Brevet FR3079141A1 — publié le 20190927

Description Title of Invention: uses of curcumin derivative Technical Field [0001] This invention relates to uses of a curcumin derivative. More specifically, the invention relates to uses of a curcumin derivative in preventing and/or treating Alzheimer's disease. Background Art [0002] Curcumin is a major component of Curcumin longa, generically known as turmeric, which belongs to the ginger family of perennial plants that grows naturally in India and other parts of Asia. The rhizome of turmeric can be crushed into a yellow powder and then used in Indian delicacies such as curries in Southeast Asia. The major components of turmeric extract are named curcuminoids, which contain about 75% curcumin (diferuloyl methane), 15% demethoxycurcumin, and 10% bisdemethoxycurcumin . Recent studies also indicate that the active ingredient of turmeric is curcumin [Aggarwal BB, Harikumar KB: Potential therapeutic effects of curcumin, the antiinflammatory agent, against neurodegenerative, cardiovascular, pulmonary, metabolic, autoimmune and neoplastic diseases. Int J Biochem Cell Biol 2009, 4](1):40-591. Emerging evidence shows that curcumin is effective against a wide variety of diseases like cancer, cardiovascular disease [Jagtap S, Meganathan K, Wagh V, Winkler J, Hescheler J, Sachinidis A: Chemoprotective mechanism of the natural compounds, epigallocatechin-3-0-gallate, quercetin and curcumin against cancer and cardiovascular diseases. Curr Med Chem 2009, ]6(12):1451-1462, obesity , liver disease , inflammatory disease and aging , through multiple molecular targets [ Taylor RA, Leonard MC: Curcumin for inflammatory bowel disease: a review of human studies. Altern Med Rev 2011, 16(2):152-156, 22; Shehzad A, Rehman G, Lee YS: Curcumin in inflammatory diseases. Biofactors 2013, 39(1):69-771. [0003] Three major components of turmeric extract, curcumin (diferuloyl methane), demethoxycurcumin, and bisdemethoxycurcumin, are identified and named curcuminoids . Although the beneficial effects of curcumin have been suggested by a series of epidemiological investigations, animal experiments, and cell line studies, the low bioavailability and rapid metabolism of curcumin have hindered the therapeutic development of curcumin for human diseases. [0004] Alzheimer' s disease (AD) is a progressive neurodegenerative disease gaining prevalence worldwide due to increasing lifespan. It has been estimated that the total number of individuals with AD will grow to approximately 74.7 million by 2030 [ Ijaopo EO: Dementia-related agitation: a review of non-phannacological inter- 2 ventions and analysis of risks and benefits of phannacotherapy. Translational psychiatry 2017, 7(10):e12501, and the cost of healthcare will rise to $1 trillion annually by 2050 [Alzheimer's A: 2015 Alzheimer's disease facts and figures. Alzheimer's & dementia: the journal of the Alzheimer's Association 2015, 11(3):332-3841. Although more than 100 trials have been conducted in the past decades, few agents are expected to reach market mainly because of inadequate understanding of the complex pathogenesis of AD, and the lack of reliable diagnostic tests or biomarkers to choose the target population. Thus, there is urgent demand for development of therapeutic drugs for AD patients to slow down, arrest, or reverse the progressive decline of cognitive function, especially at the mild cognitive impairment or early stage of AD. The current FDA-approved drugs for AD patients are cholinesterase inhibitor and NMDA antagonist, which act nonspecifically and can only alleviate the symptoms with high adverse events. Summary of Invention [0005] The present disclosure demonstrates for the first time that a curcumin derivative, TML-6, can treat and/or prevent Alzheimer's disease by regulating the expression of ApoE through PPARa and RXRa response elements in the APOE promoter. [0006] The present disclosure is to provide a method for preventing and/or treating Alzheimer's disease in a subject in need of such treatment, comprising administrating to said subject an effective amount of TML-6 or a pharmaceutically acceptable salt, solvate, hydrate, isotopologue, or prodrug of TML-6, and optionally a pharmaceutically acceptable carrier or excipient, [Chem. 1] TML-6. [0007] In one embodiment, Alzheimer's disease is treated and/or prevented by regulating apolipoprotein E (ApoE) expression, promoting beta-amyloid (A13) clearance, inhibiting amyloid precursor protein (APP) synthesis, and/or inhibiting inflammation in brain regions in a subject in need of such treatment. [0008] The present disclosure also provides a method for identifying a candidate compound which may increase ApoE expression, promote Ai3clearance, inhibit APP synthesis, 3 and/or inhibit inflammation in brain regions, which comprises contacting the candidate compound with AP-2 or PPARa/RXRa binding site of ApoE promoter region and determining if the candidate compound binds to AP-2 or PPARa/RXRa binding site of ApoE promoter region, wherein binding indicates that the candidate compound may increase ApoE expression, promote A13 clearance, inhibit APP synthesis, and/or inhibit inflammation in brain regions. [0009] The present disclosure is described in detail in the following sections. Other charac- teristics, purposes and advantages of the present invention can be found in the detailed description and daims. Brief Description of Drawings [0010] [fig.1] shows