Lipid blends

Abstract

The present invention relates to blends comprising lipids such as fatty acids and / or sphingolipids, wherein said blends comprising between about 0.5-30 wt% of C22-C38 fatty alcohols, and compositions comprising the lipid blends. The present invention also provides a method for the large-scale production of said blends of lipids, wherein rice bran wax (RBW) is utilized as the source of fatty acids.

Description

[0001] Description

[0002] Lipid Blends

[0003] Filed of the invention

[0004] The present invention relates to blends of lipids comprising fatty acids and / or ceramides, and to a method for the production thereof.

[0005] Background

[0006] Sphingolipids are an important class of polar lipids mainly found on the surface of eukaryotic cells. Sphingolipids are structurally characterized by a sphingoid base backbone and by M-acyl moiety deriving from a fatty acid. Sphingolipids can be divided into different classes such as, ceramides, and glycosphingolipids.

[0007] Sphingolipids are involved in diverse biological processes and play important structural and functional roles such as cell-cell recognition, communication, and intercellular adhesion. Particularly, glycosphingolipids, such as gangliosides are found in the brain and play roles in neurological diseases, whereas ceramides are the main constituent of the stratum corneum lipid layer and have a major role in the water-retaining properties of the epidermis, as well as in the barrier function of the skin.

[0008] Accordingly, sphingolipids hold great potential as therapeutics and cosmetics.

[0009] There are over 1000 different ceramide types in human skin. Ceramides are / V-acylated sphingoid bases lacking additional head groups at the 1-position of the sphingoid base backbone, and wherein the A / -acyl group of ceramides typically derives from a fatty acid. The high diversity of ceramides arises from their molecular structure, which may be composed by various combinations of a sphingoid bases and acyl moieties. The diversity and complexity of ceramide molecules create a challenge for their low-cost industrial production, which typically comprises separate procedures to synthesize each of the components of the ceramide molecule, and wherein, each procedure, often comprising multiple steps.

[0010] Progress has been made for the large-scale production of spingoid bases, which can be used as building blocks for synthesis of ceramides (W02009150022, WO2013023878, WO2017033463, WO201923897).

[0011] Spingoid bases, like phytosphingosine, its derivatives, and its ceramides thereof, are now commercially available, (e.g. Skinmimics® by Evonik). However, most of the commercially available ceramides comprise the acyl moiety of palmitic or stearic acid (Cis or Cis acyl, respectively), whereas natural occurring ceramides typically contain longer acyl moieties.

[0012] Ceramides containing longer acyl moieties (i.e. > C20acyl), as well as longer fatty acids (i.e. > C20), are not widely commercialized, however, they are in big demand for cosmetic and therapeutic applications. In fact, it has been reported that skin enriched with blends comprising ceramides carrying long acyl moieties (i.e. > C20acyl) and / or long chain fatty acids (i.e. > C2o) has an improved barrier function as described in Smeden et al., J. Invest. Dermatol 2014, 134:1238-1245.

[0013] Natural sources of fatty acids are plant oils and waxes. Fatty acids may also be produced via biotechnology, however biotech production is still limited to medium-chain length fatty acids, like Ci6- Cig fatty acids.

[0014] Plant waxes, in particular Rice Bran Wax (RBW), have a unique composition of very long chain esters, and therefore could be utilized as a source of long chain fatty acids, such as C22-C3s fatty acids. Recently, it has been described an industrially applicable technology for fractionation of plant waxes (in particular, Carnauba wax) to obtain a population of very long chain fatty acids (> C24) comprising more than 60 wt% of C26.C33 fatty acids (W02022124600). This reference, however, does not describe a method of synthesis of ceramides using the obtained fatty acids.

[0015] Production of ceramides from spingoid bases can be done chemically or enzymatically.

[0016] Chemical or enzymatic processes for the production of ceramides are typically based on the M-acylation of sphingoid bases, which may comprise the use of acylating agents such as acyl chloride (W00172701 Al), activating agents such as l-ethyl-3-(3'-dimethylaminopropyl)carbodiimide hydrochloride (EDC) (Skolova et al, Biochim Biophys Acta Biomembr. 2017, 1859, 824-834, W00172701 Al), or enzymes such as lipases (WO1994026919A1, US2011077302A1). Alternatively, esters of acids have been utilized for the A / -acylation of phytosphingosine and dihydrosphingosine (EP2757090A1, EP2796444A1).

[0017] Still, at present, neither very long chain fatty acids, nor ceramides comprising long chain acyl moieties are commercially available, and their industrial-scale production represents a challenge.

[0018] Brief Description of the Figures

[0019] Figure 1: Schematic diagram of the method for the production of a blend of lipids.

[0020] Summary of the invention

[0021] In a first aspect the present invention relates to a method for the production of a blend of lipids from defatted and saponified rice bran wax (dsRBW), wherein said dsRBW comprising 50 wt% of fatty alcohols, said blend comprising:

[0022] - about 70-99.5 wt% of lipids of formula (1): wherein

[0023] R1is a C7-C33 alkyl, preferably a C13-C29 alkyl,

[0024] R2is selected from -OH, and / or -NHR3, wherein R3is a moiety of formula (2): wherein

[0025] W is hydrogen or a glycosyl moiety,

[0026] R4is hydrogen, aryl, or a substitute or unsubstituted C1-C50 alkyl, preferably a substituted or unsubstituted C1-C17 alkyl, more preferably a substituted or unsubstituted C13-C17 alkyl,

[0027] R5is hydrogen or -OR7, wherein R7is selected from hydrogen, a substituted or unsubstituted Ci-Ce alkyl, or a substituted or unsubstituted C2-C6acyl, the bond - may be a double or a single bond when R5is hydrogen, or is a single bond when

[0028] R5is -OR7,

[0029] Rsis hydrogen, a substituted or unsubstituted Ci-Cg alkyl, or a substituted or unsubstituted Ci- Csacyl, preferably hydrogen, and

[0030] - about 0.5-30 wt% of fatty alcohols; and wherein said method comprising a step of reducing the fatty alcohol content of the dsRBW or a derivative thereof, to a content of around 0.5-30 wt%.

[0031] In a second aspect the present invention relates to a blend of lipids comprising lipids of formula (1): wherein

[0032] R1is a C7-C33 alkyl, preferably a C13-C29 alkyl,

[0033] R2is selected from -OH, and / or -NHR3, wherein R3is a moiety of formula (2): wherein

[0034] W is hydrogen or a glycosyl moiety,

[0035] R4is hydrogen, aryl, or a substituted or unsubstituted C1-C50 alkyl, preferably a substituted or unsubstituted C1-C17 alkyl, more preferably a substituted or unsubstituted C13-C17 alkyl, R5is hydrogen or -OR7, wherein R7is selected from hydrogen, a substituted or unsubstituted Ci-C6alkyl, or a substituted or unsubstituted C2-C3acyl, the bond - may be a double or a single bond when R5is hydrogen, or is a single bond when R5is -OR7,

[0036] Rsis hydrogen, a substituted or unsubstituted Ci-Cg alkyl, or a substituted or unsubstituted Ci-C6acyl, preferably hydrogen, wherein said blend of lipids comprising about 0.5-30 wt% of a C22-C38 fatty alcohol.

[0037] In a third aspect the present invention relates to a cosmetic or therapeutic composition comprising the lipid blends of the second aspect.

[0038] In a fourth aspect, the present invention relates to use of a lipid blend or composition comprising thereof for maintaining physiologically normal skin barrier function or improving a deteriorated skin barrier function in an individual.

[0039] Detailed Description of Invention

[0040] The present invention relates to a blend comprising lipids such as fatty acids and / or sphingolipids, wherein a majority of said lipids either represented or comprising long-chain fatty acids, such as C22and C24 fatty acids (behenic and lignoceric acids, respectively). In one preferred embodiment, the blend comprises at least 50 wt% of lipids that are either representing or comprising long-chain fatty acids, such as C22 and C24fatty acids. According to the invention, the lipid blends also comprise between about 0.5- 30 wt% of C22-C3s fatty alcohols.

[0041] The present invention also provides a method for the large-scale production of said blend of lipids, wherein rice bran wax (RBW) is utilized as the source of fatty acids.

[0042] It is understood that the terms and definitions described throughout the specification relate to all aspect and embodiments of the invention, unless specified otherwise. The term "a" grammatically is a singular, but it may as well mean the plural of e.g., the intended compound. For example, a skilled person would understand that in the expression "a lipid of formula (1)", the provision of not only one single a lipid of formula (1) but of a variety of lipids of formula (1) is meant.

[0043] The terms "about" or "around" or "approximately" may be used interchangeably, and mean being close to the value or range following these terms, as understood by one of ordinary skill in the art and include a deviation up to 10 % of the values or ranges that follow.

[0044] The term "at least" means an unlimited range of values starting from the indicated value or, in case of wt% range, a range starting from the indicated value and up to 100 wt%.

[0045] The term "blend" or "composition" or "mixture" or "mix" mean a combination of different elements, e.g. molecules, in which the component elements are individually distinct; in some embodiments the component elements could be identical, in other embodiments they could be different by one or more structural features.

[0046] The terms "derivative" or "derivate" may be used interchangeably and mean a compound that is synthesised from a structurally similar compound by a chemical reaction with the replacement of one atom or group of atoms. For example - ester is a derivative of carboxylic acid.

[0047] As used herein, the term "population" means several (two or more) identical or structurally different entities present simultaneously in one and the same place , e.g. one or more molecules in a composition.

[0048] The term "cosmetic" relates to improving in appearance, in particular, in appearance of keratinous tissue, i.e. human skin and hair.

[0049] The term "therapeutic" relates to healing of disease or pathologic condition.

[0050] As used herein the term "treat" means in different embodiments either to cosmetically address deterioration in appearance of keratinous tissue, or to therapeutically address a pathologic condition or disease with the objective of improving or stabilising an outcome in the person being treated, or addressing an underlying need, or prophylactically preventing development a pathologic condition or disease by maintaining normal / healthy condition. "Treating" and "treatment" have grammatically corresponding meanings. Treating includes Preferably, both cosmetic and therapeutic treating are topical.

[0051] The term "topical treatment" means treatment applied to body surfaces such as the skin.

[0052] Throughout the specification "wt%" is meant the weight of the named substance contained in the 100 g of the named composition e.g., blend comprising at least about 50 wt% of C22 and C24 fatty acids means that 100 g of the blend typically contains at least about 50 g of C22 and C24 fatty acids. In formulas representing a moiety, or a group such as for example the alkyl of formula (2) the symboljuuv™ means a point of attachment to another group or atom.

[0053] As used herein, the various functional groups or substituents represented will be understood to have a point of attachment at the functional group or atom having the dash (-). For example, in the case of -OR7it will be understood that the point of attachment is the oxygen atom. If a group is listed without a dash, then the attachment point is indicated by the plain and ordinary meaning of the recited group.

[0054] The skilled person would understand that when speaking of position C-l, C-2, C-3, C-4, C-5 etc., reference is herein always made to the respective carbon atoms of lysosphingolipids of formula (5), the lipid of formula (6), or the moiety of formula (2). Positions C-l, C-2, C-3, C-4, C-5 may also be referred to as 1-position, 2-position, 3-position, 4-position, and 5-position respectively.

[0055] The skilled person will understand that in formulas showing a specific compound, like for example formulas (2), (5), or (6) unless the chemical formula expressly describes a carbon atom having a particular stereochemical configuration, the formula is intended to cover compounds where such a stereocenter has an R or an S configuration, or wherein a double bond has a cis or a trans configuration.

[0056] As used herein, the term "alkyl" refers to an acyclic straight or branched hydrocarbyl group having 1-50 carbon atoms which may be saturated or contain one or more double and / or triple bonds (so, forming for example an alkenyl or an alkynyl), and / or which may be substituted or unsubstituted, as herein further described. Typically, the term alkyl refers to a straight acyclic hydrocarbyl group having 1-50 carbons, which may be substituted or unsubstituted.

[0057] As used herein, the term "aryl" refers to an aromatic cyclic hydrocarbyl group having 5-14 ring carbon atoms, which may be mono- or polycyclic, which may contain fused rings, preferably 1 to 3 fused or unfused rings, and which may contain one or more heteroatoms, and / or which may be substituted or unsubstituted, as herein further described. Examples of "aryl" include, but are not limited to, phenyl, naphtyl, anthracyl, phenantryl, pyrrolyl, imidazolyl, thiophenyl, furanyl, oxazolyl, thiazolyl, pyridinyl, pyrimidinyl, pyrazinyl, triazinyl, and benzofuranyl, each of which may be substitute or unsubstituted. Typically, the term "aryl" refers to a substituted or unsubstituted phenyl.

[0058] As used herein, the term "acyl" refers to a group derived by the removal of one or more hydroxyl group from an oxoacid, preferably from a carboxylic acid. The acyl group according to the present invention is typically a saturated or unsaturated C2-C34 acyl, which may be substitute or unsubstituted.

[0059] As used herein, the term "substituted” means that the group in question is substituted with a group which typically modifies the general chemical characteristics of the group in question. The substituents can be used to modify characteristics of the molecule, such as molecule stability, molecule solubility and the ability of the molecule to form crystals. The person skilled in the art will be aware of other suitable substituents of a similar size and charge characteristics, which could be used as alternatives in a given situation.

[0060] In connection with the terms "alkyl", "aryl", and "acyl the term substituted means that the group in question is substituted one or several times, preferably 1 to 3 times, with group(s) selected from hydroxy (which when bound to an unsaturated carbon atom may be present in the tautomeric keto form), oxo, Ci-C6-alkoxy (i.e. Ci-C3-alkyl-oxy), C2-C6-alkenyloxy, carboxy, oxo, Ci-C6-alkoxycarbonyl, Ci-C3- alkylcarbonyl, formyl, aryl, aryloxycarbonyl, aryloxy, arylamino, arylcarbonyl, heteroaryl, heteroarylamino, heteroaryloxycarbonyl, heteroaryloxy, heteroarylcarbonyl, amino, mono- and di-(Ci- C6-alkyl)amino, carbamoyl, mono- and di-(Ci-C6-alkyl)aminocarbonyl, amino-Ci-C6-alkyl-aminocarbonyl, mono- and di-(Ci-C6-alkyl)amino-Ci-C6-alkyl-aminocarbonyl, Ci-C6-alkylcarbonylamino, cyano, guanidino, carbamido, Ci-C6-alkyl-sulphonyl-amino, aryl-sulphonyl-amino, heteroaryl-sulphonyl-amino, Ci-C6- alkanoyloxy, Ci-Cs-alkyl-sulphonyl, Ci-Cs-alkyl-sulphinyl, Ci-Cs-alkylsulphonyloxy, nitro, Ci-Cg-alkylthio, halogen, where any alkyl, alkoxy, and the like representing substituents may be substituted with hydroxy, Ci-C6-alkoxy, C2-C6-alkenyloxy, carboxy, Ci-C6-alkylcarbonylamino, halogen, Ci-C6-alkylthio, Ci- C6-alkyl-sulphonyl-amino, or guanidino.

[0061] In connection with the term "alkyl" the term "substituted" preferably means that the group in question is substituted one or several times, preferably 1 to 3 times, with group(s) selected from a hydroxyl group, an alkoxy group, an acyloxy group, an acylamido group, a thiol, a thioether or a phosphorus- containing functional group.

