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WO2013024028A1 - Composés et compositions pour le traitement de protéinopathies - Google Patents

Composés et compositions pour le traitement de protéinopathies Download PDF

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Publication number
WO2013024028A1
WO2013024028A1 PCT/EP2012/065687 EP2012065687W WO2013024028A1 WO 2013024028 A1 WO2013024028 A1 WO 2013024028A1 EP 2012065687 W EP2012065687 W EP 2012065687W WO 2013024028 A1 WO2013024028 A1 WO 2013024028A1
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Prior art keywords
group
ghb
optionally substituted
sodium
above formula
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Inventor
Michel Maitre
Jean-Jacques Bourguignon
Martine Schmitt
Christian Klein
Ayikoe Guy MENSAH-NYAGAN
Chantal MATHIS
Jean-Christophe CASSEL
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Institut National de la Sante et de la Recherche Medicale INSERM
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Institut National de la Sante et de la Recherche Medicale INSERM
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/191—Carboxylic acids, e.g. valproic acid having two or more hydroxy groups, e.g. gluconic acid
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid 
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
    • A61K31/19—Carboxylic acids, e.g. valproic acid
    • A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/40—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil
    • A61K31/403—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with one nitrogen as the only ring hetero atom, e.g. sulpiride, succinimide, tolmetin, buflomedil condensed with carbocyclic rings, e.g. carbazole
    • A61K31/404—Indoles, e.g. pindolol
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/4164—1,3-Diazoles
    • A61K31/4172—Imidazole-alkanecarboxylic acids, e.g. histidine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/41—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having five-membered rings with two or more ring hetero atoms, at least one of which being nitrogen, e.g. tetrazole
    • A61K31/42—Oxazoles
    • A61K31/421—1,3-Oxazoles, e.g. pemoline, trimethadione
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/44—Non condensed pyridines; Hydrogenated derivatives thereof
    • A61K31/4412—Non condensed pyridines; Hydrogenated derivatives thereof having oxo groups directly attached to the heterocyclic ring
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/435—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom
    • A61K31/47—Quinolines; Isoquinolines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/55—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having seven-membered rings, e.g. azelastine, pentylenetetrazole
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00—Drugs for disorders of the nervous system
    • A61P25/28—Drugs for disorders of the nervous system for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • G01N33/5058—Neurological cells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/28—Neurological disorders
    • G01N2800/2814—Dementia; Cognitive disorders
    • G01N2800/2821—Alzheimer

Definitions

  • the present invention relates to compounds and compositions for treating proteinopathies.
  • Proteinopathies arise from the abnormal accumulation of protein material, often forming plaques, within the nervous system.
  • proteinopathies refers to these deleterious modifications.
  • Several diseases namely in aged people, are proteinopathies that affect various organs (brain, pancreas, heart... etc).
  • Most neurodegenerative diseases including Alzheimer's disease, belong to this class of pathological alterations and show specific protein deposits in the brain. These deposits are in most cases toxic for neurons and synaptic connections.
  • cognitive and mnesic deficits and/or neurological motor or sensitive disturbances are installed, with morphologic and functional modifications in several brain areas.
  • Alzheimer's disease accumulation of ⁇ -peptides (1 -40 and 1 -42) in the brain is thought to be the primitive molecular event that induces amylo ' id deposits, Tau phosphorylation and neuronal death.
  • This accumulation of ⁇ peptides in the brain has been shown to play a key role in the development of Alzheimer's disease. Accordingly, the prevention of this accumulation or the degradation of these ⁇ amyloid peptides is considered as potentially effective strategies in the treatment or prevention of Alzheimer's disease.
  • Several results are in favor of a decrease in ⁇ -peptides degradation in sporadic Alzheimer's disease.
  • proteolysis and elimination are under the control of proteolytic enzymes, namely metalloproteinases from the neuronal plasmic membranes.
  • proteolytic enzymes namely metalloproteinases from the neuronal plasmic membranes.
  • neprilysin enzyme a zinc metalloproteinase
  • neuroinflammation and neurodegeneration displayed beneficial roles in neuroinflammation and neurodegeneration, in particular those produced by the neurotoxic and inflammatory ⁇ amyloid peptide, by being involved in amyloid degrading processes
  • El-Amouri et al. demonstrated that over-expression of neprilysin protected neuronal cells from ⁇ peptide in vitro. Accordingly, the use of compounds increasing the activity of neprilysin could be very useful in the treatment of Alzheimer's disease.
  • the aim of the present invention is thus to provide new compounds for the treatment of proteinopathy.
  • the present invention therefore relates to compounds chosen from GHB, GHB derivatives and GHB structurally-related compounds thereof, including their isosteres, prodrugs, homologues and bioprecursors or a pharmaceutically acceptable salt thereof, for use in the treatment of a proteinopathy.
  • GHB ⁇ -hydroxybutyrate
  • GHB was also shown to be usable as a drug in several approved indications (narcolepsy/cataplexia (Gamma hydroxybutyrate (Xyrem) for narcolepsy. (2002) Med. Lett. Drugs. Ther. 44:103-105), anesthesia (Campbell (1983) Anaaesthesia 38:1 105-1 106), sleep induction (Kothare et al. (2007) Neurology 68:1 157- 1 158) and treatment of alcohol withdrawal (Nava et al. (2007) Am. J. Drug Alcohol Abuse 33:379-392)) because of its GABA-mimetic profile when its brain concentration reaches several hundred micromolar (Williams et al. (1995) Neuroscience 66:133-141 ).
  • the present invention arises from the unexpected finding by the inventors that GHB, GHB derivatives and GHB structurally-related compounds, induces in vivo an increase of neprilysin expression and reduces the level of ⁇ peptides in Alzheimer's disease mice models.
  • the present invention demonstrates the role of GHB and related compounds in inducing neprilysin activity in cell neuronal culture and/or in the brain of mice treated per os. These compounds provide a new way to promote the elimination of amylo ' id plaques and slow down the appearance of cognitive deficits in Alzheimer's disease. They are also useful for neuroprotective purposes in several proteinopathies, but also in acute pathologies (strokes, ischemic trauma) affecting the survival of brain cells.
  • the present invention also relates to a method of treatment and/or of prevention of a proteinopathy comprising the administration of a therapeutically effective amount of GHB, GHB derivatives or GHB structurally-related compounds, including their isosteres, prodrugs, homologues and bioprecursors thereof, or a pharmaceutically acceptable salt thereof, to a subject in need thereof.
  • the present invention also relates to a pharmaceutical composition comprising
  • GHB GHB, GHB derivatives or GHB structurally-related compounds, including their isosteres, prodrugs, homologues and bioprecursors thereof, or a pharmaceutically acceptable salt thereof, and a compound suitable for the treatment of said proteinopathy, optionally in association with a pharmaceutically acceptable carrier.
  • the present invention also concerns the in vitro use of GHB, GHB derivatives or GHB structurally-related compounds, including their isosteres, prodrugs, homologues and bioprecursors thereof, or a pharmaceutically acceptable salt thereof, for increasing the removal of ⁇ -amyloid peptides and/or decreasing the accumulation of ⁇ -amyloid peptides.
  • Another aspect of the present invention concerns the in vitro use of GHB, GHB derivatives or GHB structurally-related compounds, including their isosteres, prodrugs, homologues and bioprecursors thereof, for increasing the expression and/or activity of neprylisin.
  • a “proteinopathy” refers to a disease due to abnormal accumulation, in the nervous system, of substances, in particular protein material, deleterious for neurons and/or synapses.
  • the proteinopathy according to the invention is in particular a neurodegenerative proteinopathy.
  • the proteinopathy according to the invention is due to abnormal accumulation of protein material in the central nervous system.
  • Proteinopathies include for example Alzheimer disease, Lewy body dementia, amyotrophic lateral sclerosis and Huntington's disease.
  • ⁇ -amyloid peptides examples include in particular ⁇ -amyloid peptides, oc-synuclein and huntingtin.
  • ⁇ -amyloid peptide refers to a peptide of 39 ⁇ 43 amino acids that appears to be the main constituent of amyloid plaques in the brains of Alzheimer's disease patients. These plaques are composed of a tangle of regularly ordered fibrillar aggregates called amyloid fibers.
  • ⁇ is formed after sequential cleavage of the amyloid precursor protein (APP), a transmembrane glycoprotein of undetermined function, by ⁇ -, ⁇ - or ⁇ -secretases.
  • APP amyloid precursor protein
  • ⁇ protein is generated by successive action of the ⁇ and ⁇ secretases.
  • the ⁇ secretase cleaves within the transmembrane region of APP and can generate a number of isoforms of 39-43 amino acid residues in length. The most common isoforms are ⁇ 40 and ⁇ 42.
  • the proteinopathy according to the invention is a neurodegenerative proteinopathy.
  • the proteinopathy according to the invention is a ⁇ -amyloid proteinopathy.
  • ⁇ -amyloid proteinopathies are well known from those skilled in the art and include in particular Alzheimer 's disease.
  • the proteinopathy according to the invention is Alzheimer's disease.
  • the proteinopathy according to the invention is selected from early-onset Alzheimer's disease, late-onset Alzheime'sr disease or familial Alzheimer's disease.
  • neurodegenerative proteinopathies are caused by accumulation of protein material in the brain, forming protein deposits or plaques.
  • Alzheimer's disease is caused by accumulation of ⁇ -amyloid peptides in the brain, specifically, in the grey matter of the brain, forming ⁇ -amyloid peptide deposits or plaques.
  • ⁇ -amyloid plaques are also called "senile plaques”.
  • GHB GHB, GHB derivatives, GHB structurally-related compounds, including their isosteres, prodrugs homologues and bioprecursors thereof
  • GHB derivative refers to a compound which possesses a structure similar to GHB, consisting in a carboxylic acid and an alcohol function separated by three carbon atoms, wherein at least one of the three carbon atoms may be further substituted.
  • a “GHB structurally-related compound” refers to a compound which presents a common minimal structural fragment of the GHB, namely an anion as a carboxylate and a hydrogen bond acceptor donor system OH, wherein the number of carbon atoms between the carboxylate and the hydrogen bond system may vary.
