharmaceutical composition, comprising a compound as defined above. The pharmaceutical compositions according to the invention advantageously comprise a vehicle or excipient as well as, if necessary, other active ingredients. Thus, the compounds according to the invention can be used alone or in combination (s), in particular with other agents or active principles on certain disorders of the nervous system. The active ingredients can be used simultaneously (and in this case packaged together or separately), or separately or spaced over time.

 The compounds or compositions according to the invention can be administered in different ways and in different forms. Thus, they can be injected by the systemic route, such as for example by the intravenous, intramuscular, subcutaneous, trans-dermal, intra-arterial, intracerebral, etc. route, or administered by the oral route, the oral, intravenous routes, intramuscular and subcutaneous being preferred.

 For injections, the compounds are generally packaged in the form of liquid suspensions, which can be injected using syringes or infusions, for example. In this regard, the compounds are generally dissolved in saline, physiological, isotonic, buffered solutions, etc., compatible with pharmaceutical use and known to those skilled in the art. Thus, the compositions can contain one or more agents or vehicles chosen from dispersants, solubilizers, stabilizers, preservatives, etc. Agents or vehicles which can be used in liquid and / or injectable formulations are in particular methylcellulose, hydroxymethylcellulose, carboxymethylcellulose, polysorbate 80, mannitol, gelatin, lactose, vegetable oils, acacia, etc.

 The compounds can also be administered in the form of gels, oils, tablets, suppositories, powders, capsules, capsules, etc., optionally by means of

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formes galéniques ou de dispositifs assurant une libération prolongée et/ou retardée. Pour ce type de formulation, on utilise avantageusement un agent tel que la cellulose, des carbonates ou des amidons.

dosage forms or devices ensuring prolonged and / or delayed release. For this type of formulation, an agent such as cellulose, carbonates or starches is advantageously used.

 It is understood that the flow rate and / or the dose administered can be adapted by a person skilled in the art depending on the patient, the pathology concerned and its stage of development, the mode of administration, etc. Typically, the compounds are administered in doses which can vary between 0.1 μg and 100 mg / kg of body weight, more generally from 0.01 to 10 mg / kg, typically between 0.1 and 10 mg / kg. In addition, repeated administrations can be performed, if necessary.

 The present invention can be used for the treatment or management, for preventive or curative purposes, of pathologies of the nervous system, in particular of neurodegenerative pathologies, even more preferably of pathologies linked to the presence of cleaved forms of the APP protein (in particular of the amyloid peptide Ass), specifically of Alzheimer's disease. The term treatment means, within the meaning of the invention, the treatment of symptoms, the reduction of the progression or symptoms, the reduction in the development or the appearance of the pathology, the improvement in the quality and / or the duration of life, etc.

 Thus, a particular object of the invention resides in the use of a compound as defined above for the preparation of a composition intended for the treatment of pathologies of the nervous system, in particular of neurodegenerative pathologies, even more preferably of pathologies linked to the presence of cleaved forms of the APP protein. The invention is particularly suitable for the treatment of Alzheimer's disease.

 Another subject of the invention resides in a method of treatment of pathologies of the nervous system, in particular of neurodegenerative pathologies, even more preferably of pathologies linked to the presence of cleaved forms of the APP protein, comprising the administration to a subject of an effective amount of a compound as defined above. The subject can be any mammal, preferably human. As indicated above, the method of the invention is particularly suitable for reducing or slowing down the appearance, progression or development of the pathology, in particular of Alzheimer's disease.

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Other aspects and advantages of the present invention will be described in detail in the following examples, which should be considered as illustrative and not limiting.

Acide 4-Nitro-homophtalique 1 Formule brute : C9H7NO6 Masse moléculaire : 225. 15 g/mol NcooH COOH

A une solution d'acide nitrique fumant à 0 C est additionné l'acide homophtalique (5g, 31mmol). Le mélange réactionnel est laissé sous agitation magnétique à température ambiante pendant 4 heures. La solution est alors jetée sur 1g de glace pilée. Le solide résultant est filtré puis séché sous pression réduite pour donner 1 (3. 88g, 88%) sous la forme d'une poudre blanche. Examples Example-Synthesis of 4-Nitro-homophthalic acid

4-Nitro-homophthalic acid 1 Crude formula: C9H7NO6 Molecular mass: 225. 15 g / mol NcooH COOH

To a fuming nitric acid solution at 0 C is added homophthalic acid (5g, 31mmol). The reaction mixture is left under magnetic stirring at room temperature for 4 hours. The solution is then thrown on 1g of crushed ice. The resulting solid is filtered and then dried under reduced pressure to give 1 (3.88g, 88%) in the form of a white powder.

