Liquid Compositions Comprising Phosphorylcholine-Tuftsin Conjugates - Patent application

Abstract

Provided herein is a fungicidal composition comprising a pharmacologic effective amount of phosphorylcholine-tuftsin conjugate, and optionally comprising a pharmacologic acceptable carrier.Also provided is a method of using the fungicidal composition, such as for treating or preventing disease or disorder related to fungal infection in a subject in need thereof.Furthermore, provided is a method of using the fungicidal composition, such as for extending the shelf life of food and / or preventing fungal contamination.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 281,690, filed November 21, 2021, the entire contents of which are incorporated herein by reference.

[0002] Technical Field The present invention is directed to novel phosphorylcholine-tuftsin conjugates and fungicidal compositions comprising same. The present invention is further directed in some embodiments thereof to the use of the phosphorylcholine-tuftsin conjugates and / or fungicidal compositions as a preservative and / or for the prevention or treatment of fungal infections and / or conditions associated therewith in a subject. [Background technology]

[0003] Many fungicides are well known in the art. These compounds have sufficient antifungal activity, but are highly toxic. Therefore, there is a need for new fungicidal compounds that are characterized by low toxicity. Furthermore, there is a need for new non-toxic compounds that can be used as spoilage inhibitors in liquid formulations and / or foods, etc.

[0004] Tuftsin-phosphorylcholine (TRS) is a bispecific small molecule with immunomodulatory activity. Tuftsin (Thr-Lys-Pro-Arg) is a natural immunomodulatory peptide produced in the spleen by enzymatic cleavage of the Fc domain of the heavy chain of IgG. Phosphorylcholine (PC) is a zwitterionic small molecule secreted by helminths that allows them to survive in the host inducing a state of immune tolerance, as well as on the surface of some bacteria and apoptotic cells. Furthermore, TRS is known for its Toll-like receptor (TLR-4) inhibitory activity. Summary of the Invention

[0005] In one aspect of the invention, there is provided a fungicidal composition comprising a fungicidally effective amount of a phosphorylcholine-tuftsin conjugate, a salt thereof, a functional derivative thereof, or any combination thereof.

[0006] In one embodiment, a phosphorylcholine-tuftsin conjugate comprises at least one phosphorylcholine moiety or derivative thereof and tuftsin or a derivative thereof covalently attached via a spacer.

[0007] In one embodiment, the spacer comprises at least two amino acids.

[0008] In one embodiment, the phosphorylcholine moiety or a derivative thereof is covalently attached to the spacer via a diazo group.

[0009] In one embodiment, the phosphorylcholine-tuftsin conjugate has Formula 1: [ka] It is expressed by:

[0010] In one embodiment, the fungicidal composition of the present invention further comprises a suitable carrier.

[0011] In one embodiment the fungicidal composition is a pharmaceutical composition comprising a pharma- ceutically acceptable carrier.

[0012] In one embodiment the w / w of the carrier within the fungicidal composition is between 10-99.99%.

[0013] In one embodiment, the fungicidal composition is in the form of a liquid formulation.

[0014] In another aspect, there is provided a method for treating or preventing a disease or disorder associated with a fungal infection in a subject in need thereof, comprising administering to said subject a therapeutically effective amount of a fungicidal composition of the present invention.

[0015] In one embodiment, the therapeutically effective amount is an amount sufficient to reduce the fungal burden in a subject or an amount sufficient to ameliorate at least one symptom associated with said disease or disorder in a subject.

[0016] In another aspect, there is provided a method of reducing or inhibiting the activity or growth of at least one fungus, comprising the step of contacting the fungus with a fungicidally effective amount of a fungicidal composition of the invention, thereby reducing the activity or reducing the growth of said fungus.

[0017] Further embodiments and the full scope of applicability of the present invention will become apparent from the detailed description provided hereinafter. It should be understood, however, that the detailed description and specific examples, while representing preferred embodiments of the invention, are given by way of illustration only, since various changes and modifications within the spirit and scope of the invention will become apparent to those skilled in the art from the detailed description.

[0018] In addition to the exemplary aspects and embodiments described above, further aspects and embodiments will become apparent by reference to review of the following detailed description.