  • a “GHB homologue”, “GHB derivative homologue” or “GHB structurally-related compound homologue” refers to a compound which comprises a carboxylic acid and an alcohol function separated by carbon atoms as in the GHB, GHB derivative or GHB structurally-related compound structure, but wherein the number of the carbon chain is lower or higher than the three carbon atoms comprised in the GHB.
  • GHB superior homologue refers to a GHB homologue as defined above, wherein the number of carbon atoms between the carboxylic acid and the alcohol function is higher than three.
  • GHB inferior homologue refers to a GHB homologue as defined above, wherein the number of carbon atoms between the carboxylic acid and the alcohol function is lower than three.
  • a “GHB isostere”, a “GHB derivative isostere” or a “GHB structurally-related compound isostere” refers to a compound which possesses a structure respectively similar to GHB, a GHB derivative or a GHB structurally-related compound, wherein the carboxylic acid function is replaced by other acid equivalent functions chosen from the group of tetrazole, isoxazole, sulfonic acid, phospshonic and phosphinic acids or wherein the alcohol function is replaced by a phenolic function, both functions being able to establish the same prototypical H bond interaction as with GHB, GHB derivative or GHB structurally-related compound.
  • a “GHB prodrug”, “GHB derivative prodrug” or “GHB structurally- related compound prodrug” refers to a compound which possesses a structure respectively a similar to GHB, a GHB derivative or a GHB structurally-related compound, wherein the carboxylic acid is replaced by an ester or an amide.
  • a “prodrug” has the capacity to be more or less rapidly hydrolysed in vivo (acid or metabolic hydrolysis), and thus produces the active species (the carboxylic moiety) after facilitation of the penetration of the lipophilic ester through the blood brain barrier.
  • a “GHB bioprecursor”, a “GHB derivative bioprecursor” or “a GHB structurally-related compound bioprecursor” refers to a compound which possesses a structure respectively similar to the GHB, GHB derivative or GHB structurally-related compound, wherein the carboxylic acid function is replaced by another functional group, such as methanol, able to provide in vivo with the critical carboxylic acid group for its specific action, after oxidative metabolic transformation.
  • the present invention relates to the GHB, including an isostere, a homologue, a prodrug or a bioprecursor thereof.
  • the present invention relates to a GHB derivative, including an isostere, a homologue, a prodrug or a bioprecursor thereof.
  • the present invention relates to a GHB structurally-related compound, including an isostere, a homologue, a prodrug or a bioprecursor thereof.
  • GHB derivatives or GHB structurally-related compounds possess specific restricted conformations.
  • the introduction of a double bond favors extended or semi-extended conformations.
  • the bridging between two positions of the GHB provides novel scaffolds with lesser conformational flexibility
  • the present invention relates to a compound chosen from GHB ( ⁇ -hydroxybutyrate), GHB derivatives or GB structurally-related compounds thereof, having the formula (I) :
  • - A represents -OR a , -NHCOR a , -NHS0 2 R a, or -CONR a R b ; wherein R a and R b are, independently of each other, chosen from the group consisting of: H, Alk, optionally substituted aryl and optionally substituted arylalkyl;
  • - W represents COOR, CONR a R b or S0 3 R , wherein R is H, alkyl or M + , wherein M + represents a pharmaceutically acceptable counter-ion;
  • R1 and R2 are, independently of each other, chosen from the group consisting of:
  • alkyl optionally substituted heteroaryl, optionally substituted aryl and optionally substituted arylalkyl;
  • R1 or R2 is absent, when a is a double bond
  • R5 and R6 are, independently of each other, chosen from the group consisting of: H, -NR a R b , alkyl or aryl; R a and R b being as defined above;
  • R5 or R6 is absent, when b or c is a double bond
  • R7 and R8 are, independently of each other, chosen from the group consisting of:
  • R1 may form, together with the carbon atom carrying them, an optionally substituted heteroaryl group or an optionally substituted aryl group;
  • - W and R7 may form, together with the carbon atom carrying them, an optionally substituted heteroaryl group
  • R1 and R3 may form, together with the carbon atom carrying them, an optionally substituted aryl group, a cycloalkyl group or a fused ring;
  • R1 and R7 may form, together with the carbon atom carrying them, an optionally substituted aryl group, an optionally substituted heteroaryl group, a heterocyclyl group or a cycloalkyl group which may contain at least one double bond;
  • R1 and R5 may form, together with the carbon atom carrying them, an optionally substituted aryl group, a cycloalkyl group or a fused ring;
  • - n is 0 or 1 ;
  • - m 0, or 1 ;
  • R1 and R7 form, together with the carbon atoms carrying them, an optionally substituted aryl group or an optionally substituted heteroaryl group, then n is 1 and m is 0, or m is 1 and n is 0;
  • A represents -OH.
  • A represents -NHCOCH 3 . According to one embodiment, in the above formula ( ), A represents -CONH 2 .
  • A represents -NHS0 2 Alk. According to one embodiment, in the above formula ( ), A may represent :
  • W represents COOH. According to one embodiment, in the above formula (I), W represents COONa. According to one embodiment, in the above formula (I), W represents COOAIk. According to one embodiment, in the above formula (I), W represents CONHAIk.
  • W represents CONHaryl. According to one embodiment, in the above formula (I), W represents CONH 2 . According to one embodiment, in the above formula (I), W represents S0 3 H.
  • W represents S0 3 Na.
  • R1 is absent, when a is a double bond.
  • R1 is H.
  • R1 is alkyl, in particular R1 is methyl.
  • R1 represents an aryl group, in particular R1 is phenyl.
  • R1 in the above formula (I) represents an aryl group, which may be substituted by two halogen, such as two chlorine atoms. In particular, the two chlorine atoms are in meta and para positions of the aromatic ring.
  • R1 in the above formula (I) represents an aryl group, which may be substituted by an alkoxy group, in particular methoxy, and/or by an arylalkyloxy group.
  • the arylalkyloxy group may be further substituted by a halogen, in particular by a chlorin atom.
  • R1 may represent :
  • R1 represents benzyl
  • said benzyl is substituted by an alkoxy group, in particular by a methoxy group.
  • R1 represents heteroaryl.
  • said heteroaryl is substituted by a benzylamide group.
  • R1 represents :
  • R2 is absent, when a is a double bond.
  • R2 is H.
  • R2 is alkyl, in particular R2 is methyl.
  • R2 represents an aryl group, in particular R2 is phenyl.
  • R2 represents an aryl group, which may be substituted by two halogens, such as two chlorine atoms. In particular, the two chlorine atoms are in meta and para positions of the aromatic ring.
  • R2 represents an aryl group, which may be substituted by an alkoxy group, in particular methoxy, and/or by an arylalkyloxy group.
  • the arylalkyloxy group is further substituted by a halogen, in particular by a chlorine atom.
  • R1 represents :
  • R2 in the above formula (I), represents benzyl.
  • said benzyl is substituted by an alkoxy group, in particular by a methoxy group.
  • R2 represents heteroaryl.
  • said heteroaryl is substituted by a benzylamide group.
  • R2 represents :
  • R3 is absent when a or b is a double bond.
  • R3 represents H.
  • R3 represents aryl
  • R3 represents -NH 2 .
  • R3 represents alkyl, in particular methyl.
  • R4 is absent when a or b is a double bond.
  • R4 represents H.
  • R4 represents aryl
  • R4 represents -NH 2.
  • R4 represents alkyl, in particular methyl.
  • R5 is absent, when b or c is a double bond.
  • R5 represents H.
  • R5 represents alkyl, in particular methyl.
  • R5 represents aryl, in particular phenyl.
  • R6 is absent, when b or c is a double bond.
  • R6 represents H.
  • R6 represents alkyl, in particular methyl.
  • R6 represents aryl, in particular phenyl.
  • R7 is absent, when c is a double bond.
  • R7 represents H.
  • R7 represents -NH 2 .
  • R7 represents NHMe. According to one embodiment, in the above formula (I), R7 represents NHBoc. According to one embodiment, in the above formula (I), R7 represents aryl, in particular phenyl. According to one embodiment, in the above formula (I), R8 is absent, when c is a double bond.
  • R8 represents H.
  • R8 represents -NH 2 .
  • R8 represents NHMe.
  • R8 represents NHBoc. According to one embodiment, in the above formula (I), R8 represents aryl, in particular phenyl.
  • a and R1 form, togethe with the carbon atoms carrying them, a heteroaryl group.
  • said heteroaryl are :
  • a and R1 form, together with the carbon atoms carrying them, a heteroaryl group which may be substituted by a substituent chosen among hydroxy, halogen, alkoxy.
  • said substituent is hydroxy.
  • a and R1 form, together with the carbon atom carrying them, an aryl group.
  • said aryl group is substituted by at least one substituent chosen from the group of amino, hydroxy, alkoxy and halogen.
  • a and R1 form together with the carbon atom carrying them, the following group :
  • W and R7 form, together with the carbon atoms carrying them, a heteroaryl group.
  • the heteroaryl group is an isoxazole group, which may be substituted by at least one substituent such as hydroxy.
  • said heteroaryl is :
  • R1 and R3 form, together with the carbon atoms carrying them, an aryl group.
  • said aryl group is a phenyl group.
  • R1 and R3 form, together with the carbon atoms carrying them, a cycloalkyl group.
  • said cycloalkyl group is a cyclohexyl group.
  • R1 and R3 form, together with the carbon atoms carrying them, a fused ring.
  • R1 and R3 form, together with the carbon atoms carrying them, the following fused ring:
  • R1 and R5 form, together with the carbon atoms carrying them, an aryl group, in particular a phenyl group.
  • R1 and R5 form, together with the carbon atoms carrying them, a phenyl group which may be substituted by at least one substituent, such as hydroxy.
  • R1 and R5 form, together with the carbon atoms carrying them, a cycloalkyl group.
  • said cycloalkyl group is a cyclohexyl group.
