Agents useful in the transport of lipophilic antioxidants & uses thereof

Abstract

The present invention is related to peptides useful in the prevention and / or treatment of a disease or disorder characterized by oxidative stress for which the supplementation in lipophilic antioxidant is beneficial. The invention in particular relates to pharmaceutical formulations, regimens, methods of treatment and uses thereof.

Description

[0001] AGENTS USEFUL IN THE TRANSPORT OF LIPOPHILIC ANTIOXIDANTS & USES THEREOF

[0002] Field of the Invention

[0003] The present invention relates to agents useful for the binding, transport and delivery of lipophilic antioxidants for the treatment of diseases and disorders wherein oxidative stress is involved.

[0004] Background of the Invention

[0005] Aging has become a major focus in populations all over the world because the number of elderly individuals is expected to increase. The formation of Reactive Oxygen Species (ROS) occurs normally in almost all physiological processes and is normally counteracted by antioxidants. In the absence of antioxidant activity, or under conditions of low levels of antioxidant activity excess ROS can damage the intracellular cellular organelles, cell membranes and tissues through oxidative stress. Thus, oxidative stress leads to and is implicated in a large variety of diseases such as cancer, metabolic syndrome, atherosclerosis, malaria, Alzheimer’s disease (AD), rheumatoid arthritis, neurodegenerative diseases and pre-eclampsia (Kiran et al., 2023, Journal of Laboratory Medicine, https: / / doi. org / 10.1515 labmed-2022-0108}. Antioxidants play a crucial role in the prevention and treatment of many diseases by reducing the concentration of species involved in oxidative stress, such as free radicals and reactive oxygen, and nitrogen species (ROS, NOS), generated by various environmental stimuli or during normal cellular metabolism.

[0006] Therefore, antioxidant supplementation has been investigated for its beneficial effects on Macular Pigment Optical Density and Visual Functions (Hu et al., 2024, Adv Nutr, 15(5): 100216. doi: 10.1016 / j. advnut.2024.100216), Sickle Cell Disease (SCD), cardiovascular diseases (atherosclerosis) and skin diseases but also and treatment of cancers (Avila-Roman, Garcia-Gil et al. 2021, Mar. Drugs 2021, 19, 531) and Sui et al. 2024, Foods 2024, 13, 1321). In addition this approach may be beneficial to cancer cells expressing high levels ofSRBl such as prostate and breast cancers (Mooberry 2016, Frontiers in Pharmacology vol 7 article 466). The addition of lipophilic antioxidants is also used as a strategy to inhibit oxidation and protect the nutritional integrity of edible oils (Liu et al, 2025, Food Research International Volume 200, January 2025, 115423, https: / / d0i.0rg / l 0.1016 / j.foodres.2024.115423)

[0007] Although many therapeutic approaches are under preclinical and clinical investigation for the prevention and treatment of AD (Cao et al, 2018, Molecular Neurodegeneration, 13:64) and Age-related macular degeneration (AMD) (Schlottmann, 2017, Asia-Pac J Ophthalmol 6:514579), only few treatments are approved and clinically effective. Although most tissue can acquire lipophilic antioxidants from plasma lipoproteins, delivery of these antioxidants to the retina and brain must overcome the Blood Retina (BR) or Blood Brain Barrier (BBB) and only antioxidants transported by the smallest lipoproteins, such as HDL3 or free (poorly lipidated) apolipoproteins can reach these tissues. LDL or particles of similar size do not reach these tissues.

[0008] Transport of antioxidant in the lipid phase of lipoproteins which occurs with all lipoproteins including LDL, VLDL and chylomicron is non selective. Therefore, the non-specific transport of lipophilic antioxidants by HDL such as described in Li et al, 2022, ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS, vol. 716 does not allow their direct transfer to the brain and retina.

[0009] There is therefore a need to develop efficient agents delivering lipophilic antioxidants to their target sites, in particular through the BBB or BRB.

[0010] Summary of the invention

[0011] The present invention relates to the discovery of peptides useful for the selective uptake, transport and tissue delivery of lipophilic antioxidants.

[0012] According to one aspect, the invention provides a peptide of SEQ ID NO: 1 and homologuous variants or fragments thereof, wherein said peptide, homologous variant or fragment thereof has a length of 15 to 20 amino acids.

