COMPOSITIONS FOR INCREASING BIOAVAILABILITY OF PEPTIDES OR PROTEINS AND METHOD THEREOF

Field of the present Invention

The invention relates to methods and compositions for the delivery of peptides and proteins topically, nasally, and by inhalation.

Background of the Invention

The advent of genetic engineering, especially recombinant DNA technology, has made available many biologically active peptides and proteins that previously were laboratory curiosities. Use of these biologically active peptides in medical practice is not so simple. Because of the nature of peptide bonds, they are not stable in the acidic conditions of the stomach and thus have very poor oral activity. The most common route of administering biologically active peptides is by the intravenous route which is not amenable to out-patient treatment. Rectal and buccal administration of the peptides have their own limitations resulting from the short half life of many peptides/proteins, eg., parathyrpoid hormone related peptide (PTHrP) 1-84 has a half life of 13.6 minutes. Other major hurdles to effective therapeutic use is proteolytic degradation and a lack of cellular internalization of the active protein/peptide.

Transdermal delivery of peptides and proteins face several problems. First, an inverse relationship appears to exist between absorption rate and molecular weight, limiting the absorption of peptides and proteins as these are typically high molecular weight drugs. Second, transdermal delivery can be hindered by the polarity of the peptide or protein. High polarity can significantly attenuate the rate of drug delivery (commonly called "flux" or "permeation rate") from a transdermal drug delivery system. Third, it has long been established that proteases are present in keratinocytes to protect the body from protein allergens. Thus, when topically applied a peptide or protein will often undergo rapid enzymatic degradation rendering the peptide or protein therapeutically ineffective (see, for example, Camilla et al., J. Toxicol. Cut. & Ocular Toxicol. 21 :175 (2002)).

Nasal delivery can be used for systemic and also topical (i.e., lung) delivery of peptides and proteins. Nasal delivery systems often involve aerosol formulations with a compressed gas carrier and administered via metered-dose inhalation (MDI). In this approach particular size plays a vital role in absorption and stability. For systemic delivery, the peptide/protein must diffuse through the pulmonary epithelia. Peptide and protein therapeutics are often water soluble and stabilized by formulation in saline. However, only four commercial peptidic drugs are formulated for nasal administration because poor diffusion limits systemic circulation (see, for example, A.E. Pontiroli, Advanced Drug Delivery Reviews 29:81 (1998)).

Therefore, there is a need to find new methods and compositions for the delivery of peptides and proteins especially by transdermal and nasal methods. Objects of the present Invention

The main object of the present invention is to develop compounds for increasing bio-availability of peptides and proteins.

Another main object of the present invention is to develop compositions for increasing bio-availability of peptides and proteins.

Yet another object of the present invention is to develop methods for increasing bio-availability of peptides and proteins.

Still another object of the present invention is to develop processes for preparing such compositions. Statement of the present invention

The present invention relates to a compound of formula I: (A)-(L)-(B) I wherein

(B) is a hydrophobic moiety;

(L) is a linker which which covalently links saccharide (A) and hydrophobic moiety (B); and

(A) is a saccharide which is unsubstituted at the hydroxyl group of the anomeric position; and corresponding compositions; processes for preparing the compositions and methods thereof. Detailed description of the present invention

We have discovered that glycoses can be derivatized with a hydrophobic moiety to form an amphipathic agent to enhance the bioavailability of peptides and proteins delivered topically and nasally. The anomeric position of the saccharide in the amphipathic agent undergoes a spontaneous and reversible Schiff base condensation with any peptide or protein bearing a free amino group (e.g., lysine side-chain ε-amino and terminal α-amino groups) to form a Schiff base conjugate. The resulting peptide or protein conjugate can be, for example, applied topically, where it is transported across the stratum corneum. Subsequent in vivo hydrolysis of the Schiff base conjugate yields the therapeutic peptide or protein.

Accordingly, in a first aspect, the invention features a compound of formula I:

(A)-(L)-(B)

I

In formula I, (B) is a hydrophobic moiety; (L) is a linker which which covalently links saccharide (A) and hydrophobic moiety (B); and (A) is a saccharide which is unsubstituted at the hydroxyl group of the anomeric position.

The hydrophobic moiety can be any moiety having a LogP of greater than 2.0. Desirably, the hydrophobic moiety is selected from tocopherol, kojic acid, ursolic acid, boswellic acid, azelaic acid, hydroquinol, retinoic acid, TBHQ, glabrene, glabridene, isoliquiritigenin, 4,4'-dihydroxy biphenyl, polyethoxylated fatty acids, alcohol-oil transesterifϊcation products, polyglycerized fatty acids, propylene glycol fatty acid esters, mixtures of propylene glycol esters and glycerol esters, mono- and diglycerides, sterol and sterol derivatives, polyethylene glycol sorbitan fatty acid esters, polyethylene glycol alkyl ethers, polyethylene glycol alkyl phenols, polyoxyethylene-polyoxypropylene block

copolymers, sorbitan fatty acid esters, lower alcohol fatty acid esters, and ionic surfactants.

Any saccharide can be used in the compounds of the invention. Desirably, the saccharide is selected from ribose, lyxose, xylose, arabinose, glucose, fructose, maltose, lactose, mannose, galactose, glucose, glucosamine, a disaccharide, a polysaccharide, and derivatives thereof.

The linker (L) which covalently links the hydrophobic moiety and the saccharide can be described, for example, by formula X:

-(zV(YV(z ² )s-(Ri4)-(z ³ )t-(YV(z ⁴ )p-

X

In formula X, each of Z ¹ , Z ² , Z ³ , and Z ⁴ is, independently, selected from O, S, and NR ₁₆ ; R ₁₆ is hydrogen or an alkyl group; each of Y ¹ and Y ² is, independently, selected from carbonyl, thiocarbonyl, sulphonyl, or phosphoryl groups; each of o, p, s, t, u, and v is, independently, 0 or 1; u + v is 1 or 2; and R ₁₄ is a linear or branched alkyl of 1 to 10 carbon atoms, a linear or branched heteroalkyl of 1 to 10 atoms, a linear or branched alkene of 2 to 10 carbon atoms, a linear or branched alkyne of 2 to 10 carbon atoms, an aromatic residue of 5 to 10 carbon atoms, a cyclic system of 3 to 10 atoms, - (CH ₂ CH ₂ O)C ₁ CH ₂ CH ₂ - in which q is 1 to 4, or a chemical bond linking -(Z ¹ ) _O -(Y ¹ ) _U -(Z ² ) _S - to -(Z ³ ) _t -(Y ² )v-(Z ⁴ )p-. Desirably, the hydrophobic moiety and saccharide are linked by a phosphodiester group.

In an embodiment of the above aspect, the compound of the invention is further described formulas II or III:

ORn (IV)

In formulas II, III, and IV X is F, OR ₁₀ , SR ₁₀ , or NHR ₁₀ and each of R ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ , R ₁₂ , and R ₁₃ is, independently, selected from H, a saccharide, and

O

— P-OB

I OH wherein OB is a radical of a hydrophobic moiety. Desirably, the hydrophobic moiety is selected from tocopherol, kojic acid, ursolic acid, boswellic acid, azelaic acid, hydroquinol, retinoic acid, TBHQ, glabrene, glabridene, isoliquiritigenin, 4,4'-dihydroxy biphenyl, and lipidated ascorbic acid. Desirably, only one OfOfR ₂ , R ₃ , R ₄ , R ₅ , R ₆ , R ₇ , R ₈ , R ₉ , R ₁₀ , R ₁₁ , R ₁₂ , and R ₁₃ is a phosphodiester linking the saccharide to a hydrophobic group.

The invention also features a pharmaceutical composition including a compound of the invention condensed with a peptide or protein.

The invention further features a pharmaceutical composition including (i) a compound of the invention and (ii) a peptide or protein.

The pharmaceutical composition can be formulated for topical administration. Desirably, the pharmaceutical composition is formulated as a cream, foam, paste, lotion, gel, stick, spray, patch, or ointment. The pharmaceutical composition can also be formulated for nasal administration or for inhalation (i.e., for systemic circulation).

In any of the above pharmaceutical compositions the peptide or protein can be an epidermal growth stimulant, epidermal growth suppressor, skin depigmenting agent; hair growth stimulant, hair growth suppressor, antiproliferative or immune suppressor. Desirably, the peptide or protein is selected from PTH, PTHrP, and melanogenosis- inhibiting peptideselected from Glu-Asp-Tyr-His-Ser-Leu-Tyr-Gln-Ser-His-Leu (SEQ ID NO. 1), Ser-Gly-Gly-Tyr-Leu-Pro-Pro-Leu (SEQ ID NO. 2), His-Ser-C(O)-NH-(CH2)k- CH2-C(O)-NH-Ser-His, wherein k is 2-10 (SEQ ID NO. 3); SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNF (PTHrP (1-34)) (SEQ ID NO. 4); LMHNLGKHLNSMERVEWLRKKLQDVHNF (PTHrP (7-34)) (SEQ ID NO.5); epidermial growth factor (EGF); melanocyte stimulating hormone (MSH); corticotropin- releasing hormone (CRH); tumor necrosis factor-related apoptosis-inducing ligand (TRAIL); fibroblast growth factor (FGF), human growth hormone (HRH), human growth hormone stimulating hormone (GHRH), and growth hormone inhibiting hormone (GHIH).

The invention also features a method for increasing the bioavailability of a peptide or protein administered to a mammal. The method includes the step of combining the peptide or protein with a compound of the invention to form a condensation product and administering the product to the mammal. The product can be administered topically, nasally, or to the lungs of the mammal.

By "hydrophobic moiety" is meant a lipophilic group having a LogP (octanol/water) greater than 2 (signifying that it is 100 fold more soluble in n.octanol).

By "saccharide" is meant an aldose or a ketose, either as a monosaccharide or part of a disaccharide or polysaccharide that is capable of Schiff-base condensation with an amino group of a peptide or protein. Glycation reactions are known to be initiated by reversible Schiff-base (aldimine or ketimine) addition reactions with lysine side-chain ε- amino and terminal α-amino groups. Typically, sugars initially react in their open-chain (not the predominant pyranose and furanose structures) aldehydo or keto forms with lysine side chain ε-amino and terminal α-amino groups through reversible Schiff base condensation. Saccharides include glycose, glycosamine, aldohexoses, ketohexoses, aldopentose, ketopentose, disaccharides, polysaccharides of 3-20 saccharide units, and deoxy and fluorinated derivatives thereof.

As used herein, "unsubstituted at the hydroxyl group of the anomeric position" is meant any aldose or ketose capable of forming an open-chain structure.

By "saccharide derivative" is meant halide (e.g., fluorinated), alkanoate, sulfate, and/or phosphate derivatives of any saccharides described herein.

As used herein, "increasing bioavailability" refers to an increase in the amount of peptide or protein delivered transdermally, or nasally, or by inhalation to a mammal for a prodrug condensation product of the invention in comparison to the same peptide or protein which is not modified as described herein and administered under the same conditions. The amount of peptide or protein delivered nasally, to the skin, or to the lungs of a mammal can be determined by pharmacokinetic analysis, e.g. radiotracer methods.