  • R1 and R5 form, together with the carbon atoms carrying them, a fused ring.
  • R1 and R5 form, together with the carbon atoms carrying them, the following fused ring:
  • R1 and R7 form, together with the carbon atoms carrying them, an aryl group, in particular a phenyl group.
  • R1 and R7 form, together with the carbon atoms carrying them, a heteroaryl group.
  • R1 and R7 form, together with the carbon atoms carrying them, a quinoline or a pyridine.
  • R1 and R7 form, together with the carbon atoms carrying them, a heteroaryl group.
  • said heteroaryl may be further substituted by at least one substituent, such as hydroxy or a chlorine atom.
  • R1 and R7 form, together with the carbon atoms carrying them, a heteroaryl chosen from the group consisting of :
  • R1 and R7 form, together with the carbon atoms carrying them, a heterocyclyl group, in particular a pyrrolidine.
  • R1 and R7 form, together with the carbon atoms carrying them, a cycloalkyl group which may contain at least one double bond.
  • R1 and R7 form, together with the carbon atoms carrying them, the following compound :
  • n is 0 or 1 .
  • m is 0 or 1 .
  • a, b and c are, independently of each other, a single or a double bond.
  • the present invention also relates to compounds having the formula (II): wherein :
  • R9 is chosen from the group consisting of : H, Alk, optionally substituted aryl, optionally substituted arylakyl;
  • R10 is absent when d is a double bond, or R10 is chosen from the group consisting in: H, Alk and aryl;
  • - p is 0 or 1 ;
  • - X represents CR c R d when d is a single bond; or X represents CR C when d is a double bond;
  • R c and R d are chosen, each independently, from H, -NR a R b and aryl, wherein R a and R b are defined as above in formula (I).
  • A represents -OH.
  • A represents -NHCOCH 3
  • A represents -CONH 2 .
  • W represents COOH
  • W represents COONa
  • W represents COOAIk.
  • W represents CONHAIk.
  • W represents CONHaryl
  • W represents CONH 2 .
  • W represents S0 3 H.
  • W represents S0 3 Na.
  • d is a single or a double bond.
  • R9 represents H.
  • R9 represents Alk, in particular methyl. According to one embodiment, in the above formula (II), R9 represents aryl, in particular phenyl.
  • R9 represents phenyl which may be substituted by at least one substituent, such as halogen, in particular a chlorine atom.
  • R9 represents phenyl substituted by two chlorine atoms, in particular in meta and para positions of the aromatic ring.
  • R9 represents arylalkyi, in particular benzyl.
  • R9 represents arylalkyi which may be substituted by an alkoxy group, such as methoxy.
  • R9 represents a benzyl group which is substituted in para position of the ring by a methoxy.
  • a preferred group of compounds of the present invention is constituted by compounds having above formula (II), wherein R10 is absent when d is a double bond.
  • Another preferred group of compounds of the invention is constituted by compounds having above formula (II), wherein R10 is H.
  • R10 represents Alk, in particular methyl.
  • R10 represents aryl, in particular phenyl.
  • X represents CH 2 , when d is a single bond.
  • X represents CHNH 2 when d is a single bond.
  • X represents CH when d is a double bond.
  • X represents CR C , when d is a double bond, wherein R c represents aryl, in particular phenyl.
  • p is 0 or 1 .
  • q is 0 or 1 .
  • the present invention also relates to compounds having the formula (III):
  • - r is 0 or 1 ;
  • - s is 0 or 1 ;
  • R12, R13, R14 are, independently of each other, chosen from the group consisting of : H, alkyl and aryl;
  • R1 1 and R12 may form together with the carbon atoms carrying them, an aryl group, a fused ring or a cycloalkyl group;
  • R1 1 and R13 may form together with the carbon atoms carrying them, an optionally substituted aryl group, an optionally substituted heteroaryl group, a cycloalkyl which may contain at least one unsaturation, or a heterocyclyl group
  • R1 1 and R14 may form together with the carbon atoms carrying them, an optionally substituted aryl group, an optionally substituted heteroaryl group, a cycloalkyl which may contain at least one double bond, or a heterocyclyl group;
  • W represents COOH. According to one embodiment, in the above formula (III), W represents COONa.
  • W represents COOAIk. According to one embodiment, in the above formula (I), W represents CONHAIk. According to one embodiment, in the above formula (I), W represents CONHaryl. According to one embodiment, in the above formula (I), W represents CONH 2 .
  • R1 1 and R12 form together with the carbon atoms carrying them, an aryl group, in particular a phenyl group.
  • R1 1 and R12 form together with the carbon atoms carrying them, a cycloalkyl group, in particular a cyclohexyl group.
  • R1 1 and R12 form together with the carbon atoms carrying them, a fused ring.
  • R1 1 and R12 form with the carbon atoms carrying them, a cycloheptyl ring which is fused with a phenyl.
  • R1 1 and R13 form together with the carbon atoms carrying them, an aryl group, in particular a phenyl group.
  • R1 1 and R13 form together with the carbon atoms carrying them, an aryl group, in particular a phenyl.
  • said phenyl is substituted by hydroxy.
  • R1 1 and R13 form together with the carbon atoms carrying them, a heteroaryl group.
  • said heteroaryl is quinoline.
  • R1 1 and R13 y form together with the carbon atoms carrying them, a heteroaryl group, such as quinoline, which may be substituted by a hydroxy or a chlorine atom.
  • a heteroaryl group such as quinoline
  • said heteroaryl group is substituted by two chlorine atoms.
  • R1 1 and R13 form together with the carbon atoms carrying them, a cycloalkyl which may contain at least one double bond, in particular a cyclohexene.
  • R1 1 and R13 form together with the carbon atoms carrying them, a heterocyclyl group, in particular a pyrrolidine.
  • R1 1 and R14 form together with the carbon atoms carrying them, an aryl group, in particular a phenyl group.
  • R1 1 and R14 form together with the carbon atoms carrying them, an aryl group, in particular a phenyl group.
  • R1 1 and R14 form together with the carbon atoms carrying them, a phenyl group which may be substituted by a hydroxy.
  • R1 1 and R14 form together with the carbon atoms carrying them, a heteroaryl group, such as a quinoline.
  • R1 1 and R14 form together with the carbon atoms carrying them, a heteroaryl group, such as a quinoline, which may be substituted by a hydroxy or a chlorine atom.
  • a heteroaryl group such as a quinoline
  • said heteroaryl group is substituted by two chlorine atoms.
  • R1 1 and R14 form together with the carbon atoms carrying them, a cycloalkyl which may contain at least one double bond, in particular a cyclohexene.
  • R1 1 and R14 form together with the carbon atoms carrying them, a heterocyclyl group, in particular a pyrrolidine.
  • the present invention also relates to compounds having the formula (IV):
  • B represents a heteroaryl group, -NHCOAlk, or -CONR a R b ;
  • R a and R b being, independently of each other, chosen from the group consisting of: H, Alk, optionally substituted aryl and optionally substituted arylalkyl;
  • R15 and R16 are, independently of each other, chosen from the group consisting of : H and -NR a R b ; or R15 and R16 may form together with the carbon atoms carrying them, a cycloalkyl group;
  • B represents a heteroaryl group, chosen from the group consisting of indole or imidazole, which may be substituted by hydroxy.
  • B represents:
  • B represents -NHCOMe.
  • B represents -CONH 2 .
  • B represents -CONHR b , wherein R b represents an optionally substituted aryl.
  • said aryl group is phenyl, which may be substituted by two chlorine atoms.
  • t is 0, 1 or 2.
  • R15 is H.
  • R16 is H.
  • R16 is -NH 2 .
  • R15 and R16 may form together with the carbon atoms them, a cycloalkyl group, in particular a cyclohexyl group.
  • the present invention also relates to compounds having the formula (V):
  • - A represents - OR a or -NHCOAIk .
  • R a and R b being, independently of each other, chosen from the group consisting of: H and Alk;
  • R17 and R18 are, independently of each other, chosen from the group consisting of : H, alkyl, optionally substituted aryl, optionally substituted arylalkyl, or optionally substituted heteroaryl;
  • - R is chosen from: H, Alk or M + , wherein M + represents a pharmaceutically acceptable counter-ion.
  • A represents -OH.
  • A represents -NHCOMe.
  • R17 represents H.
  • R17 represents alkyl, in particular methyl.
  • R17 represents aryl, in particular phenyl.
  • R17 is a phenyl group, substituted by alkoxy, in particular methoxy, and/or by an arylalkyloxy which may be further substituted on the aryl ring by a chlorine atom.
  • R17 represents:
  • R17 represents arylalkyi, in particular benzyl.
  • said arylalkyi may be substituted by alkoxy, in particular methoxy.
  • R17 represents a heteroaryl group.
  • said heteroaryl is substituted, in particular by a benzylamide.
  • R17 represents:
  • R is H.
  • R is Na
  • R is Alk
  • u is 1 or 2.
  • the present invention also relates to compounds having the formula (VI):
  • R19 and R20 are, independently of each other, chosen from the group consisting of: H and aryl. According to one embodiment, in the above formula (VI), R19 is H.
  • R19 is aryl, in particular phenyl.
  • R20 is H.
  • R20 is aryl, in particular phenyl.
  • the present invention also relates to compounds having the formula (VII)
  • - R is H, Alk or M + , wherein M + represents a pharmaceutically acceptable counter- ion;
  • R21 , R22 and R23 are, independently of each other, chosen from the group consisting of: H, alkyl, optionally substituted aryl, optionally substituted arylalkyi and - NR a R b , R a and R b being as defined above in formula (I).
  • R is H.
  • R is Na
  • R is Alk. According to one embodiment, in the above formula (VII), R21 is H.
  • R21 is alkyl, in particular methyl.
  • R21 is aryl, in particular phenyl.
  • R21 is aryl, optionally substituted by halogen.
  • said aryl is phenyl which may be substituted by two chlorine atoms.
  • R21 is benzyl. According to one embodiment, in the above formula (VII), R21 is benzyl, which may be substituted, in particular by alkoxy, such as methoxy.