Rf (methanol 1 / dichloromethane 1) = 0.6 H-NMR (250 MHz, DMSO) ô: 8. 43 (d, 1H, J = 2. 4Hz, CH = C-COOH), 8. 18 (dd, 1H, J = 2. 4Hz, J '= 8.4Hz, N02-C-CH = CH), 7. 48 (d, 1H, J = 8.4Hz, N02-C-CH = CH), 3. 93 (s, 2H, CHJ IR: 1681, 1513, 1412, 1340, 902 cm- 'Mass spectrum (GT-FAB> O): (M + H) 226 Mp: 217OC Log P (n-octanol / H2O) = 1. 11 0, 33 Example 2-Synthesis of 2- (2-Methoxy-ethoxycarbonylmethyl) -5-nitro-benzoic acid

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Acide 2- (2-Methoxy-ethoxycarbonylmethyl)-5-nitro-benzoïque 2 Formule brute : C12H13N07 O'NlXl Masse moléculaire : 283, 24 g/mol 1 0 1 1

A une solution de 1 (2. 19g, 9. 73mmol) dans du 2-Méthoxyéthanol (20ml), on ajoute de l'acide sulfurique (95%, 150 l, 2. 76mmol). Le mélange réactionnel est chauffé à 100oC, sous agitation magnétique, pendant 2 heures. 10ml d'AcOEt sont ajouté au mélange et la solution est extraite avec 3xl0ml d'une solution de NaHC03 5%. Les phases aqueuses sont réunies et acidifiées par une solution d'acide citrique jusqu'à obtenir un pH=3. Cette solution est alors extraite avec 3x15ml d'AcOEt, les phases organiques sont réunies et séchées sur sulfate de magnésium, filtrées et évaporées sous pression réduite. On obtient ainsi 2 (2.40g, 87%) sous la forme d'une poudre blanche.

2- (2-Methoxy-ethoxycarbonylmethyl) -5-nitro-benzoic acid 2 Crude formula: C12H13N07 O'NlXl Molecular mass: 283, 24 g / mol 1 0 1 1

To a solution of 1 (2.19 g, 9.73 mmol) in 2-methoxyethanol (20 ml), sulfuric acid (95%, 150 l, 2. 76 mmol) is added. The reaction mixture is heated to 100 ° C., with magnetic stirring, for 2 hours. 10ml of AcOEt are added to the mixture and the solution is extracted with 3xl0ml of a 5% NaHC03 solution. The aqueous phases are combined and acidified with a citric acid solution until a pH = 3 is obtained. This solution is then extracted with 3x15ml of AcOEt, the organic phases are combined and dried over magnesium sulfate, filtered and evaporated under reduced pressure. 2 (2.40 g, 87%) are thus obtained in the form of a white powder.

Rf (methanol 2 / dichloromethane 14) = 0.70
H NMR (250 MHz, CD30D) #: 8.99 (d, 1H, J = 2.51Hz, CH = C-COOH), 8.52 (dd, IH,
J = 2. 50Hz, J '= 8. 41Hz, N02-C-CH = CH), 7.77 (d, IH, J = 8.43Hz, N02-C-CH = CH), 4.38 (s, 2H, CH-COO), 4.38 (t, 2H, COOCH) , 3.76 (t, 2H, J = 4.63Hz, CH2-OMe), 3.52 (s,
3H, OCH3)
Mass spectrum (NOBA-FAB> 0): (M + H) = 284
Mp: 55 C
Log P (n-octanol / H2O) = 1.58 0.37 Example 3-Synthesis of 4-Chloro-3- (2-methoxy-ethoxy) -7-nitro-isochromen-1-one

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4-Chloro-3- (2-methoxy-ethoxy)-7-nitro-isochromen-l-one 3 Formule brute : C12HIOC1N06 Masse moléculaire : 299. 66 g/mol 02N 1 10 0 0 15C ! CI