[0019] Unless otherwise defined, all technical and / or scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs.Although methods and materials similar or equivalent to those described herein may be used in the practice or testing of embodiments of this invention, exemplary methods and / or materials are described below.In case of conflict, the present specification, including definitions, will take precedence.In addition, materials, methods, and examples are merely examples and are not necessarily intended to be limiting. [Brief description of the drawings]

[0020] [Figure 1]FIG. 1 is a graph depicting the fermentation activity of aqueous yeast (S. cerevisiae) suspensions as a function of concentration of the phosphorylcholine-tuftsin conjugate of Formula 1 compared to a control suspension containing no phosphorylcholine-tuftsin conjugate. [Diagram 2] FIG. 2 is a graph depicting the fermentation activity of aqueous yeast (S. cerevisiae) suspensions as a function of concentration of the phosphorylcholine-tuftsin conjugate of Formula 2 compared to a control suspension containing no phosphorylcholine-tuftsin conjugate. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0021] The present invention, in some embodiments thereof, relates to a fungicidal composition comprising a phosphorylcholine-tuftsin conjugate (hereinafter "TRS") of formula 1, and a method of treating a medical condition (e.g., a disease or disorder) in a subject associated with a mycosis by administering to a subject in need thereof a therapeutically effective amount of the fungicidal composition. In further embodiments thereof, the present invention relates to an antifungal (or anti-mold) composition for use in preventing or reducing the fungal load in a composition (e.g., an aqueous composition or a food substance).

[0022] The present invention, in some embodiments thereof, is based on the surprising discovery that the phosphorylcholine-tuftsin conjugates disclosed herein were capable of substantially inhibiting yeast function (fermentation) in an aqueous suspension comprising yeast and nutrients (e.g., sucrose).

[0023] Before describing at least one embodiment of the invention in detail, it is to be understood that the invention is not necessarily limited in its application to the details set forth in the following description or illustrated by way of examples, as the invention is capable of other embodiments or of being practiced or carried out in various ways.

[0024] Phosphorylcholine-Tuftsin conjugate In one aspect of the present invention, there is provided (i) a phosphorylcholine-tuftsin conjugate, (ii) a functional derivative thereof, comprising any salt or any combination of (i) and (ii), wherein the phosphorylcholine-tuftsin conjugate is represented by Formula 2: [ka] It is expressed by:

[0025] In some embodiments, functional derivatives include any phosphorylcholine-tuftsin conjugate having at least 80%, at least 85%, at least 90%, at least 95%, at least 97%, or between 80-99%, between 85-99%, between 90-99%, or between 95-99% structural similarity to the phosphorylcholine-tuftsin conjugate of formula 2 and characterized by antifungal activity as disclosed herein below.

[0026] In some embodiments, the term "structural similarity" refers to the fingerprint similarity between two molecules. The term "fingerprint similarity" is well understood by those skilled in the art. In some embodiments, fingerprint similarity is calculated based on circular fingerprint, substructure key-based fingerprint, and / or topological or path-based fingerprint.

[0027] Exemplary circular fingerprints include, but are not limited to, Molprint 2D, ECFP (or Morgan fingerprint), FCFP, etc. In some embodiments, the term "structural similarity" as used herein is calculated by the Morgan fingerprint.

[0028] In some embodiments, the functional derivative has formula 3: [ka] It is expressed by:

[0029] In some embodiments, the functional derivative has formula 4: [ka] or Equation 5: [ka] (wherein R includes any salt or deprotonated amine thereof as permitted by valence) [ka] and TIFF2024542317000023.tif2214) It is expressed by:

[0030] In some embodiments, the phosphorylcholine-tuftsin conjugate and / or functional derivatives of the phosphorylcholine-tuftsin conjugate include any salts thereof, wherein the salts are pharma- ceutically acceptable salts.

[0031] As used herein, the term "pharmaceutically acceptable salt" refers to a compound of the present invention or an active metabolite or salt thereof that is administered, directly or indirectly, to a subject, e.g., a human, after administration. It refers to any non-toxic salt of the compound of the present invention that is capable of providing a residue. For example, the term "pharmaceutical acceptable" can mean approved by a federal or state government regulatory agency for use in animals, more particularly in humans, or listed in the United States Pharmacopeia or other generally recognized pharmacopeia.