[0062] The term "glycosyl moiety" when used herein is defined to encompass a moiety derived from a monosaccharide or from an oligosaccharide (more than one monosaccharide units), wherein the anomeric carbon of the monosaccharide or the anomeric carbon at the reducing end of the oligosaccharide is engaged in a glycosidic bond with another chemical entity, and the bond, if not further specified, may be an alpha or a beta glycosidic bond. A glycosyl moiety having more than one monosaccharide unit may represent a linear or a branched structure.

[0063] The monosaccharide unit can be any 5-9 carbon atom sugar, comprising aldoses (e.g. D-glucose, D- galactose, D-mannose, D-ribose, D-arabinose, L-arabinose, D-xylose, etc.), ketoses (e.g. D-fructose, D- sorbose, D-tagatose, etc.), deoxysugars (e.g. L-rhamnose, L-fucose, etc.), deoxy-aminosugars (e.g. N- acetylglucosamine, N-acetylmannosamine, N-acetylgalactosamine, etc.), uronic acids, ketoaldonic acids (e.g. sialic acid). The monosaccharide unit can form different cyclic structures such as pyranose (sixmembered) cyclic structures or furanose (five-membered) cyclic structures.

[0064] The glycosyl moieties according to the present invention may be illustrated in the following style: Ga I pi-4G Icl-, wherein the dash (-) represents the point of attachment of the glycosyl moiety and wherein the glycosyl moiety may be linked via an alpha or a beta glycosidic bond, preferably a beta glycosidic bond.

[0065] The term "fatty alcohol", as used herein, refers to a C4-C38 acyclic aliphatic primary alcohol, preferably to a C22-C38 acyclic aliphatic primary alcohol. The fatty alcohol may be linear or branched, saturated or unsaturated.

[0066] In the context of the present invention the term "lipid" refers to hydrophobic or amphiphilic molecules such as for example fatty acids, sphingolipids, glycosphingolipids, phospholipids etc.

[0067] Lipids according to the present invention are typically represented by a lipid of formula (1): wherein

[0068] R1is a C7-C33 alkyl, preferably a C13-C29 alkyl,

[0069] R2is selected from -OH, and / or -NHR3, wherein R3is a moiety of formula (2):

[0070] (2), wherein

[0071] W is hydrogen or a glycosyl moiety,

[0072] R4is hydrogen, aryl, or a substituted or unsubstituted C1-C50 alkyl, preferably a substituted or unsubstituted C1-C17 alkyl, more preferably a substituted or unsubstituted C13-C17 alkyl,

[0073] R5is hydrogen or -OR7, wherein R7is selected from hydrogen, a substituted or unsubstituted Ci-Cs alkyl, or a substituted or unsubstituted C2-C6acyl, the bond - may be a double or a single bond when R5is hydrogen, or is a single bond when R5is -OR7, Rsis hydrogen, a substituted or unsubstituted Ci-C3alkyl, or a substituted or unsubstituted Ci-Cg acyl, preferably hydrogen.

[0074] In some embodiments, R2of the lipid of formula (1) is -OH. Accordingly, in some embodiments, the lipid of formula (1) is a fatty acid of formula (3): wherein R1is a C7-C33 alkyl, preferably a C13-C29 alkyl,

[0075] In some embodiments, the present invention describes a method for the production of a blend comprising a C8-C34 non-hydroxy fatty acid ( N), preferably C14-C30 non hydroxy fatty acid (N).

[0076] In some embodiments, R2of the lipid of formula (1) is -NHR3, wherein R3is a moiety of formula (2). Accordingly in some embodiments, the lipid of formula (1) is a lipid of formula (6):

[0077] (6), wherein

[0078] R1is a C7-C33 alkyl, preferably a C14-C30 alkyl,

[0079] W is hydrogen or a glycosyl moiety,

[0080] R4is hydrogen, aryl, or a substituted or unsubstituted C1-C50 alkyl, preferably a substituted or unsubstituted C1-C17 alkyl, more preferably a substituted or unsubstituted C13-C17 alkyl,

[0081] R5is hydrogen or -OR7, wherein R7is selected from hydrogen, a substituted or unsubstituted Ci-Cs alkyl, or a substituted or unsubstituted C2-C6acyl, the bond - may be a double or a single bond when R5is hydrogen, or is a single bond when R5is -OR7,

[0082] Rsis hydrogen, a substituted or unsubstituted Ci-C3alkyl, or a substituted or unsubstituted Ci-C6acyl, preferably hydrogen. In some embodiments, for the moiety of formula (2), for the lysosphingolipid of formula (5) and the for the lipid of formula (6) R4is a saturated unsubstituted Cu-Ci? alkyl, R5is -OH, R6is hydrogen, and the bond - is a single bond.

[0083] In some embodiments, for the moiety of formula (2), for the lysosphingolipid of formula (5) and for the lipid of formula (6) R4is a saturated unsubstituted C13-C17 alkyl, R5and R6are hydrogen, and the bond - is a single bond.

[0084] In some embodiments, for the moiety of formula (2), for the lysosphingolipid of formula (5) and for the lipid of formula (6) R4is a saturated unsubstituted C13-C17 alkyl, R5and R6are hydrogen, and the bond - is a double bond.

[0085] In some preferred embodiments, for the moiety of formula (2), for the lysosphingolipid of formula (5) and for the lipid of formula (6) W is hydrogen. Accordingly, in some preferred embodiments, the lipid of formula (1) is a lipid of formula (6), wherein the lipid of formula (6) is a ceramide.

[0086] In some embodiments, the lipid of formula (6) is a ceramide selected from the group consisting of CER[NP], CER[NDS], CER[NS],or CER[NH],

[0087] In some embodiments, the present invention describes a method for the production of a blend comprising a ceramide carrying the acyl moiety of a C8-C34 non hydroxy fatty acids (N), preferably of a Cu-Cao non hydroxy fatty acids (N).

[0088] In some embodiments, the present invention describes a method for the production of a blend comprising a C8-C34 non hydroxy fatty acids (N), and a ceramide carrying the acyl moiety of a C8-C34 non hydroxy fatty acid (N).

[0089] In some embodiments, the present invention describes a method for the production of a blend comprising a C14-C30 non hydroxy fatty acids (N), and a ceramide carrying the acyl moiety of a C14-C30 non hydroxy fatty acid (N).

[0090] In some embodiments, the present invention describes a method for the production of a blend comprising CER[N (8-34:0) P(18)].

[0091] In some embodiments, the present invention describes a method for the production of a blend comprising CER[N (14-30:0) P(18)].

[0092] In some embodiments, the present invention describes a method for the production of a blend comprising CER[N (8-34:0) DS(18)].

[0093] In some embodiments, the present invention describes a method for the production of a blend comprising CER[N (14-30:0) DS(18)]. In some embodiments, the present invention describes a method for the production of a blend comprising CER[N (8-34:0) S(18)].

[0094] In some embodiments, the present invention describes a method for the production of a blend comprising CER[N (14-30:0) S(18)].

[0095] In some embodiments, the present invention describes a method for the production of a blend comprising CER[N (8-34:0) H(18)].

[0096] In some embodiments, the present invention describes a method for the production of a blend comprising CER[N (14-30:0) H(18)].

[0097] In some embodiments, the present invention describes a method for the production of a blend comprising CER[N (8-34:0) P(18)] and CER[N (8-34:0) DS( 18)].

[0098] In some embodiments, the present invention describes a method for the production of a blend comprising CER[N (14-30:0) P(18)] and CER[N (14-30:0) DS(18)].

[0099] In some embodiments, the present invention describes a method for the production of a blend comprising a C8-C34 non hydroxy fatty acid and CER[N (8-34:0) P( 18)] .

[0100] In some embodiments, the present invention describes a method for the production of a blend comprising a C14-C30 non hydroxy fatty acid and CER[N (14-30:0) P( 18)].

[0101] In some embodiments, the present invention describes a method for the production of a blend comprising a Cg-C34 non hydroxy fatty acid and CER[N (8-34:0) DS(18)].

[0102] In some embodiments, the present invention describes a method for the production of a blend comprising C14-C30 non hydroxy fatty acids, CER[N (14-30:0) P(18)] and CER[N (14-30:0) DS(18)].

[0103] Ceramides denote, in the context of the present invention, naturally occurring ceramides, analogues thereof or derivatives thereof. Preferred ceramides are those naturally occurring in humans. Naturally occurring human ceramides (CERs) include, but are not limited to, CER[NS], CER[AS], CER[EOS], CER[NH], CER[AH], or CER[EOH], CER[NP], CER[AP], or CER[EOP], CER[NDS], CER[ADS], or CER[EODS], wherein letters in brackets refer to the shorthand nomenclature developed by Motta et al., Biochim Biophys Acta., 1993, 1182:147-151 and expanded by Rabionet et al., Biochim Biophys Acta, 2014, 1841:422-434, and by Masukawa et al., Journal of Lipid Research, 2008, 49, 1466-1476. The letters N, A, and EO represent non-hydroxy fatty acids (N), alpha-hydroxy fatty acids (A), and omega-linoleoyloxy fatty acids (EO), respectively, wherein the number of fatty acid carbons and unsaturations may be expressed in parentheses following the letters of N, A, E, and O. The letters, S, H, P, and DS represent o-erythro- sphingosine (S), 6-hydroxy-D-eryt ro-sphingosine (H), D-r / bo-phytosphingosine (P), D-erythro- dihydrosphingosine (DS), respectively, wherein the number of sphingoid carbons may be expressed in parenthesis following the letters S, H, P, and DS. Ceramides, CER[NDS], CER[ADS], or CER[EODS], may also be referred to as CER[NG], CER[AG], or CER[EOG], respectively, wherein the letter G represents the INCI name for D-erythro-dihydrosphingosine.

[0104] In some embodiments, for the moiety of formula (2), for the lysosphingolipid of formula (5) and for the lipid of formula (6) W is a glycosyl moiety. Accordingly, in some embodiments, the lipid of formula (1) is a lipid of formula (6), wherein the lipid of formula (6) is a glycosphingolipid.

[0105] In some preferred embodiments, W is a glycosyl moiety selected from the group consisting of Glcl-, Gall-, Gaipi-4Glcl-.

[0106] In some embodiments, the present invention describes a method for the production of a blend comprising a glycosphingolipid carrying the acyl moiety of C8-C34 non hydroxy fatty acids [N(8-34:0)].

[0107] In some embodiments, for the lipid of formula (6) W is Glcl-.

[0108] In some embodiment, the lipid of formula (6) is a glycosphingolipid selected from the group consisting of GIcCE R[N(8-34:0) P(18)], GlcCER[N(8-34:0) DS(18)], GlcCER[N(8-34:0) S(18)],or GlcCER[N (8-34:0) H(18)].

[0109] In some embodiment, the lipid of formula (6) is a glycosphingolipid selected from the group consisting of GlcCER[N(14-30:0) P(18)], GlcCER[N(14-30:0) DS(18)], GlcCER[N(14-30:0) S(18)],or GlcCER[N (14-30:0) H(18)].

[0110] In some embodiments, for the lipids of formula (6) W is Gall-.

[0111] In some embodiment, the lipid of formula (6) is a glycosphingolipid selected from the group consisting of GalCER[N(8-34:0) P(18)], GalCER[N(8-34:0) DS(18)], GalCER[N(8-34:0) S(18)],or GalCER[N(8-34:0) H(18)].

[0112] In some embodiment, the lipid of formula (6) is a glycosphingolipid selected from the group consisting of GalCER[N(14-30:0) P(18)], GalCER[N(14-30:0) DS(18)], GalCER[N(14-30:0) S( 18)], or GalCER[N(14-30:0) H(18)].

[0113] In some embodiments, for the lipid of formula (6) W is Gaipi-4Glcl-.

[0114] In some embodiment, the lipid of formula (6) is a glycosphingolipid selected from the group consisting of LacCER[N(8-34:0) P(18)], LacCER[N(8-34:0) DS(18)], LacCER[N(8-34:0) S(18)],or LacCER[N(8-34:0) H(18)].

[0115] In some embodiment, the lipid of formula (6) is a glycosphingolipid selected from the group consisting of LacCER[N(14-30:0) P(18)], LacCER[N(14-30:0) DS(18)], LacCER[N(14-30:0) S(18)], or LacCER[N(14-30:0) H(18)]. In some embodiments, the present invention describes a method for the production of a blend comprising a C8-C34 non hydroxy fatty acid (N), and a glycosphingolipid carrying the acyl moiety of a C8- C34 non hydroxy fatty acids (N).

[0116] In some embodiments, the present invention describes a method for the production of a blend comprising a C14-C30 non hydroxy fatty acid (N), and a glycosphingolipid carrying the acyl moiety of a CM- C30 non hydroxy fatty acids (N).

[0117] The term "lysosphingol ipid" when used herein refers to a sphingolipid which lacks the amide-linked fatty acid at the C-2 position of the sphingoid base backbone. Suitable lysosphingolipids, for use in the context of the present invention are sphingoid bases, glycosylated sphingoid bases, and analogs thereof and are preferably represented by a lysosphingolipid of formula (5): wherein

[0118] W is H or a glycosyl moiety,

[0119] R4is hydrogen, aryl, or a substituted or unsubstituted C1-C50 alkyl, preferably a substituted or unsubstituted C1-C17 alkyl, more preferably a substituted or unsubstituted C13-C17 alkyl,

[0120] R5is hydrogen or -OR7, wherein R7is selected from hydrogen, a substituted or unsubstituted Ci-Cg alkyl, or a substituted or unsubstituted C2-C6acyl, the bond - may be a double or a single bond when R5is hydrogen, or is a single bond when R5is -OR7,

[0121] Rsis hydrogen, a substituted or unsubstituted Ci-C5alkyl, or a substituted or unsubstituted Ci-C6acyl, preferably hydrogen.

[0122] In some embodiments, for the lysosphingolipid of formula (5) R4is a saturated unsubstituted C13-C17 alkyl, R5is -OH, R6is hydrogen, and the bond - is a single bond.

[0123] In some embodiments, for the lysosphingolipid of formula (5) R4is a saturated unsubstituted C13-C17 alkyl, R5and Rsare hydrogen, and the bond - is a single bond.

[0124] In some embodiments, for the lysosphingolipid of formula (5) R4is a saturated unsubstituted C13-C17 alkyl, R5and Rsare hydrogen, and the bond - is a double bond. In some embodiments, for the lysosphingolipid of formula (5) W is H. Accordingly, in some embodiments the lysosphingolipid of formula (5) is a sphingoid base.

[0125] In some embodiments, the lysosphingolipid of formula (5) is a sphingoid base selected from the group consisting of D-erytbro-sphingosine [S(18)], 6-hydroxy-D-erytbro-sphingosine [H(18J], D-ribo- phytosphingosine [P(18)], D-erytbro-dihydrosphingosine [DS(18)].

[0126] In some preferred embodiments, the lysosphingolipid of formula (5) is D-r / bo-phytosphingosine [P(18)].

[0127] In some preferred embodiments, the lysosphingolipid of formula (5) is D-erytbro-dihydrosphingosine [DS(18)].