A une solution de 2 (2. 4g, 8. 49mmol) dans du toluène (50ml), on ajoute PC15 (4. 4g, 21.22mmol). Le mélange réactionnel est porté à reflux pendant 15 heures. Le solvant est évaporé sous pression réduite et 10ml d'eau sont ajoutés au résidu. La solution hétérogène est portée à reflux pendant 1 heure et le mélange est extrait avec 2x20ml d'AcOEt. Les phases organiques sont réunies et lavées avec 3x10ml de brine, puis séchées sur Sulfate de magnésium et évaporées sous vide. Le résidu est recristallisé à chaud dans le méthanol pour donner 3 (1.77mg, 69%) sous la forme d'une poudre jaune.

4-Chloro-3- (2-methoxy-ethoxy) -7-nitro-isochromen-l-one 3 Crude formula: C12HIOC1N06 Molecular mass: 299. 66 g / mol 02N 1 10 0 0 15C! THIS

To a solution of 2 (2.4g, 8.49mmol) in toluene (50ml), PC15 (4.4g, 21.22mmol) is added. The reaction mixture is brought to reflux for 15 hours. The solvent is evaporated under reduced pressure and 10 ml of water are added to the residue. The heterogeneous solution is brought to reflux for 1 hour and the mixture is extracted with 2x20ml of AcOEt. The organic phases are combined and washed with 3 × 10 ml of brine, then dried over magnesium sulphate and evaporated under vacuum. The residue is recrystallized hot from methanol to give 3 (1.77 mg, 69%) in the form of a yellow powder.

OCH) OL) RMN 13C (250MHz, CDCI3) 8 : 157. 488,155. 788,149. 891,143. 143,129. 819,
126.388, 123.683, 116.947, 90.406, 70.508, 69.991, 59.340
IR : 2919,1737, 1102,843 cm-'
Spectre de masse (NOBA-FAB > O) : (M+H) += 300
Mp : 127 C
Log P (n-octanol/H2O) = 2.25 # 0,75 Exemple 4-Synthèse de 7-Amino-4-chloro-3-(2-methoxy-ethoxy)-isochromen-7-one Rf (ethyl acetate 7 / dichloromethane 3 / hexane 12) = 0.37 1 H NMR (250 MHz, CDCl) 8: 8.95 (d, 1H, J = 2.32Hz, CH = C-COO-), 8.45 ( dd, IH,
J = 2.37Hz, J '= 8. 97Hz, N02-C-CH = CH), 7.78 (d, 1H, J = 8.95Hz, N02-C-CH = CH), 4.54 (t, 2H, J = 4. 40Hz, O-CH2-CH2-OMe ), 3.71 (t, 2H, J = 4.44Hz, -CH2-OMe), 3.37 (s, 3H,

OCH) OL) 13C NMR (250MHz, CDCI3) 8: 157. 488,155. 788.149. 891.143. 143.129. 819,
126.388, 123.683, 116.947, 90.406, 70.508, 69.991, 59.340
IR: 2919.1737, 1102.843 cm- '
Mass spectrum (NOBA-FAB> O): (M + H) + = 300
Mp: 127 C
Log P (n-octanol / H2O) = 2.25 # 0.75 Example 4-Synthesis of 7-Amino-4-chloro-3- (2-methoxy-ethoxy) -isochromen-7-one

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7-Amino-4-chloro-3- (2-methoxy-ethoxy)-isochromen-l-one 4 Formule brute : CH, 2CINO4 Masse moléculaire : 269. 68 g/mol H2N o ooMe

A une solution de 3 (500g, 1. 66mmol) dans du THF (30ml), on ajoute le palladium activé sur charbon (10% Pd, 50mg). Le mélange réactionnel est hydrogéné à température ambiante pendant 6 heures puis filtré sur célite. Le solvant est évaporé sous pression réduite et le résidu est purifié par chromatographie sur colonne de silice (acétate d'éthyle 9/ cyclohexane 12) pour donner 4 (447mg, 100%) sous la forme d'une poudre jaune.