[0032] Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. describe pharma-ceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19. Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. These salts can be prepared in situ during the final isolation and purification of the compounds. Acid addition salts can be prepared by 1) reacting the purified compound in its free base form with a suitable organic or inorganic acid, and 2) isolating the salt thus formed.

[0033] Non-limiting examples of pharma- ceutically acceptable salts include, but are not limited to, acetate, trifluoroacetate, aspartate, benzenesulfonate, benzoate, bicarbonate, carbonate, halide (e.g., bromide, chloride, iodide, fluoride), bitartrate, citrate, salicylate, stearate, succinate, sulfate, tartrate, decanoate, edetate, fumarate, gluconate, and lactate salts, or any combination thereof.

[0034] Further examples of pharma- ceutically acceptable non-toxic acid addition salts are salts of amino groups formed with inorganic acids such as hydrochloric, hydrobromic, phosphoric, sulfuric, and perchloric acids, or with organic acids such as acetic, oxalic, maleic, tartaric, citric, succinic, or malonic acid, or by using other methods used in the art, such as ion exchange.

[0035] Other pharma- ceutically acceptable salts include adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecyl sulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, glycolate, gluconate, glycolate, hemisulfate, heptanoate, hexanoate, hydrochloride, hydrobromide, hydroiodide, 2-hydroxy-ethanesulfonate, and the like. Examples of salts that may be used include esters, lactobionates, lactates, laurates, lauryl sulfates, malates, maleates, malonates, methanesulfonates, 2-naphthalenesulfonates, nicotinates, nitrates, oleates, oxalates, palmitates, palmoates, pectinates, persulfates, 3-phenylpropionates, phosphates, picrates, pivalates, propionates, salicylates, stearates, succinates, sulfates, tartrates, thiocyanates, p-toluenesulfonates, undecanoates, and valerates.

[0036] Base addition salts can be prepared by 1) reacting the purified compound in its acid form with an appropriate organic or inorganic base and 2) isolating the salt thus formed. Salts derived from appropriate bases include alkali metal (e.g., sodium, lithium, and potassium), alkaline earth metal (e.g., magnesium and calcium), ammonium, and N+(C1-4 alkyl)4 salts. The present invention also contemplates the quaternization of any basic nitrogen-containing groups of the compounds disclosed herein. Water- or oil-soluble or dispersible products may be obtained by such quaternization.

[0037] Further pharma- ceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and ammonium cations, formed, where appropriate, using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkylsulfonates and arylsulfonates, etc. Other acids and bases, while not themselves pharma- cetically acceptable, may be used in the preparation of salts which are useful as intermediates to obtain the compounds of the invention and their pharma- ceutical acceptable acid or base addition salts.

[0038] In some embodiments, the phosphorylcholine-tuftsin conjugates described herein are chiral compounds (i.e., have asymmetric carbon atoms). In some embodiments, diastereomers, geometric isomers, and individual isomers are encompassed within the scope of the present invention. In some embodiments, the phosphorylcholine-tuftsin conjugates described herein are in the form of a racemic mixture. In some embodiments, the phosphorylcholine-tuftsin conjugates are in the form of a single enantiomer or a single diastereomer.

[0039] In some embodiments, the phosphorylcholine-tuftsin conjugate is in the form of a single enantiomer having an enantiomeric purity of greater than 70%. In some embodiments, the chiral compound is in the form of a single enantiomer having an enantiomeric purity of greater than 80%. In some embodiments, the chiral compound is in the form of a single enantiomer having an enantiomeric purity of greater than 90%. In some embodiments, the chiral compound is in the form of a single enantiomer having an enantiomeric purity of greater than 95%.

[0040] In some embodiments, the phosphorylcholine-tuftsin conjugate has formula 2A: [ka] It is expressed by:

[0041] In some embodiments, the phosphorylcholine-tuftsin conjugate has formula 3A: [ka] It is expressed by:

[0042] In some embodiments, the phosphorylcholine-tuftsin conjugate containing an unsaturated bond is in the form of a trans-isomer or a cis-isomer. In some embodiments, the phosphorylcholine-tuftsin conjugate comprises a mixture of cis- and trans-isomers, as described hereinabove.

[0043] In some embodiments, the phosphorylcholine-tuftsin conjugates described herein may exist in solvated forms, including unsolvated and hydrated forms. In general, solvated forms are equivalent to unsolvated forms and are included within the scope of the present invention. Certain compounds of the present invention may exist in polycrystalline or amorphous forms. In general, all physical forms are equivalent for the uses contemplated by the present invention and are intended to be within the scope of the present invention.