[0013] According to another aspect, the invention provides a pharmaceutical composition comprising at least one peptide of the invention and at least one pharmaceutically acceptable carrier, diluent or excipient thereof.

[0014] According to one aspect, the invention provides a peptide of SEQ ID NO: 1 for use in the prevention and / or treatment of diseases or disorders involving oxidative stress.

[0015] According to another aspect, the invention provides a use of a peptide of SEQ ID NO: 1 for the prevention and / or treatment of diseases involving oxidative stress.

[0016] According to another aspect, the invention provides a method of preventing and / or treating diseases or disorders involving oxidative stress.

[0017] Description of the figures

[0018] Figure 1 shows the hydrophobic interaction between the ligand, zeaxanthin displayed as stick and ball and the target sequence (SEQ ID NO: 2) shown as ribbon, with both phenylalanine (F57 and F71) clearly visible. Dotted lines indicate hydrophobic interactions between ligand and target (ribbon).Figure 2 shows beta-cryptoxanthin displayed as stick and ball and the target sequence (SEQ ID NO: 2) as ribbon, with both phenylalanine (F57 and F71) clearly visible. Dotted lines indicate hydrophobic interactions between ligand and target (ribbon).

[0019] Figure 3 shows lutein displayed as stick and ball and the target sequence (SEQ ID NO: 2) displayed as ribbon, with both phenylalanine (F57 and F71) clearly visible. Dotted lines indicate hydrophobic interactions between ligand and target (ribbon).

[0020] Figure 4 shows A: the name and smiles and B: the structures of compounds used in docking assays from the examples.

[0021] Detailed description

[0022] The amino acids of the peptid can be D or L or a combination thereof but should not compromise the formation of an alpha-helices (no D / L mixture within the same helix).

[0023] The term “hydrophobic residue” according to the invention refers to any natural or unnatural amino acid known to interact favorably by hydrophobic interactions such as Valine, Leucine, Isoleucine, Methionine, Phenylalanine and Tyrosine or Tryptophan. Examples of unnatural amino acids include cyclobutyl alanine (Cba) which has properties similar to Vai, cyclohexyl alanine (Cha), halogenated or otherwise substituted aromatic amino acids (Phe, Tyr, Trp), alternative heterocyclic rings such as pyridyl alanine.

[0024] The term “any natural or unnatural amino acid” according to the invention refers to the canonical amino acids and their modifications (alternative side chains, alternative stereochemistry).

[0025] The term “cationic residue” refers to any natural or un-natural amino acid which has a cation on its side chain or a basic amine on the side chain that is protonated at physiological pH (i.e. cationic).

[0026] The term “anionic residue” refers to any natural or un-natural amino acid that has a an anion on its side chain or a carboxylic acid on the side chain that is deprotonotated at physiological pH (i.e. anionic).

[0027] The term “neutral residue with an hydrophilic side chain” refers to any natural or unnatural amino acid with a side chain containing heteroatoms but no charges such as Apargine, Glutamine, Serine, threonine.

[0028] The term “fragment” refers to a peptide comprising a portion of a peptide sequence corresponding to contiguous amino acids of a polypeptide set forth herein, including all intermediate lengths and variants thereof according to the invention.The term “variant” applies to both a polypeptide and an amino acid. A polypeptide "variant," as the term is used herein, is a peptide or a polypeptide substantially homologous to the referenced peptide sequence, but which has an amino acid sequence different from that of the referenced. Such variants may be synthetically generated, for example, by modifying one or more of the above polypeptide sequences of the invention described herein using any of a number of techniques well known in the art. Substantially homologous means a variant amino acid sequence which is identical to the referenced peptide sequence with the exception of the substitution of a few amino acids wherein indicated, e.g. 1, 2, 3, 4, or 5 amino acids amongst the defined variable amino acids of the referenced polypeptide sequences. The identity or homology of two amino acid sequences or of two nucleic acid sequences can be determined by visual inspection and / or mathematical calculation, or more easily by comparing sequence information using known computer program used for sequence comparison such as Clustal package version 2.1.

[0029] A conservative, or homologous, variant may comprise a sequence having at least one conservatively substituted amino acid.