7-Amino-4-chloro-3- (2-methoxy-ethoxy) -isochromen-l-one 4 Crude formula: CH, 2CINO4 Molecular mass: 269. 68 g / mol H2N o ooMe

To a solution of 3 (500g, 1.66mmol) in THF (30ml), the palladium activated on carbon (10% Pd, 50mg) is added. The reaction mixture is hydrogenated at room temperature for 6 hours and then filtered through celite. The solvent is evaporated under reduced pressure and the residue is purified by chromatography on a silica column (ethyl acetate 9 / cyclohexane 12) to give 4 (447 mg, 100%) in the form of a yellow powder.

Rf (Ethyl acetate 9 / hexane 12) = 0.39 tH NMR (250 MHz, CDCl3): 7.48 (d, 1H, J = 8.57 Hz, NH2-C-CH = CH), 7. 38 (d, 1H, J = 2.42Hz, NH2-C-CH = C-C02), 05.05 (dd, 1H, J = 2.47Hz, J '= 8.57Hz, NH2-C-CH = CH ), 4.39 (t, 2H, J = 4.52Hz, O-CHrCHOMe), 3.68 (t, 2H, J = 4.51Hz ,. O-CH2-CH-OMe), 3.38 (s , 3H, 0-CHJ IR: 1799, 1748cm '' Mass spectrum (NOBA-FAB> O): (M + H) + = 270 Mp: 127OC Log P (n-octanol / H2O) = 1. 37 0, 75 Example 5-Synthesis of [4-Chloro-3- (2-methoxy-ethoxy) -1-oxo-77 isochromen-7-yl] carbamic acid adamantan-1-yl ester

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4-Chloro-3- (2-methoxy-ethoxy)-l-oxo-lH-isochromen-7-yl]-carbamicacid adamantan-1-yl ester 5 Formule brute : CHCINOo Masse moléculaire : 447. 91 g/mol s /7 ! i 1 Cl

A une solution de 4 (50g, 0. 22mmol) dans du THF (1. 25ml), on ajoute le l-Adamantyl fluoroformate (225mg, 2. 1mmol) et la TEA (1401, Immol). Le mélange réactionnel est laissé sous agitation magnétique à température ambiante pendant 3jours. Le solvant est évaporé sous pression réduite et le résidu est solubilisé dans 10ml d'AcOEt. La solution organique est alors lavée avec 2x10ml d'une solution de NaHC03 5%, puis avec 2x10ml de Brine et enfin avec 2x10ml d'acide citrique puis elle est séchée sur sulfate de magnésium et évaporée sous pression réduite. Le résidu est purifié par chromatographie sur colonne de silice (acétate d'éthyle 6/cyclohexane 12) pour donner 5 (50mg, 50%) sous la forme d'une poudre jaune.

4-Chloro-3- (2-methoxy-ethoxy) -l-oxo-1H-isochromen-7-yl] -carbamicacid adamantan-1-yl ester 5 Crude formula: CHCINOo Molecular mass: 447. 91 g / mol s / 7! i 1 Cl

To a solution of 4 (50g, 0.22mmol) in THF (1.25ml), l-Adamantyl fluoroformate (225mg, 2.1mmol) and TEA (1401, Immol) are added. The reaction mixture is left under magnetic stirring at room temperature for 3 days. The solvent is evaporated off under reduced pressure and the residue is dissolved in 10 ml of AcOEt. The organic solution is then washed with 2 × 10 ml of a 5% NaHCO 3 solution, then with 2 × 10 ml of Brine and finally with 2 × 10 ml of citric acid, then it is dried over magnesium sulfate and evaporated under reduced pressure. The residue is purified by chromatography on a silica column (ethyl acetate 6 / cyclohexane 12) to give 5 (50 mg, 50%) in the form of a yellow powder.

Rf (Ethyl acetate 6 / hexane 12) = 0.59 H NMR (250 MHz, CDC): 7.93 (d, IH, J = 2.25Hz, NH-C-CH = C-COO), 7.73 (dd, IH , J = 8.69Hz, NH-C = CH-CH), 7.47 (d, IH, J = 8.74Hz, NH-C = CH-CH), 6.59 (s, 1H, -
NH), 4.31 (t, 2H, J = 4.45Hz, O-CH-CH2-OMe), 3.58 (t, 2H, J = 4.48Hz, O-CH2-CH-
OMe), 3.27 (s, 3H, OCH3)
IR: 3335.2912, 1820.1775, 1732.16651, 1064 cm- '
Mass spectrum (NOBA-FAB> O): (M + H) + = 449
MP: 140OC
Log P (n-octanol / H2O) = 5.14 # 0.75 Example 6-Inhibition of the production of AP peptide The activity of the compounds on the production of Ap peptide is measured according to the protocol of