[0128] In some embodiments, the lysosphingolipid of formula (5) is a mixture of D-r / bo-phytosphingosine [P(18)] and D-erytbro-dihydrosphingosine [DS(18)].

[0129] In some embodiments, for the lysosphingolipid of formula (5) W is a glycosyl moiety. Accordingly, in some embodiments the lysosphingolipid of formula (5) is a glycosylated sphingoid base.

[0130] In some preferred embodiments, W is a glycosyl moiety selected from the group consisting of Glcl-, Gall-, Gaipi-4Glcl-.

[0131] In some embodiments, W is Glcl-. Accordingly, in some embodiment, the lysosphingolipid of formula (5) is a glycosylated sphingoid base selected from the group consisting of Glc[P(18)], Glc[DS( 18)], Glc[S(18)],or Glc[H(18)].

[0132] In some embodiments, W is Gall-. Accordingly, in some embodiment, the lysosphingolipid of formula (5) is a glycosylated sphingoid base selected from the group consisting of Gal[P(18)], Gal[DS(18)], Gal[S(18)],or Gal[H(18)].

[0133] In some embodiments, W is Gaipi-4Glcl-. Accordingly, in some embodiment, the lysosphingolipid of formula (5) is a glycosylated sphingoid base selected from the group consisting of Lac [P(18)], Lac[DS(18)], Lac[S(18)],or Lac[H(18)].

[0134] Lysosphingolipids for use in the context of the present invention are preferably obtained via synthetic and / or biotechnological approaches such as those described in WO 2021170624 A2, or in WO2019238970 Al, WO2022158993 Al, or by Sarmientos et al., Eur. J. Biochem. 1986, 160,527-535.

[0135] In some embodiments, the lysosphingolipid of formula (5) may be produced or utilized in the form of a salt, preferably in the form of pharmaceutical acceptable salts.

[0136] In some embodiments, the salt of lysosphingolipids of formula (5) may be formed from the following acids: hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, polyphosphoric acid, acetic acid, camphor sulfonic acid, p-toluene sulfonic acid, methane sulfonic acid, trifluoromethanesulfonic acid, perchloric acid. In some embodiments, the acid is selected from hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, polyphosphoric acid, perchloric acid.

[0137] In some preferred embodiments, the acid is hydrochloric acid.

[0138] The term "C8-C34 triazine-based acylating agent(s)" as used herein refers to an activated derivate of a carboxylic acid, wherein the acidic hydroxyl group of the carboxylic acid is converted into a good leaving group via the replacement of the hydrogen atom with a 1,3,5-triazynyl group. Suitable triazine-based acylating agents for use in the context of the present invention are for example those represented by formula (7), or a combination thereof: wherein represents a conjugated system of bonds such that either two or three double bonds are present in the ring;

[0139] Y is selected from C(O-C(=O)R1), or C(=O);

[0140] Xais selected from N, NR8, or N(C(=O)R1);

[0141] Xbis selected from N, or NR8;

[0142] Z is selected from C(=O), or C(OR8); and provided that: when Y is C(O-C(=O)R1), Z is C(OR8), Xaand Xbare N, and three double bonds are present in the ring, or when Y is C(O-C(=O)R1), Z is C(=O), one of Xaand Xbis N and the other group is NR8, and two double bonds are present in the ring, or when Y is C(=O), Z is C(OR8), Xais N(C(=O)R1), Xbis N, and two double bonds are present in the ring; and wherein

[0143] R1is C7-C33 alkyl, preferably a C13-C29 alkyl,

[0144] R8is selected from methyl, ethyl, 2,2,2-trifluoroethyl, and substituted or unsubstituted benzyl, preferably methyl.

[0145] The expression "one of Xaand Xbis N and the other group is NR8," means that when Xais N the other group Xbis NR8, or when Xbis N the other group Xais NR8. Suitable triazine-based acylating agents for use in the context of the present invention carry one acyl group, such as a C8-C34 acyl group, preferably such as a C14-C30 acyl group. Triazine-based acylating agents carrying one acyl group can form several isomeric structures resulting from the migration of substituents on different positions of the 1,3,5-triazine ring. Depending on conditions such as temperature, solvent, and / or the use of a certain reagent such as a certain base, these isomeric structures can be isolated in a pure form or obtained as an isomeric mixture.

[0146] Accordingly, in the context of the present invention, the term "combination of triazine-based acylating agents" refers to a mixture comprising triazine isomeric structures which differ only in the position of the substituents on the triazine ring and can all serve as acylating agents.

[0147] In some embodiments, the triazine-based acylating agent of formula (7) is a triazine-based acylating agent of formula (8), (9), (10), or (11): wherein

[0148] R1and R8are as defined as for the triazine-based acylating agent of formula (7).

[0149] In some embodiments, the triazine-based acylating agent of formula (7) is a combination comprising triazine-based acylating agents of formulas (8), (9), (10) and (11), and wherein each of said triazinebased acylating agent is present, in said combination, in the amount from about 1% to about 99%.

[0150] In some embodiments, the triazine-based acylating agent of formula (7) is a combination comprising triazine-based acylating agents of formulas (8) and (10), and wherein each of said triazines is present, in said combination, in the amount from about 1% to about 99%.

[0151] In some embodiments, the triazine-based acylating agent of formula (7) is a combination comprising triazine-based acylating agents of formulas (8) and (10), and wherein the triazine-based acylating agents of formulas (8) is present, in said combination, in the amount from about 5% to about 75%.

[0152] In some embodiments, the triazine-based acylating agent of formula (7) is a combination comprising triazine-based acylating agents of formulas (8) and (10), and wherein the triazine-based acylating agents of formulas (9) is present, in said combination, in the amount from about 5% to about 75%. In some embodiments, the triazine-based acylating agent of formula (7) is a combination comprising triazine-based acylating agents of formulas (9) and (11), and wherein each of said triazines is present, in said combination, in the amount from about 1% to about 99%.

[0153] In some embodiments, the triazine-based acylating agent of formula (7) is a combination comprising triazine-based acylating agents of formulas (9) and (11), and wherein the triazine-based acylating agents of formulas (8) is present, in said combination, in the amount from about 5% to about 50%.

[0154] In some embodiments, the triazine-based acylating agent of formula (7) is a combination comprising triazine-based acylating agents of formulas (9) and (11), and wherein the triazine-based acylating agents of formulas (10) is present, in said combination, in the amount from about 5% to about 50%.

[0155] For those embodiments, describing a combination of triazine-based acylating agents, the amount of the triazine-based acylating agents comprised in the combination is typically determined via1H NMR spectroscopy.

[0156] In a typical procedure: i. characteristic signals deriving from each of the triazine-based acylating agents of the combination are identified, ii. the signals are integrated, iii. the ratio of the integrals is calculated and utilized to the define the amount of the different triazine-based acylating agents of the combination.

[0157] The person skilled in the art will understand that the amount of the triazine-based acylating agents of the combination may represent a molar ratio or mol %, or a weight ratio or wt% . The person skilled in the art will also understand that the molar ratio or mol %, or the weight ratio or wt% of the triazinebased acylating agents of the combination, may vary over time due interconversion between the different isomeric structures of the triazine-based acylating agent.

[0158] In some preferred embodiments, R8of the compound of formula (4) and of triazine-based acylating agents of formulas (8)-( 11) is methyl.

[0159] The triazine-based acylating agents according to the present invention may be utilized or produced in different polymorphic forms. Polymorphic forms as referred to herein can include crystalline and amorphous forms as well as solvate and hydrate forms, which can be further characterized as follows: i. Crystalline forms have different arrangements and / or conformations of the molecules in the crystal lattice. ii. Amorphous forms consist of disordered arrangements of molecules that do not possess a distinguishable crystal lattice. iii. Solvates are crystal forms containing either stoichiometric or non-stoichiometric amounts of a solvent. If the incorporated solvent is water, the solvate is commonly known as a hydrate.

[0160] The present invention provides a method for the production of a blend of lipids comprising lipids of formula (1), from defatted and saponified rice bran wax (dsRBW), wherein said dsRBW comprising 50 wt% of fatty alcohols, and wherein the method comprising a step of reducing the fatty alcohol content of the dsRBW, or a derivative thereof, to a content of around 0.5-30 wt%. dsRBW may be obtained by methods known to the skilled person such as that described by Vali et al., JAOCS 2005, 82, 57-64, or such as that described below in Examples 1 and 2.

[0161] The step of reducing the fatty alcohol content is typically performed via solvent extraction, wherein the solvent extraction may be conducted in in a continuous mode (e.g. Soxhlet extraction), or in a batch mode, and by using one or more extraction solvent(s).

[0162] Solvents suitable for use in the context of the present invention may be polar or apolar solvents, in particular one can mention Ci-Cio alkyl esters (e.g. ethyl or butyl acetate), Ci-Cio aliphatic nitriles (e.g. acetonitrile), Cs-Cio hydrocarbons (e.g. n-heptane or cyclohexane), or supercritical fluids (e.g. supercritical CO2). In some embodiments, the one or more extraction solvent(s) are selected from ethyl acetate, n-heptane, cyclohexane, acetonitrile, or supercritical CO2.

[0163] In some embodiments, the extraction solvent is ethyl acetate.

[0164] In some embodiments, the extraction solvent is supercritical CO2.

[0165] In some embodiments, the extraction is conducted in a continuous mode, wherein dsRBW, or a derivative thereof, is contacted with an extraction solvent in a Soxhlet extractor at reflux temperature. The extraction is continued for a period of time sufficient to extract most of the fatty alcohols from the dsRBW, or the derivative thereof. Typically, the extraction is continued for about 1-6 hours, preferably the extraction is continued for about 4-6 hours. Accordingly in some preferred embodiments, the extraction is continued for about 4, 4.5, 5, 5.5, or 6 hours. The solid residue enriched with fatty acid salts, or derivatives thereof, is collected for further processing (e.g. solvent washes, drying, and / or further synthetic steps).

[0166] In some embodiments the extraction is conducted in a batch mode, wherein the dsRBW, or a derivative thereof, is contacted with an extraction solvent at a temperature ranging from room temperature to reflux temperature, for a period of time sufficient to extract most of the fatty alcohols from the dsRBW, or the derivative thereof. Typically, the extraction is continued for about 1-4 hours, preferably between about 1-2 hours. Accordingly, in some preferred embodiments, the extraction is continued for about 1, 1.5, or 2 hours. The solvent extract, containing the fatty alcohols is separated from the extraction slurry by filtration or centrifugation. The residue enriched with fatty acid salts, or the derivatives thereof is collected for further processing (e.g. solvent washes, drying and / or further synthetic steps). The dsRBW, or the derivative thereof, may be subjected to several extraction cycle(s), as described above.

[0167] The solvent extract obtained after the continuous extraction, or the batch extraction is enriched with fatty alcohols.

[0168] Fatty alcohols hold potential as therapeutics, cosmetics, and functional food. Therefore, it is of interest to recover the fatty alcohols after extraction.

[0169] Fatty alcohols can be easily recovered from the solvent extract by subjecting the solvent extract to further processing.

[0170] Typically, in those embodiments where ethyl acetate is used as the extraction solvent, O-acetylated derivatives of the fatty alcohols are formed during the extraction. Accordingly, in some embodiments, the solvent extract comprising the fatty alcohols and their O-acetylated derivatives is subjected to an acid treatment.

[0171] The acid treatment of the solvent extract is typically performed via the use of an inorganic acid or an organic acid, preferably with an inorganic acid. Acids suitable for use in the context of the present invention include but are not limited to hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, polyphosphoric acid, camphor sulfonic acid, p-toluene sulfonic acid, methane sulfonic acid, trifluoromethanesulfonic acid, perchloric acid.

[0172] In some embodiments the acid is selected from the group consisting of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, polyphosphoric acid, perchloric acid.

[0173] In some preferred embodiments, the acid is hydrochloric acid.

[0174] In some embodiments the acid may be utilized in the form of an aqueous solution.

[0175] It will be appreciated that the amount of acid used in the reaction is dependent upon a variety of factors such as the identity of the reactants (including the identity of the acid), and the reaction conditions that are employed. It is therefore understood that amount of acid may be optimally and independently chosen for each reaction.

[0176] Typically, the acid treatment is performed in a solvent, preferably a polar solvent.

[0177] In some embodiments, the solvent is methanol.

[0178] The acid treatment is typically performed at a temperature between about 60° C and about 65° C. Accordingly, in some embodiments, the acid treatment is performed at a temperature of about 60° C, 51° C, 62° C, 63° C, 64° C, or 65° C.

[0179] The components of the acid treatment may be combined in any order, and it will be appreciated that the order of combining the reactants may be adjusted as needed.

[0180] For example, the acid may be added to the solvent extract. As another example, the solvent extract may be added to a solution of the acid. The acid, the solvent extract, as well as any other reagent used during the acid treatment may be added to the reaction either as a solid, a liquid, or dissolved in a solvent, and in any quantities and manner effective for the intended result of the reaction.

[0181] In some embodiments, the fatty alcohols are precipitated from the reaction of mixture of the acid treatment. The precipitation of the fatty alcohols may be achieved for example via partial removal of the reaction solvent by evaporation, i.e. concentrating the reaction mixture, or via the addition of another solvent to the reaction mixture, or via changes of temperature or pressure, or via addition of other solutes, or combinations of these.

[0182] In some embodiments, the precipitation of the fatty alcohols is achieved via cooling the reaction mixture of the acid treatment to a temperature from about 0° C to about 30° C, preferably from about 10° C to about 25° C. Accordingly, in some preferred embodiments, the precipitation of the salt is achieved via cooling the reaction mixture to a temperature of about 10° C, 11° C, 12° C, 13° C, 14° C, 15° C, 16° C, 17° C, 18° C, 19° C, 20° C, 21° C, 22° C, 23° C, 24° C, or 25° C.

[0183] Following the procedure described above, for the processing of the solvent extract, a fatty alcohol blend comprising 55-65 wt% of C22-C36 fatty alcohols is obtained.

[0184] In some embodiments, the extraction of dsRBW, or a derivative thereof, is performed via super critical liquid CO2(scrCO2) extraction.

[0185] The scrCO2extraction is a known technique for extraction of oily, waxy, and fatty material from rice bran, as described by Garcia A., et al. JAOCS 1996, 73, 1127-1131, by Moreira et al., The Journal of Supercritical Fluids 2023, 192, 105786, and in CN102994215A. These references, however, do not describe the scrCO2extraction of blends deriving from the saponification of defatted rice bran wax (dsRBW).

[0186] According to the present invention, the dsRBW, or the derivative thereof, could be extracted in a supercritical fluid equipment using carbonic anhydride at high pressures (250-500 bar) and at a temperature ranging from about 40 to about 120°C. The extraction process typically takes 1-4 h. The dsRBW, or the derivative thereof, enriched with the fatty acid salts, or derivatives thereof is collected for further processing (e.g. further synthetic steps)

[0187] In some embodiments, the scrCO2extraction is performed in the presence of a co-solvent.

[0188] The solvent extraction typically results in the reduction of the C22-C38 fatty alcohol content from about 50 wt% to a content of about 0.5-30 wt%. Accordingly, in some embodiments, the fatty alcohol content may be reduced to a content of about 0.5-5 wt% , or to a content of about 5-10 wt% , or to a content of about 10-20 wt% , or to a content of about 20-30 wt% .