[0030] A "conservative substitution" or “homologous variant” is one in which an amino acid is substituted for another amino acid that has similar properties, such that one skilled in the art of peptide chemistry would expect the secondary structure and hydropathic nature of the polypeptide to be substantially unchanged (e.g., having similar physiochemical characteristics). Modifications may be made in the structure of the polynucleotides and polypeptides of the present invention and still obtain a functional molecule that encodes a variant or derivative polypeptide with desirable characteristics according to the invention. In making such changes, the hydropathic index, polarity, charge, solubility, hydrophobicity, hydrophilicity and / or the amphipathic nature of the amino acids are considered. The importance of the hydropathic amino acid index in conferring interactive biologic function on a protein is generally understood in the art (Kyte, et al., 1982, J. Mol. Biol., 157: 105- 131). Examples of conservative substitutions include substitution of one aliphatic residue for another, such as He, Vai, Leu, or Ala for one another, or substitutions of one polar residue for another, such as between Lys and Arg; Glu and Asp; or Gin and Asn. Other such conservative substitutions, for example, substitutions of entire regions having similar hydrophobicity characteristics, are well known (Kyte, et al, 1982, supra). For example, a "conservative amino acid substitution" may involve a substitution of a native amino acid residue with a non-native residue such that there is little or no effect on the polarity or charge of the amino acid residue at that position. Desired amino acid substitutions (whether conservative or non-conservative) can be determined by those skilled in the art at thetime such substitutions are desired. Exemplary amino acid substitutions are presented in Table 1 below. The term "variant" also includes a peptide or polypeptide substantially homologous to the referenced peptide sequence, but which has an amino acid sequence different from that of the referenced sequence because one, two, three, four or five amino acids amongst the defined non-variable amino acids of the reference polypeptide sequences have been chemically modified or substituted by amino acids analogues. This term also includes glycosylated polypeptides or other residues that have been post-translationally modified, in which case, the number of post-translationally modified amino acids is unlimited.

[0031] Table 1

[0032]

[0033] Polypeptides of the invention are prepared using any of a variety of well-known synthetic and / or recombinant techniques.

[0034] The term “pharmaceutically acceptable” refers to a carrier comprised of a material that is not biologically or otherwise undesirable.

[0035] The term “carrier” refers to any components present in a pharmaceutical formulation other than the active agent and thus includes diluents, binders, lubricants, disintegrants, fillers, coloring agents, wetting, or emulsifying agents, pH buffering agents, preservatives and the like.

[0036] The term “diseases or disorders involving oxidative stress” refers to diseases or disorders wherein excessive reactive oxygen species (ROS) damages the essential biomolecules, leadingto cellular malfunction and neurodegeneration. Several neurological disorders, including Alzheimer's, Parkinson's, Amyotrophic lateral sclerosis, multiple sclerosis, and ischemic stroke, are associated with oxidative stress. In particular, diseases or disorders involving oxidative stress according to the invention are diseases or disorders for which the delivery of lipophilic antioxidants is beneficial such as AD, AMD, diabetes, cancers and Sickle Cell Disease.

[0037] The term “lipophilic” antioxidants, refers to lipophilic substances having antioxidant properties, those encompass liposoluble antioxidants such as tocopherols, tocotrienols and xanthophylls carotenoids.

[0038] The term “carotenoids” refers to one of the most widespread groups of pigments with more than 600 identified in nature. In plants, cyclization of lycopene either leads to the formation of beta-carotene or alpha-carotene and its derivative xanthophylls, beta-cryptoxanthin, zeaxanthin, astaxanthin, violaxanthin and lutein. Carotenes are characterized by cyclization at one or both ends whereas xanthophylls are formed by the introduction of oxygen. According to a particular aspect, carotenoids can be used as liposoluble vitamins according to the invention. In a further particular embodiment, carotenoids are selected from xanthophylls, lutein, zeaxanthin and analogues thereof including and isomers / enanti omers thereof (e.g. meso-zeaxanthin, isozeaxanthin, (3S,3'S,all-E)-zeaxanthin), alloxazeaxanthin, zeaxanthin diglucoside, beta-cryptoxanthin, astaxanthin, sapotexanthin, and violaxanthin).