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Nigel J. Clarke et al. (1998) Febs Letters 430, 419-423 (appareil LC-MS Waters équipé d'un système electrospray-résonance quadrupolaire ZMD 2000) ou de Christie et al. (1999) J. of Neurochemistry, 73,195-204. Les composes 4 et 5 sont actifs et possèdent la capacité d'inhiber la production de peptide Ap.

Nigel J. Clarke et al. (1998) Febs Letters 430, 419-423 (LC-MS Waters device equipped with a ZMD 2000 quadrupole electrospray-resonance system) or by Christie et al. (1999) J. of Neurochemistry, 73, 195-204. Compounds 4 and 5 are active and have the capacity to inhibit the production of Ap peptide.

Example 7-Non-inhibition of the development of the embryo.

sont refermés et replacés à 37 C pour effectuer un autre point d'observation 4 jours après l'injection. L'activité des composés est comparée à celle de composés de référence MG132 (Calbiochem 474790) et MW167 (Enzyme System DFK-167). Les résultats sont présentés dans le tableau ci-dessous. Eggs (JA57) are freshly laid (max 2 days) or kept at 15 ° C from laying. The eggs are incubated for 60 hours then an opening is made in the shell where the embryo has been located by gravity. A 1 mM solution of the test substance is injected into the embryo. Then, the eggs are closed and the development is allowed to continue at 37 C. 24 hours after the injection, the eggs are opened and the embryos observed, the toxicity of the substance is then examined, then the eggs

are closed and replaced at 37 C to make another observation point 4 days after the injection. The activity of the compounds is compared with that of reference compounds MG132 (Calbiochem 474790) and MW167 (Enzyme System DFK-167). The results are presented in the table below.

24 hour survival 4 day survival
MW 167 0/5 0 MG132 0/9 0 Compound 11/15 6/15 Compound 10/12 8/12
The compounds of the invention are therefore much less toxic than the reference compounds on embryonic development.

Claims (18)

Y represents an oxygen, sulfur or nitrogen atom, the alkyl, alkenyl, cycloalkyl, aryl, alkoxy, thioalkoxy groups and the ring system  (C3-C15) alkoxy, (C3-C15) thioalkoxy, COR ', COOR'or CONHR', R 'being a hydrogen atom or a (Cl-C24) alkyl group, (C2-C24) alkenyl, (C3C15 ) cycloalkyl, (C5-C12) aryl, (C5-C12) aryl (CI-C24) alkyl, (C5-CI2) aryl (C2C24) alkenyl, a fatty acid, saturated or unsaturated, or an amino acid, protected or not , or, RI and R2 taken together form a monocyclic or bicyclic system having from 5 to 15 links, of which optionally 1 to 4 heteroatoms chosen from N, 0 and S, R3 represents a group (CI-C24) alkyl, (C2-C24) alkenyl, (C3-C15) cycloalkyl (C5-C12) aryl, (C5-C12) aryl (Cl-C24) alkyl, (C5-C12) aryl (C2-C24) alkenyl or a heterocyle in (C3-CI5) from 1 to 3 identical or different heteroatoms chosen from N, 0 and S, A represents a nitrogen, phosphorus atom or a CX group in which X is a halogen atom (preferably a chlorine atom) or a group OR ", R" being a hydrogen atom or a (C5-CI2) aryl, (CI-C6) alkyl or tosyle, and

 in which: - RI and R2, independently of each other, are chosen from a hydrogen atom, a (CI-C24) alkyl group ((C2-C24) alkenyl, (C3-CI5) cycloalkyl, ( C5-C12) aryl, (C5-C12) aryl, (C1-C24) alkyl, (C5-C 12) aryl (C2-C24) alkenyl,

CLAIMS 1. Compounds general formula (I) below: <Desc / Clms Page number 21>  optionally formed by RI and R2 being unsubstituted or substituted by one or more substituents preferably chosen from a halogen atom, a (Cl-C6) alkyl, (C3-C15) cycloalkyl, (C5-C12) aryl, ( C3-C15) alkoxy, (C3-C15) thioalkoxy, amino, amino (CI-CI5) (cyclo) alkyl, aminodi (CI-C15) (cyclo) alkyl, COOH, (Cl-C15) acyl, (ClCul5) acyloxy , nitro and trifluoromethyl, as well as their salts and, where appropriate, enantiomers, as inhibitors of the production of amyloid peptide Ag.