By "C _1-10 alkyl" is meant a branched or unbranched saturated hydrocarbon group, having 1 to 10 carbon atoms, inclusive. An alkyl may optionally include monocyclic, bicyclic, or tricyclic rings, in which each ring desirably has three to six members. The alkyl group may be substituted or unsubstituted. Exemplary substituents include alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio, halogen, hydroxyl, fluoroalkyl, perfluoralkyl,

amino, aminoalkyl, disubstituted amino, quaternary amino, hydroxyalkyl, carboxyalkyl, and carboxyl groups.

By "C _2-10 alkene" is meant a branched or unbranched hydrocarbon group containing one or more double bonds, desirably having from 2 to 10 carbon atoms. A C ₂ . io alkene may optionally include monocyclic, bicyclic, or tricyclic rings, in which each ring desirably has five or six members. The C _2-10 alkene group may be substituted or unsubstituted. Exemplary substituents include alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio, halogen, hydroxyl, fluoroalkyl, perfluoralkyl, amino, aminoalkyl, disubstituted amino, quaternary amino, hydroxyalkyl, carboxyalkyl, and carboxyl groups.

By "C _2-1 O alkyne" is meant a branched or unbranched hydrocarbon group containing one or more triple bonds, desirably having from 2 to 10 carbon atoms. A C _2-10 alkyne may optionally include monocyclic, bicyclic, or tricyclic rings, in which each ring desirably has five or six members. The C _2-10 alkyne group may be substituted or unsubstituted. Exemplary substituents include alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio, halogen, hydroxyl, fluoroalkyl, perfluoralkyl, amino, aminoalkyl, disubstituted amino, quaternary amino, hydroxyalkyl, carboxyalkyl, and carboxyl groups.

By "heteroalkyl" is meant a branched or unbranched alkyl group in which one or more methylenes (-CH ₂ -) are replaced by nitrogen, oxygen, sulfur, carbonyl, thiocarbonyl, phosphoryl, or sulfonyl moieties. Some examples include tertiary amines, ethers, thioethers, amides, thioamides, carbamates, thiocarbamates, phosphoramidates, ^• sulfonamides, and disulfides. A heteroalkyl may optionally include monocyclic, bicyclic, or tricyclic rings, in which each ring desirably has three to six members. The heteroalkyl group may be substituted or unsubstituted. Exemplary substituents include alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio, halogen, hydroxyl, fluoroalkyl, perfluoralkyl, amino, aminoalkyl, disubstituted amino, quaternary amino, hydroxyalkyl, carboxyalkyl, and carboxyl groups

By "C _5-10 aryl" or "aryl" is meant an aromatic group having a ring system with conjugated π electrons (e.g., phenyl, or imidazole ). The ring of the aryl group is

preferably 5 to 10 atoms. The aromatic ring may be exclusively composed of carbon atoms or may be composed of a mixture of carbon atoms and heteroatoms. Preferred heteroatoms include nitrogen, oxygen, sulfur, and phosphorous. Aryl groups may optionally include monocyclic, bicyclic, or tricyclic rings, where each ring has preferably five or six members. The aryl group may be substituted or unsubstituted. Exemplary substituents include alkyl, hydroxyl, alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio, halogen, fluoroalkyl, carboxyl, carboxyalkyl, amino, aminoalkyl, monosubstituted amino, disubstituted amino, and quaternary amino groups.

The term "cyclic system" refers to a compound that contains one or more covalently closed ring structures, in which the atoms forming the backbone of the ring are composed of any combination of the following: carbon, oxygen, nitrogen, sulfur, and phosphorous. The cyclic system may be substituted or unsubstituted. Exemplary substituents include, without limitation, alkyl, hydroxyl, alkoxy, aryloxy, sulfhydryl, alkylthio, arylthio, halogen, fluoroalkyl, carboxyl, carboxyalkyl, amino, aminoalkyl, monosubstituted amino, disubstituted amino, and quaternary amino groups.

This invention provides methods and compositions for enhancing the bioavailability of therapeutic peptides and proteins. The compounds of the invention form a reversible covalent bond with a protein or peptide to yield a prodrug that can have improved bioavailability. The compounds of the invention can also exhibit enhanced cell surface adhesion, resulting in better absorption and penetration of the lung epithelium. The compounds of the present invention have three characteristic components: a saccharide (A) covalently tethered via a linker (L) to a hydrophobic moiety (B). They are described by formula I.

(A)-(L)-(B) I

In formula I, (B) is a hydrophobic moiety; (L) is a linker which covalently links saccharide (A) and hydrophobic moiety (B); and (A) is a saccharide which is unsubstituted at the hydroxyl group of the anomeric position.

A description of how these compounds are prepared is provided below and in the examples. Saccharide (A)

The invention features amphipathic compound including a hydrophobic moiety covalently attached via a linker to a saccharide. The compounds of the invention undergo reversible Schiff base condensation with the amino group of a peptide or protein. Accordingly, the anomeric hydroxyl group of the saccharide is, necessarily, unsubstituted so that ring opening and Schiff base condensation (i.e., the N-glycosylation of the anomeric position of the saccharide) will occur.

The saccharide is a glycose chosen from aldo or keto pentoses, hexoses and polysaccharides with two to twenty glycose units. The glycose units can contain one or more amine functionality.

Suitable monosaccharides include, but are not limited to, any of several simple open or closed chain sugars (in the L or D configuration), typically having 5 or 6 carbons (a pentose monosaccharide or a hexose monosaccharide), as well as 7 carbons (heptose monosaccharide). Included are sugar derivatives in which the ring oxygen atom has been replaced by carbon, nitrogen or sulfur, amino sugars in which a hydroxyl substituent on the simple sugar is replaced with an amino group or sugars having a double bond between two adjacent carbon atoms, (e.g. glucosamine, 5-thio-D-glucose, nojirimycin, deoxynojirimycin, 1,5-anhydro-D-sorbitol, 2,5-anhydro-D-mannitol, 2-deoxy-D- galactose, 2-deoxy-D-glucose, 3-deoxy-D-glucose, allose, arabinose, arabinitol, fucitol, fucose, galactitol, glucitol, iditol, lyxose, mannitol, levo-rhamnitol, 2-deoxy-D-ribose, ribose, ribitol, ribulose, rhamnose, xylose, xylulose, allose, altrose, fructose, galactose, glucose, gulose, idose, levulose, mannose, psicose, sorbose, tagatose, talose, galactal, glucal, fucal, rhamnal, arabinal, xylal, valienamine, validamine, valiolamine, valiol, valiolon, valienol, valienone, glucuronic acid, galacturonic acid, N-acetylneuraminic acid, gluconic acid D-lactone, galactonic acid .gamma.-lactone, galactonic acid .delta.- lactone, mannonic acid .gamma.-lactone, D-altro-heptulose, D-manno-heptulose, D-

glycero-D-manno-heptose, D-glycero-D-gluco-heptose, D-allo-heptulose, D-altro-3- heptulose, D-glycero-D-manno-heptitol, D-glycero-D-altro-heptit- ol and the like), The hydroxyl groups on monosaccharides may optionally be replaced with hydrogen (deoxyglycoses), alkoxy (e.g. 2-O-methyl-D-fructose), alkanoate or halogen groups (especially fluorine atoms). Included are sulfate and/or phosphate derivatives of monosaccharides as defined herein.

Suitable oligosaccharides include, but are not limited to, carbohydrates having from 2 to 10 or more monosaccharides linked together. The constituent monosaccharide unit may be, for example, a pentose monosaccharide, or a hexose monosaccharide.

The Linker (L)

The linker component (L) of the present invention provides a molecular skeleton covalently linking (A) and (B). That is, a linear, cyclic, or branched molecular skeleton, with pendant groups which bind covalently with (A) and (B).

Thus, the linking of (A) with (B) is achieved by covalent means, involving bond formation with one or more functional groups located on (A) and (B). Examples of chemically reactive functional groups which may be employed for this purpose include, without limitation, amino, hydroxyl, sulfhydryl, carboxyl, carbonyl, carbohydrate groups, vicinal dials, thioethers, 2-aminoalcohols, 2-aminothiols, guanidinyl, imidazolyl, and phenolic groups.

The covalent linking of (A) with (B) may therefore be effected using a linker (L) which contains reactive moieties capable of reaction with such functional groups present in (A) and (B). The product of this reaction is a linkage group which contains the newly formed bonds linking (L) with (A) and (L) with (B). For example, a hydroxyl group of (A) may react with a carboxylic acid group of (L), or an activated derivative thereof, vide infra, resulting in the formation of an ester linkage group.

Examples of moieties capable of reaction with sulfhydryl groups include α- haloacetyl compounds of the type XCH ₂ CO- (where X=Br, Cl or I), which show

particular reactivity for sulfhydryl groups, but which can also be used to modify imidazolyl, thioether, phenol, and amino groups as described by Gurd, Methods Enzymol. 11:532, 1967. N-Maleimide derivatives are also considered selective towards sulfhydryl groups, but may additionally be useful in coupling to amino groups under certain conditions. Reagents such as 2-iminothiolane (Traut et al., Biochemistry 12:3266, 1973), which introduce a thiol group through conversion of an amino group, may be considered as sulfhydryl reagents if linking occurs through the formation of disulphide bridges.

Examples of reactive moieties capable of reaction with amino groups include, for example, alkylating and acylating agents. Representative alkylating agents include: (i) α-haloacetyl compounds, which show specificity towards amino groups in the absence of reactive thiol groups and are of the type XCH ₂ CO- (where X=Cl, Br or I), for example, as described by Wong Biochemistry 24:5337, 1979;

(ii) N-maleimide derivatives, which may react with amino groups either through a Michael type reaction or through acylation by addition to the ring carbonyl group, for example, as described by Smyth et al., J Am. Chem. Soc. 82:4600, 1960 and Biochem. J. 91:589, 1964;

(iii) aryl halides such as reactive nitrohaloaromatic compounds;

(iv) alkyl halides, as described, for example, by McKenzie et al., J. Protein Chem. 7:581, 1988;

(v) aldehydes and ketones capable of Schiff s base formation with amino groups, the adducts formed usually being stabilized through reduction to give a stable amine; (vi) epoxide derivatives such as epichlorohydrin and bisoxiranes, which may react with amino, sulfhydryl, or phenolic hydroxyl groups;

(vii) chlorine-containing derivatives of s-triazines, which are very reactive towards nucleophiles such as amino, sufhydryl, and hydroxyl groups;

(viii) aziridines based on s-triazine compounds detailed above, e.g., as described by Ross, J Adv. Cancer Res. 2:1, 1954, which react with nucleophiles such as amino groups by ring opening;

(ix) squaric acid diethyl esters as described by Tietze, Chem. Ber. 124:1215, 1991; and (x) α-haloalkyl ethers, which are more reactive alkylating agents than normal alkyl halides because of the activation caused by the ether oxygen atom, as described by Benneche et al, Eur. J. Med. Chem. 28:463, 1993. Representative amino-reactive acylating agents include: (i) isocyanates and isothiocyanates, particularly aromatic derivatives, which form stable urea and thiourea derivatives respectively; (ii) sulfonyl chlorides, which have been described by Herzig et al., Biopolymers

2:349, 1964; (iii) acid halides;

(iv) active esters such as nitrophenylesters or N-hydroxysuccinimidyl esters; (v) acid anhydrides such as mixed, symmetrical, or N-carboxyanhydrides; (vi) other useful reagents for amide bond formation, for example, as described by M. Bodansky, Principles of Peptide Synthesis, Springer- Verlag, 1984; (vii) acylazides, e.g. wherein the azide group is generated from a preformed hydrazide derivative using sodium nitrite, as described by Wetz et a\., Anal. Biochem. 58:347, 1974; and

(viii) imidoesters, which form stable amidines on reaction with amino groups, for example, as described by Hunter and Ludwig, J Am. Chem. Soc. 84:3491, 1962.