[0189] In some embodiments, the step of reducing the alcohol content is performed on a derivative of dsRBW. Accordingly, in some embodiments, the method comprising the steps of:

[0190] - providing defatted and saponified rice bran wax (dsRBW), wherein said dsRBW comprising 50 wt% of fatty alcohols ,

[0191] - reacting the dsRBW with a Ci-C6alcohol in the presence of an acid, therby obtaining a dsRBW derivative, wherein said derivative comprising C8-C34fatty acid esters, preferably C14-C30 fatty acid esters, and about 50 wt% of fatty alcohols,

[0192] - reducing the fatty alcohol content of said derivative thereby obtaining a blend of fatty acid esters comprising about 70-99.5 wt% of C8-C34fatty acid esters, preferably Ci4-C30fatty acid esters, and about 0.5-30 wt% of fatty alcohols.

[0193] Derivative of dsRBW according to the present invention are typically obtained via reacting dsRBW with an alcohol in the presence of an acid, thereby obtaining a dsRBW derivative comprising C8-C34, preferably Ci4-C3o fatty acid esters and between about 0.5-30 wt% of fatty alcohols.

[0194] Typically, dsRBW and the alcohol are reacted in the presence of an inorganic acid or an organic acid, preferably with an inorganic acid. Acids suitable for use in the context of the present invention include but are not limited to hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, polyphosphoric acid, camphor sulfonic acid, p-toluene sulfonic acid, methane sulfonic acid, trifluoromethanesulfonic acid, perchloric acid.

[0195] In some embodiments the acid is selected from the group consisting of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, polyphosphoric acid, perchloric acid.

[0196] In some preferred embodiments, the acid is hydrochloric acid.

[0197] In some embodiments the acid may be utilized in the form of an aqueous solution.

[0198] In some embodiments, the acid may be utilized in the form of an alcoholic solution.

[0199] It will be appreciated that the amount of acid used in the reaction is dependent upon a variety of factors such as the identity of the reactants (including the identity of the acid), and the reaction conditions that are employed. It is therefore understood that amount of acid may be optimally and independently chosen for each reaction.

[0200] Alcohols suitable for use in the context of the present invention, include but are not limited to C1-C7 alcohols, or mixtures of C1-C7 alcohols.

[0201] In some embodiments, the alcohol is selected from methanol, ethanol, 1-propanol, isopropanol, 1- butanol, or isobutanol.

[0202] In some preferred embodiments, the alcohol is methanol.

[0203] The dsRBW, the alcohol, and the acid are typically reacted at a temperature from about 50° C to about 60° C. Preferably at a temperature from about 60°C to about 65° C. Accordingly, in some preferred embodiments, the reaction is performed at a temperature of about 60° C, 61° C, 62° C, 63° C, 64° C, or 65°C.

[0204] The reaction between the dsRBW and the alcohol results in the formation of a dsRBW derivative, wherein said derivative comprises C8-C34 fatty acid esters, preferably C14-C30 fatty acid esters, and about 50 wt% of fatty alcohols.

[0205] In some embodiments, the extraction is performed on dsRBW, thereby obtaining a blend comprising C8- C3 fatty acid salts, preferably C14-C30 fatty acid salts, and wherein said blend may further comprise between about 0.5-30 wt% of fatty alcohol.

[0206] Accordingly, in some embodiments, the method comprising the steps of:

[0207] - providing defatted and saponified rice bran wax (dsRBW), wherein said dsRBW comprising 50 wt% of fatty alcohols ,

[0208] - reducing the fatty alcohol content of said dsRBW thereby obtaining a blend comprising about 70-99.5 wt% of C8-C3 fatty acid salts, preferably C14-C30 fatty acid salts and between about 0.5- 30 wt% of fatty alcohols.

[0209] In some embodiments, the fatty alcohol content of the blend comprising the fatty acid salts is about 0.5- 5 wt% .

[0210] In some embodiments, the fatty alcohol content of the blend comprising the fatty acid salts is about 5- 10 wt% .

[0211] In some embodiments, the fatty alcohol content of the blend comprising the fatty acid salts is about 10- 20 wt% .

[0212] In some embodiments, the fatty alcohol content of the blend comprising the fatty acid salts is about 20- 30 wt% .

[0213] In some embodiments, the blend comprising fatty acid salts has a content of at least 50 wt% of C22 and C2 fatty acid salts. Preferably, the ratio between said fatty acid salts is from about 1:10 to about 10:1.

[0214] In some embodiments, the blend comprising the fatty acid salts has a content of at least 50 wt% of C22 and C24 fatty acid salts, and wherein the ratio between said fatty acid salts is from about 1:2.5.

[0215] In some embodiments, the method further comprising a step of treating with an acid the blend comprising the fatty acid salts, thereby obtaining a blend comprising fatty acids of formula (3): wherein R1is a C7-C33 alkyl, preferably a C13-C29 alkyl, and between about 0.5-30 wt% of fatty alcohols.

[0216] Accordingly, in some embodiments, the present invention describes a method for the production of a blend of lipids comprising a lipid of formula (1), the method comprising the steps of:

[0217] - providing defatted and saponified rice bran wax (dsRBW), wherein said dsRBW comprising 50 wt% of fatty alcohols,

[0218] - reducing the fatty alcohol content of said dsRBW thereby obtaining a blend comprising about 70-99.5 wt% of C8-C34 fatty acid salts, preferably C14-C30 fatty acid salts and between about 0.5- 30 wt% of fatty alcohols,

[0219] - treating with an acid the blend comprising the fatty acid salts, thereby obtaining a blend comprising about 70-99.5 wt% of fatty acids of formula (3), and about 0.5-30 wt% of fatty alcohols.

[0220] The acid treatment, of the blend comprising the fatty acid salts, is typically performed via reacting the blend comprising the fatty acid salts, with an inorganic acid or an organic acid, preferably with an inorganic acid. Acids suitable for use in the context of the present invention include but are not limited to hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, polyphosphoric acid, camphor sulfonic acid, p-toluene sulfonic acid, methane sulfonic acid, trifluoromethanesulfonic acid, perchloric acid.

[0221] In some embodiments the acid is selected from the group consisting of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, polyphosphoric acid, perchloric acid.

[0222] In some preferred embodiments, the acid is hydrochloric acid.

[0223] In some embodiments the acid may be utilized in the form of an aqueous solution.

[0224] It will be appreciated that the amount of acid used in the reaction is dependent upon a variety of factors such as the identity of the reactants (including the identity of the acid), and the reaction conditions that are employed. It is therefore understood that amount of acid may be optimally and independently chosen for each reaction.

[0225] Typically, the acid treatment is performed in a solvent, preferably a polar solvent.

[0226] In some embodiments, the solvent is water.

[0227] The acid treatment is typically performed at a temperature between about 50 °C and about 60 °C.

[0228] Accordingly, in some embodiments, the acid treatment is performed at a temperature of about 50 °C, 51 °C, 52 °C, 53 °C, 54 °C, 55 °C, 56 °C, 57 °C, 58 °C, 59 °C, or 60 °C.

[0229] The components of the acid treatment may be combined in any order, and it will be appreciated that the order of combining the reactants may be adjusted as needed. For example, the acid may be added to a solution of the blend comprising the fatty acid salts. As another example, the blend comprising the fatty acid salts may be added to a solution of the acid.

[0230] The acid, the blend comprising the fatty acid salts, as well as any other reagent used during the acid treatment may be added to the reaction either as a solid, a liquid, or dissolved in a solvent, and in any quantities and manner effective for the intended result of the reaction.

[0231] The acid treatment of the blend comprising the fatty acid salts, results in the formation of a blend comprising fatty acids of formula (3).

[0232] In some embodiments, the blend comprising the fatty acids of formula (3) is precipitated from the reaction of mixture of the acid treatment. The precipitation may be achieved for example via partial removal of the reaction solvent by evaporation, i.e. concentrating the reaction mixture, or via the addition of another solvent to the reaction mixture, or via changes of temperature or pressure, or via addition of other solutes, or combinations of these.

[0233] In some embodiments, the precipitation of the blend comprising the fatty acids of formula (3) is achieved via cooling the reaction mixture of the acid treatment to a temperature from about 0° C to about 30° C, preferably from about 10 °C to about 25 °C. Accordingly, in some preferred embodiments, the precipitation is achieved via cooling the reaction mixture to a temperature of about 10° C, 11° C, 12° C, 13° C, 14° C, 15° C, 16° C, 17° C, 18° C, 19° C, 20° C, 21° C, 22° C, 23° C, 24° C, or 25° C.

[0234] The acid treatment of the blend comprising the fatty acid salts, results in the formation of a blend comprising 70-99.5 wt% of fatty acids of formula (3), and about 0.5-30 wt% , of fatty alcohol.

[0235] In some embodiments, the fatty alcohol content of the blend comprising the fatty acids of formula (3) is about 0.5-5 wt% .

[0236] In some embodiments, the fatty alcohol content of the blend comprising the fatty acids of formula (3) is about 5-10 wt% .

[0237] In some embodiments, the fatty alcohol content of the blend comprising the fatty acids of formula (3) is about 10-20 wt% .

[0238] In some embodiments, the the fatty alcohol content of the blend comprising the fatty acids of formula (3) is about 20-30 wt% .

[0239] In some embodiments, the blend comprising the fatty acids of formula (3) has a content of at least about 50 wt% , of fatty acids of formula (3) wherein R1is a C2ialkyl and of fatty acids of formula (3) wherein R1is a C23alkyl, and wherein the ratio between said fatty acids is from about 1:10 to about 10:1.

[0240] In some embodiments, the blend comprising the fatty acids of formula (3) has a content of at least about 50 wt% , of fatty acids of formula (3) wherein R1is a C2ialkyl and of fatty acids of formula (3) wherein R1is a C23alkyl. Preferably, the ratio between said fatty acids is about 1:2.5. In some embodiments, the method further comprising a step of reacting the blend comprising the fatty acids of formula (3) with an alcohol in the presence of an acid, thereby obtaining a blend comprising a C8-C34 fatty acid ester, preferably C14-C30 fatty acid ester.

[0241] Accordingly, in some embodiments, the present invention describes a method for the production of a blend of lipids comprising lipids of formula (1), the method comprising the steps of:

[0242] - providing defatted and saponified rice bran wax (dsRBW), wherein said dsRBW comprising 50 wt% of fatty alcohols,

[0243] - reducing the fatty alcohol content of said dsRBW thereby obtaining a blend comprising about 70-99.5 wt% of C8-C34 fatty acid salts, preferably C14-C30 fatty acid salts, and about 0.5-30 wt% of fatty alcohols,

[0244] - treating with an acid the blend comprising the fatty acid salts, thereby obtaining a blend comprising about 70-99.5 wt% of fatty acids of formula (3), and about 0.5-30 wt% of fatty alcohols,

[0245] - reacting the blend comprising the fatty acids of formula (3) with an alcohol in the presence of an acid, thereby obtaining a blend comprising about 70-99.5 wt% of C8-C34 fatty acid esters, preferably of C14-C30 fatty acid esters, and about 0.5-30 wt% of fatty alcohols.

[0246] Typically, the blend comprising the fatty acids of formula (3) and the alcohol are reacted in the presence of an inorganic acid or an organic acid, preferably with an inorganic acid. Acids suitable for use in the context of the present invention include but are not limited to hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, polyphosphoric acid, camphor sulfonic acid, p-toluene sulfonic acid, methane sulfonic acid, trifluoromethanesulfonic acid, perchloric acid.

[0247] In some embodiments the acid is selected from the group consisting of hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, polyphosphoric acid, perchloric acid.

[0248] In some preferred embodiments, the acid is hydrochloric acid.

[0249] In some embodiments the acid may be utilized in the form of an aqueous solution.

[0250] In some embodiments, the acid may be utilized in the form of an alcoholic solution.

[0251] It will be appreciated that the amount of acid used in the reaction is dependent upon a variety of factors such as the identity of the reactants (including the identity of the acid), and the reaction conditions that are employed. It is therefore understood that amount of acid may be optimally and independently chosen for each reaction.

[0252] Alcohols suitable for use in the context of the present invention, include but are not limited to C1-C7 alcohols, or mixtures of C1-C7 alcohols.

[0253] In some embodiments, the alcohol is selected from methanol, ethanol, 1-propanol, isopropanol, 1- butanol, or isobutanol. In some preferred embodiments, the alcohol is methanol.

[0254] The blend comprising the fatty acids of formula (3), the alcohol, and the acid are typically reacted at a temperature from about 50° C to about 60° C. Preferably at a temperature from about 60° C to about 65° C. Accordingly, in some preferred embodiments, the reaction is performed at a temperature of about 60° C, 61° C, 62° C, 63° C, 64° C, or 65° C.

[0255] The reaction between the blend comprising the fatty acids of formula (3) and the alcohol, results in the formation of a blend comprising fatty acid esters.

[0256] In some embodiments, the blend comprising fatty acid esters is precipitated from the reaction mixture. The precipitation may be achieved for example via partial removal of the reaction solvent by evaporation, i.e. concentrating the reaction mixture, or via the addition of another solvent to the reaction mixture, or via changes of temperature or pressure, or via addition of other solutes, or combinations of these.

[0257] In some embodiments, the precipitation of the blend of fatty acid esters is achieved via cooling the reaction mixture of the acid treatment to a temperature from about 0° C to about 30° C, preferably from about 10 °C to about 25 °C. Accordingly, in some preferred embodiments, the precipitation is achieved via cooling the reaction mixture to a temperature of about 10° C, 11° C, 12° C, 13° C, 14° C, 15° C, 16° C, 17° C, 18° C, 19° C, 20° C, 21° C, 22° C, 23° C, 24° C, or 25° C.

[0258] The reaction between the alcohol and the blend comprising fatty acids of formula (3) results in the formation of a blend comprising about 70-99.5 wt% of C8-C34 fatty acid ester, preferably a C14-C30 fatty acid ester, and about 0.5-30 wt% , of fatty alcohol.

[0259] In some embodiments, the fatty alcohol content of the blend comprising the fatty acid esters is about 0.5-5 wt% .

[0260] In some embodiments, the fatty alcohol content of the blend comprising the fatty acid esters is about 5- 10 wt% .

[0261] In some embodiments, the fatty alcohol content of the blend comprising the fatty acid esters is about 10-20 wt% .

[0262] In some embodiments, the fatty alcohol content of the blend comprising the fatty acid esters is about 20-30 wt% .

[0263] In some embodiments, the blend comprising the fatty acid esters has a content of at least about 50 wt% , of C22 and a of C24 fatty acid esters, and wherein the ratio between said fatty acid esters is from about 1:10 to about 10:1.

[0264] In some embodiments, the blend comprising the fatty acid esters has a content of at least about 50 wt% , of C22 and a of C24 fatty acid esters, and wherein the ratio between said fatty acid esters is about 1:2.5. In some embodiments, the method further comprising a step of reacting the blend comprising the fatty acids of formula (3) with a triazine of formula (4): wherein

[0265] R8is selected from methyl, ethyl, 2,2,2-trifluoroethyl, and substituted or unsubstituted benzyl, preferably methyl, in the presence of an organic base, thereby obtaining a blend comprising triazine-based acylating agents of formula (7), and about 0.5-30 wt% of fatty alcohols.

