A peptide composition and uses thereof

Abstract

Disclosed herein are compositions / pharmaceutical compositions and kits comprising a peptide and a stabilizer; wherein the peptide comprises an ester and is between 2 and 10 amino acids long; and wherein the stabilizer is selected from: a diol, a polyol, a mono-saccharide, a di- saccharide, an oligo-saccharide, and a poly-saccharide, including any combination thereof.

Description

A PEPTIDE COMPOSITION AND USES THEREOFCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of priority of IL Patent Application No. 304603, filed on July 19, 2023. The contents of the above application are all incorporated by reference as if fully set forth herein in their entirety.FIELD OF THE INVENTION

[0002] The present invention relates generally to the field of peptide-based formulation and is further directed to solid or aqueous compositions comprising a peptide and a stabilizer.BACKGROUND OF THE INVENTION

[0003] The utilization of peptides as therapeutically active ingredients in pharmaceutical therapy has been emerging, since they can facilitate targeting intracellular protein-protein interaction, which is almost impossible using small molecule-based drugs.

[0004] However, aqueous peptide solutions have limited solution stability, especially solutions containing peptides with a limited water solubility. In most cases, aqueous peptide solutions are only stable for up to 24 hours at room temperature.

[0005] Therefore, there is an unmet need for improving stability of aqueous solutions containing poorly water-soluble peptides.SUMMARY OF THE INVENTION

[0006] In one aspect of the invention, there is provided a composition comprising: (i) a peptide, including any enantiomer thereof, a salt thereof, a hydrate or a solvate thereof, or any combination thereof; and (ii) a stabilizer; wherein: the peptide is between 2 and 10 amino acids long; the peptide comprises an ester; and wherein the stabilizer is selected from: a diol, a polyol, a mono-saccharide, a di-saccharide, an oligo-saccharide, and a polysaccharide, including any combination thereof.

[0007] In some embodiments, the ester is a carboxy terminal ester.

[0008] In some embodiments, the composition is an aqueous solution.

[0009] In some embodiments, the aqueous solution comprises citric acid and / or citrate salt.

[0010] In some embodiments, the peptide is present at a concentration of up to 40 weight % within the aqueous solution.

[0011] In some embodiments, the composition is stable at a temperature between 2 and 40 °C, for at least 72h.

[0012] In some embodiments, the peptide and the stabilizer are present within the composition at a molar ratio of between 1:1 and 10:1.

[0013] In some embodiments, the peptide comprises a structure represented by Formula 1:, wherein R is C1-C5 alkyl.

[0014] In some embodiments, the peptide is L-Leucyl-L-Leucine ester, a salt thereof, a hydrate or a solvate thereof, or any combination thereof.

[0015] In some embodiments, R is methyl.

[0016] In some embodiments, the composition is in a form of a powder and is characterized by a water content of below 5%w / w.

[0017] In another aspect, there is provided a pharmaceutical composition, comprising: (i) a peptide, including any enantiomer thereof, a salt thereof, a hydrate or a solvate thereof, or any combination thereof; (ii) a stabilizer; and (iii) a pharmaceutically acceptable carrier; wherein the peptide is between 2 and 10 amino acids long and the peptide comprises an ester; and wherein the stabilizer is selected from: a diol, a polyol, a mono-saccharide, a disaccharide, an oligo-saccharide, and a poly-saccharide, including any combination thereof.

[0018] In some embodiments, the pharmaceutically acceptable carrier is an aqueous solution.

[0019] In another aspect, there is provided a kit comprising: (i) a peptide, including any enantiomer thereof, a salt thereof, a hydrate or a solvate thereof, or any combination thereof; and (ii) a stabilizer; wherein the peptide is between 2 and 10 amino acids long and the peptide comprises an ester; and wherein the stabilizer is selected from: a diol, a polyol, a mono-saccharide, a di-saccharide, an oligo-saccharide, and a poly-saccharide, and any combination thereof.

[0020] In some embodiments, the peptide and the stabilizer are in a form of a mixture.

[0021] In some embodiments, the peptide and the stabilizer are stored in separate containers.

[0022] In some embodiments, the kit further comprises an aqueous citrate buffer.

[0023] In some embodiments, the peptide and the stabilizer are present within the kit at a molar ratio of between 1:1 and 10:1.

[0024] In some embodiments, the peptide and the aqueous citrate buffer are present within the kit at a weight ratio of between 1:500 and 1: 10,000.

[0025] In some embodiments, the kit comprising instructions for mixing the peptide, the stabilizer and the aqueous citrate buffer to obtain the composition of the invention.DETAILED DESCRIPTION OF THE PRESENT INVENTION

[0026] In the following detailed description, numerous specific details are set forth in order to provide a thorough understanding of the invention. However, it will be understood by those skilled in the art that the present invention may be practiced without these specific details. In other instances, well-known methods, procedures, and components have not been described in detail so as not to obscure the present invention.

[0027] According to one aspect of the invention, there is provided a composition comprising a peptide and a stabilizer; wherein the peptide is characterized by a water solubility below 200 mg / ml at a temperature between 20 and 30°C and a pH between 5 and 8; and wherein the stabilizer is selected from: a diol, a polyol, a mono-saccharide, a disaccharide, an oligo-saccharide, and a poly-saccharide, including any combination thereof.

[0028] According to another aspect of the invention, there is provided a composition comprising a peptide and a stabilizer; wherein the peptide is a low soluble peptide characterized by a water solubility below 200 mg / ml at a temperature between 20 and 30°C and a pH between 5 and 8; and wherein the stabilizer is selected from: a diol, a polyol, a mono-saccharide, a di-saccharide, an oligo-saccharide, and a poly-saccharide, including any combination thereof.

[0029] In some embodiments, the low soluble peptide is characterized by a water solubility of at most 200 mg / ml, at most 150 mg / ml, at most 100 mg / ml, at most 40 mg / ml, at most 35 mg / ml, at most 30 mg / ml, at most 20 mg / ml, at most 25 mg / ml, at most 15 mg / ml, at most 10 mg / ml, including any range between at a temperature between 20°C and 30°C and at a pH between 5 and 8, between 5 and 6, or between 6 and 7.5. In some embodiments, the low soluble peptide is characterized by a water solubility as disclosed above, and is further characterized by stability of an aqueous peptide solution of below 72h, below 48h, or below 24h, when stored at 20-30°C; and wherein the aqueous peptide solution consists essentially of the peptide and water or an aqueous buffer and is devoid of the stabilizer disclosed herein.

[0030] In some embodiments, the peptide is characterized by a water solubility between about 1 mg / ml and about 200 mg / ml, between about 1 mg / ml and about 150 mg / ml, between about 1 mg / ml and about 130 mg / ml, between about 1 mg / ml and about 100 mg / ml, between about 1 mg / ml and about 40 mg / ml, between about 1 mg / ml and about 30 mg / ml, between about 1 mg / ml and about 25 mg / ml, between about 5 mg / ml and about 40 mg / ml, between about 5 mg / ml and about 30 mg / ml, or between about 5 mg / ml and about 25 mg / ml, including any range between at a temperature between 20°C and 30°C and at a pH between 5 and 8, between 5 and 6, or between 6 and 7.5.

[0031] The term “water solubility” refers to the ability of the compound to undergo complete dissolution in an aqueous solution (e.g., water or an aqueous buffer, such as citrate buffer), so as to form a solution substantially devoid of a particulate matter (i.e., undissolved aggregates or solid particles of the compound); and wherein the aqueous solution is substantially devoid of organic solvents (e.g., DMSO, ethanol, etc.). In some embodiments, the particulate matter has an average particle size above 100 nm, or above 200nm. In some embodiments, the particulate matter in the solution is tested immediately after dissolution or at a time period ranging between 1 and 48h, between 1 and 24h, or between 1 and 72h after dissolution, including any range between.

[0032] The presence of particulate matter in the solution can be determined by various methods, such as by DLS, light microscope, or by visual inspection. Additionally, substantially devoidness of a particulate matter can be assessed by filtering the solution via a 0.22 micron-filter and calculating the content (e.g. w / w concentration) of the compound of interest in the flatted solution, as compared to unfiltered solution (e.g. by HPLC), wherein the solution is considered to be substantially devoid of a particulate matter when the content of the filtered solution is above 90%, or above 95% relative to the content of the unfiltered solution.

[0033] In some embodiments, the aqueous solution substantially devoid of organic solvents has a v / v concertation of an organic solvent up to 1%, up to 0.5%, up to 0.1%, up to 100 ppm, up to 10 ppm, or up to 1 ppm, including any range between. In some embodiments, the aqueous solution is completely devoid of organic solvents (i.e., containing an amount of an organic solvent below the detection limit of a GC machine). In some embodiments, the water solubility of the peptide refers to the solubility of the protonated amine form of the peptide (i.e., whereas at least one amino group of the peptide is in a protonated form -NH3+, such as HC1 salt of the peptide).