[0044] The term "solvate" refers to a complex of variable stoichiometry (e.g., di-, tri-, tetra-, penta-, hexa-, etc.) formed by a solute (a conjugate as described herein) and a solvent, whereby the solvent does not interfere with the biological activity of the solute. Suitable solvents include, for example, ethanol, acetic acid, and the like.

[0045] The term "hydrate" refers to a solvate as defined hereinabove, wherein the solvent is water.

[0046] Unless otherwise stated, the structures described herein are also meant to include all isomeric (e.g., enantiomeric, diastereomeric, geometric, conformational, and rotational) forms of the structure. For example, the R and S configurations of each asymmetric center, (Z) and (E) double bond isomers, and (Z) and (E) conformational isomers are included in the present invention. As will be understood by those skilled in the art, substituents can freely rotate around any rotatable bond. Thus, single stereochemical isomers as well as mixtures of enantiomeric, diastereomeric, geometric, conformational, and rotational isomers of the compounds of the present invention are within the scope of the present invention.

[0047] Unless otherwise stated, all tautomeric forms of the phosphorylcholine-tuftsin conjugates of the present invention are within the scope of the present invention.

[0048] In some embodiments, the phosphorylcholine-tuftsin conjugates of the invention are single compounds characterized by a chemical purity of at least 90%, at least 92%, at least 95%, at least 96%, at least 97%, at least 99%, at least 99.5%, between 90-99%, between 92-95%, between 92-97%, between 92-99% (including any range therebetween). Chemical purity may be determined by analytical HPLC, GC, or LC-MS.

[0049] In some embodiments, the phosphorylcholine-tuftsin conjugates of the invention contain less than 0.5%, less than 0.1%, less than 0.001% of phosphorylcholine-tuftsin conjugate of formula 1. In some embodiments, the phosphorylcholine-tuftsin conjugates of the invention lack detectable traces of phosphorylcholine-tuftsin conjugate of formula 1, where detectable is determined by standard analytical methods such as nuclear magnetic resonance (NMR), high performance liquid chromatography (HPLC), and mass spectrometry (MS), gas chromatography mass spectrometry, liquid chromatography mass spectrometry (GC-MS, LC-MS) and similar methods used by those skilled in the art to assess such purity.

[0050] In some embodiments, the phosphorylcholine-tuftsin conjugates of the invention comprise multiple (eg, 2, 3, 4, or 5) chemically distinct compounds (eg, multiple conjugates of Formulas 1-5).

[0051] fungicidal composition In another aspect of the present invention, a composition is provided that comprises one or more of the phosphorylcholine-tuftsin conjugates of the present invention and a carrier.In some embodiments, the carrier is a liquid carrier.In some embodiments, the carrier is a pharmaceutically acceptable carrier.In some embodiments, the composition is a pharmaceutical composition or a fungicide composition.

[0052] In another aspect of the invention, there is provided a fungicidal composition comprising one or more phosphorylcholine-tuftsin conjugates of the invention. In some embodiments, the fungicidal composition comprises a fungicidally effective amount of one or more phosphorylcholine-tuftsin conjugates of the invention (i.e., conjugates of formulas 1-5). In some embodiments, the fungicidal composition comprises a fungicidally effective amount of a phosphorylcholine-tuftsin conjugate of formula 1. In some embodiments, the fungicidal composition comprises a fungicidally effective amount of a phosphorylcholine-tuftsin conjugate of formula 2. In some embodiments, the fungicidal composition is devoid of a fungicide that is not a phosphorylcholine-tuftsin conjugate. In some embodiments, the fungicidal composition consists essentially of one or more phosphorylcholine-tuftsin conjugates of the invention as a fungicidally active ingredient. In some embodiments, the fungicidal composition consists essentially of one or more phosphorylcholine-tuftsin conjugates of the invention and a carrier.

[0053] In some embodiments, the fungicidal composition comprises a plurality (e.g., 2, 3, 4, 5, 10, including any range therebetween) of phosphorylcholine-tuftsin conjugates of the present invention, and at least one phosphorylcholine-tuftsin conjugate is present in the fungicidal composition in a fungicidally effective amount.