  • R22 is H.
  • R22 is alkyl, in particular methyl.
  • R22 is aryl, in particular phenyl.
  • R23 is H.
  • R23 is -NH 2 .
  • R23 is -NHBoc.
  • the present invention also relates to compounds having the formula (VIII):
  • - R is H, Alk or M + , wherein M + represents a pharmaceutically acceptable counter-ion;
  • - R24 being H or -NR a R b ;
  • - X being absent or chosen from the group comprising: alkoxy, hydroxy and halogen.
  • R is H.
  • R is Na
  • R is Alk
  • R24 is H.
  • R24 is -NH 2 .
  • X is -OH.
  • v is 0 or 1 .
  • the present invention also relates to compounds having the formula (IX):
  • - f is a single or a double bond.
  • f is single.
  • f is double.
  • X is absent.
  • X is halogen.
  • the present invention also relates to compounds having the formula (X):
  • X is absent.
  • X is OH.
  • OH is in position 8 of the quinoline ring.
  • X is CI.
  • OH is in position 7 of the quinoline ring.
  • X is CI.
  • CI is in positions 5 and 7 of the quinoline ring.
  • the present invention also relates to compounds having the formula (XI):
  • X is absent. According to one embodiment, in the above formula (XI), X is halogen.
  • the present invention also relates to compounds having the formula (XII):
  • - R is: H, Alk or M + , wherein M + represents a pharmaceutically acceptable counter-ion.
  • X is absent. According to one embodiment, in the above formula (XII), X is halogen. According to one embodiment, in the above formula (XII), y is 1 .
  • y is 2.
  • R is H.
  • R is Na
  • R is Alk
  • the present invention also relates to compounds having the formula (XIII):
  • X is absent.
  • the present invention also relates to compounds having the formula (XIV):
  • M + represents a pharmaceutically acceptable counter-ion.
  • y is 1 .
  • R is H.
  • R is Na
  • R is Alk
  • the compounds chosen from gamma-hydroxybutyrate (GHB), GHB derivatives, and GHB structurally-related compounds thereof, or a pharmaceutically acceptable salt thereof, for use in the treatment of a proteinopathy are:
  • the compounds herein described may have asymmetric centers.
  • Compounds of the present invention containing an asymmetrically substituted carbon atom may be isolated in optically active or racemic forms. It is well-known in the art how to prepare optically active forms, such as by resolution of racemic forms or by synthesis from optically active starting materials. All chiral, diastereomeric, racemic forms and all geometric isomeric forms of a compound are intended, unless the stereochemistry or the isomeric form is specifically indicated.
  • Pharmaceutically acceptable salts of the compounds as defined above include those derived from pharmaceutically acceptable inorganic and organic acids and bases.
  • suitable acid salts include acetate, adipate, alginate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, citrate, camphorate, camphorsulfonate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptanoate, glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2- hydroxyethanesulfonate, lactate, maleate, malonate, methanesulfonate, 2- naphthalenesulfonate, nicotinate, nitrate, palmoate, pectinate, persulfate, 3- phenylpropionate, phosphate,
  • Salts derived from appropriate bases include alkali metal (e.g., sodium), alkaline earth metal (e. g., magnesium), ammonium and N-(alkyl) 4 + salts.
  • alkali metal e.g., sodium
  • alkaline earth metal e. g., magnesium
  • ammonium e.g., ammonium
  • N-(alkyl) 4 + salts e.g., ammonium
  • This invention also encompasses the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. Water or oil-soluble or dispersible products may be obtained by such quaternization.
  • Salt forms of the compounds as described above can be amino acid salts of carboxy groups (e.g., L- arginine, -lysine,-histidine salts).
  • a "pharmaceutically acceptable counter-ion” refers to an ion which has a charge opposite to the substance to which it is associated and which is pharmaceutically acceptable.
  • alkyl or “Alk” means a saturated or unsaturated aliphatic hydrocarbon group which may be straight or branched having 1 to 12 carbon atoms in the chain. Preferred alkyl groups have 1 to 6 carbon atoms in the chain.
  • alkoxy refers to an -O-alkyl radical.
  • aryl refers to an aromatic monocyclic or bicyclic hydrocarbon ring system having 5 to 10 carbons, wherein any ring atom capable of substitution may be substituted by a substituent.
  • aryl moieties include, but are not limited to, phenyl, naphthyl, and anthracenyl.
  • arylalkyl or “aralkyl” refer to an alkyl moiety in which an alkyl hydrogen atom is replaced by an aryl group (possibly substituted).
  • arylalkyl or “aralkyl” include benzyl.
  • arylalkyloxy or "-O-arylalkyl” refer to an -O-alkyl moiety in which an alkyl hydrogen atom of the alkyl moiety is replaced by an aryl group (possibly substituted).
  • arylalkyloxy or "-O-arylalkyl” include, but is not limited to, -O-benzyl.
  • cycloalkyi as employed herein includes saturated cyclic, bicyclic, tricyclic, or polycyclic hydrocarbon groups having 5 to 20 carbons, wherein any ring atom capable of substitution may be substituted by a substituent.
  • cycloalkyi moieties include, but are not limited to, cyclohexyl and cycloheptyl.
  • fused ring refer to bicyclic ring system consisting of a cycloalkyi group having 5 to 20 atoms fused with an aryl group having 5 to 20 atoms.
  • fused ring moiety include, but is not limited to a cycloheptyl moiety fused with a phenyl ring.
  • halo refers to the atoms of the group 17 of the periodic table (halogens) and includes in particular fluorine, chlorine, bromine, and iodine atom.
  • heterocyclyl refers to a nonaromatic 3-8 membered monocyclic, 7-12 membered bicyclic, having 1 -3 heteroatoms if monocyclic, 1 -6 heteroatoms if bicyclic, said heteroatoms selected from O, N, or S (e. g. , carbon atoms and 1 -3, 1 -6, or 1 -9 heteroatoms of N, O, or S if monocyclic, or bicyclic, respectively), wherein any ring atom capable of substitution may be substituted by a substituent.
  • heteroaryl refers to an aromatic 5-8 membered monocyclic, 8-12 membered bicyclic, having 1 -3 heteroatoms if monocyclic, 1 -6 heteroatoms if bicyclic, said heteroatoms selected from O, N, or S (e.g., carbon atoms and 1 -3, 1 -6, or 1 -9 heteroatoms of N, O, or S if monocyclic or bicyclic, respectively), wherein any ring atom capable of substitution may be substituted by a substituent.
  • substituted refers to a group "substituted” on an alkyl, cycloalkyl, heterocyclyl, aryl, aralkyl or heteroaryl group at any atom of that group.
  • Suitable substituents include, without limitation, alkyl, alkenyl, alkynyl, alkoxy, halo, hydroxy, cyano, nitro, amino, S0 3 H, sulphate, phosphate, perfluoroalkyl, perfluoroalkoxy, methylenedioxy, ethylenedioxy, carboxyl, oxo, thioxo, imino (alkyl, aryl, aralkyl), S(0) n alkyl (where n is 0-2), S(0) n aryl (where n is 0-2), S(0) n heteroaryl (where n is 0-2), S(0) n heterocyclyl (where n is 0- 2), amine (mono-,
  • treating means reversing, alleviating, inhibiting the progress of, or preventing the disorder or condition to which such term applies, or one or more symptoms of such disorder or condition.
  • a method of treatment and/or of prevention of a proteinopathy comprising the administration of a therapeutically effective amount of
  • GHB, GHB derivatives or GHB structurally-related compounds including their isosteres, prodrugs, homologues and bioprecursors thereof, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, as defined above to a subject in need thereof is also included in the present invention.
  • the compounds according to the invention may be administered for example by the oral route, by inhalation, or by the parenteral route (in particular by intravenous injection).
  • the compounds may be in the form of injectable solutions and suspensions, conditioned in ampoules or flasks.
  • the forms for parenteral delivery are conventionally obtained by mixing the compounds according to the invention with buffers, stabilizers, preservatives, solubilizing agents, isotonic agents and slurrying agents. According to known techniques, these mixtures can then be sterilized and conditioned in the form of intravenous injections.
  • One of skill in the art may use organic phosphate salts-based buffers as buffer.
  • slurrying agents include methylcellulose, acacia and sodium carboxymethylcellulose.
  • stabilizers include sodium sulphite and sodium metasulphite, and examples of preservatives include sodium p-hydroxybenzoate, sorbic acid, cresol and chlorocresol.
  • the amount of compound administered naturally depends on the administration route, the size and/or weight of the subject.
  • a "therapeutically effective amount” refers to an amount of a compound that confers a therapeutic effect on the treated subject.
  • the therapeutic effect may be objective (i.e., measurable by some test or marker) or subjective (i.e., the subject gives an indication of or feels an effect).
  • a "subject” denotes a human or non-human mammal, such as a rodent (rat, mouse, rabbit), a primate (chimpanzee), a feline (cat), a canine (dog).
  • the subject is human.
  • GHB GHB, GHB derivatives or GHB structurally-related compounds, including their isosteres, prodrugs, homologues and bioprecursors thereof, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, is defined above, for use in the treatment and/or prevention of a proteinopathy as defined above, increases the removal of ⁇ -amyloid peptides and/or decreases the accumulation of ⁇ -amyloid peptides in the brain.
  • GHB, GHB derivative or GHB structurally-related compound including their isosteres, prodrugs, homologues or bioprecursors thereof, as defined above, for use in the treatment and/or prevention of a proteinopathy as defined above, induces the removal of ⁇ -amyloid peptide deposits and/or prevents the formation of ⁇ -amyloid peptide deposits in the brain.
  • the GHB, GHB derivatives or GHB structurally-related compounds including their isosteres, prodrugs, homologues and bioprecursors thereof, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, as defined above, is for use in the prevention of the formation of senile plaques.
  • the present invention also concerns the in vitro use of GHB, GHB derivatives or GHB structurally-related compounds, including their isosteres, prodrugs, homologues and bioprecursors thereof, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, as defined above, for increasing the removal of ⁇ -amyloid peptides and/or decreasing the accumulation of ⁇ -amyloid peptides.