[0034] In some embodiments, the peptide is between 2 and 10 amino acids long, between 2 and 8 amino acids long, between 2 and 6 amino acids long, between 2 and 3 amino acids long, or between 2 and 4 amino acids long, or is 2, 3, 4, 5, 6, 7, 8, 9, 10 amino acids long, including any range or value in between. In some embodiments, the peptide is a therapeutically active peptide. In some embodiments, the peptide is the sole pharmaceutically active agent in the composition of the invention.

[0035] As used herein, the term “peptide”, including any enantiomer thereof, any salt thereof, any hydrate or any solvate thereof, or any combination thereof refers to a polymer or an oligomer of amino acid residues bound to each other via a peptide bond.

[0036] The term "peptide" as used herein encompasses native peptides, liner or cyclic peptides, peptide derivatives such as beta peptides, peptidomimetics (typically including non-peptide bonds or other synthetic modifications) and the peptide analogs peptoids and semi-peptoids or any combination thereof. In another embodiment, the term “peptide” applies to amino acid polymers in which at least one amino acid residue is an artificial chemical derivative or analog of a corresponding naturally occurring amino acid.

[0037] The term "derivative" or "chemical derivative" includes any chemical derivative of the polypeptide having one or more residues chemically derivatized by reaction on the side chain or on any functional group within the peptide. Such derivatized molecules include, for example, peptides bearing one or more protecting groups (e.g., side chain protecting group(s) and / or N-terminus protecting groups), and / or peptides in which free amino groups have been derivatized to form amine hydrochlorides, p-toluene sulfonyl groups, carbobenzoxy groups, t-butyloxycarbonyl groups, acetyl groups or formyl groups. Free carboxyl groups may be derivatized to form amides thereof, salts, alkyl esters such as methyl and ethyl esters or other types of esters or hydrazides. Free hydroxyl groups may be derivatized to form O-acyl or O-alkyl derivatives. The imidazole nitrogen of histidine may be derivatized to form N-im-benzylhistidine. Also included as chemical derivatives are those peptides, which contain one or more naturally occurring amino acid derivatives of the twenty standard amino acid residues. For example: 4-hydroxyproline may be substituted for proline; 5-hydroxy lysine may be substituted for lysine; 3 -methylhistidine may be substituted for histidine; homoserine may be substituted or serine; and Dab, Daa, and / or ornithine (O) may be substituted for lysine. The term "derivative" or "chemical derivative" further includes an alkyl ester of the peptide, such as carboxy -terminal alkyl ester.

[0038] In addition, a peptide derivative can differ from the natural sequence of the peptide of the invention by chemical modifications including, but are not limited to, terminal-NH2 acylation, acetylation, or thioglycolic acid amidation, and by amidation of the terminal and / or side-chain carboxy group, e.g., with ammonia, methylamine, and the like. Peptides can be either linear, cyclic, or branched and the like, having any conformation, which can be achieved using methods known in the art.

[0039] The term "amino acid" as used herein encompasses amino acid residue(s) bound to each other via at least one peptide bond. Included within this term are naturally occurring amino acids (i.e. val, ala, arg, asn, asp, cys, glu, gin, gly, his, met, ile, leu, lys, phe, pro, ser, thr, trp, and tyr) , protected amino acids (e.g., comprising one or more protecting groups at the carboxyl, at the amine, and / or at the side chain of the amino acid), unusual, non-naturally occurring amino acids (such as D-amino acids), as well as amino acids which are known to occur biologically in free or combined form but usually do not occur in proteins. Included within this term are modified and unusual amino acids, such as those disclosed in, for example, Roberts and Vellaccio (1983) The Peptides. 5: 342-429. Modified, unusual or non- naturally occurring amino acids include, but are not limited to, D-amino acids, hydroxylysine, 4-hydroxyproline, N-Cbz-protected aminovaleric acid (Nva), ornithine (O), aminooctanoic acid (Aoc), 2,4-diaminobutyric acid (Abu), homoarginine, norleucine (Nle), N-methylaminobutyric acid (MeB), 2-naphthylalanine (2Np), aminoheptanoic acid (Ahp), phenylglycine, P-phenylproline, tert-leucine, 4-aminocyclohexylalanine (Cha), N-methyl- norleucine, 3,4-dehydroproline, N,N-dimethylaminoglycine, N-methylaminoglycine, 4- aminopipetdine-4-carboxylic acid, 6-aminocaproic acid, trans-4- (aminomethyl) - cyclohexanecarboxylic acid, 2-, 3-, and 4- (aminomethyl) - benzoic acid, 1- aminocyclopentanecarboxylic acid, 1 -aminocyclopropanecarboxylic acid, cyano-propionic acid, 2-benzyl-5- aminopentanoic acid, Norvaline (Nva), 4-O-methyl-threonine (TMe), 5- O-methyl-homoserine (hSM), tert-butyl-alanine (tBu), cyclopentyl-alanine (Cpa), 2-amino- isobutyric acid (Aib), N-methyl-glycine (MeG), N-methyl- alanine (MeA), N-methyl- phenylalanine (MeF), 2-thienyl-alanine (2Th), 3-thienyl-alanine (3Th), O-methyl-tyrosine (YMe), 3-Benzothienyl-alanine (Bzt) and D-alanine (DAI).

[0040] In some embodiments, the term “peptide bond” refers to a secondary amide represented by Formulatertiary amide represented by Formula B:, wherein R is an amino acid side chain, C1-C5 alkyl, or a proline residue. In some embodiments, the term “peptide bond” encompasses Formula A and / or B, wherein R is a natural amino acid side chain.

[0041] In some embodiments, a number of the amino acid residues within the peptide is between 1 and 9, between 1 and 7, between 1 and 5, and between 1 and 3, or 2, including any range in between. In some embodiments, the peptide is a di-peptide, a tri-peptide, or a tetra-peptide. In some embodiments, the peptide is a linear or cyclic di-peptide or a linear or cyclic tri-peptide. In some embodiments, the peptide comprises a hydrophobic amino acid selected from gly, ala, leu, iso, val, pro, met, phe, tyr and trp. In some embodiments, the peptide is devoid of polar and / or charged amino acid, such as ser, thr, asn, gin, asp, glu, arg, his and lys.

[0042] In some embodiments, the peptide further comprises an ester. In some embodiments, the ester is a carboxy-terminal ester. In some embodiments, the C-terminus of the peptide, at least one side chain carboxy group of the peptide or both is / are esterified. In some embodiments, the carboxy terminal amino acid is an esterified amino acid. In some embodiments, the ester refers to a C1-C10 alkyl ester, or a C1-C10 substituted alkyl ester comprising 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms wherein each of the carbon atoms is optionally substituted by a substituent. In some embodiments, the alkyl ester comprises up to 5 carbon atoms (e.g., 1, 2, 3, 4, or 5 carbon atoms). In some embodiments, the ester is a methyl ester.

[0043] The term “substituent” is as disclosed herein.

[0044] In some embodiments, there is provided a composition comprising a peptide, including any enantiomer thereof, a salt thereof, a hydrate or solvate thereof, or any combination thereof; and a stabilizer; wherein the peptide is between 2 and 10 amino acids long, the peptide further comprises an ester (i.e. an esterified amino acid); and wherein thestabilizer is selected from: a diol, a polyol, a mono-saccharide, a di-saccharide, an oligosaccharide, and a poly-saccharide, including any combination thereof.

[0045] In some embodiments, the ester is a carboxy-terminal ester. In some embodiments, the C-terminus of the peptide, at least one side chain carboxy group of the peptide or both is / are esterified. In some embodiments, the carboxy terminal amino acid is an esterified amino acid. In some embodiments, the ester refers to a C1-C10 alkyl ester, or a C1-C10 substituted alkyl ester comprising 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 carbon atoms wherein each of the carbon atoms is optionally substituted by a substituent. In some embodiments, the alkyl ester comprises up to 5 carbon atoms (e.g., 1, 2, 3, 4, or 5 carbon atoms).

[0046] In some embodiments, the peptide is a low-soluble peptide, as disclosed hereinabove.

[0047] In some embodiments, the peptide is represented by Formula 1 :, including any salt thereof, any tautomer thereof, any stereoisomer thereof, any hydrate or hydrate thereof, or any combination thereof, wherein R is C1-C5 alkyl.

[0048] As used herein, the term “C1-C5 alkyl” refers to any linear or branched alkyl chain comprising between 1 and 5, between 1 and 3, between 1 and 4, or 1, 2, 3, 4 or 5 carbon atoms, including any range therebetween. In some embodiments, C1-C5 alkyl comprises any of methyl, ethyl, propyl, iso-propyl, butyl, pentyl, iso-pentyl, tert-butyl, or any combination thereof. In some embodiments, C1-C5 alkyl is a substituted C1-C5 alkyl. In some embodiments, C 1-C5 alkyl as described herein further comprises an unsaturated bond, wherein the unsaturated bond is located at 1st, 2nd, 3rd, 4th, or 5th, position of the C1-C5 alkyl.