[0054] In some embodiments, the fungicidal composition comprises a fungicidally effective amount of a phosphorylcholine-tuftsin conjugate of formula 1, and further comprises a phosphorylcholine-tuftsin conjugate of formula 2, and optionally comprises one or more functional derivatives of the phosphorylcholine-tuftsin conjugate of formula 2.

[0055] In some embodiments, the fungicidal composition comprises a fungicidally effective amount of a phosphorylcholine-tuftsin conjugate of formula 2, and further comprises a phosphorylcholine-tuftsin conjugate of formula 1, and optionally comprises one or more functional derivatives of the phosphorylcholine-tuftsin conjugate of formula 2.

[0056] In some embodiments, the carrier is a liquid carrier (e.g., a flowable carrier, e.g., an aqueous carrier, a semi-solid carrier, a semi-liquid carrier). In some embodiments, the carrier is a pharma- ceutically acceptable carrier.

[0057] In some embodiments, the fungicidal composition is a liquid composition. In some embodiments, the fungicidal composition comprises a phosphorylcholine-tuftsin conjugate, and / or a salt thereof, and / or a functional derivative thereof (e.g., a peptide having a similar structure and characterized by antifungal activity). In some embodiments, the fungicidal composition is a pharmaceutical composition comprising a phosphorylcholine-tuftsin conjugate and a pharma- ceutically acceptable carrier. In some embodiments, the fungicidal composition is an anti-mold composition comprising a phosphorylcholine-tuftsin conjugate and a suitable (e.g., food-grade) carrier.

[0058] In some embodiments, the salt is a pharma- ceutically acceptable salt. In some embodiments, the salt is a cosmetically acceptable salt. In some embodiments, the salt is a food grade salt.

[0059] In some embodiments, the fungicidally effective amount is a preservative effective concentration of one or more of the phosphorylcholine-tuftsin conjugates of the present invention in the fungicidal composition. In some embodiments, the terms "preservative effective concentration" and "preservative effective amount" are used interchangeably herein and refer to a concentration of phosphorylcholine-tuftsin conjugate sufficient for the preservative activity of the fungicidal compositions disclosed herein.

[0060] In some embodiments, a fungicidally effective amount comprises a concentration of one or more phosphorylcholine-tuftsin conjugates of the present invention within a fungicidal composition of at least 1 nM, at least 10 nM, at least 100 nM, at least 500 nM, at least 1 μM, at least 2 μM, at least 5 μM, at least 10 μM, at least 15 μM, at least 20 μM, at least 30 μM, at least 40 μM, at least 50 μM, at least 75 μM, at least 100 μM, at least 150 μM, at least 200 μM, at least 400 μM, at least 500 μM, at least 600 μM, at least 1,000 μM, or any range or value therebetween. Each possibility represents a separate embodiment of the present invention.

[0061] In some embodiments, a fungicidally effective amount comprises a concentration of one or more phosphorylcholine-tuftsin conjugates of the present invention within a fungicidal composition of at least 1 nM, at least 10 nM, at least 100 nM, at least 500 nM, at least 1 μM, at least 2 μM, at least 5 μM, at least 10 μM, at least 15 μM, at least 20 μM, at least 30 μM, at least 40 μM, at least 50 μM, at least 75 μM, at least 100 μM, at least 150 μM, at least 200 μM, at least 400 μM, at least 500 μM, at least 600 μM, at least 1,000 μM, or any range or value therebetween. Each possibility represents a separate embodiment of the present invention.

[0062] In some embodiments, a fungicidally effective amount comprises a concentration of one or more phosphorylcholine-tuftsin conjugates of the invention (e.g., conjugates of Formula 2) within the fungicidal composition of at least 1 ppm, at least 5 ppm, at least 10 ppm, at least 100 ppm, at least 500 ppm, at least 1000 ppm, or any range or value therebetween. Each possibility represents a separate embodiment of the present invention.

[0063] In some embodiments, a fungicidally effective amount comprises a concentration of one or more phosphorylcholine-tuftsin conjugates of the invention (e.g., conjugates of Formula 1) within the fungicidal composition of at least 1 ppm, at least 5 ppm, at least 10 ppm, at least 100 ppm, at least 500 ppm, at least 1000 ppm, or any range or value therebetween. Each possibility represents a separate embodiment of the present invention.