Aldehydes and ketones may be reacted with amines to form Schiffs bases, which may advantageously be stabilized through reductive animation. Alkoxylamino moieties readily react with ketones and aldehydes to produce stable alkoxamines, for example, as described by Webb et al., in Bioconjugate Chem. 1:96, 1990.

Examples of reactive moieties capable of reaction with carboxyl groups include diazo compounds such as diazoacetate esters and diazoacetamides, which react with high specificity to generate ester groups, for example, as described by Herriot, Adv. Protein Chem. 3:169, 1947. Carboxylic acid modifying reagents such as carbodiimides, which react through O-acylurea formation followed by amide bond formation, may also be employed.

It will be appreciated that functional groups in the saccharide (A) and/or the hydrophobic moiety (B) may, if desired, be converted to other functional groups prior to reaction, for example, to confer additional reactivity or selectivity. Examples of methods useful for this purpose include conversion of amines to carboxylic acids using reagents such as dicarboxylic anhydrides; conversion of amines to thiols using reagents such as N- acetylhomocysteine thiolactone, S-acetylmercaptosuccinic anhydride, 2-iminothiolane, or thiol-containing succinimidyl derivatives; conversion of thiols to carboxylic acids using reagents such as α-haloacetates; conversion of thiols to amines using reagents such as ethylenimine or 2-bromoethylamine; conversion of carboxylic acids to amines using reagents such as carbodiimides followed by diamines; and conversion of alcohols to thiols using reagents such as tosyl chloride followed by transesterification with thioacetate and hydrolysis to the thiol with sodium acetate.

Typically, the linker will include two or more reactive moieties, as described above, connected by a spacer element. The presence of such a spacer permits bifunctional linkers to react with specific functional groups within (A) and (B), resulting in a covalent linkage between these two compounds. The reactive moieties in a linker (L) may be the same (homobifunctional linker) or different (heterobifunctional linker, or, where several dissimilar reactive moieties are present, heteromultifunctional linker), providing a diversity of potential reagents that may bring about covalent attachment between (A) and (B).

Spacer elements typically consist of chains which effectively separate (A) and (B) by a linear or branched alkyl of 1 to 10 carbon atoms, a linear or branched heteroalkyl of 1 to 10 atoms, a linear or branched alkene of 2 to 10 carbon atoms, a linear or branched alkyne of 2 to 10 carbon atoms, an aromatic residue of 5 to 10 carbon atoms, a cyclic system of 3 to 10 atoms, or -(CH ₂ CH ₂ O) _n CH ₂ CH ₂ -, in which n is 1 to 4.

For example, an amide linkage can be formed using glucosamine or glucuronic acid. Furthermore, the amphipathic agent can have two hydropohilic heads by using a bifunctional hydrophobic moiety, such as the dicarboxylic acid azalaic acid. The linkage

can also be an amide by using long chain dicarboxylic acids or diamines using glucosamine or glucuronic acid as desired.

Typically, the linkage group will be as simple as a phosphodiester or phosphoroamidate linkage, which offers the advantage of being stable towards hydrolytic degradation. Hydrophobic Moiety (B)

The hydrophobic moiety component (B) of the present invention can be selected from any lipophilic group having one or more heteroatoms to which a linker (L) can be attached.

Hydrophobic moieties that can be used in the compositions and methods of the invention include, without limitation, tocopherol, kojic acid, ursolic acid, boswellic acid, azelaic acid, hydroquinol, retinoic acid, TBHQ, glabrene, glabridene, isoliquiritigenin, 4,4'-dihydroxy biphenyl, lipidated ascorbic acid (such as ascorbyl palmitate or laurate) and compounds belonging to the following classes: polyethoxylated fatty acids, alcohol- oil transesterification products, polyglycerized fatty acids, propylene glycol fatty acid esters, mixtures of propylene glycol esters and glycerol esters, mono- and diglycerides, sterol and sterol derivatives, polyethylene glycol sorbitan fatty acid esters, polyethylene glycol alkyl ethers, polyethylene glycol alkyl phenols, polyoxyethylene- polyoxypropylene block copolymers, sorbitan fatty acid esters, lower alcohol fatty acid esters, and ionic surfactants. Commercially available examples for each class of hydrophobic moiety are provided below. These may be modified as described above for reaction with a saccharide to form an amphipathic compound of the invention.

Polyethoxylated fatty acids can be used as hydrophobic moieties amphipathic compounds of the invention. Examples of commercially available polyethoxylated fatty acid monoester surfactants include: PEG 4-100 monolaurate (Crodet L series, Croda), PEG 4-100 monooleate (Crodet O series, Croda), PEG 4-100 monostearate (Crodet S series, Croda, and Myrj Series, Atlas/ICI), PEG 400 distearate (Cithrol 4DS series, Croda), PEG 100, 200, or 300 monolaurate (Cithrol ML series, Croda), PEG 100, 200, or

300 monooleate (Cithrol MO series, Croda), PEG 400 dioleate (Cithrol 4DO series, Croda), PEG 400-1000 monostearate (Cithrol MS series, Croda), PEG-I stearate (Nikkol MYS-IEX, Nikko, and Coster Kl, Condea), PEG-2 stearate (Nikkol MYS-2, Nikko), PEG-2 oleate (Nikkol MYO-2, Nikko), PEG-4 laurate (Mapeg® 200 ML, PPG), PEG-4 oleate (Mapeg® 200 MO, PPG), PEG-4 stearate (Kessco® PEG 200 MS, Stepan), PEG-5 stearate (Nikkol TMGS-5, Nikko), PEG-5 oleate (Nikkol TMGO-5, Nikko), PEG-6 oleate (Algon OL 60, Auschem SpA), PEG-7 oleate (Algon OL 70, Auschem SpA), PEG-6 laurate (Kessco® PEG300 ML, Stepan), PEG-7 laurate (Lauridac 7, Condea), PEG-6 stearate (Kessco® PEG300 MS, Stepan), PEG-8 laurate (Mapeg® 400 ML, PPG), PEG-8 oleate (Mapeg® 400 MO, PPG), PEG-8 stearate (Mapeg® 400 MS, PPG), PEG-9 oleate (Emulgante A9, Condea), PEG-9 stearate (Cremophor S9, BASF), PEG-IO laurate (Nikkol MYL-IO, Nikko), PEG-IO oleate (Nikkol MYO-10, Nikko), PEG-12 stearate (Nikkol MYS-10, Nikko), PEG-12 laurate (Kessco® PEG 600 ML, Stepan), PEG-12 oleate (Kessco® PEG 600 MO, Stepan), PEG-12 ricinoleate (CAS # 9004-97-1), PEG-12 stearate (Mapeg® 600 MS, PPG), PEG-15 stearate (Nikkol TMGS-15, Nikko), PEG-15 oleate (Nikkol TMGO-15, Nikko), PEG-20 laurate (Kessco® PEG 1000 ML, Stepan), PEG-20 oleate (Kessco® PEG 1000 MO, Stepan), PEG-20 stearate (Mapeg® 1000 MS, PPG), PEG-25 stearate (Nikkol MYS-25, Niklco), PEG-32 laurate (Kessco® PEG 1540 ML, Stepan), PEG-32 oleate (Kessco® PEG 1540 MO, Stepan), PEG-32 stearate (Kessco® PEG 1540 MS, Stepan), PEG-30 stearate (Myrj 51), PEG-40 laurate (Crodet L40, Croda), PEG-40 oleate (Crodet O40, Croda), PEG-40 stearate (Emerest® 2715, Henkel), PEG-45 stearate (Nikkol MYS-45, Nikko), PEG-50 stearate (Myrj 53), PEG-55 stearate (Nikkol MYS-55, Nikko), PEG-100 oleate (Crodet O-100, Croda), PEG-100 stearate (Ariacel 165, ICI), PEG-200 oleate (Albunol 200 MO, Taiwan Surf.), PEG-400 oleate (LACTOMUL, Henkel), and PEG-600 oleate (Albunol 600 MO, Taiwan Surf.). Amphipathic compounds according to the invention may include one or more of the polyethoxylated fatty acids above.

Alcohol-oil transesterification products may also be used as hydrophobic moieties amphipathic compounds of the invention. Examples of commercially available alcohol- oil transesterification products include: PEG-3 castor oil (Nikkol CO-3, Nikko), PEG-5, 9, and 16 castor oil (ACCONON CA series, ABITEC), PEG-20 castor oil, (Emalex C-20, Nihon Emulsion), PEG-23 castor oil (Emulgante EL23), PEG-30 castor oil (Incrocas 30, Croda), PEG-35 castor oil (Incrocas-35, Croda), PEG-38 castor oil (Emulgante EL 65, Condea), PEG-40 castor oil (Emalex C-40, Nihon Emulsion), PEG-50 castor oil (Emalex C-50, Nihon Emulsion), PEG-56 castor oil (Eumulgin® PRT 56, Pulcra SA), PEG-60 castor oil (Nikkol CO-60TX, Nikko), PEG-100 castor oil, PEG-200 castor oil (Eumulgin® PRT 200, Pulcra SA), PEG-5 hydrogenated castor oil (Nikkol HCO-5, Nikko), PEG-7 hydrogenated castor oil (Cremophor WO7, BASF), PEG-IO hydrogenated castor oil (Nikkol HCO-IO, Nikko), PEG-20 hydrogenated castor oil (Nikkol HCO-20, Nikko), PEG-25 hydrogenated castor oil (Simulsol® 1292, Seppic), PEG-30 hydrogenated castor oil (Nikkol HCO-30, Nikko), PEG-40 hydrogenated castor oil (Cremophor RH 40, BASF), PEG-45 hydrogenated castor oil (Cerex ELS 450, Auschem Spa), PEG-50 hydrogenated castor oil (Emalex HC-50, Nihon Emulsion), PEG-60 hydrogenated castor oil (Nikkol HCO-60, Nikko), PEG-80 hydrogenated castor oil (Nikkol HCO-80, Nikko), PEG-100 hydrogenated castor oil (Nikkol HCO-100, Nikko), PEG-6 corn oil (Labrafil® M 2125 CS, Gattefosse), PEG-6 almond oil (Labrafil® M 1966 CS, Gattefosse), PEG-6 apricot kernel oil (Labrafil® M 1944 CS, Gattefosse), PEG-6 olive oil (Labrafil® M 1980 CS, Gattefosse), PEG-6 peanut oil (Labrafil® M 1969 CS, Gattefosse), PEG-6 hydrogenated palm kernel oil (Labrafil® M 2130 BS, Gattefosse), PEG-6 palm kernel oil (Labrafil® M 2130 CS, Gattefosse), PEG-6 triolein (Labrafil® M 2735 CS, Gattefosse), PEG-8 corn oil (Labrafil® WL 2609 BS, Gattefosse), PEG-20 corn glycerides (Crovol M40, Croda), PEG-20 almond glycerides (Crovol A40, Croda), PEG-25 trioleate (TAGAT® TO, Goldschmidt), PEG-40 palm kernel oil (Crovol PK-70), PEG-60 corn glycerides (Crovol M70, Croda), PEG-60 almond glycerides (Crovol A70, Croda), PEG-4 caprylic/capric triglyceride (Labrafac®