[0049] In some embodiments, the peptide comprises a mixture of diastereomers. In some embodiments, the peptide comprises a single diastereomer. In some embodiments, the peptide comprises a mixture of enantiomers (e.g., a racemic mixture). In some embodiments, the peptide is enriched with an enantiomer of interest. In some embodiments, the peptide is a single enantiomer. In some embodiments, the peptide is a single diastereomer.

[0050] In some embodiments, the peptide is represented by Formula 2:, including any salt thereof, any tautomer thereof, any hydrate or solvate thereof, or any combination thereof, wherein R is as described above. In some embodiments, the salt is a pharmaceutically acceptable salt.

[0051] In some embodiments, R is methyl.

[0052] In some embodiments, the composition comprises a plurality of peptides, wherein each of the plurality of peptides as described herein. In some embodiments, each of the plurality of peptides differs in its chemical structure and / or chirality. In some embodiments, the plurality of peptides consist of chemically distinct peptides. In some embodiments, the plurality of peptides consist of peptides having the same chemical composition and / or the same chemical structure or chirality. In some embodiments, each of the peptides in the composition has the same amino acid sequence. In some embodiments, the peptides in the composition have at least 2 distinct amino acid sequences. In some embodiments, the peptides in the composition are in a form of a racemic mixture. In some embodiments, the peptides in the composition are in a form of a single enantiomer. In some embodiments, the peptides in the composition are characterized by an enantiomeric purity of between 70% and 100%, between 90% and 100%, between 80% and 99%, or between 90 and 95%, including any range in between.

[0053] In some embodiments, the stabilizer is selected from a diol, a polyol, a monosaccharide, a di-saccharide, an oligo-saccharide, or a poly-saccharide, including any combination thereof.

[0054] In some embodiments, non-limiting examples of the stabilizer are Sucrose, Lactose , Maltose, Trehalose, Cellobiose, Chitobiose, Kojibiose, Nigerose, Isomaltose, P,P- Trehalose, a,P-Trehalose, Sophorose, Laminaribiose, Gentiobiose, Turanose, Trehalulose, Maltulose, Leucrose, Isomaltulose, Gentiobiulose, Mannobiose, Melibiose, Allolactose, Melibiulose, Lactulose, Rutinose, Rutinulose, Xylobiose, Mannitol, Sorbitol, P- cyclodextrin, Raffinose, Nigerotriose, Maltotriose, Melezitose, Maltotriulose, Kestose, ethylene glycol, glycerin, polyethylene glycol, propylene glycol, or any combination thereof. In some embodiments, the stabilizer is devoid of a polymeric stabilizer. In some embodiments, the stabilizer is a small molecule having a MW below 1000 Da, or below 500 Da.

[0055] In some embodiments, the stabilizer comprises a di-saccharide (e.g., lactose, trehalose, etc.). In some embodiments, the stabilizer is trehalose.

[0056] In some embodiments, the stabilizer a tri-saccharide (e.g., kestose, raffinose, etc.). In some embodiments, the stabilizer is raffinose.

[0057] In some embodiments, the stabilizer is capable of increasing water solubility of the low soluble peptide, as compared to the same pristine peptide devoid of the stabilizer. In some embodiments, the stabilizer is capable of forming stable hydrogen bonds with the low soluble peptide in an aqueous solution, thereby improving or enhancing water solubility of the low soluble peptide.

[0058] In some embodiment, a weight ratio between the peptide and the stabilizer within the composition of the invention is between 1:1 and 10:1, between 1:1 and 8:1, between 1:1 and 6:1, between 1:1 and 4:1, between 1:1 and 1:2, between 1:5 and 1:10, or between 1:6 and 1:9, including any range in between.

[0059] In some embodiments the composition of the invention is capable of increasing water solubility of the low soluble peptide compared to the peptide. In some embodiments, the water solubility of the low soluble peptide within the composition of the invention compared to the same pristine peptide is increased by at least 20%, at least 50%at least 2, at least 5, at least 10, at least 15, or at least 20-fold, including any range in between.

[0060] In some embodiments, the low soluble peptide within the composition of the invention is characterized by water solubility of up to 600 g / L, up to 500 g / L, up to 400 g / L, up to 300 g / L, between 40 g / L and 400 g / L, between 50 g / L and 400 g / L, between 50 g / L and 300 g / L, between 50 g / L and 450 g / L, between 50 g / L and 500 g / L, including any range in between, wherein the water solubility is measured at a pH of about 5.5 and at a temperature between 20 and 25C.

[0061] In some embodiments, the composition of the invention is devoid of a surfactant, and / or a dispersant. In some embodiments, the composition of the invention is devoid of a lipid. In some embodiments, the composition of the invention is devoid of an additional stabilizer, which is not the stabilizer of the invention. In some embodiments, the composition of the invention is devoid of an additional peptide, which is not the low soluble peptide disclosed herein.

[0062] In some embodiments, the composition of the invention is for use in treating a disease in a subject in need thereof. In some embodiments, the disease is an ocular or retinal degeneration disease. In some embodiments, the composition of the invention is aformulation, formulated for ocular administration, such as in a form of eye drops or by intravitreal injection.Solution

[0063] In some embodiments, the composition of the invention is in a form of a stable solution (also referred herein as “solution”). In some embodiments, the solution is a clear solution. In some embodiments, the solution is devoid of a color (i.e., visible color). In some embodiments, the solution of the invention is a homogenous solution. In some embodiments, clearness is determined by visual observation.

[0064] In some embodiments, the stable solution is substantially devoid of particulate matter such as aggregates, precipitate, crystals or undissolved matter, as disclosed above. Various methods for the detection of aggregates / particles in the solution are as disclosed above. In some embodiments, solution stability is determined by visual inspection. In some embodiments, the solution is devoid of solid particles with an average particle size of above 10 nm, above 20 nm, above 30 nm, above 50 nm, above 100 nm, or above 200 nm, including any range in between.

[0065] In some embodiments, the solution of the invention is stable at a temperature between 2°C and 50°C, between 2°C and 40°C, between 2°C and 25°C, between 10°C and 30°C, between 5°C and 40°C, between 20°C and 30°C, between 20°C and 27°C, or between 5 and 35°C, including any range or value in between.

[0066] In some embodiments, the solution of the invention is stable for a period of time of at least 72h, at least 48h, or at least 26h, or between 10 and 90h, between 20 and 80h, between 20 and 75h, between 20 and 72h, including any range in between.

[0067] In some embodiments, the solution of the invention is stable at a temperature between 2°C and 40°C, between 2°C and 25°C, between 10°C and 30°C, between 5°C and 40°C, between 20°C and 30°C, between 20°C and 27°C or between 5 and 35°C, including any range or value in between, for a period of time as disclosed hereinabove.

[0068] In some embodiments, the solution of the invention is characterized by a prolonged stability, as compared to a control (i.e., the same solution devoid of the stabilizer of the invention). In some embodiments, prolonged stability is by at least 20%, at least 30%, at least 50%, at least 70%, at least 100%, at least 150%, at least 200%, at least 300%, at least 500%, including any range between.

[0069] In some embodiments, the solution is an aqueous solution. In some embodiments, the aqueous solution is substantially devoid of organic solvents, as disclosed above. In some embodiments, the aqueous solution is completely devoid of an organic solvent.

[0070] In some embodiments, the solution is dilutable up to 1000-fold, wherein upon dilution the resulting solution is substantially devoid of particular matter and is characterized by at least the same solution stability as the solution of the invention.

[0071] In some embodiments, the solution is characterized by a pH between 5 and 8, between 5 and 7, between 5 and 6.5, between 5.1 and 6.3, between 5 and 6, between 5.2 and 6, between 5 and 5.8, between 5.3 and 5.7, including any range in between. In some embodiments, the solution is an aqueous buffered solution. In some embodiments, the aqueous buffered solution is characterized by a pH between 5 and 8, between 5 and 7, between 5 and 6.5, between 5.1 and 6.3, between 5 and 6, between 5.2 and 6, between 5 and 5.8, between 5.3 and 5.7, including any range in between.

[0072] Examples of aqueous solutions include but are not limited to buffers (e.g., phosphate buffer, bicarbonate buffer, etc.), salt (e.g., inorganic salt) solution or saline solutions.

[0073] In some embodiments, the aqueous solution is a buffered solution. In some embodiments, the aqueous solution comprises citrate buffer. In some embodiments, the aqueous solution is a citrate buffered solution.

[0074] In some embodiments, the solution of the invention is devoid of liquid particles (e.g., micelles, liposomes, nanodroplets, etc.). In some embodiments, the peptide is not encapsulated in a liquid particle. In some embodiments, the solution of the invention is devoid of emulsion, suspension, or a dispersion.

[0075] In some embodiments, the water content of the solution is at least 60%, at least 70%, at least 80%, at least 90%, at least 96%, at least 97%, or at least 98% by weight of the aqueous solution, including any range or value in between. In some embodiments, the solution comprises water as the sole solvent.