[0266] Accordingly, in some embodiments, the present invention describes a method for the production of a blend comprising a lipid of formula (1), the method comprising the steps of:

[0267] - providing defatted and saponified rice bran wax (dsRBW), wherein said dsRBW comprising 50 wt% of fatty alcohols,

[0268] - reducing the fatty alcohol content of said dsRBW thereby obtaining a blend comprising about 70-99.5 wt% of C8-C34 fatty acid salts, preferably C14-C30 fatty acid salts, and about 0.5-30 wt% of fatty alcohols,

[0269] - treating with an acid the blend comprising the fatty acid salts, thereby obtaining a blend comprising about 70-99.5 wt% of fatty acids of formula (3), and about 0.5-30 wt% of fatty alcohols,

[0270] - reacting the blend comprising the fatty acids of formula (3) with a triazine of formula (4) in the presence of an organic base thereby obtaining a blend comprising C8-C34 triazine-based acylating agents, preferably C14-C30 triazine-based acylating agents and about 0.5-30 wt% of fatty alcohols.

[0271] Typically, the step of reacting the blend comprising the fatty acids of formula (3) with the triazine of formula (4) is performed in the presence of an organic base.

[0272] In some embodiments, the organic base is selected from 4-methylmorpholine, 1,4- diazabicyclo[2.2.2]octane, preferably 4-methylmorpholine.

[0273] Typically, the steps of reacting the blend comprising the fatty acids of formula (3) with the compound of formula (4) is performed in a non-halogenated solvent. The non-halogenated solvent is preferably selected from a ketone, an alcohol, or an aliphatic hydrocarbon. In some embodiments, the non-halogenated solvent is a ketone selected from acetone, diethyl ketone, methyl isobutyl ketone, or butan-2-one, preferably acetone. In some embodiments, when the solvent is a ketone, water may be added to the reaction mixture. In some embodiments, the non-halogenated solvent is an alcohol selected from methanol, ethanol, propanol, isopropanol, butanol, isobutanol, preferably methanol, or a mixture thereof. In some embodiments, the non-halogenated solvent is an aliphatic hydrocarbon selected from petroleum ether, hexane, or an isomeric mixture thereof, n-heptane, or an isomeric mixture thereof, octane, or an isomeric mixture thereof. In some embodiments, the non-halogenated solvent is a mixture of two alcohols, and wherein the mixture of two alcohols is preferably selected from a mixture of methanol and ethanol, methanol and propanol, methanol and isopropanol, methanol and butanol, or methanol and isobutanol, preferably a mixture of methanol and ethanol.

[0274] In some embodiments, the step of reacting the blend comprising the fatty acids of formula (3) with the triazine of formula (4), is performed at a temperature between about 30° C and about 100° C, preferably between about 30° C and about 55° C.

[0275] The step of reacting the blend comprising the fatty acids of formula (3) with the triazine of formula (4) results in the formation of a blend comprising C8-C34 triazine-based acylating agents, preferably C14-C30 triazine-based acylating agents.

[0276] In some embodiments, the blend comprising the triazine-based acylating agent is precipitated from the reaction. The precipitation may be achieved for example via partial removal of the reaction solvent by evaporation, i.e. concentrating the reaction mixture, or via the addition of another solvent to the reaction mixture, or via changes of temperature or pressure, or via addition of other solutes, or combinations of these.

[0277] In some embodiments, the precipitation is performed at a temperature between about -20 °C and about 25 °C, preferably at a temperature between about -10 °C and about 25 °C, even more preferably at a temperature between about 5 °C and about 25 °C.

[0278] The reaction between the blend of fatty acids of formula (3) with and triazine of formula (4) results in the formation of a blend comprising C8-C34 triazine-based acylating agents, preferably C1 -C30 triazinebased acylating, and wherein said blend further comprising about 20-30 wt% , of fatty alcohols.

[0279] In some embodiments, the fatty alcohol content of the blend comprising the triazine-based acylating agents is about 0.5-5 wt% .

[0280] In some embodiments, the fatty alcohol content of the blend comprising the triazine-based acylating agents is about 5-10 wt% . In some embodiments, the fatty alcohol content of the blend comprising the triazine-based acylating agents is about 10-20 wt% .

[0281] In some embodiments, the fatty alcohol content of the blend comprising the triazine-based acylating agents is about 20-30 wt% .

[0282] In the context of the present invention, C8-C34triazine-based acylating agents may be represented by a triazine-based acylating agent of formula (7).

[0283] In some embodiments, the blend comprising the triazine-based acylating agent has a content of at least about 50 wt% of a triazine-based acylating agent of formula (7) wherin R1is a C2ialkyl and of a triazinebased acylating agent of formula (7) wherein R1is a C23alkyl, and wherein the ratio between said triazine-based acylating agents is from about 1:10 to about 10:1.

[0284] In some embodiments, the blend comprising the triazine-based acylating agent has a content of at least about 50 wt% of a triazine-based acylating agent of formula (7) wherein R1is a C2ialkyl and of a triazine-based acylating agent of formula (7) wherein R1is a C23alkyl, and wherein the ratio between said triazine-based acylating agents is about 1:2.5.

[0285] In some embodiments, the method further comprising a step of reacting a lysosphingolipid of formula (5) with the blend comprising the fatty acid esters in the presence of a base, thereby obtaining a blend of lipids of formula (6), and 0.5-30 wt% of fatty alcohols.

[0286] Accordingly, in some embodiments, the present invention describes a method for the production of a blend comprising lipids of formula (5), the method comprising the steps of:

[0287] - providing defatted and saponified rice bran wax (dsRBW), wherein said dsRBW comprising 50 wt% of fatty alcohols,

[0288] - reducing the fatty alcohol content of said dsRBW thereby obtaining a blend comprising about 70-99.5 wt% of C8-C34fatty acid salts, preferably Ci4-C30fatty acid salts, and about 0.5-30 wt% of fatty alcohols,

[0289] - treating with an acid the blend comprising the fatty acid salts, thereby obtaining a blend comprising about 70-99.5 wt% of fatty acids of formula (3), and about 0.5-30 wt% of fatty alcohols,

[0290] - reacting the blend comprising the fatty acids of formula (3) with an alcohol in the presence of an acid, thereby obtaining a blend comprising about 70-99.5 wt% of C8-C34fatty acid esters, preferably of Ci4-C30fatty acid esters, and about 0.5-30 wt% of fatty alcohols.

[0291] - reacting a lysosphingolipid of formula (5), or a salt thereof:

[0292] (5), wherein

[0293] W is hydrogen or a glycosyl moiety,

[0294] R4is hydrogen, aryl, or a substituted or unsubstituted Ci-Csoalkyl, preferably a substituted or unsubstituted C1-C17 alkyl, more preferably a substituted or unsubstituted C13-C17 alkyl,

[0295] R5is hydrogen or -OR7, wherein R7is selected from hydrogen, a substituted or unsubstituted Ci-

[0296] Cg alkyl, or a substituted or unsubstituted C2-C6 acyl, the bond - may be a double or a single bond when R5is hydrogen, or is a single bond when R5is -OR7,

[0297] R6is hydrogen, a substituted or unsubstituted Ci-C3alkyl, or a substituted or unsubstituted Ci-C3acyl, preferably hydrogen, with the blend comprising the fatty acid esters in the presence of a base, thereby obtaining a blend comprising lipids of formula (6): wherein

[0298] R1is a C7-C33 alkyl, preferably a C13-C29 alkyl,

[0299] W, R4, R5and Rsare as defined as for the lysosphingolipid of formula (5).

[0300] Typically, the lysosphingolipid of formula (5) and the blend comprising the fatty acid esters are reacted in the presence of a base such as an alkoxide, an amine, a carbonate, or a bicarbonate.

[0301] In some embodiments, the lysosphingolipid of formula (5) and the blend comprising the fatty acid esters are reacted in the presence of an amine, wherein the amine is preferably selected from triethylamine, / V, / V-diisopropylethylamine, and pyridine. In some embodiments, the lysosph ingo li pid of formula (5) and the blend comprising the fatty acid esters are reacted in the presence of a carbonate, wherein the carbonate is preferably selected from Na2CO3, K2CO3, CaCO3, Li2CO3, (NH4)2CO3.

[0302] In some embodiments, the lysosphingolipid of formula (5) and the blend comprising the fatty acid esters are reacted in the presence of a bicarbonate, wherein the bicarbonate is preferably selected from NaHCO3, KHCO3, Ca(HCO3)2, LiHCO3, NH4HCO3.

[0303] In some preferred embodiments, the lysosphingolipid of formula (5) and the blend comprising the fatty acid esters are reacted in the presence of an alkoxide, and wherein the alkoxide is an alkoxide of formula (12):

[0304] R9-O“X+

[0305] (12), wherein

[0306] R9is a Ci-C4alkyl, preferably selected from methyl, ethyl, propyl, isopropyl, butyl, or isobutyl, more preferably selected from methyl, or ethyl;

[0307] X+is a cation selected from Na+, K+, Li+or NH4+, preferably Na+.

[0308] In some more preferred embodiments, the lysosphingolipid of formula (5) and the blend comprising the fatty acid esters are reacted in the presence of sodium methoxide.

[0309] The base may be used in catalytic amounts, equimolar amounts or in excess.

[0310] In some embodiments, the lysosphingolipid of formula (5) is in the free-base form and the base is used in a catalytic amount from about 0.1 to about 0.5 molar equivalents based on the amount of the lysosphingolipid.

[0311] In some embodiments, the lysosphingolipid of formula (5) is in a salt form and the base is used in an amount from about 1.0 to about 1.7 molar equivalents based on the amount of the lysosphingolipid.

[0312] In some preferred embodiments, the lysosphingolipid of formula (5) is in a salt form and the base is used in an amount from about 1.2 to 1.3 molar equivalents based on the amount of the lysosphingolipid.

[0313] In some embodiments, the lysosphingolipid, the blend comprising the fatty acid esters, and the base are reacted in a polar solvent such as methanol, ethanol, propanol, isopropanol, butanol, or isobutanol.

[0314] In some preferred embodiments, the reaction is performed in methanol. In some embodiments the reaction is performed in a mixture of one or more polar solvents, such as a mixture of methanol and ethanol, methanol and propanol, methanol and isopropanol, methanol and butanol, methanol and isobutanol, or the mixture of water and an aliphatic alcohol. In some embodiments, the reaction is performed in acetonitrile.

[0315] In some embodiments, the lysosphingolipid, the blend comprising the fatty acid esters, and the base are reacted in C5-Cio hydrocarbon solvent, preferably wherein the hydrocarbon solvent is heptane.

[0316] In some embodiments, the reaction is performed solvent-free.

[0317] The lysosphingolipid, the blend comprising the fatty acid esters, and the base are typically reacted at a temperature from about 50° C to about 125° C. Preferably at a temperature from about 60° C to about 65° C. Accordingly, in some preferred embodiments, the reaction is performed at a temperature of about 50° C, 51° C, 62° C, 63° C, 64° C, or 65° C.

[0318] The components of the reaction may be combined in any order, and it will be appreciated that the order of combining the reactants may be adjusted as needed.

[0319] For example, the base may be added to a solution of the lysosphingolipid and the blend comprising the fatty acid esters. As another example, a solvent may be added to a flask containing the lysosphingolipid and the blend comprising the fatty acid esters, followed by the base.

[0320] The lysosphingolipid, the blend comprising the fatty acid esters, and the base, as well as any other reagent used during the reaction may be added to the reaction either as a solid or dissolved in a solvent, and in any quantities and manner effective for the intended result of the reaction.

[0321] In some embodiments, the method further comprising a step of reacting a lysosphingolipid of formula (5) with the blend comprising the triazine-based acylating agents in the presence of a base, thereby obtaining a blend comprising lipids of formula (5), and 0.5-30 wt% of fatty alcohols.

[0322] Accordingly, in some embodiments, the present invention describes a method for the production of a blend comprising lipids of formula (5), the method comprising the steps of:

[0323] - providing defatted and saponified rice bran wax (dsRBW), wherein said dsRBW comprising 50 wt% of fatty alcohols,

[0324] - reducing the fatty alcohol content of said dsRBW thereby obtaining a blend comprising about 70-99.5 wt% of C8-C34fatty acid salts, preferably C14-C30 fatty acid salts, and about 0.5-30 wt% of fatty alcohols,

[0325] - treating with an acid the blend comprising the fatty acid salts, thereby obtaining a blend comprising about 70-99.5 wt% of fatty acids of formula (3), and about 0.5-30 wt% of fatty alcohols, reacting the blend comprising the fatty acids of formula (3) with a triazine of formula (4) in the presence of an organic base thereby obtaining a blend comprising C8-C34 triazine-based acylating agents, preferably C14-C30 triazine-based acylating agents and about 0.5-30 wt% of fatty alcohols.

[0326] - reacting a lysosphingolipid of formula (5), or a salt thereof: wherein

[0327] W is hydrogen or a glycosyl moiety,

[0328] R4is hydrogen, aryl, or a substituted or unsubstituted Ci-C5oalkyl, preferably a substituted or unsubstituted C1-C17 alkyl, more preferably a substituted or unsubstituted C13-C17 alkyl,

[0329] R5is hydrogen or -OR7, wherein R7is selected from hydrogen, a substituted or unsubstituted Ci-

[0330] Cs alkyl, or a substituted or unsubstituted C2-C6 acyl, the bond ~ ~ ~ may be a double or a single bond when R5is hydrogen, or is a single bond when R5is -OR7,

[0331] R6is hydrogen, a substituted or unsubstituted Ci-C6alkyl, or a substituted or unsubstituted Ci-C3acyl, preferably hydrogen, with the blend comprising the triazine-based acylating agents, thereby obtaining a blend comprising lipids of formula (6): wherein

[0332] R1is a C7-C33 alkyl, preferably a C13-C29 alkyl,

[0333] W, R4, R5and Rsare as defined as for the lysosphingolipid of formula (5).

[0334] Typically, the lysosphingolipid and the blend comprising the triazine-based acylating agents are reacted in a polar solvent such as methanol, ethanol, propanol, isopropanol, butanol, or isobutanol. In some preferred embodiment, the reaction is performed in methanol. In some embodiment, the reaction is performed in a mixture of one or more polar solvents, such as a mixture of methanol and ethanol, methanol and propanol, methanol and isopropanol, methanol and butanol, methanol and isobutanol, or methanol and water.

[0335] In some embodiments, the lysosphingolipid and the blend comprising the triazine-based acylating agents are reacted in the presence of a base such as NaOH, KOH, LiOH, Ca(OH)2, triethylamine, N,N- diisopropylethylamine, and pyridine. In some preferred embodiments, the base is selected from NaOH, or KOH.

[0336] The reaction between the lysosphingolipid and the blend comprising the triazine-based acylating agents is typically performed at a temperature from about 25° C to about 65° C. Accordingly, in some embodiments, the reaction is performed at a temperature of about 25° C, 26° C , 27° C, 28° C, 29° C, 30° C, 31° C, 32° C, 33° C, 34° C, 35° C, 36° C, 37° C, 38° C, 39° C, 40° C, 41° C, 42° C, 43° C, 44° C, 45° C, 46° C, 47° C, 48° C, 49° C, 50° C, 51° C, 52° C, 53° C, 54° C, 55° C, 56° C, 57° C, 58° C, 59° C, 60° C, 61° C, 62° C, 63° C, 64° C, or 65° C. Preferably, the reaction is performed at temperature from about 45° C to about 55° C.