Hydro, Gattefosse), PEG-8 caprylic/capric glycerides (Labrasol, Gattefosse), PEG-6 caprylic/capric glycerides (SOFTIGEN®767, HuIs), lauroyl macrogol-32 glyceride (GELUCIRE 44/14, Gattefosse), stearoyl macrogol glyceride (GELUCIRE 50/13, Gattefosse), mono, di, tri, tetra esters of vegetable oils and sorbitol (SorbitoGlyceride, Gattefosse), pentaerythrityl tetraisostearate (Crodamol PTIS, Croda), pentaerythrityl distearate (Albunol DS, Taiwan Surf), pentaerythrityl tetraoleate (Liponate PO-4, Lipo Chem.), pentaerythrityl tetrastearate (Liponate PS-4, Lipo Chem.), pentaerythrityl tetracaprylate tetracaprate (Liponate PE-810, Lipo Chem.), and pentaerythrityl tetraoctanoate (Nikkol Pentarate 408, Nikko). Also included as oils in this category of surfactants are oil-soluble vitamins, such as vitamins A, D, E, K, 7-dehydrocholesterol, stigmasterol and cholesterol etc. Thus, derivatives of these vitamins, such as tocopheryl PEG-1000 succinate (TPGS, available from Eastman), are also suitable surfactants. Amphipathic compounds according to the invention may include one or more of the alcohol-oil transesterification products above.

Polyglycerized fatty acids may also be used as hydrophobic moieties amphipathic compounds of the invention. Examples of commercially available polyglycerized fatty acids include: polyglyceryl-2 stearate (Nikkol DGMS, Nikko), polyglyceryl-2 oleate (Nikkol DGMO, Nikko), polyglyceryl-2 isostearate (Nikkol DGMIS, Nikko), polyglyceryl-3 oleate (Caprol® 3GO, ABITEC), polyglyceryl-4 oleate (Nikkol Tetraglyn l-O, Nikko), polyglyceryl-4 stearate (Nikkol Tetraglyn 1-S, Nikko), polyglyceryl-6 oleate (Drewpol 6-1-0, Stepan), pblyglyceryl-10 laurate (Nikkol Decaglyn 1-L, Nikko), polyglyceryHO oleate (Nikkol Decaglyn 1-0, Nikko), polyglyceryl-10 stearate (Nikkol Decaglyn 1-S, Nikko), polyglyceryl-6 ricinoleate (Nikkol Hexaglyn PR- 15, Nikko), polyglyceryl-10 linoleate (Nikkol Decaglyn 1-LN, Nikko), polyglyceryl-6 pentaoleate (Nikkol Hexaglyn 5-0, Nikko), polyglyceryl-3 dioleate (Cremophor GO32, BASF), polyglyceryl-3 distearate (Cremophor GS32, BASF), polyglyceryl-4 pentaoleate (Nikkol Tetraglyn 5-0, Nikko), polyglyceryl-6 dioleate (Caprol® 6G20, ABITEC), polyglyceryl- 2 dioleate (Nikkol DGDO, Nikko), polyglyceryl-10 trioleate (Nikkol Decaglyn 3-0,

Nikko), polyglyceryl-10 pentaoleate (Nikkol Decaglyn 5-0, Nikko), poly glyceryl- 10 septaoleate (Niklcol Decaglyn 1-0, Nikko), polyglyceryl-10 tetraoleate (Caprol® 10G4O, ABITEC), polyglyceryl-10 decaisostearate (Niklcol Decaglyn 10-IS, Nikko), polyglyceryl-101 decaoleate (Drewpol 10-10-O, Stepan), polyglyceryl-10 mono, dioleate (Caprol® PGE 860, ABITEC), and polyglyceryl polyricinoleate (Polymuls, Henkel). Amphipathic compounds according to the invention may include one or more of the polyglycerized fatty acids above.

In addition, propylene glycol fatty acid esters may be used as hydrophobic moieties amphipathic compounds of the invention. Examples of commercially available propylene glycol fatty acid esters include: propylene glycol monocaprylate (Capryol 90, Gattefosse), propylene glycol monolaurate (Lauroglycol 90, Gattefosse), propylene glycol oleate (Lutrol OP2000, BASF), propylene glycol myristate (Mirpyl), propylene glycol monostearate (LIPO PGMS, Lipo Chem.), propylene glycol hydroxystearate, propylene glycol ricinoleate (PROPYMULS, Henkel), propylene glycol isostearate, propylene glycol monooleate (Myverol P-06, Eastman), propylene glycol dicaprylate dicaprate (Captex® 200, ABITEC), propylene glycol dioctanoate (Captex® 800, ABITEC), propylene glycol caprylate caprate (LABRAFAC PG, Gattefosse), propylene glycol dilaurate, propylene glycol distearate (Kessco® PGDS, Stepan), propylene glycol dicaprylate (Nikkol Sefsol 228, Nikko), and propylene glycol dicaprate (Nikkol PDD, Nikko). Amphipathic compounds according to the invention may include one or more of the propylene glycol fatty acid esters above.

Mixtures of propylene glycol esters and glycerol esters may also be used as hydrophobic moieties amphipathic compounds of the invention. One preferred mixture is composed of the oleic acid esters of propylene glycol and glycerol (Arlacel 186). Examples of these surfactants include: oleic (ATMOS 300, ARLACEL 186, ICI), and stearic (ATMOS 150). Amphipathic compounds according to the invention may include one or more of the mixtures of propylene glycol esters and glycerol esters above.

Further, mono- and diglycerides may be used as hydrophobic moieties amphipathic compounds of the invention. Examples of commercially available mono- and diglycerides include: monopalmitolein (C16:l) (Larodan), monoelaidin (C18:l) (Larodan), monocaproin (C6) (Larodan), monocaprylin (Larodan), monocaprin (Larodan), monolaurin (Larodan), glyceryl monomyristate (C 14) (Nikkol MGM, Nikko), glyceryl monooleate (Cl 8:1) (PECEOL, Gattefosse), glyceryl monooleate (Myverol, Eastman), glycerol monooleate/linoleate (OLICINE, Gattefosse), glycerol monolinoleate (Maisine, Gattefosse), glyceryl ricinoleate (Softigen® 701, HuIs), glyceryl monolaurate (ALDO® MLD, Lonza), glycerol monopalmitate (Emalex GMS-P, Nihon), glycerol monostearate (Capmul® GMS, ABITEC), glyceryl mono- and dioleate (Capmul® GMO- K, ABITEC), glyceryl palmitic/stearic (CUTINA MD-A, ESTAGEL-Gl 8), glyceryl acetate (Lamegin® EE, Grunau GmbH), glyceryl laurate (Imwitor® 312, HuIs), glyceryl citrate/lactate/oleate/linoleate (Imwitor® 375, HuIs), glyceryl caprylate (Imwitor® 308, HuIs), glyceryl caprylate/caprate (Capmul® MCM, ABITEC), caprylic acid mono- and diglycerides (Imwitor® 988, HuIs), caprylic/capric glycerides (Imwitor® 742, HuIs), Mono-and diacetylated monoglycerides (Myvacet® 9-45, Eastman), glyceryl monostearate (Aldo® MS, Arlacel 129, ICI), lactic acid esters of mono and diglycerides (LAMEGIN GLP, Henkel), dicaproin (C6) (Larodan), dicaprin (ClO) (Larodan), dioctanoin (C8) (Larodan), dimyristin (C 14) (Larodan), dipalmitin (C 16) (Larodan), distearin (Larodan), glyceryl dilaurate (C 12) (Capmul® GDL, ABITEC), glyceryl dioleate (Capmul® GDO, ABITEC), glycerol esters of fatty acids (GELUCIRE 39/01, Gattefosse), dipalmitolein (C 16:1) (Larodan), 1,2 and 1,3-diolein (Cl 8:1) (Larodan), dielaidin (Cl 8:1) (Larodan), and dilinolein (Cl 8:2) (Larodan). Amphipathic compounds according to the invention may include one or more of the mono- and diglycerides above.

Sterol and sterol derivatives may also be used as hydrophobic moieties amphipathic compounds of the invention. Examples of commercially available sterol and sterol derivatives include: cholesterol, sitosterol, lanosterol, PEG-24 cholesterol ether (Solulan C-24, Amerchol), PEG-30 cholestanol (Phytosterol GENEROL series, Henkel),

PEG-25 phytosterol (Nikkol BPSH-25, Nikko), PEG-5 soyasterol (Nikkol BPS-5, Nildco), PEG-IO soyasterol (Nikkol BPS-IO, Nikko), PEG-20 soyasterol (Nikkol BPS-20, Nikko), and PEG-30 soyasterol (Nikkol BPS-30, Nikko). Amphipathic compounds according to the invention may include one or more of the sterol and sterol derivatives above.

Polyethylene glycol sorbitan fatty acid esters may also be used as hydrophobic moieties amphipathic compounds of the invention. Examples of commercially available polyethylene glycol sorbitan fatty acid esters include: PEG-IO sorbitan laurate (Liposorb L- 10, Lipo Chem.), PEG-20 sorbitan monolaurate (Tween® 20, Atlas/ICI), PEG-4 sorbitan monolaurate (Tween® 21, Atlas/ICI), PEG-80 sorbitan monolaurate (Hodag PSML-80, Calgene), PEG-6 sorbitan monolaurate (Nikkol GL-I, Nikko), PEG-20 sorbitan monopalmitate (Tween® 40, Atlas/ICI), PEG-20 sorbitan monostearate (Tween® 60, Atlas/ICI), PEG-4 sorbitan monostearate (Tween® 61, Atlas/ICI), PEG-8 sorbitan monostearate (DACOL MSS, Condea), PEG-6 sorbitan monostearate (Nikkol TS 106, Nikko), PEG-20 sorbitan tristearate (Tween® 65, Atlas/ICI), PEG-6 sorbitan tetrastearate (Nikkol GS-6, Nikko), PEG-60 sorbitan tetrastearate (Nikkol GS-460, Nikko), PEG-5 sorbitan monooleate (Tween® 81, Atlas/ICI), PEG-6 sorbitan monooleate (Nikkol TO- 106, Nikko), PEG-20 sorbitan monooleate (Tween® 80, Atlas/ICI), PEG-40 sorbitan oleate (Emalex ET 8040, Nihon Emulsion), PEG-20 sorbitan trioleate (Tween® 85, Atlas/ICI), PEG-6 sorbitan tetraoleate (Nikkol GO-4, Nikko), PEG-30 sorbitan tetraoleate (Nikkol GO-430, Nikko), PEG-40 sorbitan tetraoleate (Nikkol GO-440, Nikko), PEG-20 sorbitan monoisostearate (Tween® 120, Atlas/ICI), PEG sorbitol hexaoleate (Atlas G- 1086, ICI), polysorbate 80 (Tween® 80, Pharma), polysorbate 85 (Tween® 85, Pharma), polysorbate 20 (Tween® 20, Pharma), polysorbate 40 (Tween® 40, Pharma), polysorbate 60 (Tween® 60, Pharma), and PEG-6 sorbitol hexastearate (Nikkol GS-6, Nikko). Amphipathic compounds according to the invention may include one or more of the polyethylene glycol sorbitan fatty acid esters above.