[0076] In some embodiments, the peptide content within the solution of the invention is up to 40 wt%, up to 35 wt%, up to 30 wt%, or up to 25 wt%, including any range in between. In some embodiments, a weight percentage of the peptide within the solution is between 0.5 and 40 wt%, between 1 and 40 wt%, between 5 and 15 wt%, between 1 and 10 wt%, between 1 and 20 wt%, between 5 and 25 wt%, between 15 and 25 wt%, between 25 and 35 wt%, between 20 and 40 wt%, or between 5 and 20 wt%, including any range in between.

[0077] In some embodiments, a weight percentage of the stabilizer within the solution is of at least 1 wt%, at least 3 wt%, or at least 5 wt%, including any range or value in between. In some embodiments, a weight percentage of the stabilizer within the composition is between 1 and 40 wt%, between 1 and 50 wt%, between 1 and 35 wt%, between 1 and 33 wt%, between 3 and 7 wt%, or between 3 and 15 wt%, between 1 and 10 wt%, between 3 and 38 wt%, including any range in between.

[0078] In some embodiment, a weight ratio between the peptide and the stabilizer within the solution is between 1:1 and 10:1, between 1:1 and 8:1, between 1:1 and 6:1, between 1:1 and 4:1, between 1:1 and 1:2, between 1:5 and 1:10, or between 1:6 and 1:9, including any range in between.

[0079] In some embodiments, the solution of the invention consists essentially of the peptide the stabilizer, and an aqueous buffer, wherein the peptide and the stabilizer are as disclosed herein. In some embodiments, at least 93%, at least 95%, at least 97%, at least 99%, or between 90 and 99%, between 95 and 99%, between 95 and 100% by weight of the solution of the invention consists of the peptide, the stabilizer, and an aqueous buffer solution, wherein the peptide and the stabilizer are as disclosed herein. In some embodiments, the solution of the invention is a citrate buffered solution, having a pH between 5 and 7, or between 5 and 6.Powder

[0080] In some embodiments, the composition of the invention is a solid composition. In some embodiments, the composition is a solid at a temperature below 100°C. In some embodiments, the composition of the invention is a powderous composition. In some embodiments, the composition of the invention is a powderous bulk material.

[0081] In some embodiments, the powder is a homogenous powder. In some embodiments, the powder is a homogenous mixture of the entire constituents of the powderous composition. In some embodiments, the homogenous powder is referred to a material which cannot be easily separated into individual constituents (e.g., the peptide and the stabilizer). In some embodiments, the entire bulk volume of the homogenous powder has essentially the same concentration (up to a deviation of 10%) of the entire constituents of the powderous composition.

[0082] The terms “powderous composition” and “powder” are used herein interchangeably.

[0083] In some embodiments, the powder is flowable. In some embodiments, the powder is a free flowable powder.

[0084] In some embodiments, the powder is a white powder. In some embodiments, the powder is a yellowish (light yellow) powder.

[0085] In some embodiments, the powder is a dry powder characterized by a water content below 5 wt%, below 4 wt%, or below 3 wt%, including any range or value in between. In some embodiments, the dry powder is lyophilizate. In some embodiments, the water content of the dry powder is between 0.001 and 5 wt%, between 0.001 and 0.01 wt%, between 0.001 and 0.1 wt%, between 0.1 and 5 wt%, between 0.1 and 4 wt%, between 0.1 and 4 wt%, between 0.1 and 3 wt%, between 0.1 and 2 wt%, between 0.1 and 1 wt%, between 1 and 5 wt%, between 2 and 5 wt%, including any range or value in between.

[0086] In some embodiment, a weight ratio between the peptide and the stabilizer within the powder is between 1:1 and 10:1, between 1:1 and 8:1, between 1:1 and 6:1, between 1:1 and 4: 1 , or between 1 : 1 and 1 :2, including any range in between.

[0087] In some embodiments, a weight percentage of the peptide within the powder of the invention is between 10 and 90 wt%, between 45 and 65 wt%, between 45 and 85 wt%, between 45 and 55 wt%, between 50 and 55 wt%, or between 50 and 60 wt%, between 45 and 75 wt%, including any range in between.

[0088] In some embodiments, a weight percentage of the stabilizer within the powder of the invention is between 10 and 55 wt%, between 10 and 90 wt%, between 45 and 60 wt%, between 45 and 55 wt%, between 50 and 55 wt%, or between 50 and 60 wt%, including any range in between.

[0089] In some embodiments, the powder of the invention further comprises a buffering agent. In some embodiments, a weight ratio between the buffering agent and the peptide within the powder of the invention is 1 :200 and 1 : 10,000, between 1 :200 and 1 : 800, between 1:200 and 1:600, between 1:200 and 1:400, between 1:400 and 1:600, or between 1:500 and 1:600, including any range in between. In some embodiments, a weight ratio between the buffering agent and the peptide within the powder of the invention is 100:1 and 1:100, between 50:1 and 1:50, between 20:1 and 1:20, between 20:10000 and 20:1000, between 20:500 and 10:500, including any range in between. In some embodiments, the powder of the invention comprises an amount of the buffering agent sufficient to obtain a buffered solution upon dilution of the powder of the invention, wherein dilution is so as to obtain a therapeutically effective amount of the peptide in the resulting buffered solution.

[0090] In some embodiments, the buffering agent is a salt of a weak acid. In some embodiments, the weak acid is a weak organic acid. In some embodiments, the weak acid comprises any of: citric acid, formic acid, acetic acid, lactic acid, trichloroacetic acid, benzoic acid, and dihydrogen phosphate. In some embodiments, the weak acid salt is a pharmaceutically acceptable salt of the weak acid.

[0091] In some embodiments, the buffering agent is or comprises a citrate salt (e.g. sodium citrate).

[0092] In some embodiments, the powder consists essentially of the peptide, the stabilizer and the buffering agent (e.g. a weak acid salt); wherein a weight ratio between the peptide and the stabilizer within the powder is as described hereinabove, and wherein a weight ratio between the buffering agent and the peptide within the powder is between 100: 1 and 1 : 100. In some embodiments, at least 90%, at least 95%, at least 97%, at least 98%, at least 99% or between 90 and 99.9% by dry weight of the powder consists of the peptide, the stabilizer and the buffering agent. In some embodiments, at least 90%, at least 95%, at least 97%, at least 98%, at least 99% or between 90 and 99.9% by dry weight of the powder consists of the peptide, trehalose and the buffering agent. In some embodiments, at least 90%, at least 95%, at least 97%, at least 98%, at least 99% or between 90 and 99.9% by dry weight of the powder consists of the peptide, trehalose and the weak acid salt (e.g. citrate salt).

[0093] In some embodiments, the powder is a free flowable powder. Methods for measuring powder flowability are well known to a skill in the art such as, rheometer for powder.

[0094] In some embodiments, the powder is stable. In some embodiments, the powder of the invention is stable at a temperature up to 70°C, up to 60°C, up to 50 °C, up to 40°C, including any range or value in between. In some embodiments, the powder of the invention is stable for a period of time of up to 5y, up to 3y, up to ly, up to 6m, including any range in between. A powder is referred to as stable, when it maintains at least 90%, at least 95%, at least 97% by weight of the initial concentration of the pharmaceutically active agent.

[0095] In some embodiments, the powder of the invention is water soluble. In some embodiments, the powder of the invention is characterized by a water solubility of at least 1 g / L, at least 10 g / L, at least 50 g / L, at least at least 100 g / L, at least 200 g / L, at least 300 g / L, or up to 900g / L, up to 800 g / L, up to 500 g / L or up to 750 g / L, including any range between, wherein water solubility is determined at a temperature and pH values disclosed hereinabove.

[0096] In some embodiments, the powder of the invention is characterized by a water solubility of between 10 g / L and 800 g / L, between 100 g / L and 400 g / L, between 10 g / L and 200 g / L, between 50 g / L and 150 g / L, between 50 g / L and 250 g / L, between 150 g / L and 250 g / L, between 200 g / L and 400 g / L, or between 50 and 400 g / L, including any range in between.

[0097] In some embodiments, the powder of the invention consists essentially of the peptide the stabilizer, and a buffering agent. In some embodiments, at least 93%, at least 95%, at least 97%, at least 99%, or between 90 and 99%, between 95 and 99%, between 95 and 100% by dry weight of the powder of the invention consist of the peptide, the stabilizer and the buffering agent. In some embodiments, between 93% and 100%, between 93% and 95%, between 93% and 97%, or between 95% and 99% by dry weight of the powder of the invention consists of the peptide, the stabilizer, and optionally the buffering agent.

[0098] In some embodiments, the powder of the invention is completely water soluble. In some embodiments, upon reconstitution of the powder of the invention the resulting aqueous solution is characterized by peptide recovery of at least 90%, at least 95%, at least 97%, or between 90 and 97%, including any range between.

[0099] As used herein, the term “peptide recovery” encompasses recovery relative to the theoretical concentration of the peptide (based on the amount of dry powder and the amount of an aqueous solution used for reconstitution). Recovery can be determined by HPLC.Pharmaceutical composition

[0100] According to another aspect there is provided a pharmaceutical composition, comprising: (i) the peptide; (ii) the stabilizer; and (iii) a pharmaceutically acceptable carrier; wherein the peptide and the stabilizer are as disclosed herein.