[0337] Accordingly, in some preferred embodiments the reaction is performed at a temperature of 45° C, 46° C, 47° C, 48° C, 49° C, 50° C, 51° C, 52° C, 53° C, 54° C, or 55° C.

[0338] The components of the reaction may be combined in any order, and it will be appreciated that the order of combining the reactants may be adjusted as needed.

[0339] For example, the lysosphingolipid may be added to a solution of the blend comprising the triazine-based acylating agents. As another example the blend comprising the triazine-based acylating agents may be added to a solution of the lysosphingolipid. As yet another example, a solvent may be added to a flask containing the lysosphingolipid and the blend comprising the triazine-based acylating agents.

[0340] The lysosphingolipid and the blend comprising the triazine-based acylating agents, as well as any other reagent used during the reaction may be added to the reaction either as a solid or dissolved in a solvent, and in any quantities and manner effective for the intended result of the reaction.

[0341] The reaction between the blend comprising the fatty acid esters and the lysosphingolipid of formula (5), or the reaction between the blend comprising the triazine-based acylating agents and the lysosphingolipid of formula (5) results in the selective / V-acylation of the amino group at the C-2 carbon atom of the lysosphingolipid of formula (5), thereby producing a blend comprising lipids of formula (6), and wherein said blend may further comprise between about 0.5-30 wt% of fatty alcohols.

[0342] In some embodiments, the fatty alcohol content of the blend comprising the lipids of formula (6) is about 0.5-5 wt% .

[0343] In some embodiments, the fatty alcohol content of the blend comprising the lipids of formula (6) is about 5-10 wt% . In some embodiments, the fatty alcohol content of the blend comprising the lipids of formula (6) is about 10-20 wt% .

[0344] In some embodiments, the fatty alcohol content of the blend comprising the lipids of formula (6) is about 20-30 wt% .

[0345] In some embodiments, the blend comprising the lipids of formula (6) has a content of at least about 50 wt% of a lipid of formula (6) wherein R1is a C21 alkyl and of a lipid of formula (6) wherein R1is a C23 alkyl, and wherein the ration between said lipids is from about 1:10 to about 10:1.

[0346] In some embodiments, the ratio between the lipid of formula (6) wherein R1is a C2ialkyl and of the lipid of formula (5) wherein R1is a C23 alkyl in about 1:2.5.

[0347] In some embodiments, the present invention relates to a blend of lipids comprising lipids of formula (1): wherein

[0348] R1is a C7-C33 alkyl, preferably a C13-C29 alkyl,

[0349] R2is selected from -OH, and / or -NHR3, wherein R3is a moiety of formula (2):

[0350] (2), wherein

[0351] W is hydrogen or a glycosyl moiety,

[0352] R4is hydrogen, aryl, or a substituted or unsubstituted C1-C50 alkyl, preferably a substituted or unsubstituted C1-C17 alkyl, more preferably a substituted or unsubstituted C13-C17 alkyl,

[0353] R5is hydrogen or -OR7, wherein R7is selected from hydrogen, a substituted or unsubstituted Ci-Cs alkyl, or a substituted or unsubstituted C2-C6acyl, the bond - may be a double or a single bond when R5is hydrogen, or is a single bond when R5is -OR7,

[0354] Rsis hydrogen, a substituted or unsubstituted Ci-C3alkyl, or a substituted or unsubstituted Ci-C6acyl, preferably hydrogen, wherein said blend may further comprise between about 0.5-30 wt% of fatty alcohols.

[0355] In some embodiments, R2of the lipid formula (1) is -OH. Accordingly, in some embodiments, the lipid of formula (1) is a fatty acid of formula (3):

[0356] (3), wherein

[0357] R1is a C7-C33 alkyl, preferably a C13-C29 alkyl.

[0358] In some embodiments, the present invention relates to a blend comprising a fatty acid of formula (3), wherein said blend comprising at least about 50 wt% of a fatty acid of formula (3) wherein R1is a C2ialkyl and of a fatty acid of formula (3) wherein R1is a C23alkyl, and wherein the ratio between the fatty acid of formula (3) wherein R1is a C21 alkyl and the fatty acid of formula (3) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5-30 wt% of fatty alcohols.

[0359] In some embodiments, the present invention relates to a blend comprising lipids of formula (3), wherein said blend comprising at least about 50 wt% of a fatty acid of formula (3) wherein R1is a C21 alkyl and of a fatty acid of formula (3) wherein R1is a C23alkyl, and wherein the ratio between the fatty acid of formula (3) wherein R1is a C2ialkyl and the fatty acid of formula (3) wherein R1is a C23alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5-5 wt% of fatty alcohols.

[0360] In some embodiments, the present invention relates to a blend comprising lipids of formula (3), wherein said blend comprising at least about 50 wt% of a fatty acid of formula (3) wherein R1is a C2ialkyl and of a fatty acid of formula (3) wherein R1is a C23alkyl, and wherein the ratio between the fatty acid of formula (3) wherein R1is a C2ialkyl and the fatty acid of formula (3) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 20-30 wt% of fatty alcohols.

[0361] In some embodiments, the ratio between the fatty acid of formula (3) wherein R1is a C2ialkyl and the fatty acid of formula (3) wherein R1is a C23 alkyl is about 1:2.5. In some embodiments, the lipids of formula (3) are C8-C34 non-hydroxy fatty acid. C8-C34 Non-hydroxy fatty acids may also be referred to as N(8-34:0).

[0362] In some embodiments, the fatty acids of formula (3) are C14-C30 non-hydroxy fatty acid. C14-C30 Non- hydroxy fatty acid may also be referred to as N(14-30:0).

[0363] In some embodiments, R2of the lipids of formula (1) is -NHR3, wherein R3is a moiety of formula (2). Accordingly in some embodiments, the lipids of formula (1) are lipids of formula (6):

[0364] (6), wherein

[0365] R1is a C7-C33 alkyl, preferably a C13-C29 alkyl,

[0366] W is hydrogen or a glycosyl moiety,

[0367] R4is hydrogen, aryl, or a substituted or unsubstituted C1-C50 alkyl, preferably a substituted or unsubstituted C1-C17 alkyl, more preferably a substituted or unsubstituted C13-C17 alkyl,

[0368] R5is hydrogen or -OR7, wherein R7is selected from hydrogen, a substituted or unsubstituted Ci-C6alkyl, or a substituted or unsubstituted C2-C6 acyl, the bond - may be a double or a single bond when R5is hydrogen, or is a single bond when R5is -OR7,

[0369] Rsis hydrogen, a substituted or unsubstituted Ci-C3alkyl, or a substituted or unsubstituted Ci-C6acyl, preferably hydrogen.

[0370] In some embodiments, for the moiety of formula (2), and for the lipids of formula (6) R4is a saturated unsubstituted C13-C17 alkyl, R5is -OH, R6is hydrogen, and the bond - is a single bond.

[0371] In some embodiments, for the moiety of formula (2) and for the lipids of formula (6) R4is a saturated unsubstituted C13-C17 alkyl, R5and R6are hydrogen, and the bond - is a single bond.

[0372] In some embodiments, for the moiety of formula (2) and for the lipids of formula (6) R4is a saturated unsubstituted C13-C17 alkyl, R5and Rsare hydrogen, and the bond - is a double bond.

[0373] In some embodiments, the present invention relates to a blend comprising lipids of formula (6), wherein said blend comprising at least about 50 wt% of a lipid of formula (6) wherein R1is a C21 alkyl and of a lipid of formula (6) wherein R1is a C23 alkyl, and wherein the ratio between the lipid of formula (6) wherein R1is a C2ialkyl and the lipid of formula (6) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5-30 wt% of fatty alcohols.

[0374] In some embodiments, the present invention relates to a blend comprising lipids of formula (6), wherein said blend comprising at least about 50 wt% of a lipid of formula (6) wherein R1is a C21 alkyl and of a lipid of formula (6) wherein R1is a C23 alkyl, and wherein the ratio between the lipid of formula (6) wherein R1is a C21 alkyl and the lipid of formula (6) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5-5 wt% of fatty alcohols.

[0375] In some embodiments, the present invention relates to a blend comprising lipids of formula (6), wherein said blend comprising at least about 50 wt% of a lipid of formula (6) wherein R1is a C21 alkyl and of a lipid of formula (6) wherein R1is a C23 alkyl, and wherein the ratio between the lipid of formula (6) wherein R1is a C21 alkyl and the lipid of formula (6) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 20-30 wt% of fatty alcohols.

[0376] In some embodiments, the ratio between the lipid of formula (6) wherein R1is a C21 alkyl and the lipid of formula (6) wherein R1is a C23 alkyl is from about 1:2.5.

[0377] In some embodiments, for the lipids of formula (6), W is H. Accordingly, in some embodiments, the lipids of formula (6) are ceramides.

[0378] In some embodiments, the lipid of formula (6) is a ceramide selected from the group consisting of ceramides of formula (13), (14), (15), or (16), or a mixture thereof: wherein

[0379] R1is a C7-C33 alkyl, preferably a C13-C30 alkyl. In some embodiments, the present invention relates to a blend comprising ceramides of formula (13), wherein said blend comprising at least about 50 wt% of a ceramide of formula (13) wherein R1is a C2ialkyl and of a ceramide of formula (13) wherein R1is a C23alkyl, and wherein the ratio between the ceramide of formula (13) wherein R1is a C2ialkyl and the ceramide of formula (13) wherein R1is a C23alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5- 30 wt% of a C22-C38fatty alcohol.

[0380] In some embodiments, the present invention relates to a blend comprising ceramides of formula (13), wherein said blend comprising at least about 50 wt% of a ceramide of formula (13) wherein R1is a C2ialkyl and of a ceramide of formula (13) wherein R1is a C23alkyl, and wherein the ratio between the ceramide of formula (13) wherein R1is a C2ialkyl and the ceramide of formula (13) wherein R1is a C23alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5- 5 wt% of a C22-C3s fatty alcohol.

[0381] In some embodiments, the present invention relates to a blend comprising ceramides of formula (13), wherein said blend comprising at least about 50 wt% of a ceramide of formula (13) wherein R1is a C2ialkyl and of a ceramide of formula (13) wherein R1is a C23alkyl, and wherein the ratio between the ceramide of formula (13) wherein R1is a C2ialkyl and the ceramide of formula (13) wherein R1is a C23alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 20- 30 wt% of a C22-C38fatty alcohol.

[0382] In some preferred embodiments, the ratio between the ceramide of formula (13) wherein R1is a C2ialkyl and the ceramide of formula (13) wherein R1is a C23alkyl is about 1:2.5.

[0383] In some embodiments, the present invention relates to a blend comprising ceramides of formula (14), wherein said blend comprising at least about 50 wt% of a ceramide of formula (14) wherein R1is a C23alkyl and of a ceramide of formula (14) wherein R1is a C23alkyl, and wherein the ratio between the ceramide of formula (14) wherein R1is a C2ialkyl and the ceramide of formula (14) wherein R1is a C23alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5- 30 wt% of a C22-C38 fatty alcohol.

[0384] In some embodiments, the present invention relates to a blend comprising ceramides of formula (14), wherein said blend comprising at least about 50 wt% of a ceramide of formula (14) wherein R1is a C2ialkyl and of a ceramide of formula (14) wherein R1is a C23alkyl, and wherein the ratio between the ceramide of formula (14) wherein R1is a C2ialkyl and the ceramide of formula (14) wherein R1is a C23alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5- 5 wt% of a C22-C3s fatty alcohol.

[0385] In some embodiments, the present invention relates to a blend comprising ceramides of formula (14), wherein said blend comprising at least about 50 wt% of a ceramide of formula (14) wherein R1is a C23 alkyl and of a ceramide of formula (14) wherein R1is a C23 alkyl, and wherein the ratio between the ceramide of formula (14) wherein R1is a C2ialkyl and the ceramide of formula (14) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 20- 30 wt% of a C22-C38 fatty alcohol.

[0386] In some embodiments, the ratio between the ceramide of formula (14) wherein R1is a C21 alkyl and the ceramide of formula (14) wherein R1is a C23 alkyl is about 1:2.5.

[0387] In some embodiments, the present invention relates to a blend comprising ceramides of formula (15), wherein said blend comprising at least about 50 wt% of a ceramide of formula (15) wherein R1is a C21 alkyl and of a ceramide of formula (15) wherein R1is a C23 alkyl, and wherein the ratio between the ceramide of formula (15) wherein R1is a C2ialkyl and the ceramide of formula (15) wherein R1is a C23alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5- 30 wt% of a C22-C38 fatty alcohol.

[0388] In some embodiments, the present invention relates to a blend comprising ceramides of formula (16), wherein said blend comprising at least about 50 wt% of a ceramide of formula (16) wherein R1is a C2ialkyl and of a ceramide of formula (16) wherein R1is a C23 alkyl, and wherein the ratio between the ceramide of formula (16) wherein R1is a C21 alkyl and the ceramide of formula (16) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5- 30 wt% of a C22-C38 fatty alcohol.

[0389] In some embodiments, R1of the ceramides of formula (13)-(16) is a C7-C33 non-hydroxy alkyl. Accordingly, in some embodiments, ceramides of formula (13), (14), (15), and (16) can be referred to as CER[N (8-34:0) P(18)], CER[N (8-34:0) DS(18)], CER[N (8-34:0) 5(18)], and CER[N (8-34:0) H(18)], respectively.

[0390] In some embodiments, R1of the ceramides of formula (13)-(16) is a C13-C29 non-hydroxy alkyl. Accordingly, in some embodiments, ceramides of formula (13), (14), (15), and (16) can be referred to as CER[N (14-30:0) P(18)], CER[N (14-30:0) DS(18)], CER[N (14-30:0) 5(18)], and CER[N (14-30:0) H(18)], respectively.

[0391] In some preferred embodiments, for the lipids of formula (6), W is a glycosyl moiety. Accordingly, in some preferred embodiments, the lipids of formula (6) are glycosphingolipids.

[0392] In some embodiments, the lipid of formula (6) is a glycosphingolipid selected from the group consisting of glycosphingolipids of formula (17), (18), (19), or (20), or a mixture thereof: wherein

[0393] W is a glycosyl moiety selected from the group consisting of Glcl-, Gall-, Gaipi-4Glcl-,

[0394] R1is a C7-C33 alkyl, preferably a C13-C29 alkyl.

[0395] In some embodiments, the present invention relates to a blend comprising glycosphingolipids of formula (17), wherein said blend comprising at least about 50 wt% of a glycosphingolipid of formula (17) wherein R1is a C21 alkyl and of a glycosphingolipid of formula (17) wherein R1is a C23 alkyl, and wherein the ratio between the glycosphingolipid of formula (17) wherein R1is a C21 alkyl and the glycosphingolipid of formula (17) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5-30 wt% of a C22-C38 fatty alcohol.