In addition, polyethylene glycol alkyl ethers may be used as hydrophobic moieties amphipathie compounds of the invention. Examples of commercially available polyethylene glycol alkyl ethers include: PEG-2 oleyl ether, oleth-2 (Brij 92/93, Atlas/ICI), PEG-3 oleyl ether, oleth-3 (Volpo 3, Croda), PEG-5 oleyl ether, oleth-5 (Volpo 5, Croda), PEG-IO oleyl ether, oleth-10 (Volpo 10, Croda), PEG-20 oleyl ether, oleth-20 (Volpo 20, Croda), PEG-4 lauryl ether, laureth-4 (Brij 30, Atlas/ICI), PEG-9 lauryl ether, PEG-23 lauryl ether, laureth-23 (Brij 35, Atlas/ICI), PEG-2 cetyl ether (Brij 52, ICI), PEG-IO cetyl ether (Brij 56, ICI), PEG-20 cetyl ether (BriJ 58, ICI), PEG-2 stearyl ether (Brij 72, ICI), PEG-10 stearyl ether (Brij 76, ICI), PEG-20 stearyl ether (Brij 78, ICI), and PEG-100 stearyl ether (Brij 700, ICI). Amphipathie compounds according to the invention may include one or more of the polyethylene glycol alkyl ethers above. polyethylene glycol alkyl phenols are also useful as hydrophobic moieties amphipathie compounds of the invention. Examples of commercially available polyethylene glycol alkyl phenols include: PEG- 10- 100 nonylphenol series (Triton X series, Rohm & Haas) and PEG-15-100 octylphenol ether series (Triton N-series, Rohm & Haas). Amphipathie compounds according to the invention may include one or more of the polyethylene glycol alkyl phenols above.

Polyoxyethylene-polyoxypropylene block copolymers may also be used as hydrophobic moieties amphipathie compounds of the invention. These surfactants are available under various trade names, including one or more of Synperonic PE series (ICI), Pluronic® series (BASF), Lutrol (BASF), Supronic, Monolan, Pluracare, and Plurodac. The generic term for these copolymers is "poloxamer" (CAS 9003-11-6). These polymers have the formula XX:

HO(C ₂ H ₄ O) _a (C ₃ H ₆ O) _b (C ₂ H ₄ O) _a H XX where "a" and "b" denote the number of polyoxyethylene and polyoxypropylene units, respectively. These copolymers are available in molecular weights ranging from 1000 to 15000 daltons, and with ethylene oxide/propylene oxide ratios between 0.1 and

0.8 by weight. Amphipathic compounds according to the invention may include one or more of the polyoxyethylene-polyoxypropylene block copolymers above.

Polyoxyethylenes, such as PEG 300, PEG 400, and PEG 600, may be used as hydrophobic moieties amphipathic compounds of the invention.

Sorbitan fatty acid esters may also be used as hydrophobic moieties amphipathic compounds of the invention. Examples of commercially sorbitan fatty acid esters include: sorbitan monolaurate (Span-20, Atlas/ICI), sorbitan monopalmitate (Span-40, Atlas/ICI), sorbitan monooleate (Span-80, Atlas/ICI), sorbitan monostearate (Span-60, Atlas/ICI), sorbitan trioleate (Span-85, Atlas/ICI), sorbitan sesquioleate (Arlacel-C, ICI), sorbitan tristearate (Span-65, Atlas/ICI), sorbitan monoisostearate (Crill 6, Croda), and sorbitan sesquistearate (Nikkol SS-15, Nikko). Amphipathic compounds according to the invention may include one or more of the sorbitan fatty acid esters above.

Esters of lower alcohols (C2 to C4) and fatty acids (C 8 to C 18) are suitable surfactants for use in the invention. Examples of these surfactants include: ethyl oleate (Crodamol EO, Croda), isopropyl myristate (Crodamol IPM, Croda), isopropyl palmitate (Crodamol IPP, Croda), ethyl linoleate (Nikkol VF-E, Nikko), and isopropyl linoleate (Nikkol VF-IP, Nikko). Amphipathic compounds according to the invention may include one or more of the lower alcohol fatty acid esters above.

In addition, ionic surfactants may be used as hydrophobic moieties amphipathic compounds of the invention. Examples of useful ionic surfactants include: sodium caproate, sodium caprylate, sodium caprate, sodium laurate, sodium myristate, sodium myristolate, sodium palmitate, sodium palmitoleate, sodium oleate, sodium ricinoleate, sodium linoleate, sodium linolenate, sodium stearate, sodium lauryl sulfate (dodecyl), sodium tetradecyl sulfate, sodium lauryl sarcosinate, sodium dioctyl sulfosuccinate, sodium cholate, sodium taurocholate, sodium glycocholate, sodium deόxycholate, sodium taurodeoxycholate, sodium glycodeoxycholate, sodium ursodeoxycholate, sodium chenodeoxycholate, sodium taurochenodeoxycholate, sodium glyco cheno deoxycholate, sodium cholylsarcosinate, sodium N-methyl taurocholate, egg yolk phosphatides,

hydrogenated soy lecithin, dimyristoyl lecithin, lecithin, hydroxylated lecithin, lysophosphatidylcholine, cardiolipin, sphingomyelin, phosphatidylcholine, phosphatidyl ethanolamine, phosphatidic acid, phosphatidyl glycerol, phosphatidyl serine, diethanolamine, phospholipids, polyoxyethylene-10 oleyl ether phosphate, esterification products of fatty alcohols or fatty alcohol ethoxylates, with phosphoric acid or anhydride, ether carboxylates (by oxidation of terminal OH group of, fatty alcohol ethoxylates), succinylated monoglycerides, sodium stearyl fumarate, stearoyl propylene glycol hydrogen succinate, mono/diacetylated tartaric acid esters of mono- and diglycerides, citric acid esters of mono-, diglycerides, glyceryl-lacto esters of fatty acids, acyl lactylates, lactylic esters of fatty acids, sodium stearoyl-2-lactylate, sodium stearoyl lactylate, alginate salts, propylene glycol alginate, ethoxylated alkyl sulfates, alkyl benzene sulfones, α-olefin sulfonates, acyl isethionates, acyl taurates, alkyl glyceryl ether sulfonates, sodium octyl sulfosuccinate, sodium undecylenamideo-MEA-sulfosuccinate, hexadecyl triammonium bromide, decyl trimethyl ammonium bromide, cetyl trimethyl ammonium bromide, dodecyl ammonium chloride, alkyl benzyldimethylammonium salts, diisobutyl phenoxyethoxydimethyl benzylammonium salts, alkylpyridinium salts, betaines (trialkylglycine), lauryl betaine (N-lauryl,N,N-dimethylglycine), and ethoxylated amines (polyoxyethylene-15 coconut amine). For simplicity, typical counterions are provided above. It will be appreciated by one skilled in the art, however, that any bioacceptable counterion may be used. For example, although the fatty acids are shown as sodium salts, other cation counterions can also be used, such as, for example, alkali metal cations or ammonium. Amphipathic compounds according to the invention may include one or more of the ionic surfactants above. Peptides and Proteins

The amphipathic compounds of the invention are useful for the administration of a peptide or protein to a mammal. Proteins are generally defined as consisting of 100 amino acid residues or more; peptides are less than 100 amino acid residues. Unless otherwise stated, the term protein, as used herein, refers to both proteins and peptides.

The proteins may be produced, for example, by isolation from natural sources, recombinantly, or through peptide synthesis. Examples include growth hormones, such as human growth hormone and bovine growth hormone; enzymes, such as DNase, proteases, urate oxidase, alronidase, alpha galactosidase, and alpha glucosidase; antibodies, such as trastuzumab (Genentech), oprelvekin (Genetics Institute), muromonab-CD3 (Ortho Biotech), infliximab (Centocor), abciximab (Eli Lilly), ritiximab (Genentech), basiliximab (Novartis), palivizumab (Medlmmune), thymocyte globulin (SangStat), cetuximab (ImClone), and daclizumab (Hoffman-La Roche); poetins, such as erythropoietin (e.g., epoetin, Amgen) and thrombopoietin; cytokines, such as TNF-alpha; interferons, such as interferon alpha and interferon beta; angiogenic factors; growth factors, including vascular endothelial growth factor (VEGF), endothelial cell growth factor (ECGF), epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), and platelet derived growth factor (PDGF); clotting factors, such as factor IV, factor VIII, and factor Vila; thyrotropin alfa; tissue plasminogen activator; glucocere- brosidase; etanercept (Immunex); pegademase bovine (Enzon); colony stimulating factor (GM-CSF); follicle-stimulating hormone (FSH); luteinizing hormone (LH); prolactin; relaxin; somatotropin-releasing hormones; tachykinins; thyroid-stimulating hormone (TSH); differentiation factors; colony-stimulating factors; ceredase; gibberellins; auxins; rhIGF-I/rhIGFBP-3 (the recombinant protein complex of insulin-like growth factor-I (IGF-I) and its most abundant binding protein, insulin like growth factor binding protein- 3 (IGFBP-3));and analogs thereof. The BAS can be a trinectin, a protein binding scaffold based on a domain of a naturally occurring plasma protein called fibronectin.

Exemplary peptides that can be used in the compositions and methods of the invention include, without limitation, adrenocorticotropic hormone (ACTH), β- amyloid(l-40), agouti peptide, agouti-related peptide, anaphylatoxins, CASH (Cortical Androgen-Stimulating Hormone), diabetes associated peptide, gliadorphin, insulin, α- & β-lactorphin, g-melanocyte stimulating hormone-like peptide, neuropeptide P, peptide histidine isoleucine (PHI), collagenase-1 and stromelysin-1 inhibitors (including those

descsribed in U.S. Patent Nos. 5,932,579, 5,929,278, and 5,840,698), erythropoietin peptide agonists (including those described in U.S. Patent Nos. 5,986,047, 5,830,851, 5,773,569), follicle stimulating hormone antagonists (including those described in U.S. Patent No.6,426,357), human neutrophil elastase inhibitors (including those described in U.S. Patent No. 5,663,143, PCT Publication No. WO 03/066824, WO 92/15605, and WO 96/20278, and European Patent No. 132593 IAl), kallikrein inhibitors (including those described in U.S. Patent Nos. 6,333,402, 6,057,287, 5,994,125, and 5,795,865), selectin binding peptides (including those described in U.S. Patent Nos. 5,728,802, 5,648,458, and 5,643,873), all of the peptides listed in Table 1, and analogs thereof. Exemplary commercially available peptides and their analogs are listed in Table 1, followed by their respective BACHEM catalogue number.