[0039] The terms “tocopherols” and “tocotrienols” refer to agents having vitamin E activity. There are eight naturally occurring vitamin E isoforms, alpha-, beta-, gamma- and delta-tocopherol and alpha-, beta-, gamma- and del ta-tocotri enol. They are all potent liposoluble antioxidants, capable of neutralizing free radicals directly by donating hydrogen from its chromanol ring. Alpha-tocopherol is regarded as the dominant form in vitamin E as the alpha-tocopherol transfer protein in the liver binds mainly alpha-tocopherol, thus preventing its degradation. According to a particular aspect, “tocopherols” and “tocotrienols” can be used as additional liposoluble vitamins according to the invention.

[0040] It is well recognized that hydrosoluble antioxidant such as vitamin C, glutathione, N-acetyl Cysteine and ions such as Zinc and those may be used to increase or maintain the antioxidant potential of liposoluble antioxidant.

[0041] As used herein, “treatment” and “treating” and the like generally mean obtaining a desired pharmacological and physiological effect. The effect may be prophylactic in terms of preventing or partially preventing a disease, symptom or condition thereof and / or may be therapeutic in terms of a partial or complete cure of a disease, condition, symptom or adverseeffect attributed to the disease. The term “treatment” as used herein covers any treatment of a disease in a mammal, particularly a human, and includes: (a) preventing the disease from occurring in a subject which may be predisposed to the disease but has not yet been diagnosed as having it; (b) inhibiting the disease, i.e., arresting its development; or relieving the disease, i.e., causing regression of the disease and / or its symptoms or conditions.

[0042] According to a particular aspect, the efficacy of a treatment or a use according to the invention can be monitored through the sustained increase of plasma levels of lipophilic antioxidant by measuring the level of biomarkers of oxidative stress (for example antioxidants, oxidized lipids, malondialdehyde (MDA), glutathione, catalase etc,) in total RBC and plasma. RBC rheological properties are also expected to be improved.

[0043] The term “subject” as used herein refers to mammals. For examples, mammals contemplated by the present invention include humans and the like.

[0044] Peptides according to the invention

[0045] According to a particular aspect is provided a peptide having the following sequence SEQ ID NO: 1 or a homologous variant or a fragment thereof, wherein said peptide, homologous variant or fragment thereof has a length of 15 to 20 amino acids.

[0046] According to a particular aspect is provided a peptide of SEQ ID NO: 1 wherein Xi is a Serine or a homologous variant thereof.

[0047] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein X2is a Threonine or a homologous variant thereof.

[0048] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein X4 is a Serine or a homologous variant.

[0049] According to another particular aspect is provided a peptide of SEQ ID NO: 1, wherein X5is an Lysine or a homologous variant thereof.

[0050] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein X6is a Leucine or a homologous variant thereof.

[0051] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein X7is a Arginine or a homologous variant thereof.

[0052] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein X8is a Glutamic Acid or a homologous variant thereof.

[0053] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein X9is a Glutamine or a homologous variant thereof.

[0054] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein X10 is a Leucine or a homologous variant thereof.According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein Xu is a Glycine or a homologous variant thereof.

[0055] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein Xi2is a Proline or or a homologous variant thereof.

[0056] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein Xi3is a Valine or a homologous variant thereof.

[0057] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein Xi4is a Threonine or a homologous variant thereof.

[0058] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein Xi5is a Glutamine or a homologous variant thereof.

[0059] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein Xi6is a Glutamic Acid or a homologous variant thereof.

[0060] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein Xi8is absent or present and when present is a Tryptophane or a homologous variant thereof.

[0061] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein Xi9is absent or present and when present is an Aspartic Acid or a homologous variant thereof.

[0062] According to another particular aspect is provided a peptide of SEQ ID NO: 1 wherein X2o is absent or present and when present is an Asparagine or a homologous variant thereof.

[0063] According to a particular aspect is provided a peptide having a sequence of SEQ ID NO: 2 or a homologous variant or a fragment thereof, wherein said peptide, homologous variant or fragment thereof has a length of 15 to 20 amino acids.

[0064] Mode of administration

[0065] Compounds and compositions of this invention may be administered or delivered in any manner including, but not limited to, orally, parenterally, transdermally, intratecally or combinations thereof. Parenteral administration includes, but is not limited to, intravenous, intra-peritoneal and subcutaneous

[0066] In a particular embodiment, a compound according to the invention is administered systemically by injection.