[0396] In some embodiments, the present invention relates to a blend comprising glycosphingolipids of formula (17), wherein said blend comprising at least about 50 wt% of a glycosphingolipid of formula (17) wherein R1is a C21 alkyl and of a glycosphingolipid of formula (17) wherein R1is a C23 alkyl, and wherein the ratio between the glycosphingolipid of formula (17) wherein R1is a C21 alkyl and the glycosphingolipid of formula (17) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5-5 wt% of a C22-C38 fatty alcohol.

[0397] In some embodiments, the present invention relates to a blend comprising glycosphingolipids of formula (17), wherein said blend comprising at least about 50 wt% of a glycosphingolipid of formula (17) wherein R1is a C21 alkyl and of a glycosphingolipid of formula (17) wherein R1is a C23 alkyl, and wherein the ratio between the glycosphingolipid of formula (17) wherein R1is a C21 alkyl and the glycosphingolipid of formula (17) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 20-30 wt% of a C22-C38 fatty alcohol. In some embodiments, the ratio between the glycosphingolipid of formula (17) wherein R1is a C2ialkyl and the glycosphingolipid of formula (17) wherein R1is a C23 alkyl is about 1:2.5.

[0398] In some embodiments, the present invention relates to a blend comprising glycosphingolipids of formula (18), wherein said blend comprising at least about 50 wt% of a glycosphingolipid of formula

[0399] (18) wherein R1is a C21 alkyl and of a glycosphingolipid of formula (18) wherein R1is a C23 alkyl, and wherein the ratio between the glycosphingolipid of formula (18) wherein R1is a C2ialkyl and the glycosphingolipid of formula (18) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5-30 wt% of a C22-C38 fatty alcohol.

[0400] In some embodiments, the present invention relates to a blend comprising glycosphingolipids of formula (18), wherein said blend comprising at least about 50 wt% of a glycosphingolipid of formula (18) wherein R1is a C21 alkyl and of a glycosphingolipid of formula (18) wherein R1is a C23 alkyl, and wherein the ratio between the glycosphingolipid of formula (18) wherein R1is a C21 alkyl and the glycosphingolipid of formula (18) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5-5 wt% of a C22-C38 fatty alcohol.

[0401] In some embodiments, the present invention relates to a blend comprising glycosphingolipids of formula (18), wherein said blend comprising at least about 50 wt% of a glycosphingolipid of formula

[0402] (18) wherein R1is a C21 alkyl and of a glycosphingolipid of formula (18) wherein R1is a C23 alkyl, and wherein the ratio between the glycosphingolipid of formula (18) wherein R1is a C2ialkyl and the glycosphingolipid of formula (18) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 20-30 wt% of a C22-C38 fatty alcohol.

[0403] In some embodiments, the ratio between the glycosphingolipid of formula (18) wherein R1is a C21 alkyl and the glycosphingolipid of formula (18) wherein R1is a C23 alkyl is about 1:2.5.

[0404] In some embodiments, the present invention relates to a blend comprising glycosphingolipids of formula (19), wherein said blend comprising at least about 50 wt% of a glycosphingolipid of formula

[0405] (19) wherein R1is a C21 alkyl and of a glycosphingolipid of formula (19) wherein R1is a C23 alkyl, and wherein the ratio between the glycosphingolipid of formula (19) wherein R1is a C21 alkyl and the glycosphingolipid of formula (19) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5-30 wt% of a C22-C38 fatty alcohol.

[0406] In some embodiments, the present invention relates to a blend comprising glycosphingolipids of formula (20), wherein said blend comprising at least about 50 wt% of a glycosphingolipid of formula

[0407] (20) wherein R1is a C2ialkyl and of a glycosphingolipid of formula (20) wherein R1is a C23 alkyl, and wherein the ratio between the glycosphingolipid of formula (20) wherein R1is a C21 alkyl and the glycosphingolipid of formula (20) wherein R1is a C23 alkyl is from about 1:10 to about 10:1, and wherein said blend may further comprise between about 0.5-30 wt% of a C22-C38 fatty alcohol. According to the invention, all the above embodiments of blends comprising least about 50 wt% , preferably comprise more than 50 wt% of the compound(s) specified in each embodiment, such as about 60 wt% or more, e.g. around 65-75 wt%, around 70-80 wt%, around 75-85 wt%, around 80-90 wt% , around 95-100 wt%.

[0408] In some embodiments, for the glycosphingolipids of formula (17)-(20) R1is a C7-C33 non-hydroxy alkyl, and W is Glcl-. Accordingly, in some preferred embodiments, the glycosphingolipids of formula (17), (18), (19), and (20) can be referred to as GlcCER[N (8-34:0) P(18)], GlcCER[N (8-34:0) DS(18)], GlcCER[N (8-34:0) 5(18)], and GlcCER[N (8-34:0) H(18)], respectively.

[0409] In some embodiments, for the glycosphingolipids of formula (17)-(20) R1is a C13-C29 non-hydroxy alkyl, and W is Glcl-. Accordingly, in some preferred embodiments, the glycosphingolipids of formula (17), (18), (19), and (20) can be referred to as GlcCER[N (14-30:0) P(18)], GlcCER[N (14-30:0) DS(18)], GlcCER[N (14-30:0) S(18)], and GlcCER[N (14-30:0) H(18)], respectively.

[0410] In some embodiments, for the glycosphingolipids of formula (17)-(20) R1is a C7-C33 non-hydroxy alkyl, and W is Gall-. Accordingly, in some preferred embodiments, the glycosphingolipids of formula (17), (18), (19), and (20) can be referred to as GalCER[N (8-34:0) P(18)], GalCER[N (8-34:0) DS(18)], GalCER[N (8-34:0) S(18)], and GalCER[N (8-34:0) H(18)], respectively.

[0411] In some embodiments, for the glycosphingolipids of formula (17)-(20) R1is a C13-C29 non-hydroxy alkyl, and W is Gall-. Accordingly, in some preferred embodiments, the glycosphingolipids of formula (17), (18), (19), and (20) can be referred to as GalCER[N (14-30:0) P(18)], GalCER[N (14-30:0) DS(18)], GalCER[N (14-30:0) 5(18)], and GalCER[N (14-30:0) H(18)], respectively.

[0412] In some embodiments, for the glycosphingolipids of formula (17)-(20) R1is a C7-C33 non-hydroxy alkyl, and W is Gaipi-4Glcl-. Accordingly, in some preferred embodiments, the glycosphingolipids of formula (17), (18), (19), and (20) can be referred to as LacCER[N (8-34:0) P(18)], LacCER[N (8-34:0) DS(18)], LacCER[N (8-34:0) 8(18)], and LacCER[N (8-34:0) H(18)], respectively.

[0413] In some embodiments, for the glycosphingolipids of formula (17)-(20) R1is a C13-C29 non-hydroxy alkyl, and W is Gaipi-4Glcl-. Accordingly, in some preferred embodiments, the glycosphingolipids of formula (17), (18), (19), and (20) can be referred to as LacCER[N (14-30:0) P(18)], LacCER[N (14-30:0) DS(18)], LacCER[N (14-30:0) S(18)], and LacCER[N (14-30:0) H(18)], respectively.

[0414] The of fatty alcohol content of the blends of the present invention may be quantified via techniques known to the skilled person such as HPLC analysis or1H NMR spectroscopy.

[0415] In some embodiments, the present invention relates to a cosmetic composition comprising the blend according to the present invention. In particular, a cosmetic composition comprising the blend of lipids any one defined in the above embodiments and in the claims, or a combination thereof. According to the invention, blend of lipids provided by the invention, are biologically active compounds and may be beneficial for use in cosmetic compositions aimed for

[0416] Improving skin complexion

[0417] Improving skin radiance

[0418] Improving skin tone

[0419] Reducing premature skin ageing

[0420] Wrinkle smoothing

[0421] Improving skin firmness,

[0422] Increasing volume of the skin,

[0423] Improving skin homogeneity, Increasing skin hydration.

[0424] Cosmetic use according to the invention includes reducing and / or ameliorating signs of skin aging such as reducing or ameliorating one or more of the following signs on the skin: wrinkles, thinning, decrease of elasticity,

[0425] - sagging, fine lines, hyperkeratosis, dry skin, loss of skin volume, stretch marks, sensitivity.

[0426] Without being bound to a theory, treatment of skin of mammals, especially, human skin, with the lipids comprised in blends of the invention has been associated with activation of biological pathways in skin cells relating to

[0427] Maintaining and / or improving skin barrier function;

[0428] Diminishing effects of the exposure of skin to environmental pollutants and unhealthy lifestyle;

[0429] Stimulating of skin cell regeneration ;

[0430] Boosting the production of energy in mitochondria of skin cells;

[0431] Maintaining and improving skin cell adhesion and communication;

[0432] Maintaining heathy skin microbiome and preventing or recovering thereof from dysbiosis. In one embodiment, the lipid blends described herein could also be beneficially used for cosmetic improvement of hair.

[0433] Cosmetic compositions comprising a lipid blend of the invention may comprise any further active compounds that could enhance or support any of the above mentioned beneficial effects, e.g. such compounds as vitamins, anti-oxidants, oils, etc.

[0434] The cosmetic compositions can be formulated as liquid, semi-liquid (e.g. emulsions) or solid compositions following guidelines of the art.

[0435] Preferably, the cosmetic compositions are topical compositions.

[0436] In one embodiment, the invention relates to therapeutic composition comprising a lipid blend of the invention. Preferably, the blend of lipids comprised in a therapeutic composition according to the invention is not a therapeutically active compound, but a compound used for a cosmetic support of therapeutic treatment, e.g. for a cosmetic recovery of skin following the therapeutic treatment.

[0437] Examples

[0438] Working examples below describe non-limiting embodiments of the invention and are given only to illustrate the invention.

[0439] General methods and material:

[0440] 1H NMR and13C NMR was recorded with a Jeol ECZ 500R (500 MHz) spectrometer.TH and13C chemical shifts are given in ppm (6) relative to tetramethylsilane (6 = 0.00), CDCL (5 = 7.26), CD3OD (6 = 3.31), DMSO-ds (6 = 2.50) as internal standard. Thin layer chromatography (TLC) was performed with silica gel TLC-plates (Merck, Silica gel, F254) with detection by UV-absorption (254 nm) where applicable and carrying (140 °C) with ammonium molybdate (25 g / L) and cerium ammonium sulfate (10 g / L) in 10% H2SO4.

[0441] LCMS analyses was performed on a SCIEX Triple Quad™ 4500 LC-MS / MS system equipped with an Accucore aQ (150 mm x 4.6 mm, 2.6 pm) column.

[0442] HPLC analyses was performed on a Dionex Ultimate 3000 HPLC system coupled with a Corona Veo Charged Aerosol Detector using an Accucore aQ (150 mm x 4.6 mm, 2.6 pm) column.

[0443] GC analysis was performed with Agilent 8890 Gas Chromatograph coupled with a flame ionization detector (FID) and equipped with a DB-5MS Ul (30 m x 0.25 mm, 0.25 pm) column.

[0444] Example 1. Production of defatted Rice Bran Wax (RBW)

[0445] RBW was suspended in isopropanol (7 vol.) and stirred under reflux until a solution was obtained. The solution was then cooled to room temperature to form a suspension. The suspension was filtered, and the collected solid was washed with isopropanol and dried under vacuum. Example 2. Saponification of defatted RBW

[0446] Defatted RBW was suspended in isopropanol (6 vol.) and potassium hydroxide was added. The mixture was refluxed until the full saponification of RBW was obtained (reaction monitored by TLC or ^-NMR). The mixture was cooled down to a temperature of about 60° C, and acetonitrile (5-6 vol.) was added. The mixture was cooled down to room temperature to form a suspension. The suspension was filtered and the collected solid washed with acetonitrile and dried under vacuum. Following the procedure described above a blend comprising a 1:1 mixture of potassium salts of fatty acids and of fatty alcohols was obtained.

[0447] Example 3. Reduction of fatty alcohols from defatted and saponified RBW

[0448] Defatted and saponified RBW (dsRBW) was subjected to Soxhlet extraction using ethyl acetate (3-5 vol.) as the extraction solvent. The extraction was continued until most of the fatty alcohols were removed (about 4-6 hours, monitored by TLC). The remaining solid residue, enriched with fatty acid salts, was suspended in ethyl acetate or acetonitrile (3-4 vol) and stirred at room temperature for about 15-30 minutes. The suspension was filtered and, the collected solid was washed with ethyl acetate or acetonitrile and dried in vacuum.

[0449] Following the procedure described above a blend comprising 75-95 wt% of potassium salts of fatty acids was obtained.

[0450] Example 4. Production of a blend of fatty acids

[0451] The blend obtained in example 3 was added to a 1:10 mixture of HCI (37% aq.) and water. The resulting suspension was stirred at a temperature of about 50-60° C for 1-2 hours, then cooled down to room temperature and stirred for additional 1-2 hours. The suspension was filtered, the solid collected, washed with water (3-4 vol.) and dried.

[0452] Following the procedure described above a blend comprising between about 75-80 wt% of Ci6-C3o fatty acids, was obtained.

[0453] Example 5. LCMS and HPLC analysis of the blend of fatty acids

[0454] The identity and content of the fatty acids comprised in the blend obtained in example 4 were determined via LCMS and HPLC analysis.

[0455] The LCMS eluent profile consisted of solvent A: 1 L water + 2 mL formic acid + 2 mmol ammonium formate, and solvent B: 1 L MeOH + 1 L acetonitrile + 4 mL formic acid + 4 mmol ammonium formate. A gradient of 80-100% B in A was applied over 50 min, followed by an isocratic of 80% B in A for 55 min. The HPLC eluent profile consisted of solvent A: 1 L water + 2 mL formic acid + 2 mmol ammonium formate, and solvent B: 1 L MeOH + 1 L acetonitrile + 4 mL formic acid + 4 mmol ammonium formate. A gradient of 80-100% B in A was applied over 50 min, followed by an isocratic of 80% B in A for 55 min. The fatty acid content of the blend was quantified via peak area analysis using external standards. The results of the LCMS and HPLC of analysis are summarized in Table 1.

[0456] Table 1. LCMS and HPLC analysis of the blend of fatty acids

[0457] Example 6. Production of a fatty acid esters blend from the blend of fatty acids The fatty acid blend obtained in Example 4 was suspended in methanol (5 vol.), and HCI (37% aq. 0.3-0.6 vol.) was added. The reaction mixture was refluxed until the fatty acids were fully converted into their corresponding methyl esters (monitored by TLC). The reaction mixture was cooled down to room temperature and water (5 mL) was added. The obtained suspension was filtered, the solid collected, washed with a 1:1 mixture of Methanol and water (3-4 vol.) and dried in vacuum. Following the procedure described above, a blend comprising between about 75-85 wt% of C14-C30 fatty acid methyl esters was obtained.

[0458] Example 7. GC analysis of the blend of fatty acid methyl esters

[0459] The fatty acid methyl ester blend obtained in Example 6 was characterized via GC analysis.

[0460] The GC conditions were as follow: The initial oven temperature was 60° C for 1 min, then programmed to 180° C at a rate of 40° C / min. and a hold time of 5 minutes, then programmed to 320° C at a rate of 10°C / min. and a hold time of 17 min. The FID detector temperature was 280° C. A constant flow of Helium was applied with a rate of 2 mL / min. The fatty acid methyl ester content of the blend was quantified via peak area analysis using external standards. The results of the GC analysis are summarized in Table 2.