Table 1 Pe tides

Table 1 continued

Table 1 continued

Table 1 continued

Table 1 continued

Table 1 continued

Table 1 continued

Table 1 continued α-Endorphin (H-2695) β-Endorphin (H-2700) Acetyl-β-Endoφhin (H-1115)

Endorphins β-Endoφhin (6-31) (H-4024) β-Endorphin (18-31) (H-5686) β-Endorphin (27-31) (H-5170) β-Endorphin (30-31) (G-2080) δ-Endorphin (H-2710) γ-Endorphin (H-2725)

Endothelin Azepane- 1 -carbonyl-Leu-D-Trp(For)-D-Trp-OH (H-4914) Antagonists Cyclo(-D-Asp-Pro-D-Ile-Leu-D-Trp) (H-3008)

Cyclo(-D-Glu-Ala-D-allo-Ile-Leu-D-Trp) (H-8405)

Cyclo(-D-Ser-Pro-D-Val-Leu-D-Tφ) (H-3064)

Cyclo(-D-Tφ-D-Asp-Pro-D-Val-Leu) (H-1252)

N-cis-2,6-Dimethylpiperidinocarbonyl-b-tBu-Ala-D-Tφ(l-me thoxycarbonyl)-D-

NIe-OH (H-2492)

Endothelin-1 (11-21) (H-1658)

Acetyl-(D-Tφ' ⁶ )-Endothelin-1 (16-21) (H-8850)

Dynoφhin A (1-7) (H-2660) (Phe ⁷ )-Dynoφhin A (1-7) (H-5150)

Enkephalins and (Phe ⁷ )-Dynoφhin A (1-7) amide (H-5155) Proenkephalins Dynoφhin A (1-6) (H-2665) Dynoφhin A (1-13) (H-2625) Gluten Exoφhin B5 (H- 1666) Leu-Enkephalin (H-2740) Leu-Enkephalin (sulfated) (H-2760) (Ala ² )-Leu-Enkephalin (H- 1276) (D-Ala ² )-Leu-Enkephalin (H-2750) (Des-Tyr^-Leu-Enkephalin (N-1175) (3,5-Dibromo-Tyr ¹ )-Leu-Enkephalin (H-2575) Boc-Leu-Enkephalin (A-2440) Leu-Enkephalin amide (H-2745) (D-Ala ² )-Leu-Enkephalin amide (H-2755) (D-Ala ² )-Leu-Enkephalin-Arg (H-3276) (Boc-Tyr ¹ ,D-Ala ² )-Leu-Enkephalin-Lys (A-2435) Leu-Enkephalin-Lys (H-2765)

(D-Cys(tBu) ² ,Thr(tBu) ⁶ )-Leu-Enkephalin-Thr (H-8170) (3,5-Diiodo-Tyr ¹ ,D-Thr ² )-Leu-Enkephalin-Thr (H-2615) (D-Ser ² )-Leu-Enkephalin-Thr (H-2770) (D-Thr ² )-Leu-Enkephalin-Thr (H-2775) Met-Enkephalin (H-2785) (Des-Tyr')-Met-Enkephalin (N-1180) (Gly°)-Met-Enkephalin (H-2850) (Met(O) ⁵ )-Enkephalin (H-5160)

Table 1 continued

ACEP-I (H- 1646)

Ac-Phe-Nle-Arg-Phe-NH ₂ (H- 1055)

AF-I (H-3338)

FMRFamide AF-2 (H- 1642) Peptides H-Asn-Arg-Asn-Phe-Leu-Arg-Phe-NH ₂ (H-1364)

H-Asp-Arg-Asn-Phe-Leu-Arg-Phe-NH ₂ (H-1362)

H-Leu-Ser-Ser-Phe-Val-Arg-Ile-NH ₂ (H-1644)

H-Met-Arg-Phe-OH (H-2965)

Met-Enkephalin-Arg-Phe (H-2830)

Met-Enkephalin-Arg-Phe amide (H-2835)

H-Nle-Arg-Phe-NH ₂ (H-2970)

Orphan GPCR SP9155 Agonist P518 (H-5984)

H-Phe-Leu-Arg-Phe-NH ₂ (H-2985)

H-Phe-Met-Arg-Phe-NH ₂ (H-2975)

H-D-Phe-Met-Arg-Phe-NH ₂ (H-3346)

H-Phe-D-Met-Arg-Phe-NH ₂ (H-3344)

H-Phe-Met-Arg-D-Phe-NH ₂ (H-3342)

H-Phe-Met-D-Arg-Phe-NH ₂ (H-7185)

H-Pro-Asp-Val-Asp-His-Val-Phe-Leu-Arg-Phe-NH ₂ (H-8040)

Pyr-Asp-Pro-Phe-Leu-Arg-Phe-NH ₂ (H-9260)

SCPA (H-6925 )

SCPB (H-3005 )

H-Thr-Asn-Arg-Asn-Phe-Leu-Arg-Phe-NH ₂ (H-9265)

H-Trp-Nle-Arg-Phe-NH ₂ (H-3000)

H-Tyr-Phe-Met-Arg-Phe-NH ₂ (H-2980)

Galanins and Galanin (H-8230)

Galanin Message (Abz-GlyVGalanin (l-lO)-Lys(retro-m-nitro-Tyr-H) amide (M-2365) Associated Galanin (1-13)-Bradykinin (2-9) amide (H-1346) Peptides (GMAP) Galanin (1-13)-Mastoparan (H-4188)

Galanin (1-13)-Neuropeptide Y (25-36) amide (H-3374)

Galanin (l-13)-Pro-Pro-(Ala-Leu-)2Ala amide (H-2576)

Galanin (1-13)-Spantide I (H-2578)

Galanin (1-13)-Substance P (5-11) amide (H-1312)

(Ala ⁶ ,D-Tφ ⁸ ,L-alaninol ¹⁵ )-Galanin (1-15) (H-4066)

(D-Thr ⁶ ,D-Trp ^s ' ⁹ ,L-alaninol ^I5 )-Galanin (1-15) (H-1576)

Galanin (1-19) (H-5754)

(D-Trp ² )-Galanin (1-29) (H-4122)

Galanin Message Associated Peptide (1-41) amide (H-6780)

Galanin Message Associated Peptide (1-41) amide (H-7615)

Galanin Message Associated Peptide (16-41) amide (H-9725)

Galanin Message Associated Peptide (25-41) amide (H-9520)

Galanin Message Associated Peptide (44-59) amide (H-7715)

Galnon (B-3645)

Table 1 continued

Table 1 continued

Table 1 continued

able 1 continued

Table 1 continued

Table 1 continued

Table 1 continued

Table 1 continued

Table 1 continued

Table 1 continued

(Ala ^2S5 )-Calmodulin-Dependent Protein Kinase II (281-302) (H-3246)

Calmodulin-Dependent Protein Kinase II (281-309) (H-3254)

Protein Kinase Calmodulin-Dependent Protein Kinase II (290-309) (H-9365) Related Peptides cAMP-Dependent Protein Kinase Inhibitor-α (5-22) amide (H-3222) (cont.) cAMP-Dependent Protein Kinase Inhibitor-α (5-24) (H-5950)

Cyclo(-Gly-Tyr(PO ₃ H ₂ )-Val-Pro-Met-Leu) (H-2062)

Ephrin-A2-Selective YSA-Peptide (H-5894 )

H-Gln-Arg-Arg-Gln-Arg-Lys-Ser-Arg-Arg-Thr-Ile-OH (H-9685)

H-Gly-Arg-Gly-Leu-Ser-Leu-Ser-Arg-OH (H-7405)

H-Gly-Ile-2-Nal-Trp-His-His-Tyr-OH (H-4084)

(Cys°)-GTP-Binding Protein Gsa (28-42) (H-5788 )

Hl-7 (H-1805 )

Kemptamide (M-2505 )

Kemptide (M-1510 )

(Trp>Kemptide (M-1525 )

(Val ⁶ ,Ala ⁷ )-Kemptide (M-1515 )

H-Leu-Arg-Arg-Arg-Arg-Phe-D-Ala-Phe-Cys(NPys)-NH ₂ (H-3696)

H-Lys-Arg-Glu-Leu-Val-Glu-Pro-Leu-Thr-Pro-Ser-Gly-Glu-Ala -Pro-Asn-Gln-Ala-

Leu-Leu-Arg-OH (H-3242)

H-Lys-Arg-Thr-Leu-Arg-OH (M- 1945)

H-Lys-Lys-Arg-Ala-Ala-Arg-Ala-Thr-Ser-Asn-Val-Phe-AIa-NH ₂ (H-3252)

Malantide (H-3262 )

MAPKK2 (1-16) (H-5778 )

Myelin Basic Protein (4-14) (H- 1072)

Acetyl-(Gln ⁴ )-Myelin Basic Protein (4-14) (H-3238)

Myristoyl-Arg-Lys-Arg-Thr-Leu-Arg-Arg-Leu-OH (N-1310)

Myristoyl-Lys- Arg-Thr-Leu-Arg-OH (N- 1305)

Myristoyl-Phe-Ala-Arg-Lys-Gly-Ala-Leu-Arg-Gln-OH (N- 1370)

Neurogranin (28-43) (H-1554 ) p60 v-src (137-157) (H-8535 )

Peptide ε (H-3236 )

H-Phe-Ala-Arg-Lys-Gly-Ala-Leu-Arg-Gln-OH (N-1375)

H-Phe-Lys-Lys-Ser-Phe-Lys-Leu-NH ₂ (H- 1638)

Phosphorylase Kinase β-Subunit Fragment (420-436) (H-1968 )

PKI-tide (H-3234) pp60 c-src (521-533) (H-3256 ) pp60 c-src (521-533) (phosphorylated) (H-3258 )

H-Pro-Leu-Ser-Arg-Thr-Leu-Ser-Val-Ala-Ala-Lys-Lys-OH (H-9375)

Protein Kinase C (19-31) (H-3232)

(Ser25)-Protein Kinase C (19-31) (H-3286)

Protein Kinase C (19-36) (H-9370)

Protein Kinase C (530-558) (H-8045)

S6 Phosphate Acceptor Peptide (H-9380 )

Syntide 2 (H-9385 )

Table 1 continued

Table 1 continued

Table 1 continued

Table 1 continued

Miscellaneous Buccalin (H-9235) Peptides Bursin (H-5920)

Chromostatin (H-8475)

Corticostatin (H-9045)

Dermaseptin (H- 1294)

Diazepam Binding Inhibitor (DBI) (H-6760)

Elcatonin (H-2247)

Enterostatin (H-6405)

Epidermal Mitosis Inhibiting Pentapeptide (H-6770)

Follicular Gonadotropin-Releasing Peptide (H-6775)

Gastric Inhibitory Polypeptide (H-5645)

Granuliberin-R (H-6800)

Seminal Plasma Inhibin (67-94) (H- 1602) (also known as β-inhibin)

Kentsin (H-3840) (also known as Contraceptive Tetrapeptide)

Magainin I (H-6565)

Magainin II (H-6570)

Metorphamide (H-6855) β-Neuroprotectin (N- 1340)

Pancreastatin (33-49) (H-5905)

Pancreastatin (H-6165)

Proctolin (N-1015)

Rigin (H-6920)

Systemin (H-8675)

Thyroid releasing hormone (TRH) (H-4915), (also known as Protirelin)

Urotensin II (H-4768)