[0064] In some embodiments, the concentration of one or more phosphorylcholine-tuftsin conjugates of the invention within the fungicidal composition is between 5 ppm and 10 (w / w)%, between 5 ppm and 99 (w / w)%, between 5 ppm and 90 (w / w)%, between 5 ppm and 95 (w / w)%, between 5 ppm and 50 (w / w)%, between 5 ppm and 60 (w / w)%, between 5 ppm and 80 (w / w)%, between 5 ppm and 5 (w / w)%, between 5 ppm and 2 (w / w)%, between 0.5 and 10 (w / w)%, between 0.5 and 90 (w / w)%, between 0.5 and 50 (w / w)%, between 5 ppm and 0.5 (w / w)% (including any range therebetween).

[0065] Throughout the specification, "fungicidal composition" is further meant to refer to a formulation. As used herein, the term "formulation" refers to a vehicle composition in the form of a solution, emulsion, lotion, cream, gel, etc., optionally further comprising a physiologically acceptable carrier and / or excipient, and optionally other chemical ingredients, such as cosmetically, cosmeceutical, or pharma- ceutical active agents (e.g., drugs). A formulation may optionally include a carrier, and optionally further active agents and / or additives.

[0066] In some embodiments, the fungicidal composition is a semi-solid or gel at a temperature between 5 and 95° C. In some embodiments, the fungicidal composition is a flowable composition. In some embodiments, the fungicidal composition is a liquid composition or a flowable composition.

[0067] In some embodiments, the w / w concentration of acceptable carriers within the fungicidal composition is between 50-99%, between 60-99%, between 70-99%, between 80-99%, between 90-99%, between 50-70%, between 70-90% (including any range therebetween).

[0068] In some embodiments, the fungicidal composition is a preservative composition. In some embodiments, the preservative composition of the present invention is an aqueous solution with the active (or antimicrobial) ingredient substantially or completely dissolved therein. In some embodiments, the fungicidal composition of the present invention is a ready-to-use composition or is in the form of a dilutable concentrate.

[0069] In some embodiments, the preservative compositions or phosphorylcholine-tuftsin conjugates of the present invention are characterized by bactericidal or preservative activity in food substances (eg, liquid, solid, or semi-solid food substances).

[0070] In some embodiments, the concentration of one or more phosphorylcholine-tuftsin conjugates of the invention in the fungicidal composition is sufficient to substantially reduce or prevent pathogen formation or pathogen growth on or in an edible substance (e.g., a foodstuff) for at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 30 days, at least 60 days (including any range or value therebetween). Pathogen formation or pathogen growth may be determined visually or by performing a microbiological test. In some embodiments, the concentration of one or more phosphorylcholine-tuftsin conjugates of the invention in the fungicidal composition is sufficient to substantially prevent spoilage of an edible substance (e.g., a foodstuff) for at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least 30 days, at least 60 days (including any range or value therebetween).

[0071] In some embodiments, the reduction comprises at least a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 100% reduction (including any range therebetween), with each possibility representing a separate embodiment of the present invention.

[0072] In some embodiments, the increase includes an increase of at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, 100%, 150%, 200%, 300%, 400%, 500%, 1000%, or 10000% (including any range therebetween), with each possibility representing a separate embodiment of the present invention.

[0073] In some embodiments, the preservative compositions of the invention are characterized by a fungicidal or preservative activity in a liquid composition (e.g., an aqueous pharmaceutical or cosmetic composition) or on a surface after contacting a fungicidally effective amount of the preservative composition with the liquid composition or surface under suitable conditions (e.g., a temperature of 1-40° C. and a time period of 1 minute to 14 days, including any range therebetween). In some embodiments, the phosphorylcholine-tuftsin conjugates of the invention (e.g., a conjugate of Formula 1 or Formula 2) are characterized by a fungicidal or preservative activity in a liquid composition. In some embodiments, the fungicidal or preservative activity of the phosphorylcholine-tuftsin conjugate is activity at a fungicidally effective amount of the phosphorylcholine-tuftsin conjugate in a liquid composition, the fungicidally effective amount being as described herein above (e.g., at least 5 ppm). In some embodiments, the preservative activity is measured according to the USP <51> This refers to the preservative effectiveness determined by PET (Preservative Effectiveness Testing) in accordance with the

[0074] In some embodiments, the preservative activity refers to antibacterial activity. In some embodiments, the preservative activity refers to fungicidal activity.