that TML-6 suppresses the expression of APP and phospho-NF-KB and increases the ApoE expression in Huh7 cell line. Huh7 cells were treated with different doses of curcumin and TML-6 for 24 hours. Western blotting was used to examine Alzheimer-related proteins such as APP, ApoE and inflammatory biomarker phosphorylated NFKB. [0011] [fig.21 shows that TML6 suppresses the A13 production in N2a/APPswe stable cell line. The stable over-expression of APP Swedish mutant in N2A cell line (N2a/APPswe) was treated with different doses of curcumin and TML-6. After 24h incubation, the levels of A1340, A1342 and A1342/40 ratio in the culture medium were examined by Colorimetric BetaMark Beta-Amyloid x-40 and x-42 ELISA Kit. [0012] [fig.31 shows that the promoter activity of ApoE is regulated by TML-6 through the transcriptional regulation of PPARa/RXRa and AP-2 binding site between -154 and +45 on ApoE promoter region. (A) Schematic representation of the potential binding sites on the full-length (-1061/+45) and serial deletion mutants of ApoE promoter constructs. The potential binding sites were predicted by JASPAR database (http://jaspar2016.genereg.net/). (B) The abbreviation of transcription factor binding sites and scores are shown based on prediction results of ApoE promoter. (C) Huh-7 cells were transiently co-transfected with different lengths of ApoE promoter and pRLTK plasmids. After incubation with traditional curcumin or TML-6, the luciferase reporter activities of the cell lysates were measured. (D) Upper panel shows the putative binding sites of PPARa/RXRa and AP-2 between -154 and+45 on ApoE promoter. AP2 binding sites locate at -120 to -111 and -86 to -76, and PPARa/RXRa binding sites locate at -125 to -114 and -84 to -73 of -154/+45 ApoE promoter region. Two dotted- square boxes showe the distal and proximal binding sites of PPARa/ 4 RXRa and AP-2 on ApoE promoter, respectively. Lower panel indicates the mutations of PPARa/RXRa and AP-2 binding sites by site-directed mutagenesis. (E) Huh-7 cells were co-transfected with mutants of ApoE promoter and pRL-TK plasmids. After incubation with TML-6 for 24h, the luciferase reporter activities of the cell lysates were measured. (F) The mutant form of ApoE promoter (-154/+45MdelAP2) was cotransfected with siPPARa and/or siRXRa into Huh7 cells. After incubation with TML6 for 24h, the luciferase reporter activities of the cell lysates were measured. The results are presented as folds of RLU after normalization using renilla luciferase activity. The means ± SD are derived from three independent experiments. *P REVENDICATIONS1. TML-6, or a pharmaceutically acceptable salt, solvate, hydrate, isotopologue, or prodrug of TML-6, and optionally a pharmaceutically acceptable carrier or excipient, Whem.41 for use in preventing and/or treating Alzheimer's disease in a subject in need of such treatment. 2. TML-6, or a pharmaceutically acceptable salt, solvate, hydrate, isotopologue, or prodrug of TML-6, and optionally a pharmaceutically acceptable carrier or excipient, for use according to daim 1, which is for increasing apolipoprotein E expression, promoting beta-amyloid clearance, inhibiting amyloid precursor protein synthesis, and/or inhibiting inflammation in brain regions. 3. TML-6, or a pharmaceutically acceptable salt, solvate, hydrate, isotopologue, or prodrug of TML-6, and optionally a pharmaceutically acceptable carrier or excipient, for use according to daim 2, which is for increasing apolipoprotein E expression, promoting beta-amyloid clearance, inhibiting amyloid precursor protein synthesis, and inhibiting inflammation in brain regions. 4. TML-6, or a pharmaceutically acceptable salt, solvate, hydrate, isotopologue, or prodrug of TML-6, and optionally a pharmaceutically acceptable carrier or excipient, for use according to daim 2, wherein the brain regions comprise hippocampus or other frontal regions. 5. A method for identifying a candidate compound which may increase apolipoprotein E expression, promote beta-amyloid clearance, inhibit amyloid precursor protein synthesis, and/or inhibit inflammation in brain regions, which comprises contacting the candidate compound with AP-2 or PPARa/RXRa binding site of ApoE promoter region and determining if the candidate compound binds to AP-2 or PPARa/ RXRa binding site of ApoE promoter region, wherein binding indicates 19 that the candidate compound may increase apolipoprotein E expression, promote beta-amyloid clearance, inhibit amyloid precursor protein synthesis, and/or inhibit inflammation in brain regions. 6. The method according to claim 5, wherein the AP-2 binding site of ApoE promoter region is located approximately at positions -154 to +45 counting from the transcription start site. Klaim 7] The method according to claim 5, wherein the AP-2 binding site of ApoE promoter region is located approximately at positions -120 to - 111 and -86 to -76 counting from the transcription start site. [Claim 8] The method according to claim 5, wherein the PPARa/RXRa binding site of ApoE promoter region is located approximately at positions -125 to -114 and -84 to -73 counting from the transcription start site. Klaim 9] The method according to claim 5, which comprises determining whether the candidate compound enhances expression of a reporter gene operably coupled by ApoE promoter region. Klaim 10] The method according to claim 5, wherein the candidate compounds are curcumin derivatives.