[0101] As used herein, the term “pharmaceutically acceptable carrier” refers to nontoxic, inert solid, semi-solid liquid filler, diluent, encapsulating material, formulation auxiliary of any type, or simply a sterile aqueous medium, such as saline. Some examples of the materials that can serve as pharmaceutically acceptable carriers are sugars, such as lactose, glucose and sucrose, starches such as com starch and potato starch, cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt, gelatin, talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; glycols, such as propylene glycol, polyols such as glycerin, sorbitol, mannitol and polyethylene glycol; esters such as ethyl oleate and ethyl laurate,agar; buffering agents; alginic acid; pyrogen-free water; isotonic saline, Ringer's solution; ethyl alcohol and phosphate buffer solutions; wetting agents and lubricants such as sodium lauryl sulfate, as well as coloring agents, flavoring agents, excipients, stabilizers, antioxidants, preservatives and other non-toxic compatible substances used in pharmaceutical formulations. Any non-toxic, inert, and effective carrier may be used to formulate the compositions contemplated herein. Suitable pharmaceutically acceptable carriers, excipients, and diluents in this regard are well known to those of skill in the art, such as those described in The Merck Index, Thirteenth Edition, Budavari et al., Eds., Merck & Co., Inc., Rahway, N.J. (2001); the CTFA (Cosmetic, Toiletry, and Fragrance Association) International Cosmetic Ingredient Dictionary and Handbook, Tenth Edition (2004); and the “Inactive Ingredient Guide,” U.S. Food and Drug Administration (FDA) Center for Drug Evaluation and Research (CDER) Office of Management, the contents of all of which are hereby incorporated by reference in their entirety.

[0102] In some embodiments, the presently described composition may also be contained in artificially created structures such as liposomes, ISCOMS, slow-releasing particles, and other vehicles which increase the half-life of the peptides or polypeptides in serum. Liposomes for use with the presently described peptides are formed from standard vesicle-forming lipids which generally include neutral and negatively charged phospholipids and a sterol, such as cholesterol. The selection of lipids is generally determined by considerations such as liposome size and stability in the blood. A variety of methods are available for preparing liposomes as reviewed, for example, by Coligan, J. E. et al, Current Protocols in Protein Science, 1999, John Wiley & Sons, Inc., New York, and see also U.S. Pat. Nos. 4,235,871, 4,501,728, 4,837,028, and 5,019,369.

[0103] In some embodiments, the pharmaceutically acceptable carrier is a liquid carrier. In some embodiments, the liquid carrier is an aqueous solution. In some embodiments, the liquid carrier is a buffered solution. In some embodiments, the liquid carrier is characterized by a pH value between 5 and 8, between 6 and 7.5, between 5 and 6.5, between 5.1 and 6.3, between 5 and 6, between 5.2 and 6, between 5 and 5.8, between 5.3 and 5.7, including any range in between. In some embodiments, the liquid carrier is saline solution. In some embodiments, the liquid carrier is a buffered saline solution. In some embodiments, the liquid carrier is a citrate buffered solution.

[0104] In some embodiments, the carrier comprise in total between 0.1 and 99.99%, between 0.1 and 50%, between 0.2 and 30%, between 0.1 and 70%, between 30 and 99.99%,between 50 and 99.99, by weight of the pharmaceutical compositions presented herein, including any range in between.

[0105] In some embodiments, the peptide is of pharmaceutical grade purity. In some embodiments, the entire constituents of the composition of the invention and of the pharmaceutical composition are of pharmaceutical grade purity, i.e., are characterized by a chemical purity of at least about 90%, at least about 95%, greater than 95%, or greater than 99%, and between 90 and 99.999%, between 90 and 95%, between 90 and 97%, between 95 and 99%, including any range in between. Chemical purity of a compound / compo sition can be determined by HPLC, and / or by LC / MS.

[0106] In some embodiments, the pharmaceutical composition consists essentially of the composition of the invention and the pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition consists essentially of the peptide of the invention as the therapeutically active ingredient. In some embodiments, the peptide of the invention is the only active ingredient within the pharmaceutical composition. In some embodiments, the pharmaceutical composition is devoid of additional therapeutically active ingredients.

[0107] In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of the peptide. In some embodiments, the pharmaceutical composition is a solution, wherein a therapeutically effective amount of the peptide within the solution is between about 1 and about 5% , including any range in between.

[0108] In some embodiments, the pharmaceutical composition is an ophthalmic composition. In some embodiments, the pharmaceutical composition is formulated for ocular administration. In some embodiments, the ophthalmic composition comprises an additive. In some embodiments, the additive includes, without limitation, a tonicity regulator, a preservative, an electrolyte, and / or any mixture thereof. In some embodiments, a w / w concentration of the additive within the ophthalmic composition is between 0.01 and 20%, including any range in between.

[0109] In some embodiments, the ophthalmic composition comprises an effective amount of a preservative. In some embodiments, the effective amount is a preservative effective amount. In some embodiments, the effective amount is an antimicrobial effective amount. In some embodiments, the effective amount of the preservative is determined based on a PET test, according to USP <51 > , or any alternative PET test.

[0110] In some embodiments, the preservative is or comprises a quaternary ammonium cation, such as benzalkonium (BAK) including any salt thereof, for example benzalkonium chloride. In some embodiments, the quaternary ammonium cation is a pharmaceutically acceptable compound. In some embodiments, the quaternary ammonium cation comprises a pharmaceutically acceptable counter anion. Additional preservatives (i.e., preservatives suitable for ophthalmic compositions) are known in the art.

[0111] In some embodiments, the ophthalmic composition in a liquid state is characterized by a viscosity between 1 and 150cps, when measured at 25C.

[0112] In some embodiments, the ophthalmic composition is characterized by a tonicity between 200 and 600 mOsmols / kg, between 200 and 500 mOsmols / kg, between 200 and 300 mOsmols / kg, between 300 and 350 mOsmols / kg, between 350 and 400 mOsmols / kg, between 400 and 450 mOsmols / kg, between 450 and 500 mOsmols / kg, between 500 and 550 mOsmols / kg, between 550 and 600 mOsmols / kg, including any range or value therebetween. In some embodiments, the ophthalmic composition is characterized by a tonicity between 250 and 450 mOsmols / kg.

[0113] In some embodiments, the pharmaceutical composition consists essentially of pharmaceutical grade constituents. In some embodiments, the entire constituents of the pharmaceutical composition are pharmaceutical grade compounds. In some embodiments, the pharmaceutical composition is sterile. In some embodiments, the effective amount of the preservative within the pharmaceutical composition of the invention is sufficient for maintaining the sterility of the pharmaceutical composition upon exposure thereof to non- sterile condition (e.g. ambient atmosphere comprising one or more microbes), wherein maintain is for a time period ranging between 1 and between 1 day and 2 months, between 1 day and Imonth, between 1 and 20d, between 1 and 60d, between 1 and 50d, between 10 and 60d, between 1 and lOd, between 10 and 50d, between 10 and 40d, between 10 and 30d, including any range between. The term “sterility” as used herein, refers inter alia to the microbial load of a composition (e.g., ophthalmic composition of the invention) or article according to the sterility requirements as recorded in US Pharmacopoeia, or in European Pharmacopoeia, respectively.

[0114] In some embodiments, the pharmaceutical composition is packaged in a unit dosage form. In some embodiments, the pharmaceutical composition is prepared by any of the methods well known in the art of pharmacy. In some embodiments, the unit dosage form is in the form of a tablet, capsule, lozenge, wafer, patch, ampoule, vial or pre-filled syringe.

[0115] In some embodiments, each unit dosage comprises an effective dose of the peptide of the invention. The term “effective dose” refers to a daily dosage of the composition / peptide of the invention applied to a subject, sufficient to induce to the desired biological response (e.g., reduce of a symptom, treat a disease, etc.) in the subject.

[0116] In some embodiments, the pharmaceutical composition is for use in treatment or prevention of a disease or a disorder, in a subject in need thereof.

[0117] In some embodiments, the subject is a mammal. In some embodiments, the mammal is a human. In some embodiments, the mammal is a laboratory animal. Examples of laboratory animals include, but are not limited to, mice, rats, rabbits, hamsters, dogs, cats, and monkeys. In some embodiments, the subject is in need of the composition of the invention. In some embodiments, the subject is in need of treatment.

[0118] In some embodiments, the disease is an ocular disease. In some embodiments, the disease is a ocular or retinal degeneration disease. In some embodiments, the composition of the invention is administered by ocular administration or by intraocular administration. In some embodiments, ocular administration is by intravitreal injection.

[0119] In some embodiments, the pharmaceutical composition as disclosed herein comprises a therapeutically effective amount of the composition of the invention. In some embodiments, the therapeutically effective amount refers to an amount effective to treat a disease or disorder in a mammal. The term “therapeutically effective amount” refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic or prophylactic result in a subject. The exact dosage form and regimen would be determined by the physician according to the patient's condition.Kit

[0120] According to another aspect, there is provided a kit comprising the peptide, the stabilizer and optionally the buffering agent, as disclosed herein.