  • the removal of ⁇ -amyloid peptides and/or the accumulation of ⁇ -amyloid peptides may be detected by many different ways including ⁇ -specific ELISA, the use of imaging compounds, in particular thioflavin S, thioflavin T, Congo red, and derivatives thereof such as 25 l-6, notably described in Wang et al. (2002) J Mol Neurosci 19:1 1 -16; 3 H-BTA-1 , notably described in Klunk et al. (2003) J Neurosci 15:2086-2092; and C-6-OH-BTA-1 also known as Pittsburgh Compound-B, notably described in Klunk et al. (2004) Ann Neurol 55:306-3019; that selectively bind to ⁇ in vitro and in vivo, optionally combined with PET imaging, and atomic force microscopy.
  • imaging compounds in particular thioflavin S, thioflavin T, Congo red, and derivatives thereof such as 25 l-6, notably described in Wang e
  • the present invention also relates to the in vitro use GHB, GHB derivatives or GHB structurally-related compounds, including their isosteres, prodrugs, homologues and bioprecursors thereof, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, as defined above, for increasing the expression and/or activity of neprylisin.
  • Neprylisin As used herein and well-known from the one skilled in the art, “neprylisin”, “membrane metallo-endopeptidase”, “neutral endopeptidase (NEP)", “CD10”, or “common acute lymphoblastic leukemia antigen (CALLA)” refers to a zinc-dependent metalloprotease enzyme that degrades a number of small secreted peptides, most notably the ⁇ -amyloid peptide. Neprylisin is the metalloprotease the most involved in the process of degradation of ⁇ -amyloid peptides.
  • GHB, GHB derivatives or GHB structurally-related compounds including their isosteres, prodrugs, homologues and bioprecursors thereof, or a pharmaceutically acceptable salt thereof, to a subject in need thereof or a pharmaceutically acceptable salt thereof, for uses as defined above or administered as defined above, is in combination with a compound suitable for the treatment of said proteinopathy.
  • the expression "compounds suitable for the treatment of a proteinopathy” preferably refers to compounds that can be used for treating symptomatically and/or mechanistically said proteinopathy.
  • Compounds suitable for the treatment of a proteinopathy according to the invention are well-known from the one skilled in the art, and are for example disclosed in Sabbagh (2009) Am J Geriatr Pharmacother 7:167-185). They include in particular cholinesterase inhibitors such as donepezil, galantamine, huperzine A and rivastigmine, memantine, dimebolin hydrochloride, rosiglitazone, intravenous immunoglobulin and methylthioninium chloride.
  • the compound suitable for the treatment of said proteinopathy as defined above is selected from the group consisting of cholinesterase inhibitors and memantine.
  • Another object of the present invention concerns a pharmaceutical composition comprising GHB, GHB derivatives or GHB structurally-related compounds, including their isosteres, prodrugs, homologues and bioprecursors thereof, or a pharmaceutically acceptable salt thereof, to a subject in need thereof, as defined above and a compound suitable for the treatment of said proteinopathy as defined above, optionally in association with a pharmaceutically acceptable carrier.
  • the expression "pharmaceutically acceptable carrier” refers to a carrier that may be administered to a subject, together with a compound of the invention, and does not destroy the pharmacological activity thereof and is non-toxic when administered in doses sufficient to deliver a pharmaceutically effective amount of the compound.
  • Pharmaceutically acceptable carriers and vehicles that may be used in the pharmaceutical compositions according to the invention include, but are not limited to, ion exchangers, alumina, aluminium stearate, lecithin, self-emulsifying drug delivery systems (SEDDS) such as d-oc-tocopherol polyethyleneglycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tweens or other similar polymeric delivery matrices, serum proteins, such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-poly
  • Another object of the present invention is a method for the in vitro screening of GHB derivatives or GHB structurally-related compounds, including isosteres, prodrugs, homologues and bioprecursors thereof, which can be used for the treatment of a proteinopathy, comprising the steps consisting of: contacting the GHB derivatives or GHB structurally-related compounds, including isosteres, prodrugs, homologues and bioprecursors thereof to be tested with a neuronal cell; and
  • an increased level of expression of the neprilysin gene is indicative of GHB derivatives or GHB structurally-related compounds, including isosteres, prodrugs, homologues and bioprecursors thereof, which can be used for the treatment of proteinopathy, such as Alzheimer's disease.
  • a small library of GHB derivatives or GHB structurally-related compounds including their isosteres, prodrugs, homologues and bioprecursors thereof, are screened for their ability to increase neprilysin concentrations in a human neuroblastoma cell line, by reference to GHB itself.
  • the method of screening may comprise a supplementary step for comparison compared to control cells which are not contacting with the compounds to be tested.
  • the first step of the method of screening consists in contacting the GHB derivatives or GHB structurally-related compounds, including isosteres, prodrugs, homologues and bioprecursors thereof to be tested with a neuronal cell.
  • the neuronal cells are SH-SY5Y neuroblatoma cells.
  • the GHB derivatives or GHB structurally-related compounds are incorporated in the supernatant of SH-SY5Y neuroblatoma cells in presence of a solvent, such as DMSO.
  • a solvent such as DMSO.
  • 0.5% of DMSO is added.
  • the GHB derivatives or GHB structurally-related compounds are added to the cultures of neuronal cells, 24h before the harvest.
  • cells lysis are performed after 24h of incubation in a lysis buffer containing a ionic detergent, then the lysates are centrifuged, preferably at 10.000 g for 15 minutes at 4 q C. Then, the supernatants are recovered for total protein measurement.
  • the second step of the method of screening consists in determining the level of expression of the neprilysin gene by the neuronal cells. In particular, the second step consists in the measurement of neprilysin concentrations in the cells lysates.
  • the measurements of neprilysin concentrations may be carried out with a Neprilysin Duoset ELISA kit (R&D Systems Europe, Oxford, UK).
  • Serial dilutions of recombinant human neprilysin or sample diluted in 1 % PBS-BSA, or 1 % PBS-BSA alone as a control, are incubated for several hours, in particular for 2 hours at room temperature, followed by 3 further washes in wash buffer.
  • Biotinylated anti-neprilysin was added for 2 hours before another triple wash in wash buffer and incubation with streptavidin-peroxidase (1 :100) for several minutes, in particular for 20 minutes, in the dark.
  • the substrate solution (R&D Systems Europe) is added for 30 minutes in the dark.
  • the stop solution (R&D Systems Europe) is added to the substrate solution and the optical density for each well is read at 450 nm, followed by a reading at 540 nm to correct the optical imperfections in the plate.
  • the neprilysin protein levels are interpolated from the standard curve generated from serial dilutions of recombinant human neprilysin.
  • each measurement are performed in triplicate by comparison to a 100% control (200 ⁇ GHB) and a 0% control containing no drug.
  • the neprilysin inducers screened by the method of screening may have a role in the elimination of deleterious protein deposits in a great variety of proteinopathies with amylo ' id degeneration and functional alterations, affecting various organs, in particular the brain.
  • the neprilysin inducers screened by the method of screening may have neuroprotective effects in general, in particular in acute or chronic neurodegenerative pathologies.
  • GHB, and GHB structurally-related compounds such as kynurenic acid and derivatives of L-serine have neuroprotective effects.
  • the neuroprotective properties and the selective proteolysis of abnormal proteins in brain may be connected.
  • Figure 1 represents histograms displaying the fluorescent signal intensity (in arbitrary units) corresponding to neprilysin expression in the hippocampus of control mice (white histogram) and GHB treated mice (hatched histogram), obtained after chip hybridization.
  • Figure 2 represents a graph displaying the change of neprilysin activity (thick line) and neprilysin mRNA expression (fine line) over time in the hippocampus mice treated per os with GHB. 3 animals were used in triplicate per experimental point.
  • Figure 3 represents in the upper panel the result of an anti-neprilysin Western Blot performed on the hippocampus of control mice (controle) or of mice treated per os with GHB (GHB) after one hour treatment. Anti-actin Western Blot was performed as an intern control.
  • Figure 4 represents a graph displaying the daily GHB intake (in mmol, left ordinate) and the daily drink consumption (in ml, right ordinate) over time of mice treated with GHB by ingestion via drinking water.
  • Figure 5 represents pictures displaying ⁇ peptides density in control mice (control) or mice treated with GHB (GHB).
  • the ⁇ depositions were visualized by immunolabelling with anti- ⁇ peptides antibodies (left panel) or by thioflavin T staining (right panel), by macroscopy or microscopy with 1 .6 or 20X magnification.
  • Figure 6 represents histograms displaying images quantification for thioflavin T experiments described in Figure 5 on control mice (white histograms) and treated mice (hatched histograms). * : p ⁇ 0.05, Student's test.
  • Figure 7 represents in the upper panel the result of a Western Blot performed on the hippocampus of APPSWE control mice (control) or of APPSWE mice treated with GHB (GHB). ⁇ peptides 1 -40 and ⁇ peptides 1 -42 were labelled by specific antibodies.
  • Figure 8 represents histograms displaying the quantification of the dual staining of neprilysin and ⁇ peptides (ratio of surface stained by neprilysin/ surface stained by ⁇ peptides) in the hippocampus of 4 control mice (control, M1 , M2, M5 and M7) and 4 APPSWE mice treated with GHB (GHB, M3, M4, M6 and M8). 80 images were analyzed per group of mice.
  • Figure 9 represents in the upper panel the result of an anti-neprilysin Western Blot performed on the hippocampus of control mice (controle) or of APPSWE mice treated with GHB (GHB). Anti-actin Western Blot was performed as an intern control.
  • the lower panel represents histograms displaying the image analysis of this Western Blot (control: dotted histogram, GHB treated: hatched histogram). *** : significantly different from control (p ⁇ 0.001 , Student's test).
  • Figure 10 represents a graph showing the dose-effect of GHB on neprilysin (NEP) synthesis in SHSY-5Y neurons. It shows the variation of NEP protein concentration in SH- SY5Y neurons as a function of GHB concentrations in the medium.