[0067] In another particular embodiment, a compound according to the invention is administered orally.

[0068] According to a particular embodiment, a compound according to the invention can be administered at a dosage between 0.1g to 10g per day, in particular at a dosage of 1g per day. According to a particular embodiment, a peptide of the invention is to be administered in combination with a carotenoid, in particular a Xantophyll or a Carotene.The dosage administered, as single or multiple doses, to an individual will vary depending upon a variety of factors, including pharmacokinetic properties, subject conditions and characteristics (gender, age, body weight, health, and size), extent of symptoms, concomitant disease or treatments, frequency of treatment and the effect desired.

[0069] Combination

[0070] According to one embodiment of the invention, the compounds according to the invention and pharmaceutical formulations thereof can be administered alone or in combination with a coagent useful in the prevention and / or treatment of a disease or adisorder involving oxidative stress.

[0071] According to a particular aspect, is provided a combination of at least one compound of the invention in combination with a carotenoid, in particular a Xantophyll or a carotene. For example, a Xantophyll is selected from zeaxanthin and any including and isomers / enantiomers thereof, lutein, P-cryptoxanthin, and astaxanthin. For example, a carotene is P-carotene.

[0072] According to a particular embodiment, the Xantophyll is zeaxanthin and any including and isomers / enantiomers thereof.

[0073] According to a further particular aspect, a combination of the invention is a pharmaceutical composition comprising of at least a compound of the invention and at least one co-agent of the invention or a mixture thereof and a pharmaceutically acceptable carrier, diluent or excipient thereof.

[0074] According to a further particular aspect, the peptides of the invention can be used as a combination comprising at least one carotenoid. For example, those combination can be obtained by pre-incubating peptides of the invention with a Xantophyll.

[0075] For example, combination of the invention may comprise at least one peptide of the invention with at least one Xantophyll in a xanthophylls to peptide molar ratios from 1 : 1 to 10: 1.

[0076] According to a further particular aspect, a combination of the invention further comprises a hydrosoluble antioxidant.

[0077] According to a further particular aspect, the combinations of the invention can be administered as a single formulation.

[0078] According to a particular aspect, the combinations of the present invention is useful in the prevention and / or treatment of a disease or a disorder involving oxidative stress.

[0079] Pharmaceutical compositionsAccording to a particular embodiment is provided a pharmaceutical composition comprising at least one peptide of the invention and at least one pharmaceutically acceptable carrier, diluent or excipient thereof.

[0080] Pharmaceutical compositions of the invention can contain one or more agent(s) of the invention in any form described herein. Compositions of this invention may further comprise one or more pharmaceutically acceptable additional ingredient(s), such as alum, stabilizers, antimicrobial agents, buffers, coloring agents, flavoring agents, adjuvants, and the like.

[0081] The agents of the invention, together with a conventionally employed adjuvant, carrier, diluent or excipient may be placed into the form of pharmaceutical compositions and unit dosages thereof, and in such form may be employed as solids, such as tablets or filled capsules, or liquids such as solutions, suspensions, emulsions, elixirs, or capsules filled with the same, all for oral use, or in the form of sterile injectable solutions for parenteral (including subcutaneous) use. Such pharmaceutical compositions and unit dosage forms thereof may comprise ingredients in conventional proportions, with or without additional active agents or principles, and such unit dosage forms may contain any suitable effective amount of the active ingredient commensurate with the intended dosage range to be employed. Compositions according to the invention are preferably oral.