[0075] In some embodiments, a liquid composition and / or surface refers to a liquid composition or surface that is susceptible to microbial infection (e.g., an article that contains a water content of at least 10%, at least 50%, or between 10-99%, between 20-90%, between 20-80%, between 30-99% (including any range therebetween)).

[0076] In some embodiments, the fungicidal compositions of the present invention are formulated for use in liquid compositions or on surfaces susceptible to microbial growth as preservatives, hi some embodiments, the liquid compositions are selected from cosmetic articles, pharmaceutical articles, edible substances, and / or foodstuffs.

[0077] The term "antimicrobial" refers to compositions or compounds capable of preventing, inhibiting, and / or controlling the growth of microorganisms; antimicrobial compounds include bactericides, bacteriostatic agents, fungicides, fungistatic agents, algicides, and algistats. The term "antimicrobial" encompasses preservatives and sterilization agents.

[0078] In some embodiments, the term "antimicrobial activity" refers to the ability to inhibit (prevent), reduce, or delay bacterial growth, fungal growth, biofilm formation, or eradicate live bacterial cells or their spores, or fungal cells or viruses in a suspension or moist environment. In some embodiments, the term "antimicrobial activity" refers to the ability of the phosphorylcholine-tuftsin conjugates of the present invention to inhibit (prevent), reduce, or delay fungal growth, biofilm formation, or eradicate live fungal cells or their spores in a suspension or moist environment. In some embodiments, the term "antimicrobial activity" refers to the ability of a fungicidally effective amount of the phosphorylcholine-tuftsin conjugates of the present invention to inhibit (prevent), reduce, or delay fungal growth, biofilm formation, or eradicate live fungal cells or their spores in a liquid composition, organism, or moist environment. Here, a fungicidally effective amount is as described herein above (e.g., at least 5 ppm) and refers to the concentration of the phosphorylcholine-tuftsin conjugate of the present invention (e.g., a conjugate of formula 1 or 2) in a liquid composition, at a target location within a living organism, or in a moist environment.

[0079] As used herein, inhibiting or reducing or slowing the formation of microbial load refers to inhibiting or reducing or slowing the growth of microorganisms and / or eradicating some or all of the existing population of microorganisms.Thus, the formulations described herein can be used both to reduce the formation of microorganisms on or in the surface of an article, and to kill microorganisms on or in the surface of an article or living tissue.

[0080] The microorganism may be, for example, a unicellular microorganism (such as a prokaryote, archaea, bacterium, eukaryote, protist, fungus, algae, mold, yeast, euglena, protozoa, dinoflagellates, apicomplexa, trypanosoma, amoeba, etc.) or a multicellular microorganism.

[0081] In some embodiments, the microorganism comprises bacteria, such as bacterial cells, such as gram-positive and gram-negative bacteria.

[0082] In some embodiments, the gram positive bacteria are Staphylococcus aureus, Staphylococcus epidermidis, and Bacillus cereus.

[0083] In some embodiments, the gram negative bacteria is Escherichia coli, Pseudomonas aeruginosa, and Burkholderia cepacia.

[0084] In some embodiments of the invention, the microorganism is a mold, yeast, or fungus, including any combination thereof. In some embodiments of the invention, the fungus is Candida albicans. In some embodiments of the invention, the yeast is a yeast of the genus Saccharomyces.

[0085] In some embodiments, the mold is Aspergillus brasiliensis.

[0086] In some embodiments, the term "fungi," including all its grammatical forms, refers to a pathogenic fungus or yeast. In some embodiments, a pathogenic fungus refers to a human pathogen.

[0087] In some embodiments, non-limiting examples of fungi are selected from Penicillium, Magnaporthe, Ophiostoma, Cryphonectria, Fusarium, Ustilago, Alternaria, Cochliobolus, Aspergillus, Candida, Cryptococcus, Histoplasma, or any combination thereof.