  • the non-linear curve fitting is obtained with GraphPad Prism Program. Each experimental point is the mean ⁇ SEM of triplicates from three different experiments. EC 50 : 129 ⁇ . R 2: : 0.98.
  • Figure 11 represents a graph showing the increase in neurons of the neprilysin (NEP) protein concentrations as a function of kynurenic acid concentration in the cells supernatant. It shows the variation of NEP protein concentration in SH-SY5Y neurons as a function of kynurenic acid concentrations in the medium.
  • the non-linear curve fitting is obtained with GraphPad Prism Program, each experimental point being the mean ⁇ SEM of triplicates from three different experiments. EC 50: 32 ⁇ 7 ⁇ . R 2 : 0.97.
  • Figure 12 represents the neprylisin (NEP) activity as function of kynurenic concentrations in the SHSY-5Y neurons in culture.
  • the graph shows an increase in neprilysin activity : an increase in NEP/enkephalinase enzymatic activities in SH-SY5Y cells as a function of kynurenic acid concentrations in the cell medium.
  • the non-linear curve fitting is obtained with GraphPad Prism Program, each experimental point being the mean ⁇ SEM of triplicates from three different experiments.
  • EC 50 42 ⁇ 7 ⁇ .
  • R 2 0.98.
  • Figure 13 represents the increase in the neprilysin mRNA as function of kynurenic acid concentration.
  • Figure 14 represents a graph displaying spatial novelty detection in control and GHB- treated tg2576 mice at the age of 12 months (B). Results are expressed as the mean time spent exploring non-displaced objects and the time spent exploring the displaced object within the first 5 min of the retention session. Significant preference for the displaced object is obtained in both groups at 6 months, but only in GHB treated mice at 1 1 months ( * P ⁇ 0.05, displaced vs non-displaced objects, SNK test). Ordinate: exploration time (s + SEM).
  • This example demonstrates the capacity of GHB to regulate brain ⁇ peptides through modulation of proteolytic degradation and therefore its potential use in the treatment of Alzheimer's disease.
  • mice Female Swiss mice (about 20 g) were housed 3 per cage. All were kept at constant temperature (21 ⁇ 1 °C) with regular light/dark schedule (7 AM-7 PM), food and water were available ad libitum. However, animals for experiments were first starving for food during 24 h, then received an intra-gastric injection of either 200 ⁇ of a water solution of sucrose (4%, control mice) or 200 ⁇ of the same mixture containing 3 mg GHB, sodium salt (treated mice). Mice (3 mice at each point for each measurement) were killed by decapitation at various time after injection (0, 30, 60, 90 and 120 min), their brain were extracted and the hippocampi were removed rapidly.
  • mice at 4 months of age B6; SLJ-Tg (APPSWE) 2576Kha from Taconic
  • APPSWE mice at 4 months of age B6; SLJ-Tg (APPSWE) 2576Kha from Taconic
  • the APPSWE mice were allowed to drink during 5 weeks either a solution of sucrose 4 % in tap water (controls), or a solution of sucrose 4 % containing 1 .66 % GHB, Na + salt (treated).
  • mice body weight and the amount of water absorbed per animal were measured each day. On this basis, treated mice were calculated to drink about 4-5 ml of the GHB mixture in water, which represented a daily GHB intake of about 0.6 mmoles per day. During the treatment, the weight of the mice did not differ significantly from the weight of controls (about 20 ⁇ 1 g).
  • one set of APPSWE mice were killed and decapitated, hippocampi were rapidly excised and immediately used or stored at - 80 'C for protein quantification by western blot. A second set of APPSWE mice was used for quantitative immunohistochemical studies. All experiments were done by reference to APPSWE control mice.
  • mice were deeply anesthetized by intraperitoneal injection of 1 ml (5.6 % w/v) pentobarbital sodium solution (CEVA, sante animal, Libourne, France) and perfused transcardially with a fixative solution.
  • the fixative consisted of a 4% freshly depolymerized paraformaldehyde solution (Sigma-Aldrich) in 0.1 M phosphate buffer (PB; pH 7.4). This solution was chilled before being injected into anesthetized animals with a peristaltic pump at a flow rate of 25 ml/min until 1 ml/g body weight was reached.
  • brains were quickly removed and left for 1 hour at room temperature in the same fixative solution. Following fixation, brains were rinsed six times for 5 min in TBS (Tris-buffered saline, pH 7.4) to block residual aldehyde groups. Brain sections were dissected according to the Allen Brain Atlas (http://www.brain- map.org). Coronal sections (50-80 ⁇ ) were cut with a vibratome (Leica VT 1000 M) and collected in TBS.
  • TBS Tris-buffered saline, pH 7.4
  • Floating sections from APPSWE mice were washed in TBS before being processed for Thioflavin-T staining as described in Klunk et at. (2003) J. Neurosci. 23:2086-2092. Briefly, sections were washed in water and stained for 3 hours with a solution of 0.015% Thioflavin-T in PBS. Then the sections were washed in water and finally were coverslipped with Fluorosave (calbiochem Laboratories). Fluorescent sections were observed using a Leica DMIRE2 fluorescence microscope using a UV filter set.
  • TBS by the usual indirect method (Hedou et al. (2000) J. Comp. Neurol. 426:549-560). Sections and culture dishes were washed in TBS and incubated in 10% NBS in TBS for 1 hour to saturate nonspecific immunoreactive sites. After six TBS washes (5 min per wash), the sections or culture dishes were incubated overnight in the different antisera. These antibodies were neprilysin (anti-CD-10 Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA rabbit, IgG, 1 /50.) APP peptide (Anti APP 1 -4, Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA goat IgG, 1/100.).
  • antisera were used in a cocktail diluted in TBS.
  • the above dilutions were used irrespective of the number of antisera employed in each preparation. Sections were washed in six baths of TBS (2 ml for 5 min each time) and the second immunoreaction step was performed with specific secondary antisera for 4 hours at room temperature, followed by extensive washes (six washes for 5 min each time) in TBS.
  • TCS-SP2 invert equipped with a plan apo oil (40X) immersion lens. Cy3 emission was excited using the He/Ne laser 543 nm beam and Cy-5 fluorescence was obtained by excitation with an He/Ne laser 643 nm beam. The emitted light was filtered through appropriate filters (TD 488/543/633) for Cy3 (from 555-610 nm) and Cy5 (from 650-725 nm). Tissue sections were subjected to optical serial sectioning to produce images in the X-Y plane. Each optical section was scanned six times for brain sections to obtain an average image. Pictures were recorded digitally in a 512 X 512 pixel format.
  • Images were taken with a 40X objective, NA 1 .25, with an electronic zoom of around 2- or 4-fold.
  • a look-up table (glowoverglonch, Leica) ensured that the full dynamic range of the photomultipliers was used.
  • Image processing and fluorescence intensity measurements were performed on the Leica Confocal System software. Before each measurement a section series was acquired on the vertical axis in order to choose the equatorial section.
  • the sections were washed thoroughly with TBS and incubated in TBS containing 5% (v/v) NBS for 15 min.
  • the anti- neprilysin antibody anti-CD-10 Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA rabbit, IgG, 1/50.
  • APP peptide Anti APP 1 - 4, Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA goat IgG, 1 /100.
  • Sections were incubated overnight at 4 ⁇ C and after washing with TBS, they were treated with goat anti-rabbit IgG coupled to horseradish peroxidase in TBS (P.A.R.I.S.
  • the sections were mounted in glycerol/TBS (v/v) before microscopic analysis using a microscope (Leica DMRB) equipped with a digital camera (Axiocam, Zeiss, Germany). Anatomical structures were identified by direct observation using the atlas and nomenclature of the Allen Brain Atlas (http://www.brain-map.org). Light microscopic sections were examined using a brightfield microscope Leica DMRB (objectives 1 .6X to 20X). The photomicrographs were saved as tiff files and adjusted for color balance, brightness, and contrast. The images were processed using Adobe Photoshop 7.0 software (Adobe Systems, San Jose, CA) which was used to generate figure plates.
  • the RGB images were analysed by using Image J software (NIH, USA). After spliting the image into red, green and blue channels, the immunolabeling of NEP or APP was isolated by using thresholding and the thresholded area was measured. In the same time, the mean and maxima of optical density (OD) were measured for each image.
  • the OD measurement for each image is the mean gray of the pixels inside the selected area of the image, with a 256 gray scale as a reference. OD increase with the intensity of labeling between a minimum (0 : white) and a maximum (255 : black).
  • Nucleospin RNAL protocol (Macherey-Nagel, Duren, Germany). This protocol included a treatment of isolated RNA by DNase I. For each time and for each cerebral structure, three different mice were prepared. Integrity and purity of RNA were checked by spectrophotometry. Reverse transcription was performed with 1 ⁇ g RNA using Biorad iScriptTMcDNA synthesis kit. q-PCR was performed in iCycler thermal cycler (Biorad, Hercules, CA, USA) using SYBR Green dye (iQ SYBR green Supermix, Biorad).
  • the reaction mix was a makeup of 320 nM forward primer, 320 nM reverse primer, 200 nM probe, and 0.5 ⁇ cDNA template in a total reaction volume of 25 ⁇ (Benosman et al. (2007) Cell Death Differ 14:2047-2057).
  • iCycler iQ optical system software version 3.1 , Bio-Rad
  • a standard curve based on successive cDNA dilutions was performed and was used to calculate starting quantities.
  • starting quantities of genes of interest were reported to those of a housekeeping gene (RPL39) in the same plate. All samples were analyzed in triplicate, and the mean and standard deviation were calculated.
  • mice neprilysin (forward) 5'- AGGCCCTTTACGGGACTACAT-3' (SEQ ID NO: 1 ), (reverse) 5'- GCCTCCCCACAGCATTCTC-3' (SEQ ID NO: 2) and for housekeeping gene RPL39 (forward) 5'-GATCCTCGCCATGTCTTCTC-3' (SEQ ID NO: 3), (reverse) 5'- GCTTCGTTCTCCTCGAGTGT-3' (SEQ ID NO: 4).