[0121] In some embodiments, the constituents of the kit are stored in separate compartments. In some embodiments, the peptide and the stabilizer are stored in the same compartment. In some embodiments, the peptide, the stabilizer, and the buffering agent are stored in separate compartments. In some embodiments, the peptide, the stabilizer, and the buffering agent are stored in separate compartments.

[0122] In some embodiments, the peptide is in a form of a powder. In some embodiments, the stabilizer is in a form of a solution or a powder. In some embodiments, the buffering agent is in a form of a powder, or a solution.

[0123] In some embodiments, the kit further comprises a diluent. In some embodiments, the diluent is an aqueous solution. In some embodiments, the diluent is a pharmaceutically acceptable carrier. In some embodiments, the diluent is an aqueous solution.

[0124] In some embodiments, the peptide and the stabilizer are present within the kit at a weight ratio of between 1:1 and 10:1, between 1:1 and 8:1, between 1:1 and 6:1, between 1 : 1 and 4: 1 , or between 1 : 1 and 2:1, including any range in between.

[0125] In some embodiments, the peptide and the citrate buffer are present within the kit at a weight ratio of between 1:500 and 1:10,000, between 1:500 and 1:5,000. between 1:500 and 1:1,000, between 1:100 and 1:10,000, between 1:5000 and 1:10,000 or between 1:500 and 1:7,500, including any range in between.

[0126] In some embodiments, the kit further comprises instructions for mixing the constituents of kit (dry or liquid constituents) at predetermined ratio, to obtain the composition of the invention.

[0127] In some embodiments, kit further comprises instructions for mixing or contacting the constituents of kit with a predetermined amount of the diluent to obtain the solution of the invention. In some embodiments, mixing or contacting, is performed at a temperature between 10 and 50C, and accompanied by a sufficient mixing time to obtain a clear solution.

[0128] In some embodiments, the kit is for use in conjunction with the method of the invention.

[0129] According to an embodiment of the present invention, the pharmaceutical composition and / or the described herein above is packaged in a packaging material and identified in print, in or on the packaging material, for use in the treatment of a disease or disorder, as described herein.

[0130] According to another embodiment of the present invention, the pharmaceutical composition / kit is packaged in a packaging material and identified in print, in or on the packaging material, for use in monitoring a disease or disorder, as described herein.

[0131] Products of the present invention may, if desired, be presented in a pack or dispenser device, such as an U.S . Food and Drug Administration (FDA) approved kit, which may contain one or more unit dosage forms containing the disclosed composition. The pack may, for example, comprise metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accommodated by a notice associated with the container in a form prescribed by a governmental agency regulating the manufacture, use or sale ofpharmaceuticals, which notice is reflective of approval by the agency of the form of the compositions or human or veterinary administration. Such notice, for example, may be of labeling approved by the FDA for prescription drugs or of an approved product insert.Method

[0132] According to another aspect, there is provided a method for treating a disease, disorder or a condition within a subject in need thereof, comprising administering a therapeutically effective amount of the pharmaceutical composition of the invention to the subject, thereby treating the disease, disorder or the condition.

[0133] In some embodiments, administering comprises systemic administration. In some embodiments, administering comprises a local administration. In some embodiments, administering comprises intravenous administration. In some embodiments, administering comprises ocular administration. In some embodiments, administering comprises intraocular administration. In some embodiments, administering comprises an intravitreal injection.

[0134] In some embodiments, the method is for reducing at least one symptom associated with the disease or condition within the subject.

[0135] In some embodiments, the disease is an ocular disease. In some embodiments, the disease is a degenerative disease. Non limiting examples of degenerative disease include but are not limited to a retinal disease, cell senescence Familial drusen, glaucoma, Stargardt disease and Best disease.

[0136] In some embodiments, the subject is afflicted with a retinal degeneration disease. Non limiting examples of retinal degeneration diseases include but are not limited to age- related macular degeneration (AMD), wet AMD, diabetic retinopathy or retinitis pigmentosa.

[0137] As used herein, the terms “administering”, “administration”, and like terms refer to any method which, in sound medical practice, delivers a composition containing an active agent to a subject in such a manner as to provide a therapeutic effect.

[0138] The administering step may be repeated if needed. The mode of administration of the present composition depends on the form of the composition. For example, if the composition is a solution, drops of the composition may be applied to the eye, e.g., from a conventional eye dropper, or the composition may by injected into an eye.

[0139] In some embodiments, the amount (dose) of a composition to be administered will, of course, be dependent on the subject being treated, in the medical condition beingtreated for, the severity of the affliction, the manner of administration, the judgment of the prescribing physician, etc.

[0140] In some embodiments, the daily dose administered into each eye (i.e. the amount of the peptide disclosed herein per day per each eye) is between 25 and 15,000 pg, between 50 and 10,000 pg, between 100 and 10,000 pg, between 200 and 2000 pg, between 50 and 100 pg, between 100 and 200 pg, between 200 and 300 pg, between 300 and 400 pg, between 400 and 500 pg, between 500 and 600 pg, between 600 and 700 pg, between 700 and 800 pg, between 800 and 900 pg, between 900 and 1000 pg, between 1000 and 1100 pg, between 1100 and 1300 pg, between 1300 and 1500 pg, between 1500 and 1800 pg, between 1000 and 15,000 pg, between 1800 and 2000 pg, including any range or value therebetween.

[0141] In some embodiments, the composition is administered 1, 2, 3, 4, 5, 6, 8, 9, 10 times a day, including any range therebetween.Definitions

[0142] As used herein, the term “stable” refers to a clear solution, wherein the solution is devoid of precipitate, crystals and / or particles.

[0143] As used herein, the term "consists essentially of" means that the composition may include additional ingredients but only if the additional ingredients do not materially alter the basic and novel characteristics of the claimed composition. As used herein, the terms “treatment” or “treating” of a disease, disorder, or condition encompasses alleviation of at least one symptom thereof, a reduction in the severity thereof, or inhibition of the progression thereof. Treatment need not mean that the disease, disorder, or condition is totally cured. To be an effective treatment, a useful composition herein needs only to reduce the severity of a disease, disorder, or condition, reduce the severity of symptoms associated therewith, or provide improvement to a patient or subject’s quality of life.

[0144] As used herein, the term “prevention” of a disease, disorder, or condition encompasses the delay, prevention, suppression, or inhibition of the onset of a disease, disorder, or condition. As used in accordance with the presently described subject matter, the term "prevention" relates to a process of prophylaxis in which a subject is exposed to the presently described active ingredients prior to the induction or onset of the disease / disorder process. This could be done where an individual has a genetic pedigree indicating a predisposition toward occurrence of the disease / disorder to be prevented. Forexample, this might be true of an individual whose ancestors show a predisposition toward certain types of inflammatory disorders.

[0145] The term "suppression" is used to describe a condition wherein the disease / disorder process has already begun but obvious symptoms of the condition have yet to be realized. Thus, the cells of an individual may have the disease / disorder, but no outside signs of the disease / disorder have yet been clinically recognized. In either case, the term prophylaxis can be applied to encompass both prevention and suppression.

[0146] Conversely, the term "treatment" refers to the clinical application of active agents to combat an already existing condition whose clinical presentation has already been realized in a patient.

[0147] As used herein, the term “substituted” or the term “substituent” are related to one or more (e.g., 2, 3, 4, 5, or 6) substituents, wherein the substituent(s) is as described herein.

[0148] As used herein, the term “substituent“ comprises hydrogen, halogen, -NO2, -CN, -OH, -CONH2, -CONR’2, -CNNR’2, -CSNR’2, -CONH-OH, -CONH-NH2, -NHCOR, - NHCSR, -NHCNR, -NC(=O)OR, -NC(=0)NR’, -NC(=S)OR’, -NC(=S)NR’, -SO2R’, - SOR’, -SR’, -SO2OR’, -SO2N(R’)2, -NHNR’2, -NNR’, C1-C6 haloalkyl, optionally substituted C1-C6 alkyl, -NH2, -NH(C1-C6 alkyl), -N(C1-C6 alkyl)2, C1-C6 alkoxy, Cl- C6 haloalkoxy, hydroxy(Cl-C6 alkyl), hydroxy(Cl-C6 alkoxy), alkoxy(Cl-C6 alkyl), alkoxy(Cl-C6 alkoxy), C1-C6 alkyl-NR’2, C1-C6 alkyl-SR’, -CONH(C1-C6 alkyl), - CON(C1-C6 alkyl)2, -CO2H, -CO2R’, -OCOR, -OCOR’, -OC(=O)OR’, -OC(=O)NR’, - OC(=S)OR’, -OC(=S)NR’, or a combination thereof; wherein each R’ independently represents hydrogen, or is selected from the group comprising optionally substituted Cl- C10 alkyl, optionally substituted C3-C10 cycloalkyl, optionally substituted C3-C10 heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, or a combination thereof.