The specific peptides listed for each class provided in Table 1 are intended to be exemplary. The invention can be applied as well to any peptide falling within the general classifications above (e.g., any peptide or peptide analog sharing affinity for the same molecular target and administered for the same therapeutic purpose(s)). Skin Active Peptides

The peptide or protein delivered using the compositions and methods of the invention can be skin-active peptides or proteins, such as PTH, PTHrP, and melanogenosis-inhibiting peptideselected from Glu-Asp-Tyr-His-Ser-Leu-Tyr-Gln-Ser- His-Leu (SEQ ID NO. 1), Ser-Gly-Gly-Tyr-Leu-Pro-Pro-Leu (SEQ ID NO. 2), His-Ser- C(O)-NH-(CH2)k-CH2-C(O)-NH-Ser-His, wherein k is 2-10 (SEQ ID NO. 3); the hair growth suppressor SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNF (PTHrP (1- 34)) (SEQ ID NO. 4); the hair growth stimulator LMHNLGKHLNSMERVEWLRKKLQDVHNF (PTHrP (7-34)) (SEQ ID NO.5); epidermial growth factor (EGF); melanocyte stimulating hormone (MSH); corticotropin-

releasing hormone (CRH); tumor necrosis factor-related apoptosis-inducing ligand (TRAIL); or fibroblast growth factor (FGF). Formulation

The invention provides amphipathic compounds for the formulation of peptides and proteins. Topical formulations of the invention can take a variety of forms, including, without limitation, creams, foams, pastes, lotions, gels, sticks, sprays, patches, and ointments. Nasal formulations and formulations for inhalation will typically take the form of a fine powder or solution. The pharmaceutical compositions may be formulated according to conventional pharmaceutical practice (see, e.g., Remington: The Science and Practice of Pharmacy, 20th edition, 2000, ed. A.R. Gennaro, Lippincott Williams & Wilkins, Philadelphia, and Encyclopedia of Pharmaceutical Technology, eds. J. Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York).

The peptide or protein and amphipathic compound of the invention may be mixed under sterile conditions with a pharmaceutically-acceptable carrier, and with any preservatives, buffers, or propellants which may be required. Any conventional pharmacologically and cosmetically acceptable vehicles may be used. For example, the compounds may also be administered in liposomal formulations that further enhance skin penetration. Such liposomal formulations are described in U.S. Patent Nos. 5,169,637; 5,000,958; 5,049,388; 4,975,282; 5,194,266; 5,023,087; 5,688,525; 5,874,104; 5,409,704; 5,552,155; 5,356,633; 5,032,582; 4,994,213; and PCT Publication No. WO 96/40061. Examples of other appropriate vehicles are described in U.S. Patent No. 4,877,805 and EP Publication No. 0586106A1. Suitable vehicles of the invention may also include mineral oil, petrolatum, polydecene, stearic acid, isopropyl myristate, polyoxyl 40 stearate, stearyl alcohol, or vegetable oil.

The formulations can include various conventional colorants, fragrances, thickeners (e.g., xanthan gum), preservatives, emollients (e.g., hydrocarbon oils, waxes, or silicones), demulcents, solubilizing excipients, dispersants, penetration enhancers, plasticizing agents, preservatives, stabilizers, demulsifiers, wetting agents, emulsifϊers,

moisturizers, astringents, deodorants, and the like can be added to provide additional benefits and improve the feel and/or appearance of the topical preparation.

The ointments, pastes, creams and gels may further include excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.

When the formulation takes the form of a powder or spray, the formulation may further contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.

Transdermal patches can be used with the added advantage of providing controlled delivery of the peptide or protein being administered. For example, a rate controlling membrane, polymer matrix, or gel can be used to control the rate flux of peptide or protein across the skin.

It is preferred that the peptide or protein is brought in contact with the lipidated saccharide at a slightly basic pH (8-9) for a brief time such that the N-glycoside or reversible Schiff s base is formed. The formulation is readjusted to neutral pH using a buffer or acid. Care is taken when the saccharide is an aldohexose so that Amadori rearrangement can be prevented by keeping the temperature lower than 37 ⁰ C, or avoiding transition metal ions and deliberately adding sodium chloride. See, for example, Kwak et al., Amino Acids 27:85 (2004) and Robert, et al., Challenges in Taste Chemistry and Biology ACS Symposium 867:195 (2004); Herminia et al., J Phys. Org. Chem. 18:183 (2005), and Sara et al., Food Chemistry 90:257 (2005)). Therapy

The compounds of the invention can be used to treat any disease or condition for which the unconjugated partent peptide/protein can be used. Typically, the compounds of the invention are administered at levels known to be efficacious for the parent

peptide/protein, or less.

Diseases which can be treated using peptides and proteins include, for example, Melanoma, Psoriasis, Acne, Actinic keratosis, vitiligo, Skin Hyperpigmentation, Acne, Actinic Keratosis, Albinism, Baldness, Hair loss, Behcet's Syndrome, Burns, Skin rejuvenation, Skin Antiaging, Wound Healing, Eczema, Scabies, Cholesteatoma, Dermatitis Herpetiformis, Ectodermal Dysplasia, Epidermolysis Bullosa, Erythema Multiforme, Gustatory Sweating, Head Lice, Hidradenitis Suppurativa, Hives, Hirsuitism, Lupus, Nail Patella Syndrome, Pemphigoid, Pemphigus, Rosacea, Scleroderma, Seborrheic Dermatitis, Skin Cancer, Solar Keratosis, Swimmer's Itch, Warts, Xeroderma and Pigmentosum.

The following examples are to illustrate the invention. They are not meant to limit the invention in any way.

Example 1. Diacetonide Protection of Saccharides. Preparation of protected glycosyl donors with one free hydroxyl group for phosphorylation: IA. Preparation of 1 -α or β-Hydroxy-2,3,4,6-tetra-O-acetyl β-D-glucopyranoside:

Dimethlamine gas was generated from commercially available 30% aqueous solution by adding into sodium hydroxide pellets and was added to penta-acetyl glucose prepared as above. Thus in a 2 necked RB flask containing lOOg of NaOH pellets was added 200 ml of 30% dimethyl amine solution slowly through dropping funnel. The liberated dimethylamine gas was dissolved using Teflon tubing to another 2L jacketed reactor cooled to -2O ⁰ C containing 800 ml of acetonitrile. After the acetonitrile solution attains PH of 9-11, glucose pentaacetate (65 g) was added and stirred for 15 minutes at - 15°C. TLC monitoring of the reaction was done for completion of the reaction. Acetonitrile and excess dimethylamine gas were removed quickly by rotory evaporator. The resulting paste was dissolved in 500 mL ethylacetate and washed once with 200 mL of water. The organic portion was dried over magnesium sulfate and evaporated. The mixture was pure enough for further use in phosphorylation reactions.

IB. Preparation of 1- β-Hydroxy-2,3,4,6-tetra-O-acetyl-β-D-glucopyranoside: l-β-Acetyl-penta-O-acetyl-glucopyranoside was obtained from Sigma. A methanolic solution of 1-β-acetate was heated to 4O ⁰ C with imidazole (1 molar equivalent) for 26 hours and TLC monitored for the completion of the anomeric deacetylation. Methanol was removed and after water addition and extraction with ethylacetate followed by 1 N hydrochloric acid wash gave exclusive beta hydroxyl anomer in almost quantitative manner. 1 C. Preparation of 1 ,2:5,6-Di-O-cyclohexylidene-α-D-glucofuranose: lOOOg (1050 ml, 10 mol) of redistilled cyclohexanone was added to a 3 L cooled to 0 ^° C. 62.5ml of concentrated sulphuric acid was added slowly into the vigorously stirred cyclohexanone. 45Og (2.5 mol) of finely powdered dried α-D-glucose was added slowly. The jacketed reactor was allowed to reach ambient temperature with continual stirring for over a period of 8 hours. The reaction mixture becomes progressively more viscous and finally sets into a solid off-white crystalline mass. The crystalline mass was agitated with 750 ml of heptane and a solution of 124g of sodium carbonate in 375ml of water at 7O ⁰ C. Heptane layer was allowed to perculate by gravity into a Erlenmayer flask. Upon cooling the heptane layer, the crystals were filtered and the heptane was resubjected to the residue similarly. The purified l,2:5,6-di-O-cyclohexylidene-α-D- glucofuranose (once again crystallized from heptane) has m.p. 131-132°C, [α]20D -2.2° (c 1.8 in EtOH); the yield is 380g (47%). ID. Preparation of l,2:4,5-Di-O-cyclohexylidene-D-fructopyranose:

20Og (1.11 mol) of finely powdered dry D-fructose and 419g (440 ml, 4.49 mol) of ice-cooled cyclohexanone containing 30 ml of concentrated sulphuric acid in a jacketed reactor was stirred as in 3 and the reaction mixture becomes solid within 30 minutes. The mixture was left overnight at room temperature and the product was dissolved in 500ml of chloroform. The organic layer was washed with dilute aqueous sodium hydroxide, dilute hydrochloric acid and water and finally dried and evaporated. The residue has m.p. 145-156 ^° C, [α]20D -133.5°(c 1 in CHC13). The yield is 142g

(37%). 1 E. Preparation of 1 ,2 : 5,6-Di-O-isopropylidene-α-D-glucofuranose :

A suspension of 15Og (0/83 mol) of dry D-glucose, 12Og (0.83 mol) of anhydrous zinc chloride and 7.5g of phosphoric acid (88% v/v) in 1 litre of dry acetone was stirred at ambient temperature for 30 hours. Unchanged glucose was removed by filtration, and inorganic salts were precipitated by the addition of a solution of 58g of sodium hydroxide in 85ml of water. The resulting suspension was filtered, the residue washed with acetone and the acetone layer evaporated. The mass was dissolved in 200ml of water and extracted with five 100-ml portions of dichloromethane. The organic phase was dried and evaporated on rotary evaporator. Recrystallisation from light petroleum (b.p. 80-100°C) gave 7Og of product, m.p. 109-110°C, [α]20D -18.5° (c 5 in H2O). IF. Preparation of l,2:5,6-Di-O-isopropylidene-D-mannitol:

To a solution of 6Og of zinc chloride in 300ml of acetone was added 1Og of finely powdered D-mannitol. The mixture was stirred vigorously at 20°C until clear solution (2- 3 hours) and was allowed to stand for further 16 hours. The reaction mixture was then poured into a solution of 7Og of potassium carbonate in 70ml of water and 300ml of ether. The product was filtered and 100ml of 1:1 acetone-ether solution was used to wash the filtrate. The combined filtrates was evaporated to dryness on a rotary evaporator. The dry residue was successively extracted with five 250-ml portions of boiling light petroleum (b.p. 60-80°C) and the combined filtrates cooled to give the ρroduct,7.9g (55%), having m.p.l 19°C. 1 G. Preparation of 1 ,2:4,5-O-Diisopropylidene-D-galactopyranose:

To a suspension of acetone and cupric chloride (catalyst) was added galactose and the solution refluxed for 8 hours upon which time galactose was mostly dissolved. The reaction mixture was stirred for a further period of 10 hours at ambient temperature. Upon removal of acetone and extraction with ethylacetate and an aqueous wash afforded the diisopropylidene derivative in 80% yield. The product upon LC-MS examination gave molecular ion as sodium adduct 282 M ⁺ .