  • APPSWE mice hippocampus extracts (controls and GHB-treated) were analyzed by SDS-PAGE gel electrophoresis/immunoblotting with antibodies recognizing ⁇ Amyloid 1 -40 (Abeam Inc., Cambridge, MA, USA; 1 /1000), ⁇ Amyloid 1 -42 (Abeam Inc., Cambridge, MA, USA; 1 /1000), neprilysin (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA; 1/800) and actin (Abeam Inc., Cambridge, MA, USA; 1/1000) as described in Gaiddon et al. (1996) J. Neurochem. 66:2279-2286.
  • Equal amounts of protein samples were separated along with molecular weight markers (Fermentas, Inc., Hanover, MD, USA) in 10% polyacrylamide gels. Gel loading equivalence was confirmed by actin. Gels were transferred onto 0.2 ⁇ nitrocellulose membranes (Bio-Rad, Hercules, CA, USA). Species-specific immunoglobulin G horseradish peroxidase (IgG-HRP) secondary antibodies (P.A.R.I.S, Compiegne, France) were used. Blots were developed with chemiluminescent substrate (Pierce Biotechnology, Inc., Rockford, IL, USA), and autoradiography was performed utilizing BioMax MR Film (Kodak, Rochester, NY, USA). Image analysis of the blots was performed on optical density-calibrated images captured with Adobe Photoshop software (Adobe Systems Incorporated). The protein level was quantified by measuring the integrated optical density of the bands, after subtracting background by Image J software.
  • Neprilysin activity was spectrophotometrically assayed as described in El-Amouri et at. (2007) Brain Res. 1152:191 -200. Briefly, Swiss mice hippocampus were sonicated. Proteins in whole cell lysates were quantified using the BCA protein assay kit (Pierce, Rockford, IL, USA). Then, the whole cell lysates were separately incubated in a standard assay mixture consisted of 50 ⁇ g of cell lysates, 50 ⁇ Z-Ala-Ala-Leu-p-nitroanilide (Peptide Institute, Osaka, Japan) as substrate, and 50 mM MES buffer (pH 6.5) in a total volume of 100 ⁇ .
  • the reaction was initiated by addition of substrate to the assay mixture, and carried out at 37 ⁇ C for 45 min.
  • the neprilysin activity was determined from the absorbance of the liberated p-nitroanilide at 405 nm and from the decrease in the rate of digestion caused by 10 ⁇ thiorphan (Enzo Life Sciences International, Inc., formerly BIOMOL International, L.P., Madison Meeting, PA, USA), a specific inhibitor of neprilysin.
  • cDNA were synthesized with the Superscript Choice system (Invitrogen), and then biotin-labelled cRNA were obtained with the Affymetrix IVT labelling kit (Affymetrix, Santa Clara, CA). After purification, 10 ⁇ g of fragmented cRNA were hybridized to the Affymetrix GeneChip Rat Genome 230 2.0 Array (31 ,042 probe sets including >28,000 rat genes), and the chips were automatically washed and stained with streptavidin-phycoerythrin using an Affymetrix fluidics station 450. Finally, arrays were scanned at 570 nm with a resolution of 1 .56 ⁇ / ⁇ with the Gene Chip Scanner 3000 7G from Affymetrix. Raw images (CEL format files) were generated with Affymetrix GeneChip Operating Software (GCOS 1 .4). Microarray data analysis.
  • the calculation for selecting genes differentially expressed was based on data processed using the dChip algorithm (Li and Wong (2001 ) Proc. Natl. Acad. Sci. USA 98:31 -36). Data were first normalized using the rank invariant method at probe level, then the Model Based Index algorithm was used with the PM/MM for calculating gene expression indexes. The dChip algorithm also allows associating detection level (P for present, A for absent and M for marginal) to probe set. The replicate samples from the test and the reference were averaged and a Fold Change (FC) was calculated for each probeset. For any of the above steps, except for the comparative analysis, the inventors used the R affy package of Bioconductor (Irizarry and Cope. An R package for analyses of affymetrix oligonucleotide arrays. In: The analysis of Gene Expression Data: Methods and Software Ed: Purgiani, Garrett, Irizarry, Zeger, New York: Springer (2002)).
  • Stringent filtering criteria were used to identify genes whose expression level was significantly changed between two stages.
  • the inventors performed a student t-test analysis to compare treated versus non treated rats for each structure. They selected genes that met all of the following requirements: 1 ) student t-test p value ⁇ 0.05 for the three replicates of each treated versus non treated rat.
  • At least one of the values of the microarray signal must be superior to 4 for each sample group.
  • FatiGO was used to extract relevant GO terms (http://godatabase.org/) for ours groups of genes with respect to a set of genes of reference (typically the rest of genes) (Al-Shahrour et at. (2004) Bioinformatics 20:578-580). The terms were considered to be relevant by the application of a Fisher's exact test that considers the multipletesting nature of the statistical contrast performed.
  • mice In order to confirm that acute GHB administration via oral route at a dose that was spontaneously ingested in the drinking water by APPSWE mice weighting about 20 g, normal mice (Swiss albinos) were injected with 3 mg GHB-Na + dissolved in 200 ⁇ water/sucrose 4% via an intragastric cannula. Animals were killed and decapitated at time 0, 30, 60, 90 and 120 min and the expression of neprilysin protein, activity and mRNA were followed in the hippocampus of these mice.
  • mice either by intragastric administration or by spontaneous drinking each day
  • mice induced a very significant increase in neprilysin protein and activity that followed an increase in neprilysin gene transcription in hippocampus.
  • mice Female APPSWE mice (B6, SJL-Tg (APPSWE) 2576 Kha, Taconic Europe, Denmark) carry a transgene coding for the 695-aminoacid isoform of human Alzheimer ⁇ - amyloid precursor protein. They express high concentrations of the mutant ⁇ which increases with the age of the animal, associated with the development of amyloid deposits in cortex, cerebellum and hippocampus (Carter et al. (2006) Neurosci. Lett. 392:235-239; Kobayashi and Chen (2005) Genes Brain Behav. 4:173-196). Numerous studies have documented spatial, learning and memory impairments in these transgenic mice, beginning as early as age 3 months.
  • mice Forty of these mice separated into two identical groups were used.
  • the mice were obtained from Taconic (Denmark) at age 15 weeks and after one week of adaptation to the local animal facilities, half of these animals received in their drinking water a mixture of sucrose 4% + GHB-Na + 1 .66% while the rest of the colony received only tap water containing sucrose 4%.
  • the treatment was carried out during 5 weeks.
  • Daily GHB-Na + intake by the group of treated mice which ingested the drug via the drinking water was investigated. After a short period of adaptation (about 5 days) where the volume of water/GHB ingested remained low (about 2.5-3 ml per day), the volume of water absorbed increased to reach 4-5 ml/day until the end of treatment. This represented a daily intake of about 0.7 mmol GHB-Na + per animal ( Figure 4).
  • the animals were 21 weeks of age (about 5 months).
  • GHB GHB-like proteinase
  • metalloproteinase genes whose products (namely neprilysin) contribute to the clearance of ⁇ peptides in transgenic mice.
  • These peptides are the precursors of amyloid plaques that produce neuroinflammation and neurotoxicity followed in aged mice by behavioral and cognitive deficits.
  • GHB can be used to regulate deleterious brain proteins deposits and thus to treat proteinopathies, in particular Alzheimer's disease.
  • Example 2 Object recognition and spatial novelty
  • mice were tested in a spatial novelty task at the ages of 6 (7 mice per group) and 1 1 months (7 treated mice, 5 control mice). They were additionally tested in an object recognition task at the younger age. Both tasks are based on the spontaneous tendency of mice to explore preferentially displaced objects or new objects within a familiar environment.
  • the mice were tested in a plexiglas open field (52 x 52 cm) with a white floor, black walls (40 cm) and a white striped card (21 x 29.7 cm) fixed on one wall (see Figure below).
  • Completely different sets of objects (height: 13 to 24 cm; width: 5 to 9 cm) were used for each behavioral testing phase.
  • the mouse was systematically introduced in the center of the open field. Object exploration was defined as the nose pointing to the object at a distance less than 1 cm. It was recorded with stopwatches for each object. Five days of testing are necessary to complete the two object exploration tasks.
  • mice are given a 10-min habituation period to the open field in presence of 2 new objects each day (X/Y and W/Z).
  • the spatial novelty task consists of a 10-min acquisition session in presence of 3 new objects (A,B and C), a 3-min resting period in the home cage and, again, a 10-min retention session with the same 3 objects, but the object B moved at a new position.
  • 6-month old mice were also given a 10 min session in presence of the same configuration of objects except that object C was replaced by a new one (D). This object recognition task was performed in order to verify that the chronic GHB treatment interrupted 9 days earlier did not affect basic object discrimination capacities.
  • a mean exploration time per object was calculated for non-displaced/familiar objects and compared to the mean time spent exploring the displaced/new object.
  • object preference in both tasks was evaluated on the first 5 min of the retention session. Note that all objects were similarly explored when tested for spontaneous preference in preliminary studies. Data were processed using analysis of variance with repeated measure on the "Object Category” factor. Direct comparison between groups were performed with the Student Newman- Keuls (SNK) test.
  • Example 3 GHB derivatives and GHB structurally-related compounds
  • GHB Gamma-hydroxybutyrate
  • GHB represents a model for derivatives or structurally- related compounds that act as neprilysin inducers, with an increased efficacy and a reduced concentration.
  • a small library of GHB derivatives and GHB structurally-related compounds, including their isosteres, homologues, prodrugs and bioprecursor have been screened, for their ability to increase neprilysin concentrations in a human neuroblastoma cell line, by reference to GHB itself.
  • GHB derivatives or GHB structurally-related compounds were incorportated in the supernatants of SH-SY5Y neuroblastoma cells at concentration of 100 or 200 ⁇ for 24h in the presence of DMSO 0.5%.