[0082] Compositions of this invention may be liquid formulations, including, but not limited to, aqueous or oily suspensions, solutions, emulsions, syrups, and elixirs. Liquid forms suitable for oral administration may include a suitable aqueous or non-aqueous vehicle with buffers, suspending and dispensing agents, colorants, flavors and the like. The compositions may also be formulated as a dry powder for reconstitution with water or other suitable vehicle before use. Such liquid preparations may contain additives, including, but not limited to, suspending agents, emulsifying agents, non-aqueous vehicles and preservatives. Suspending agents include, but are not limited to, sorbitol syrup, methyl cellulose, fructose, glucose / sugar syrup, gelatin, hydroxyethylcellulose, carboxymethyl cellulose, aluminum stearate gel, and hydrogenated edible fats. Emulsifying agents include, but are not limited to, lecithin, sorbitan monooleate, and acacia. Non-aqueous vehicles include, but are not limited to, edible oils, almond oil, fractionated coconut oil, oily esters, propylene glycol, and ethyl alcohol. Preservatives include, but are not limited to, methyl or propyl p-hydroxybenzoate and sorbic acid. Further materials as well as processing techniques and the like are set out in Remington: The Science & Practice of Pharmacy, 23rdEdition, 2020, Ed. Adeboye Adejare, Academic Press, which is incorporated herein by reference. Solid compositions of this invention may be in the form of tablets or lozenges formulated in a conventional manner. For example, tabletsand capsules for oral administration may contain conventional excipients including, but not limited to, binding agents, fillers, lubricants, disintegrants and wetting agents. Binding agents include, but are not limited to, syrup, acacia, gelatin, sorbitol, tragacanth, mucilage of starch and polyvinylpyrrolidone. Fillers include, but are not limited to, lactose, sugar, microcrystalline cellulose, maizestarch, calcium phosphate, and sorbitol. Lubricants include, but are not limited to, magnesium stearate, stearic acid, talc, polyethylene glycol, and silica. Disintegrants include, but are not limited to, potato starch and sodium starch glycollate. Wetting agents include, but are not limited to, sodium lauryl sulfate. Tablets may be coated according to methods well known in the art.

[0083] Injectable compositions are typically based upon injectable sterile saline or phosphate-buffered saline or other injectable carriers known in the art.

[0084] Compositions of this invention may also be formulated for parenteral administration, including, but not limited to, injection or continuous infusion. Formulations for injection may be in the form of suspensions, solutions, or emulsions in oily or aqueous vehicles, and may contain formulation agents including, but not limited to, suspending, stabilizing, and dispersing agents. The composition may also be provided in a powder form for reconstitution with a suitable vehicle including, but not limited to, sterile, pyrogen-free water.

[0085] Compositions of this invention may also be formulated as a depot preparation, which may be administered by implantation or by intramuscular injection. The compositions may be formulated with suitable polymeric or hydrophobic materials (as an emulsion in an acceptable oil, for example), ion exchange resins, or as sparingly soluble derivatives (as a sparingly soluble salt, for example).

[0086] Compositions of this invention may also be formulated as a liposome preparation. The liposome preparation can comprise liposomes which penetrate the cells of interest or the stratum corneum. and fuse with the cell membrane, resulting in delivery of the contents of the liposome into the cell. Other suitable formulations can employ niosomes. Niosomes are lipid vesicles similar to liposomes, with membranes consisting largely of non-ionic lipids, some forms of which are effective for transporting agents across the stratum corneum.

[0087] The agents of this invention can also be administered in sustained release forms or from sustained release drug delivery systems. A description of representative sustained release materials can also be found in the incorporated materials in Remington’s Pharmaceutical Sciences.

[0088] PatientsIn an embodiment, subjects according to the invention are subjects suffering from a disease or disorder characterized by oxidative stress.

[0089] In another embodiment, subjects according to the invention are subjects suffering from a disease or disorder characterized by oxidative stress for which the supplementation in lipophilic antioxidant is beneficial.

[0090] In another embodiment, subjects according to the invention are subjects suffering from a disease or disorder selected from diabetes, a cancer, a cardiovascular disorder and Sickle Cell Disease.

[0091] In another embodiment, subjects according to the invention are subjects suffering from a disease or disorder characterized by increased amyloid formation and accumulation.

[0092] In another further embodiment, subjects according to the invention are subjects suffering from any neurodegenerative disorder such as AD or AMD.

[0093] Use of the invention

[0094] According to another particular aspect, the peptides of the invention are to be administered in combination with at least a co-agent as defined herein or a mixture thereof and a pharmaceutically acceptable carrier, diluent or excipient thereof.

[0095] According to a further aspect, compounds of the invention or combinations thereof are to be administered orally.

[0096] According to a further aspect, compounds of the invention or combinations thereof are to be administered by injection, in particular by parenteral, subcutaneous or intravenous administration.

[0097] According to another aspect, the invention provides a method of preventing and / or treating a disease or disorder characterized by oxidative stress for which the supplementation in lipophilic antioxidant is beneficial, said method comprising administering at least a peptide of the invention alone or in combination with at least one carotenoid, in particular a Xantophyll or a carotene, to a subject in need thereof.