IH. Preparation of 1,2:4,5-O-Dicyclohexylidene- D-galactopyranose:

Instead of acetone cyclohexanone was used similar to example 7. The product was isolated after cyclohexanone removal (yield 72%) as above. Example 2. General Procedure for the glycophosphorylation of substrates: Method 1. H-Phosphonic acid of sugar moieties. 2 A. Preparation of H-Phosphonic acid derivatives of sugar moieties:

Protected sugar as in examples 1-8 (10 mMoles) was dissolved in dry Toluene (50 mL) and freshly distilled phosphorous trichloride (25 mMoles) was added at room temperature and the mixture was cooled to 10°C under inert atmosphere. A solution of triethylamine (20 mMoles) in dry Toluene (2OmL) was added over a period of 30 minutes to the cooled and stirred mixture. The mixture was stirred and monitored for the complete disappearance of the starting material. Precipitated triethylamine hydrochloride was removed by filtration and water (1OmL) was added and sufficient amount of sodium acetate was added when cold to neutralize the pH to about 5. The toluene layer was evaporated. Upon evaporation of the solvent, the product was obtained as a gummy solid and was used immediately for the next reaction without any purification. Direction injection using LC-MS (Applied Biosystems) gave a strong negative ion spectrum as expected. The TLC after work-up also showed most of the starting material was consumed and a more polar product same as the one monitored from the reaction mixture was obtained indicating that the dichloride can be hydrolyzed readily to the H- Phosphonic acid. (The product was more water soluble with the diacetonides and the tetraacetates and thus ethyl acetate was substituted for the aqueous extraction).

Thus dicyclohexylidene derivative obtained from fructose (see below) gave a gummy product in 85% isolated yield. LC-MS showed a strong negative molecular ion at 393 amu.

Method 2: H-Phosphonic acid of aglycons.

2B. The aglycon (10 mMole) and phosphorous trichloride (25 mMol) in dry THF (75 mL) was stirred and cooled to 10°C and a solution of triethylamine (25 mMol) in dry

THF (25mL) was added over a period of 1 hour when cold. TLC monitoring of the reaction mixture maintained at room temperature showed a polar product. The reaction mixture was stirred for over 2 hours for the complete conversion. The product was isolated after addition of water and pH neutralization (5-6.5) using sodium bicarbonate, extraction with ethylacetate (3 times 50 mL). Organic portion was dried over magnesium sulfate and evaporated on a rotory evaporator to obtain the H-Phosphonic acid derivative which was pure enough for subsequent conjugation with the free sugar hydroxyl group. 2C. Preparation of Tocopherol-H-Phosphonic acid:

Imidazole (11.2g; 7 equivalent), THF (100ml) and PC13 (12mL) were mixed at O ⁰ C and stirred. Vitamin E (1Og) in 60 mL THF containing triethylamine (8.2g) was added over 20 minutes. After workup with water, ethylacetate extraction (3 times 100 mL), sodium sulfate drying and evaporation gave 14g of tocopherol-H- phosphonic acid. The product was purified by silica gel column as below.

2D. Preparation of Tocopherol-H-Phosphonic acid (without imidazolidated phosphorous):

In a 1 litre flask 200ml of THF under argon was cooled to O ⁰ C. 17.5ml of PCl ₃ was added to the above stirred mixture. 20g of Vitamin E, 45 ml of triethylamine and 50 ml of THF were combined and added slowly during 30 minutes to the tocopherol mixture. The reaction mass was warmed to 25° C and stirred for 1 h. The reaction mixture was added to 10% hydrochloric acid (5OmL) and stirred for 5 minutes. The THF layer was separated and the aqueous layer was further extracted with ethylacetate (200 ml x 3). The organic portion was combined and evaporated to afford 22 g of crude product. The product was purified by silica gel Column. Methanol-ethylacetate mixture gave pure product (16g; isolated; 69% yield). RF value: 0.25 in dichloromethane methanol (85:15). LC-MS showed the molecular ion peak at m/z 493.

2E. Coupling of H-Phosphonic acid sugar derivative with aglycon: General procedure:

H-Phosphonic acid derivative of the protected sugar was dried thoroughly by a high vacuum pump and 0.8 molar equivalent of the aglycon containing the free hydroxyl group was added in dry pyridine (10 mL per each millimole of sugar-H-Phosphonic acid). The resulting mixture was cooled to 0°C and 4 molar equivalents of pivolyl chloride was added. The mixture was stirred for a further period of 60 minutes at 0°C and 60 minutes at room temperature. To this mixture was added 1 equivalent of iodine dissolved in pyridine (or 3 equivalents of freshly prepared peracetic acid). The mixture was stirred for 10 minutes at room temperature and partitioned between water and ethylacetate. The organic portion was evaporated and purified through silica gel to obtain the clean glycophosphorylated material. Depending upon the protecting group, the deprotection is done. Ammonia gas was used for deprotecting the acetylated glycoses. Diketals were deprotected in refluxingj N hydrochloric acid/acetone mixture in almost quantitative yield.

2F. Coupling of Vitamin E H Phosphonate with 2,3,4,6-tetra-O-acetyl β-D- glucopyranoside:

In a 500ml flame dried round bottomed Flask 23.0 g of H-Phosphonate of Vitamin E and 24.35 g of 2,3,4,6- tetra-O-acetate glucopyranoside were taken. 50 ml of toulene was added and connected to the high vacuum pump for 1.5 h. 200 ml of pyridine was added and the mixture cooled to 0°C. 23.7 ml of Pivaloyl chloride was added and after 10 minutes at O ⁰ C , keep the Round Bottomed flask at room temperature and stirred for 1 hour. After 1 h 2 Ig of I ₂ in 50ml of pyridine- water mixture(9:l) was added and stirred for 45 mins.. 15% sodium thiosulphate; 300ml) was added and the mixture extracted with EtOAC (300ml x 3), organic portion dried and concentrated to afford56 g of crude product. TLC monitoring showed rf value of 0.35, DCMMeOH (85:15). LC- MS showed the molecular ions peak at m/z 839. 2G. Deacetylation of Vitamin E Glycophosphonate:

In a 1 litre flask 56g of the above crude product was dissolved in 200 ml of MeOH and 150 ml of 25% NH ₃ solution were added. The mixture was heated to 65°C and

maintained for about two hours during which time the TLC showed complete deacetylation. The mixture was evaporated to dryness under vacuum. The desired product was partitioned was obtained after a silica gel column separation using methanol- ethylacetate mixture in about 78% yield (24 g). LC-MS showed the molecular ions at m/z 671. HPLC examination showed that the product is a mixture of anomeric isomers. Example 3. Preparation of l,2:5,6-Diisopropylidene-3-O-Phosphoryl-tocopherol:

The procedure was similar to as in example F. Instead of tetra-O-acetyl glucopyranose, diketalized glucofuranose was used.

Dekatalization of l,2:5,6-Diisopropylidene-3-0-Phosphoryl-tocopherol:

IN Hydrochloric acid (5mL per mMol) in acetone was refluxed till the completion of the reaction. Trifluroacetic acid can be replaced for hydrochloric acid for room temperature dekatalization. LC-MS showed the molecular ions at m/z 671 and the product was obtained as a crystalline solid.

Example 4. Amide Derivatives of Glucosamine in General.

Retinoic acid amidated with glucosamine

Glucosamine hydrochloride (18gm) was dissolved in water (10Og) and sodium hydroxide (8gm) was added. The mixture was cooled to O ⁰ C and lauroyl chloride (22gm) was added in portions over 1 hour period. The solidified product was filtered and washed once with water, acetone and dried to afford clean material (34gms).

Example 5. Exemplary Compounds of the Invention.

The following compounds can be useful in the methods of the invention:

Glycophosphorylated ascorbic acid derivatives:

Phosphoglycosylated ursolic acid using 6-OH position of glucose.

Phosphoglycosylated retinoic acid using fructose.

Retinoic ester with fructose Kojic Acid linked to glucose at 6 position with azalaic acid as linker:

Example VI. Preparation of 3-O-n-Octo-phosphoryl-l,2:4,6-Dicyclohexylidene- glucofuranose:

In a roundbottomed flask (250 mL) was added 1,2: 4,6-dicyclohexylidene-H- Phosphonic acid (5 grams; 0.012 mole) and n.Octanol (3.2 grams; 0.024 mole) and was

dried through the Toulene (2x15 mL) times through vacuum pump to ensure complete dryness of reactants. The dried material was dissolved in dry Pyridine (25 mL) and chilled to 0°C. Pivaloyl Chloride (4.5 mL; 3 equivalents) was added at 0°C and after 5 minutes warmed to room temperature and stirred for 1 hour. After one hour at room temperature, iodine (8 grams; 2 equivalents in 20 mL of pyridine:water, 9:1) at 0°C was added and the mixture warmed to room temperature and stirred for 30 minutes more. The product was isolated after addition of thiosulfate (15 % in water 200 mL) and extracting with ethyl acetate (2x150 mL), dried over sodium sulfate and evaporated to yield the desired product (5.8 grams). LC-MS gave a strong negative molecular ion at 531 amu as expected. Example VII. Preparation of 3-O-n.Octo-phosphoryl-l,2-cyclohexylidene-glucofuranose:

Dicyclohexylidene derivative prepared above (5 grams) was dissolved in acetone (54 mL). 2N hydrochloric acid (6 mL) was added and the mixture refluxed for an hour. ,- An aliquot was taken and neutralized with triethyl amine and the sample was analyzed by LCJvIS which showed strong negative ion at 451 amu. The product was isolated after removing the solvent and lyophilizing the water. Example VIII. Phosphogylcosylated Tocopherol Lotion Containing Peptide or Protein:

A lotion containing phosphogylcosylated tocopherol was prepared using the following excipients: 0.25 part of vitamin E-phosphate-O-fructose; 2.8 parts POE-10- stearyl ether; 1.8 parts cholesterol; 4.94 parts octyl glucoside; 0.1 parts methyl paraben; 3 parts mineral oil; 17 parts coconut oil, and the remainder water. The fat-soluble materials were mixed together and warmed to 8O ⁰ C. The aqueous portion with the dissolved peptide or protein was warmed to 40°C. The aqueous was mixed with the oil portion through a three-way syringe until cooled to near room temperature. Example IX. Animal Study.

The following experiment was done using peptide-BBBL-hgs-1 (SVSEIQLMHNLGKHLNSMERVEWLRKKLQDVHNF, PTHrP (1-34)). 20 meg of peptide-BBBL-hgs-1, a hair growth suppressor, was dissolved per mL of the cream/lotion

described above. 1 niL of the cream was applied on Wistar rats of about 3 square inch area everyday and the total growth suppression of the hair was noted during a 15 day trial. The results show that there is hair growth suppression (picture 1) versus without the skin penetration enhancer (picture 2) clearly demonstrating that the skin permeation is possible for the peptide-BBBL-hgs-1. Other Embodiments

All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each independent publication or patent application was specifically and individually indicated to be incorporated by reference.

While the invention has been described in connection with specific embodiments thereof, it will be understood that it is capable of further modifications and this application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the . present disclosure that come within known or customary practice within the art to which the invention pertains and may be applied to the essential features hereinbefore set forth, and follows in the scope of the claims.

Other embodiments are within the claims.