  • the SH-SY5Y neuroblastoma cells were maintained in a humidified atmosphere of 95 % air and 5 % C0 2 at 37°C. Cells were seeded into 100 mm dishes in Dulbecco's modified Eagle's medium (DMEM) W/GLUTAMAX-I PYR (GIBCO, Invitrogen, UK) supplemented with glucose, 10 % (v/v) fetal bovine serum, antibiotics and 0.5 % dimethylsulfoxide (DMSO).
  • DMEM Dulbecco's modified Eagle's medium
  • Neprilysin Duoset ELISA kit (R&D Systems Europe, Oxford, UK) was used according to the manufacturer's guidelines. Goat anti-human neprilysin diluted in PBS (pH 7.4) without carrier protein was coated on a high-binding Costar 96-well plate (R&D Systems Europe) for 18 hours at room temperature. The plates were washed 3 times in wash buffer (R&D Systems Europe).
  • Non-specific binding of antibody was blocked by addition of reagent diluents containing 1 % bovine serum albumin (BSA) in PBS (R&D Systems Europe) for 3 hours at room temperature, then the plates were washed a further 3 times in wash buffer.
  • BSA bovine serum albumin
  • the substrate solution (R&D Systems Europe) was added for 30 minutes in the dark.
  • the stop solution (R&D Systems Europe) was added to the substrate solution and the optical density for each well was read at 450 nm, followed by a reading at 540 nm to correct the optical imperfections in the plate.
  • the neprilysin protein levels were interpolated from the standard curve generated from serial dilutions of recombinant human neprilysin (R&D Systems Europe). Each measurement was performed in triplicate by comparison to a 100% control (200 ⁇ GHB) and a 0% control containing no drug. Results were expressed as percentage of neprilysin increases in the presence of 200 ⁇ GHB (statistics with the student't test).
  • the method of screening has allowed finding 45 compounds as potential inducer of neprilysin which are presented in table 1 .
  • Table 2 shows the neprilysin production of the screened compounds by reference to the control.

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Abstract

La présente invention concerne le gamma-hydroxybutyrate (GHB) ou des dérivés de GHB ou des composés structurellement associés au GHB, pour l'utilisation dans le traitement d'une protéinopathie.
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Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2217902A1 (fr) * 1997-07-07 1999-01-07 Laboratorio Farmaceutico C.T. S.R.L. Utilisation d'acide gammahydroxybutyrique pour traiter des syndromes de demence
US20040092529A1 (en) * 2002-10-30 2004-05-13 Pfizer Inc Methods of using piperazine derivatives
EP1486488A1 (fr) * 2003-06-10 2004-12-15 Phosphosites GmbH Utilisation d'un composé de formule I pour la préparation d'une composition pharmaceutique
WO2005112906A2 (fr) * 2004-05-21 2005-12-01 Duke University Procede destine a augmenter les effets des inhibiteurs de la reabsorption de la serotonine
US20060063827A1 (en) * 2004-09-23 2006-03-23 Yu Ruey J Systemic administration of therapeutic amino acids and N-acetylamino acids
EP1650182A1 (fr) * 2003-07-15 2006-04-26 Ono Pharmaceutical Co., Ltd. Compose d'acide carboxylique ramifie et utilisation de ce compose
US20060127506A1 (en) * 2004-12-10 2006-06-15 Hebert Rolland F Compositions of S-adenosyl-L-methionine
WO2007059031A2 (fr) * 2005-11-10 2007-05-24 Massachusetts Institute Of Technology Procedes et compositions pour elever les niveaux et la liberation d’acide gamma-aminobutyrique

Patent Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CA2217902A1 (fr) * 1997-07-07 1999-01-07 Laboratorio Farmaceutico C.T. S.R.L. Utilisation d'acide gammahydroxybutyrique pour traiter des syndromes de demence
US20040092529A1 (en) * 2002-10-30 2004-05-13 Pfizer Inc Methods of using piperazine derivatives
EP1486488A1 (fr) * 2003-06-10 2004-12-15 Phosphosites GmbH Utilisation d'un composé de formule I pour la préparation d'une composition pharmaceutique
EP1650182A1 (fr) * 2003-07-15 2006-04-26 Ono Pharmaceutical Co., Ltd. Compose d'acide carboxylique ramifie et utilisation de ce compose
WO2005112906A2 (fr) * 2004-05-21 2005-12-01 Duke University Procede destine a augmenter les effets des inhibiteurs de la reabsorption de la serotonine
US20060063827A1 (en) * 2004-09-23 2006-03-23 Yu Ruey J Systemic administration of therapeutic amino acids and N-acetylamino acids
US20060127506A1 (en) * 2004-12-10 2006-06-15 Hebert Rolland F Compositions of S-adenosyl-L-methionine
WO2007059031A2 (fr) * 2005-11-10 2007-05-24 Massachusetts Institute Of Technology Procedes et compositions pour elever les niveaux et la liberation d’acide gamma-aminobutyrique

Non-Patent Citations (29)

* Cited by examiner, † Cited by third party
Title
"Gamma hydroxybutyrate (Xyrem) for narcolepsy", MED. LETT. DRUGS. THER., vol. 44, 2002, pages 103 - 105
AI-SHAHROUR ET AL., BIOINFORMATICS, vol. 20, 2004, pages 578 - 580
ANDRIAMAMPANDRY ET AL., FASEB. J., vol. 17, 2003, pages 1691 - 1693
ANDRIAMAMPANDRY ET AL., FASEB. J., vol. 21, 2007, pages 885 - 895
BENOSMAN ET AL., CELL DEATH DIFFER, vol. 14, 2007, pages 2047 - 2057
BOURGUIGNON J-J ET AL: "ANALOGOUS OF GAMMA HYDROXYBUTYRIC ACID SYNTHESIS AND BINDING STUDIES", JOURNAL OF MEDICINAL CHEMISTRY, AMERICAN CHEMICAL SOCIETY, US, vol. 31, no. 5, 1 May 1988 (1988-05-01), pages 893 - 897, XP002560423, ISSN: 0022-2623, DOI: 10.1021/JM00400A001 *
CAMPBELL, ANAAESTHESIA, vol. 38, 1983, pages 1105 - 1106
CARTER ET AL., NEUROSCI. LETT., vol. 392, 2006, pages 235 - 239
EI-AMOURI ET AL., BRAIN RES., vol. 1152, 2007, pages 191 - 200
EI-AMOURI ET AL., BRAIN. RES., vol. 1152, 2007, pages 191 - 200
GAIDDON ET AL., J. NEUROCHEM., vol. 66, 1996, pages 2279 - 2286
HEDOU ET AL., J. COMP. NEUROL., vol. 426, 2000, pages 549 - 560
IRIZARRY; COPE: "The analysis of Gene Expression Data: Methods and Software", 2002, SPRINGER, article "An R package for analyses of affymetrix oligonucleotide arrays"
KLUNK ET AL., ANN NEUROL, vol. 55, 2004, pages 306 - 3019
KLUNK ET AL., J NEUROSCI, vol. 15, 2003, pages 2086 - 2092
KLUNK ET AL., J. NEUROSCI, vol. 23, 2003, pages 2086 - 2092
KOBAYASHI; CHEN, GENES BRAIN BEHAV., vol. 4, 2005, pages 173 - 196
KOTHARE ET AL., NEUROLOGY, vol. 68, 2007, pages 1157 - 1158
LEE C-W ET AL: "Isomerization of (Z,Z) to (E,E)1-bromo-2,5-bis-(3-hydroxycarbonyl-4-h ydroxy)-styrylbenzene in strong base: probes for amyloid plaques in the brain", JOURNAL OF MEDICINAL CHEMISTRY, AMERICAN CHEMICAL SOCIETY, US, vol. 44, no. 14, 1 June 2001 (2001-06-01), pages 2270 - 2275, XP002958808, ISSN: 0022-2623, DOI: 10.1021/JM010161T *
LI; WONG, PROC. NATL. ACAD. SCI. USA, vol. 98, 2001, pages 31 - 36
MAITRE, PROG. NEUROBIOL., vol. 51, 1997, pages 337 - 361
MAMELAK ET AL: "Alzheimer' s disease, oxidative stress and gammahydroxybutyrate", NEUROBIOLOGY OF AGING, TARRYTOWN, NY, US, vol. 28, no. 9, 1 September 2007 (2007-09-01), pages 1340 - 1360, XP022165124, ISSN: 0197-4580, DOI: 10.1016/J.NEUROBIOLAGING.2006.06.008 *
MAMELAK ET AL: "Neurodegeneration, sleep, and cerebral energy metabolism: a testable hypothesis", 1 January 1997, JOURNAL OF GERIATRIC PSYCHIATRY AND NEUROLOGY, DECKER PERIODICALS, HAMILTON, CA, PAGE(S) 29 - 32, ISSN: 0891-9887, XP008148801 *
MAMELAK M ET AL: "gamma.-Hydroxybutyrate and oxidative stres", 1 January 2002, GAMMA-HYDROXYBUTYRATE : MOLECULAR, FUNCTIONAL AND CLINICAL ASPECTS / EDITED BY GODFREY TUNNICLIFF AND CHRISTOPHER D. CASH, LONDON [U.A.] : TAYLOR & FRANCIS, 2002, UK, PAGE(S) 218 - 235, ISBN: 978-0-415-28499-8, XP008148866 *
NAVA ET AL., AM. J. DRUG ALCOHOL ABUSE, vol. 33, 2007, pages 379 - 392
SABBAGH, AM J GERIATR PHARMACOTHER, vol. 7, 2009, pages 167 - 185
TURNER; NALIVAEVA, INT. REV. NEUROLBIOL., vol. 82, 2007, pages 113 - 135
WANG ET AL., J MOL NEUROSCI, vol. 19, 2002, pages 11 - 16
WILLIAMS ET AL., NEUROSCIENCE, vol. 66, 1995, pages 133 - 141

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JP2021513522A (ja) * 2018-02-02 2021-05-27 コペンハーゲン大学 急性脳損傷の治療のための化合物
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