[0098] Peptides of the invention or a pharmaceutical formulation thereof, can be administered to an individual prior to, simultaneously or sequentially with at least one further co-agent. Co-agents or pharmaceutical formulations thereof that are administered simultaneously with at least one peptide of the invention, can be administered in the same or different composition(s) and by the same or different route(s) of administration.

[0099] According to one aspect, a combination of the invention advantageously allows the transport and delivery of lipophilic antioxidants.The amyloid-P peptide (AP) can generate cytotoxic oligomers, and their accumulation is thought to underlie the neuropathologic changes found in Alzheimer’ s Disease (Are-Gomez and Schon Journal of Alzheimer’s Disease 98 (2024) 1243 1275). Xanthophylls delivered to brain tissue may interact and stabilize the Ap 13-23 helix and alter its aggregation properties of and reduce Ap toxicity. Thus xanthophylls delivered by peptides of the invention to tissues in need may be useful to prevent amyloidogenesis.

[0100] EXAMPLES

[0101] Example 1: In Silico docking of lipophilic antioxidants to a peptide of the invention Docking of various lipophilic antioxidant were performed using the SwissDock Autodock Vina Platform (Bugnon et al., 2024, Nucleic Acids Res, 52(W1):W324-W332, doi: 10.1093 / nar / gkae300.. Nucleic Acids Res. 2024; Eberhardt et al., 2021, J. Chem. Inf. Model., 61, 8, 3891 3898, https: / / doi. org / 10.1021 / acs.jcim. lc00203).

[0102] Results are given for position 1 with the lowest Gibbs value (AG in kcal / mol) i.e. the highest affinity of the ligand for the target peptide sequence (SEQ ID NO: 2). Binding affinity: Dock Vina Swiss Dock and presented in Table 2 below.

[0103] Table 2

[0104]

[0105] Figures 1-3 illustrates the interaction of a lipophilic antioxidant with a peptide of the invention. Structures of the compounds are presented on Figure 4B.

[0106] Example 2: Selectivity of a peptide of the invention compared to the known carotenoid binding protein StarD3StarD3 (StAR-related lipid-transfer protein 3; also known as MLN64) with a sequence under PDB: 519 J) was described as a high affinity binding protein for a number of carotenoids (Li et al., 2022, Arch Biochem Biophys. 15, 716; Horvath et al., 2016, Acta Cryst., F72, 609 -618) Seven selected, carotenoids and tocopherols, with measured affinities for a peptide of the invention (Table 2) were compared for their binding affinity for StarD3 (PDB: 519 J) using the same Vina Autodock platform and results are presented in Table 3 below.

[0107] Table 3

[0108]

[0109] From Table 3 it can be concluded that all 7 compounds bound efficiently to StarD3. Although chemically very different (Figure 4), no quantitative difference was measured between xanthophylls and tocopherols. This comparison clearly demonstrates that many lipophilic compounds can bind proteins such as StarD3 in a non selective manner. Indeed, alpha and gamma tocopherols are as lipophilic (logP values) as xanthophylls and they bind similarly to StarD3.

[0110] On the contrary, peptides of the invention show very high selectivity and enantioselectivity for xanthophylls and do discriminate xanthophylls, carotene and carotenoids from structurally different tocopherols (Figure 4).

[0111] Example 3: In vivo activity of a peptide of the invention

[0112] A peptide of the invention (SEQ ID NO: 2) is pre-incubated with the selected xanthophyll as described herein and is administered to wild type mice. The amount of xanthophylls is measured in plasma, lipoprotein fractions, tissues such as retina, brain, kidney and skin.