2. Compounds of general formula (I) according to claim 1, characterized in that Y represents 0 or N, preferably O. 3. Compounds of general formula (I) according to claim 1 or 2, characterized in that A represents a nitrogen atom or a CX group in which X is a halogen atom, preferably a chlorine atom or a group OR ", R" being a hydrogen atom or a (CI-C3) alkyl or tosyl group. 4. Compounds of general formula (I) according to one of claims 1 to 3, characterized in that RI is a hydrogen atom. 5. Compounds of general formula (I) according to one of claims 1 to 4, characterized in that R2 is a hydrogen atom or a COOR group in which R is an (Cl-C24) alkyl group, ( C2-C24) alkenyl, (C3-CI5) cycloalkyl, (C5-CI2) aryl, (C5C12) aryl (Cl-C24) alkyl or (C5-C12) aryl (C2-C24) alkenyl. 6. Compounds according to claim 5, characterized in that R2 is a COOR 'group in which R' is a (C3-C15) cycloalkyl group, preferably adamantyl. 7. Compounds of general formula (I) according to one of claims 1 to 6, characterized in that R3 is a substituted alkyl group, preferably an alkylalkoxy group. 8. Compounds of general formula (I) according to one of claims 1 to 6, characterized in that R3 is an alkyl group, alkenyl or aryl omega-functionalized by a function <Desc / Clms Page number 22>  alcohol, acid or amine terminal.

9. Compounds of general formula (I) as represented in claim 1, characterized in that A is a C-Cl group, RI is a hydrogen atom, R2 is a hydrogen atom or a COOR group in which R is a (C3-C15) cycloalkyl group, Y is an oxygen atom, and R3 is a substituted alkyl group, preferably an alkylalkoxy group.  10. Method for the selection, identification, characterization or qualification of compounds which inhibit the production of amyloid peptide Ap, characterized in that it comprises: a) a determination of the action of one or more test compounds on the production of Ap peptide, in particular by cleavage of APP, b) the selection of test compounds capable of inhibiting the production activity in a), c) a determination of the action of compounds selected in b) on embryonic development or the expression of metabolic pathways associated with embryonic development, and d) the selection of compounds having an inhibitory activity on the production of Ap peptide in a) greater than the inhibitory activity on embryonic development in c).  11. Method according to claim 10, characterized in that the compounds selected in d) are essentially devoid of effect or action in test c).  12. Method according to claims 10 or 11, characterized in that the inhibitory effect on the production of Ap peptide is determined by bringing the test compound (s) into contact with a mammalian cell expressing a nucleic acid encoding APP

 wild human or an isoform or mutant thereof, and determining the appearance of the cleaved form of the APP protein.  13. Method according to claim 12, characterized in that the mammalian cell <Desc / Clms Page number 23>  is a nerve cell, possibly recombinant.

14. Method according to one of claims 10 to 13, characterized in that it comprises in a) determining the action of the test compound (s) on the production of the peptide Ass40 or Ass42, in particular Ass42, from l APP.  15. Method according to one of claims 10 to 14, characterized in that step c) comprises determining the effect of the test compound (s) on the expression (or the wave of propagation) of the lunatic fringe gene to from embryos, or in a cell test of the Notch Delta function or in a test for blocking embryonic development observed on a newly embryonated egg.  16. Pharmaceutical composition characterized in that it comprises at least one compound according to one of claims 1 to 9 17. Pharmaceutical composition characterized in that it comprises at least one compound obtained by the method according to one of claims 10 to 15.  18. Use of a compound according to one of claims 1 to 9 or of a compound obtained by the method according to one of claims 10 to 15 for the preparation of a composition intended for the treatment of neurodegenerative pathologies, in particular for inhibiting the cleavage of APP in subjects, in particular for treating or inhibiting or preventing the onset or development of Alzheimer's disease.