[0149] As used herein, the term “substantially” refers to at least 85%, at least 90%, at least 92%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or between 60 and 99.9%, between 70 and 80%, between 70 and 90%, between 80 and 90%, between 90 and 95%, between 95 and 99.9%, including any range or value therebetween.

[0150] In the discussion unless otherwise stated, adjectives such as “substantially” and “about” modifying a condition or relationship characteristic of a feature or features of an embodiment of the invention, are understood to mean that the condition or characteristic is defined to within tolerances that are acceptable for operation of the embodiment for anapplication for which it is intended. Unless otherwise indicated, the word “or” in the specification and claims is considered to be the inclusive “or” rather than the exclusive or, and indicates at least one of, or any combination of items it conjoins.

[0151] It should be understood that the terms “a” and “an” as used above and elsewhere herein refer to “one or more” of the enumerated components. It will be clear to one of ordinary skills in the art that the use of the singular includes the plural unless specifically stated otherwise. Therefore, the terms “a”, “an” and “at least one” are used interchangeably in this application.

[0152] For purposes of better understanding the present teachings and in no way limiting the scope of the teachings, unless otherwise indicated, all numbers expressing quantities, percentages or proportions, and other numerical values used in the specification and claims, are to be understood as being modified in all instances by the term “about.” Accordingly, unless indicated to the contrary, the numerical parameters set forth in the following specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques.

[0153] In the description and claims of the present application, each of the verbs, “comprise”, “include”, and “have” and conjugates thereof, are used to indicate that the object or objects of the verb are not necessarily a complete listing of components, elements or parts of the subject or subjects of the verb.

[0154] Other terms as used herein are meant to be defined by their well-known meanings in the art.

[0155] Unless specifically stated or obvious from context, as used herein, the term "or" is understood to be inclusive.

[0156] Throughout this specification and claims, the word “comprise” or variations such as “comprises” or “comprising” indicate the inclusion of any recited integer or group of integers but not the exclusion of any other integer or group of integers.

[0157] As used herein, the terms "comprises", "comprising", "containing", "having" and the like can mean "includes", "including", and the like; "consisting essentially of or "consists essentially" likewise has the meaning ascribed in U.S. patent law and the term is open- ended, allowing for the presence of more than that which is recited so long as basic or novel characteristics of that which is recited is not changed by the presence of more than thatwhich is recited, but excludes prior art embodiments. In one embodiment, the terms "comprises" "comprising", and "having" are / is interchangeable with "consisting".

[0158] Although the invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications, and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations that fall within the spirit and broad scope of the appended claims.

[0159] All publications, patents and patent applications mentioned in this specification are herein incorporated in their entirety by reference into the specification, to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated herein by reference. In addition, citation, or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. To the extent that section headings are used, they should not be construed as necessarily limiting.EXAMPLESMaterials and methodsMaterials

[0160] L-Leucine, L-leucyl-, methyl ester, monohydrochloride (CAS# 6491-83-4) was purchased from Angene. Polysorbate 80 (Tween 80) was purchased from Croda. Polysorbate 20 (Tween 20), Trehalose dihydrate, Citric acid monohydrate and Sodium Hydroxide were purchased from Merck. Potassium dihydrogen phosphate was purchased from Fisher Chemical, Deionized (DI) water was purchased from Formulex, and Trisodium Citrate Dihydrate was purchased from Sigma Aldrich.Citric Buffer (0.1M, pH 5.5)

[0161] 23.3 mL of citric acid solution (21.01g citric acid in IL DI water) was mixed with 76.7 ml of sodium citrate solution (29.0 g sodium citrate in IL DI water), to obtain a 100 ml buffer citrate.Phosphate Buffer (0.1M, pH 7.4)

[0162] 50.0 mL of KH2PO4 solution (27.22 g of KH2PO4 in 1 L of DI water ) was mixed with 39.1 ml of sodium hydroxide solution (8.0 g of sodium hydroxide in 1 L of DI water), 110.9 ml of DI water was added to prepare 200 ml of buffer phosphate.EXAMPLE 1 - Solubility of an exemplary di-peptide of the invention

[0163] First, water solubility of an exemplary low soluble peptide (L-Leucyl-L-Leucine methyl ester hydrochloride) as compared to its free carboxy-counterpart (L-Leu-L-Leu-OH *HC1) was tested as disclosed hereinbelow.

[0164] 100.6 mg of L-Leu-L-Leu OH was weighed and placed in clear glass testing tube. About 800 pL of IM HC1 (about twice excess) was added by micro-pipettor and mixed well. About 4200 pL of acetonitrile to form azeotrope was added by micro-pipettor and mixed well. The mixture was heated using water bath till 80 °C to allow the liquid to evaporate. The tube with dry L-Leu-L-Leu OH* HC1 was weighed. Water was added dropwise at RT to the tube with L-Leu-L-Leu OH* HC1. The adding water was stopped when the near clear solution is observed (visual test). The solution was allowed to be mixed overnight till complete clarity. The resulting solution was weighed. The solubility of 115.3 mg L-Leu-L- Leu OH* HC1 in obtained amount (mL=l mg) of water was calculated. The calculated water solubility of L-Leu-L-Leu OH* HC1 was 35.7 mg / mL.

[0165] 100.3 mg of L-Leucyl-L-Leucine methyl ester hydrochloride (LLME*HC1) was weighed and placed in a clear glass testing tube. The tube with dry LLME* HC1 was weighed. Water was added dropwise at RT to the tube with LLME* HC1. The adding water was stopped when the near clear solution is observed (visual test). The solution was allowed to be mixed overnight till complete clarity. The resulting solution was weighed. The solubility of LLME* HC1 1 in obtained amount (mL=l mg) of water was calculated. The calculated water solubility of LLME* HC1 was 19.9 mg / mL.

[0166] Subsequently, the solubility and solution stability of LLME-HC1 in 1-10% of phosphate buffer (pH 7.4) and 1-10% of citric buffer, in the presence of Trehalose, Tween 20, Tween 80 as potential stabilizers was examined. All compositions were visually evaluated for clearness and absences of precipitation for two weeks. Exemplary compositions were made by mixing LLME-HC1 and trehalose at a temperature of between 40°C and 50°C according to Table 1.Table 1: formulation variables, and the visual stability observed.* Citric acid buffer pH 5.5; Phosphate buffer pH 7.4.

[0167] Citric acid buffer increased the peptide stability. In addition, the presence of trehalose in the formulation enabled stabilization of the peptide solution at a concentration of up to 30%w / w for a time period ranging between 2 and 14 days, depending on the trehalose: peptide ratio and peptide concentration.In addition, the formulation were filtered using a nylon filter 0.22 um, no major difference was observed, indicating sterilization can be done.

[0168] The inventors demonstrated that an aqueous solution comprising the composition of the invention has significantly increased solution stability of LLME-HC1, as compared to a similar solution devoid of the stabilizer.

[0169] Surprisingly, the inventors obtained that the presence of a surfactant significantly reduced stability of an exemplary solution of the invention.EXAMPLE 2 - Stability enhancement by various stabilizers

[0170] Exemplary stabilizers and their effect on the peptide (LLME-HC1 ) stability were tested. In brief, 0.98 g of LLME-HC1 and 0.81 g of various stabilizers were added to 8.21 g or 18.21 of an aqueous citrate buffer (pH 5.5) to obtain different solutions with a final w / w concentration of LLME-HC1 being 9.8% or 4.9%, respectively. Stability was examined by visual observation of crystal formation in the solutions. The tested stabilizers are provided in Table 2 below.Table 2:00171] Surprisingly, all the tested stabilizers increased solution stability of LLME-HC1 by at least 2 times compared to the same solution without the stabilizer. The entire tested solution were stable for at least 4 days. Form the above, trehalose and raffinose showed superior stabilizing activity.EXAMPLE 3Reconstitution of an exemplary powderous compositionPowder preparation

[0172] An exemplary powder of the invention was obtained by lyophilization using Company Freeze Dryer (at -50°C, 10Pa< and drying time of 20 to 24 hours) of an aqueous solution containing 1.18 g LLME-HC1, 1g trehalose in 10L citric buffer pH 5.5. The stock was divided into 10 vials for repetition. The lyophilized powder content is described in table 3.Table 3: lyophilized content of the tested powderous compositions00173] The inventors were able to show that the amount of LLME-HC1 within the powder corresponds to the theoretical amount of the peptide.Reconstitution

[0174] A high-pressure liquid chromatography (HPLC) assay for LLME HC1 was developed, and the amount of LLME HC1 was examined in a composition reconstituted from the lyophilized powder. Results demonstrate that upon reconstitution at least 95% recovery of the peptide has been obtained. Results of the reconstitution tests are summarized in Table 4.

[0175] Table 4: LLME-HC1 assay in reconstituted composition.The reconstitution of the exemplary powder at a final peptide concentration was tested.Surprisingly, sufficient peptide recovery was observed even at a final LLME concentration of 30%, without impairing the formulation stability.