[0113] The activity is also assessed in an animal model of Alzheimer’s Disease, (Sasaguri et al EMBO Journal, 2017,36: 2473- -2487) and in addition to the measurement of the amount of xanthophylls measured in plasma, lipoprotein fractions, tissues such as retina, brain, kidney and skin, the amount of amyloid accumulated in brain tissue and cognition are assessed as described by Lewis 2010 JBC 285, 47, 36958-36968.LIST OF SEQUENCES

[0114] Consensus sequence

[0115] SEQ ID NO: 1

[0116] X1X2FX4X5X6X7X8X9X1OX11X12X13X14X15X16FX18X19X2O

[0117] wherein

[0118] Xi is absent or present and when present is a Serine or a homologous variant thereof;

[0119] X2 is absent or present and when present is a Threonine or a homologous variant thereof; X4 is a Serine or a homologous variant thereof;

[0120] X5 is an Lysine or a homologous variant thereof;

[0121] Xe is a Leucine or a homologous variant thereof;

[0122] X7 is a Arginine or a homologous variant thereof;

[0123] Xs is a Glutamic Acid or a homologous variant thereof;

[0124] X9 is is a Glutamine or a homologous variant thereof;

[0125] X10 is a Leucine or a homologous variant thereof;

[0126] Xi 1 is a Glycine or a homologous variant thereof;

[0127] X12 is a Proline or a homologous variant thereof;

[0128] X13 is a Valine or a homologous variant thereof;

[0129] X14 is a Threonine or a homologous variant thereof;

[0130] X15 is a Glutamine or a homologous variant thereof;

[0131] Xi6 is a Glutamic Acid or a homologous variant thereof;

[0132] Xis is absent or present and when present is a Tryptophane or a homologous variant thereof; X19 is absent or present and when present is an Aspartic Acid or a homologous variant thereof; X20 is absent or present and when present is an Asparagine or a homologous variant thereof.

[0133] Peptide 1

[0134] SEQ ID NO: 2

[0135] STFSKLREQLGPVTQEFWDN

Claims

Claims1. A peptide of SEQ ID NO: 1 or a homologous variant or a fragment thereof, wherein said peptide, homologous variant or fragment thereof has a length of 15 to 20 amino acids.

2. A peptide according to claim 1, wherein said peptide is of SEQ ID NO: 2 or a homologous variant or a fragment thereof, wherein said homologous variant or a fragment thereof has a length of 15 to 20 amino acids.

3. A pharmaceutical composition comprising at least one peptide of any one of claims 1 to 2 and at least one pharmaceutically acceptable carrier, diluent or excipient thereof4. A pharmaceutical composition according to claim 3, further comprising at least one coagent selected from agent useful in the prevention and / or treatment of a disease or adisorder involving oxidative stress.

5. A pharmaceutical composition according to claim 4, wherein said at least one co-agent is a carotenoid, in particular a Xantophyll or a carotene.

6. A pharmaceutical composition according to claim 5 wherein said Xanthophyll is selected from Zeaxanthin and isomers / enanti omers thereof, P-Cryptoxanthin, and Astaxanthin.

7. A pharmaceutical composition according to claim 5 wherein said carotene is P- Carotene.

8. A pharmaceutical composition according to any one of claims 3 to 7 further comprising a hydrosoluble antioxidant.

9. A peptide according to any one of claims 1 to 2 or a formulation thereof for use in the prevention and / or treatment of a disease or a disorder involving oxidative stress.

10. A peptide for use according to claim 9, wherein said peptide is to be administered in combination with a carotenoid, in particular a Xantophyll or a carotene.

11. A peptide for use according to claim 10, wherein said Xanthophyll is selected from Zeaxanthin and isomers / enanti omers thereof, P-Cryptoxanthin, and Astaxanthin.

12. A peptide for use according to claim 10, wherein said carotene is P— Carotene.

13. A peptide for use according to any one of claims 9 to 12, wherein said a disease or a disorder involving oxidative stress is selected a disease or a disorder characterized by increased amyloid formationa and accumulation such as Alzheimer's Disease, Age- related macular degeneration, diabetes, a cancer, a cardiovascular disorder and Sickle Cell Disease.

14. A peptide for use according to any one of claims 9 to 13, wherein said a disease or a disorder involving oxidative stress is selected from Alzheimer's disease and Age-related macular degeneration.

15. A method of preventing and / or treating diseases or disorders involving oxidative stress, said method comprising administering at least a peptide according to any one of claims 1 or 2 or a pharmaceutical formulation thereof alone or in combination with at least one carotenoid, in particular a Xantophyll or a carotene, to a subject in need thereof.

16. A method according to claim 1 wherein said peptide is of SEQ ID NO: 2 or a homologous variant or a fragment thereof, wherein said homologous variant or a fragment thereof has a length of 15 to 20 amino acids.

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