[0461] Table 2. GC analysis of the blend of fatty acid methyl esters Example 8. Production of a triazine-based acylating agent blend

[0462] The fatty acid blend obtained in Example 4 was suspended in acetone (8 vol.). 4-Methylmorpholine (1.1 eq.) was added and obtained mixture was stirred at 35-40 °C for 15 min. Then, 2-chloro-4,6-dimethoxy- l,3,5-triazine(1.05 eq.) was added and the mixture was stirred at 35-40° C. After 2 h. The reaction mixture was cooled down to room temperature, water (12 vol.) was added, and the obtained suspension was filtered. The collected solid was washed with a 1:1 mixture of acetone and water (5-6 vol.) and dried in vacuum.

[0463] ‘H-NMR (500 MHz, CDCI3) 64.09 - 3.93 (m, 11H), 2.88 - 2.80 (m, 1H), 2.56 (t, J = 7.5 Hz, 2H), 2.40 (t, J = 7.5 Hz, 1H), 1.75 - 1.47 (m, 6H), 1.41 - 1.02 (m, 92H), 0.84 (t, J = 6.9 Hz, 7H).

[0464] Example 9. General procedure for the production of ceramides blends from the blend of fatty acid methyl esters

[0465] A hydrochloride salt of a sphingoid base (1 eq.) was dissolved in MeOH (10 vol.), sodium methoxide (25% in MeOH, 1.4 eq.) was added followed by the fatty acid methyl ester blend obtained in Example 6 (1.1 eq.). The reaction mixture was stirred at 60 °C until full conversion of the starting materials (monitored by TLC). Subsequently, the reaction mixture was cool down to room temperature to form a suspension. The suspension was filtered, and the obtained solid was first washed with methanol (10 vol.), then resuspended in methanol and refluxed for about 0.5-1 h. Subsequently, water was added to the suspension, and the mixture was first cooled down to room temperature and then filtered. The collected solid was washed with a 10:1 mixture of methanol and water (2 times, 2-3 vol.) and dried in vacuum.

[0466] Example 10. Production of CER[N (14-30:0) DS(18)] blend

[0467] D-erythro-Dihydrosphingosine hydrochloride (1 eq.) and the blend of fatty acid methyl esters were reacted under the conditions of Example 9 to produce a blend comprising ceramides DS(18) carrying non-hydroxy C16-C30 acyl groups.

[0468] The CER[N (14-30:0) DS( 18)] blend was characterized by LCMS analysis.

[0469] The LCMS eluent profile consisted of solvent A: 2 mL formic acid, 2mM ammonium formate in IL of water, and solvent B: 2ml formic acid, 2mM ammonium formate in 500ml Acetonitrile+500ml Methanol. A gradient of 80-100% B in A was applied over 10 min., followed by an isocratic of 100% B in A for 50 min., followed by an isocratic of 80% B in A for 40 min.

[0470] The results of the LCMS of analysis are summarized in Table 3.

[0471] Table 3. LCMS analysis of the ceramide blend

[0472] Example 11. Production of a fatty alcohol blend

[0473] The solvent extract obtained in Example 3, comprising fatty alcohols and O-acetylated fatty alcohols, was concentrated, MeOH (10 vol.) was added, followed by HCI (37% aq., 0.5-1.2 vol.). The resulting mixture was stirred at reflux until all the O-acetylated fatty alcohol derivatives were converted into the corresponding fatty alcohols (reaction monitored by TLC). Then, the reaction mixture was cooled down to RT and stirred for about 0.5-1 hour. The obtained suspension was filtered, the solid collected, washed with MeOH (3-4 vol.) and dried in vacuum.

[0474] Following the procedure described above a thirteenth blend, comprising between about 55-65 wt% of C22-C36 fatty alcohols, was obtained.

[0475] Example 12. HPLC Analysis of the fatty alcohol blend

[0476] The fatty alcohol blend obtained in Example 11 was characterized via HPLC analysis.

[0477] The HPLC eluent profile consisted of solvent A: methanol, and solvent B: acetonitrile. A gradient of 50- 93% A in B was applied over 20 min, followed by an isocratic of 50% A in B for 45 min. The fatty alcohol content of the blend was quantified via peak area analysis using external standards. The results of the HPLC analysis are summarized in Table 2.

[0478] Table 2. HPLC analysis of the alcohol blend

[0479] Example 13. Production of a fatty acid esters blend from dsRBW

[0480] Defatted and saponified RBW (dsRBW) comprising a 1:1 mixture of potassium salts of fatty acids and fatty alcohols, was suspended in methanol (10 vol.). Hydrochloric acid was added to the suspension (37% aq.), and the suspension was stirred under reflux until full conversion (monitored by TLC). The reaction mixture was cooled down to room temperature and the resulting suspension was filtered. The obtained solid was washed with methanol (3-4 vol.), dried in vacuum, and then suspended in ethyl acetate (10 vol.). The suspension was refluxed for 1-2 hours, then cooled down to room temperature and filtered. The obtained solid was washed with ethyl acetate (3-4 vl.) and dried in vacuum.

[0481] Following the procedure described above, a blend comprising between about 60-70 wt% of C14-C30 fatty acid methyl esters was obtained.

[0482] Example 14. scCCh extraction of defatted and saponified RBW

[0483] Defatted and saponified RBW (dsRBW) was extracted with a Separex@GSP Lab using carbonic anhydride and the following processing conditions: static period 15 min. at 60 °C and 350 bar, dynamic period from 60 °C and 350 bar to 80 °C and 450 bar over 150 min.

[0484] Following the procedure described above a blend comprising 70-80 wt% of potassium salts of fatty acids was obtained.

[0485] Where ranges are given, endpoints are included. Furthermore, it is to be understood that unless otherwise indicated or otherwise evident from the context and / or the understanding of one of ordinary skill in the art, values that are expressed as ranges can assume any specific value within the stated ranges in different embodiments of the invention, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise. It is also to be understood that unless otherwise indicated or otherwise evident from the context and / or the understanding of one of ordinary skill in the art, values expressed as ranges can assume any subrange within the given range, wherein the endpoints of the subrange are expressed to the same degree of accuracy as the tenth of the unit of the lower limit of the range.

[0486] The disclosure should not be seen in any way restricted to the embodiments described and a person with ordinary skill in the art will foresee many possibilities to modifications thereof.

[0487] The above-described embodiments are combinable. The following claims further set out particular embodiments of the disclosure.

Claims

CLAIMS1. A method for the production of a blend of lipids from defatted and saponified rice bran wax (dsRBW), wherein said dsRBW comprising about 50 wt% of fatty alcohols, said blend comprising:- about 70-99.5 wt% of lipids of formula (1):O R’(1) whereinR1is a C7-C33 alkyl, preferably a C13-C29 alkyl,R2is selected from -OH, and / or -NHR3, wherein R3is a moiety of formula (2):whereinW is hydrogen or a glycosyl moiety,R4is hydrogen, aryl, or a substitute or unsubstituted C1-C50 alkyl, preferably a substituted or unsubstituted C1-C17 alkyl, more preferably a substituted or unsubstituted C13-C17 alkyl,R5is hydrogen or -OR7, wherein R7is selected from hydrogen, a substituted or unsubstituted Ci-C6alkyl, or a substituted or unsubstituted C2-C6acyl, the bond - may be a double or a single bond when R5is hydrogen, or is a single bond when R5is -OR7,Rsis hydrogen, a substituted or unsubstituted Ci-Cs alkyl, or a substituted or unsubstituted Ci-Cg acyl, preferably hydrogen, and- about 0.5-30 wt% of fatty alcohols; and whereinsaid method comprising a step of reducing the fatty alcohol content of the dsRBW or a derivative thereof, to a content of around 0.5-30 wt%.

2. The method according to claim 1, wherein the step of reducing the fatty alcohol content is performed via solvent extraction.

3. The method according to claim 2, wherein the solvent is selected from the group consisting of ethyl acetate, cyclohexane, n-heptane, acetonitrile, or supercritical CO2, preferably ethyl acetate.

4. The method according to any one of claims 1 to 3, wherein the blend of lipids comprising at least 50 wt% of lipids of formula (1) wherein R1is a C2ialkyl and lipids of formula (1) wherein R1is a C23 alkyl, and wherein the ratio between said lipids is from about 1:10 to about 10:1.

5. The method according to any one of claims 1 to 4, wherein the step of reducing the fatty alcohol content is performed on dsRBW, thereby obtaining a blend comprising about 70-99.5 wt% of C8-C34 fatty acid salts, preferably C14-C30 fatty acid salts and between about 0.5-30 wt% of fatty alcohols; or- a dsRBW derivative, wherein said derivative is obtained via reacting the dsRBW with a Ci- C6alcohol in the presence of an acid, and said derivative comprises C8-C34 fatty acid esters, preferably C14-C30 fatty acid esters, and about 50 wt% of fatty alcohols, thereby obtaining a blend of fatty acid esters comprising about 70-99.5 wt% of C8-C34 fatty acid esters, preferably Ci4-C30fatty acid esters, and about 0.5-30 wt% of fatty alcohols.

6. The method according to claim 5, further comprising a step of treating the blend comprising the fatty acid salts with an acid, thereby obtaining a blend comprising about 70-99.5 wt% of fatty acids of formula (3):wherein R1is as defined for the lipids of formula (1), and about 0.5-30 wt% of fatty alcohols.

7. The method according to claim 6, further comprising a step of reacting the blend comprising the fatty acids of formula (3) with:-a Ci-Cs alcohol in the presence of an acid, thereby obtaining a blend of fatty acid esters comprising about 70-99.5 wt% of C8-C34 fatty acid esters, preferably C14-C30 fatty acid esters, and about 0.5-30 wt% of fatty alcohols; or- a triazine of formula (4):whereinR8is selected from methyl, ethyl, 2,2,2-trifluoroethyl, and substituted or unsubstituted benzyl, preferably methyl, in the presence of an organic base, thereby obtaining a blend comprising Cs-C34 triazine-based acylating agents, preferably C14- C30 triazine-based acylating agents, and about 0.5-30 wt% of fatty alcohols.

8. The method according to any one of claims 5 to 7, further comprising a step of reacting a lysosphingolipid of formula (5)or a salt thereof, wherein W, R4, R5, R6and the bond - are as defined as for the moiety of formula (2) with the blend comprising the fatty acid esters in the presence of a base, or the blend comprising the triazine-based acylating agents,thereby obtaining a blend comprising lipids of formula (6):whereinR1is as defined as for the lipids of formula (1), and W, R4, R5, R6and the bond - are as defined as for the moiety of formula (2), and about 5-30 wt% of fatty alcohols.

9. The method according to any one of claims 1 to 8, wherein the for the moiety of formula (2), the lysosphingolipid of formula (5) and the lipids of formula(6) R4is a saturated unsubstituted C13-C17 alkyl, R5is -OH, R6is hydrogen, and the bond - is a single bond.

10. The method according to any one of claims 1 to 8, wherein for the moiety of formula (2), foe the lysosphingolipid of formula (5) and for the lipids of formula (6) R4is a saturated unsubstituted C13-C17 alkyl, R5and R6are hydrogen, and the bond - is a single bond.

11. The method according to any one of claims 1 to 8, wherein the for the moiety of formula (2), for the lysosphingolipid of formula (5) and for the lipids of formula (6) R4is a saturated unsubstituted C13-C17 alkyl in the position, R5and R6are hydrogen, and the bond - is a double.

12. The method according to any one of claims 1 to 11, wherein the moiety of formula (2), for the lysosphingolipid of formula (5) and for the lipids of formula (6) W is hydrogen.

13. The method according to any one of claims 1 to 11, wherein for the moiety of formula (2), for the lysosphingolipid of formula (5) and for the lipids of formula (6) W is a glycosyl moiety selected from the group consisting of Glcl-, Gall-, Gaipi-4Glcl-.

14. A blend of lipids comprising lipids of formula (1):whereinR1is a C7-C33 alkyl, preferably a C13-C29 alkyl,R2is selected from -OH, and / or -NHR3, wherein R3is a moiety of formula (2):whereinW is hydrogen or a glycosyl moiety,R4is hydrogen, aryl, or a substituted or unsubstituted C1-C50 alkyl, preferably a substituted or unsubstituted C1-C17 alkyl, more preferably a substituted or unsubstituted C13-C17 alkyl,R5is hydrogen or -OR7, wherein R7is selected from hydrogen, a substituted or unsubstituted Ci-C6alkyl, or a substituted or unsubstituted C2-C6acyl, the bond - may be a double or a single bond when R5is hydrogen, or is a single bond when R5is -OR7,Rsis hydrogen, a substituted or unsubstituted Ci-Cg alkyl, or a substituted or unsubstituted Ci-Cs acyl, preferably hydrogen, wherein said blend of lipids comprising about 0.5-30 wt% of a C22-C38 fatty alcohol.

15. The blend according to claim 14, wherein the blend comprising about 0.5-5 wt% of fatty alcohols.

16. The blend according to claim 14, wherein the blend comprising about 20-30 wt% of fatty alcohols.

17. The blend according to any one of claims 14 to 17, wherein the blend comprises lipids of formula (3):wherein R1is as defined for the lipids of formula (1).

18. The blend according to any one of claims 14 to 17, wherein the blend comprising lipids of formula (6):wherein R1is as defined as for the lipids of formula (1), W, R4, R5, R6and the bond - are as defined as for the moiety of formula (2).

19. The blend according to claim 18, comprising lipids of formula (6) wherein R4is a saturated unsubstituted C13-C17 alkyl, R5and R6are hydrogen, and the bond - is a single bond.

20. The blend according to claim 18, comprising lipids of formula (6) wherein R4is a saturated unsubstituted C13-C17 alkyl, R5is -OH, Rsis hydrogen, and the bond - is a single bond.

21. The blend according to claim 18, comprising the lipids of formula (6) wherein R4is a saturated unsubstituted C13-C17 alkyl, R5and R6are hydrogen, and the bond - is a double bond.

22. The blend according to any one of claims 18 to 21, comprising lipids of formula (6) wherein W is hydrogen.

23. The blend according to any one of claims 18 to 21, comprising lipids of formula (6), wherein W is a glycosyl moiety selected from the group consisting of Glcl-, Gall-, Gaipi-4Glcl-.

24. The blend according to any one of claims 14 to 23, wherein the content of lipids of formula (1), (3) or(6) wherein R1is a C21 alkyl and lipids of formula (1) wherein R1is a C23 alkyl is at least about 50 wt%.

25. The blend according to any one of claims 14 to 23, wherein the ratio between the lipids of formula (1), (3) or (6) wherein R1is a C21 alkyl, and the lipids of formula (1) wherein R1is a C23 alkyl is from about 1:10 to about 10:1.

26. The blend according to any one of preceding claims 14 to 25, wherein the blend is obtainable by the method of any one of claims 1 to 13.

27. A cosmetic composition comprising a blend of lipids of any one of claims 14 to 26, or a combination thereof.

8. Use of a blend of lipids any one of claims 14 to 26, or a cosmetic composition accoding to claim27, for maintaining physiologically normal skin barrier function or improving a deteriorated skin barrier function in an individual.

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