[0176] In an additional experiment, a reconstitution of a composition comprising raffinose as the stabilizer (0.98 g LLME-HC1, 0.81 g raffinose and 18.21 citric buffer pH 5.5) has been performed. Peptide recovery above 90% was obtained in this experiment.EXAMPLE4

[0177] The inventors have compared water solubility of methyl esters of additional di- and tri-peptides in an exemplary composition of the invention or in a citrate buffer (i.e. without trehalose).

[0178] General Procedure for Determination of Di- and Tripeptides Solubility in 0.1M Citric buffer pH 5.5

[0179] Weight 100 mg of di-tripeptide methyl ester sample and place in a clear glass testing tube or bicker.

[0180] Weight the tube with a dry di-tripeptide methyl ester sample.

[0181] Add 0.1M Citric buffer solution dropwise at RT to the tube with di-tripeptide methyl ester sample. Periodically vortex the mixture.

[0182] Stop adding 0.1M Citric buffer solution when the clear solution is observed (visual test).

[0183] Weight the resulting solution.

[0184] Calculate the solubility as 100 mg of di-tripeptide methyl ester sample in the obtained amount (mL) of 0.1 M Citric buffer solution.

[0185] Results of initial solubility and solutions stability (appearance or absence of precipitate) were determined after 2, 4, 6, and 72 hours.

[0186] General Procedure for Determination of Di- and Tripeptides Solubility in 10% Trehalose - 0.1M Citric buffer pH 5.5

[0187] Weight 100 mg of di-tripeptide methyl ester sample and place in a clear glass testing tube or bicker.

[0188] Weight the tube with a dry di-tripeptide methyl ester sample.

[0189] Add 10% Trehalose in 0.1M Citric buffer pH 5.5 solution dropwise at RT to the tube with di-tripeptide methyl ester sample. Periodically vortex the mixture.

[0190] Stop adding 10% Trehalose in 0.1M Citric buffer pH 5.5 solution when the clear solution is observed (visual test).

[0191] Weight the resulting solution.

[0192] Calculate the solubility as 100 mg of di-tripeptide methyl ester sample in the obtained amount (mL) of 10% Trehalose in 0.1M Citric buffer pH 5.5 solution.

[0193] Results of initial solubility and solutions stability (appearance or absence of precipitate) were determined after 2, 4, 6, and 72 hours.

[0194] The Di- and Tripeptides Solubility in 0.1M Citric buffer pH 5.5 dissolution results are summarized in Table 5.

[0195] Table 5: Di- and Tripeptides Solubility in 0.1M Citric buffer pH 5.5

[0196] The Di- and Tripeptides Solubility in 10% Trehalose- 0.1M Citric buffer pH 5.5 dissolution results are summarized in Table 6.

[0197] Table 6. Di- and Tripeptides Solubility in 10% Trehalose-O.IM Citric buffer pH5.500198] The solubility summary and difference in % between solubility in citric buffer, pH 5.5 and 10% trehalose in citric buffer solutions is depicted in Table 7.

[0199] Table 7. solubility summary and difference in % between solubility in citric buffer, pH 5.5 and 10% trehalose in citric buffer solutions.

[0200] As shown in Table 7, the solubility of di- and tripeptides is at least 20% higher in trehalose / citric buffer solutions, compared to sole citric buffer solution.

[0201] To test the stabilizing effect of 10% trehalose in citric buffer solution, the concentrations of di- and tripeptides were made equal in both media, i.e. certain amount of trehalose solution was added to each di- or tripeptide solution. Then, the concentration of the solution was raised by slow evaporation of the water till the beginning of the peptides' precipitation was observed. The observations are summarized in Table 8.

[0202] Table 8. Observational stability of di and tripeptides in citric buffer, pH 5.5 and 10% trehalose in citric buffer solutions1- About 2-3 Hr. after gentle evaporation at 40-50°C.

[0203] To this end, peptides dissolved in the composition of the invention (e.g. an aqueous solution containing trehalose as the stabilizer and citric buffer) demonstrated both improved solubility and stability of the aqueous solution compared to citric buffer solution without the stabilizer.

[0204] Although the invention has been described in conjunction with specific embodiments thereof, it is evident that many alternatives, modifications and variations will be apparent to those skilled in the art. Accordingly, it is intended to embrace all such alternatives, modifications and variations that fall within the spirit and broad scope of the appended claims.

[0205] All publications, patents and patent applications mentioned in this specification are herein incorporated in their entirety by reference into the specification, to the same extent as if each individual publication, patent or patent application was specifically and individually indicated to be incorporated herein by reference. In addition, citation or identification of any reference in this application shall not be construed as an admission that such reference is available as prior art to the present invention. To the extent that section headings are used, they should not be construed as necessarily limiting.

Claims

CLAIMSWhat is claimed:

1. A composition comprising: (i) a peptide, including any enantiomer thereof, a salt thereof, a hydrate or a solvate thereof, or any combination thereof; and (ii) a stabilizer; wherein: said peptide is between 2 and 10 amino acids long; said peptide comprises an ester; and wherein said stabilizer is selected from: a diol, a polyol, a mono-saccharide, a disaccharide, an oligo- saccharide, and a poly-saccharide, including any combination thereof.

2. The composition of claim 1, wherein said ester is a carboxy terminal ester.

3. The composition of claim 1 or 2, wherein said composition is an aqueous solution.

4. The composition of claim 3, wherein said aqueous solution is a citrate buffered solution.

5. The composition of claim 3 or 4, wherein said peptide is present at a concentration of up to 40 weight % within the aqueous solution; and wherein said peptide is between 2 and 5 amino acid long.

6. The composition of any of claims 3 to 5, wherein said composition is stable at a temperature between 2 and 40 °C, for at least 72h.

7. The composition of any of claims 1 to 6, wherein said peptide and said stabilizer are present within said composition at a molar ratio of between 1 : 1 and 10:1.

8. The composition of any of claims 1 to 7, wherein said peptide comprises a structure represented by Formula 1 :, wherein R is C1-C5 alkyl.

9. The composition of claim 8, wherein said peptide is L-Leucyl-L-Leucine ester, a salt thereof, a hydrate or a solvate thereof, or any combination thereof.

10. The composition of claim 9, wherein said R is methyl.

11. The composition of claim 1 or 2, wherein said composition is in a form of a powder and is characterized by a water content of below 5%w / w.

12. The composition of claim 11, further comprising a buffering agent.

13. The composition of claim 12, wherein said buffering agent comprises a citrate salt.

14. The composition of any one of claims 1 to 13, wherein said stabilizer comprises trehalose.

15. A pharmaceutical composition, comprising: (i) a peptide, including any enantiomer thereof, a salt thereof, a hydrate or a solvate thereof, or any combination thereof; (ii) a stabilizer; and (iii) a pharmaceutically acceptable carrier; wherein said peptide is between 2 and 10 amino acids long and said peptide comprises an ester; and wherein said stabilizer is selected from: a diol, a polyol, a mono-saccharide, a disaccharide, an oligo- saccharide, and a poly-saccharide, including any combination thereof.

16. The pharmaceutical composition of claim 15, wherein said pharmaceutically acceptable carrier is an aqueous solution.

17. The pharmaceutical composition of claim 16, wherein said aqueous solution comprises is a buffered solution comprising a pharmaceutically acceptable buffering agent.

18. The pharmaceutical composition of claim 17, wherein said buffered solution is characterized by a pH value between 5 and 8.

19. The pharmaceutical composition of claim 17 or 18, wherein said pharmaceutically acceptable buffering agent comprises a pharmaceutically acceptable citrate salt.

20. The composition of any of claims 15 to 19, wherein said stabilizer is selected from a diol, a polyol, a mono- saccharide, a di-saccharide, an oligo-saccharide, and a poly- saccharide, including any combination thereof.

21. The composition of claim 20, wherein said di-saccharide comprises trehalose.

22. A kit comprising: (i) a peptide, including any enantiomer thereof, a salt thereof, a hydrate or a solvate thereof, or any combination thereof; and (ii) a stabilizer; wherein said peptide is between 2 and 10 amino acids long and said peptide comprises an ester; and wherein said stabilizer is selected from: a diol, a polyol, a mono-saccharide, a disaccharide, an oligo- saccharide, and a poly-saccharide, and any combination thereof.

23. The kit of claim 22, wherein said peptide and said stabilizer are in a form of a mixture.

24. The kit of claim 22, wherein said peptide and said stabilizer are stored in separate containers.

25. The kit of any one of claims 22 to 24, wherein said kit further comprises a citrate salt or an aqueous citrate buffer.

26. The kit of claim 25, wherein said peptide and said aqueous citrate buffer are present within the kit at a weight ratio of between 1:500 and 1:10,000.

27. The kit of claim 25 or 26, comprising instructions for mixing said peptide, said stabilizer and optionally said aqueous citrate buffer to obtain the composition of any one of claims 3 to 10.

28. The kit of any one of claims 22 to 27, wherein said peptide and said stabilizer are present within said kit at a molar ratio of between 1 : 1 and 10:1.

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