[0088] In some embodiments, the pathogen may be, but is not limited to, Alternaria species (e.g., Alternaria alternata, Alternaria solani); Aphanomyces species (e.g., Aphanomyces euteiches); Aspergillus species (e.g., Aspergillus niger, Aspergillus fumigatus); Atelia species (e.g., Atelia rolfsii); Aureobasidium species (e.g., Aureobasidium pullulans); Bipolaris species (e.g., Bipolaris zeicola, Bipolaris maydis); Botrytis species (e.g., Botrytis cinerea); Calonectria species (e.g., Calonectria kyotensis); Cephalosporium species (e.g., Cephalosporium maydis); Cercospora species (e.g., Cercospora medicaginis, Cercospora sojina, Colletotrichum coccodes, Colletotrichum fragariae, Colletotrichum graminicola;Coniella spp. (e.g. Coniella diplodiella);Colletotrichum spp.;Coprinopsis spp. (e.g. Coprinopsis psychromorbida);Corynespora spp. (e.g. Corynespora cassiicola;Curvularia spp. (e.g. Curvularia pallescens);Cylindrocladium spp. (e.g. Cylindrocladium crotalariae);Diplocarpon spp. (e.g. Diplocarpon earlianum);Diplodia spp. (e.g. Diplodia gossyina);Epicoccum spp. (e.g. Epicoccum nigrum);Erysiphe spp. (Erysiphe cichoracearum);Fusarium spp. (e.g. Fusarium graminearum, Fusarium oxysporum f. sp. Fragariae, Fusarium oxysporum f.sp. tuberosi, Fusarium proliferatum var. proliferatum, Fusarium solani, Fusarium verticillioides, Fusarium clumorum, Fusarium oxysporum f. sp. radicis-lycopersici, Fusarium euwallaceae); Ganoderma spp. (e.g. Ganoderma boninense); Geotrichum spp. (e.g. Geotrichum candidum); Glomerella spp. (e.g. Glomerella tucumanensis); Guignardia spp. (e.g. Guignardia bidwellii); Kabatiella spp. (e.g. Kabatiella zeae); Leptosphaerulina spp. (e.g. Leptosphaerulina briosiana);Leptotrochila species (e.g. Leptotrochila rnedicaginis);Macrophomina species (e.g. Macrophomina phaseolina);Magnaporthe species (e.g. Magnaporthe grisea, Magnaporthe oryzae);Microsphaera species (e.g. Microsphaera manshurica);Monilinia species (e.g. Monilinia fructicola);Mucor species;Mycosphaerella species (e.g. Mycosphaerella juiensis, Mycosphaerella fragariae);Nigrospora species (e.g. Nigrospora oryzae);Ophiostoma species (e.g. Ophiostoma ulmi);Penicillium species (e.g. Penicillium digitatum);Peronospora species (e.g. Peronospora manshurica);Phakopsora (e.g. Phakopsora pachyrhizi); Phoma spp. (e.g. Phoma foveata, Phoma medicaginis, Phoma tracheiphila); Phomopsis spp. (e.g. Phomopsis longicolla); Phytophthora spp. (e.g. Phytophthora cinnamomi, Phytophthora erythroseptica, Phytophthora fragariae, Phytophthora infestans, Phytophthora medicaginis, Phytophthora megaasperma, Phytophthora palmivora); Podosphaera (e.g. Podosphaera leucotricha); Pseudopeziza spp. (e.g. Pseudopeziza medicaginis); Puccinia spp. (e.g. Puccinia graminis subsp. tritici (UG99), Puccinia striiformis, Puccinia recodita, Puccinia sorghi);Pyricularia species (e.g. Pythium grisea, Pyricularia oryzae);Pythium species (e.g. Pythium ultimum, Pythium aphanidermatum);Rhizoctonia species (e.g. Rhizoctonia solani, Rhizoctonia zeae);Rosellinia species, Sclerotinia species (e.g. Sclerotinia minor; Sclerotinia sclerotiorum, Sclerotinina trifoliorum);Sclerotium species (e.g. Sclerotium rolfsii);Septoria species (e.g. Septoria glycines, Septoria lycoperski);Setomelanomma species (e.g. Setomelanomma turcica);Sphaerotheca species (e.g. Sphaerotheca macularis);Spongospora species (e.g. Spongospora subterranean);The fungus is selected from Stemphylium species, Synchytrium species (e.g., Synchytrium endobioticum), Verticillium species (e.g., Verticillium albo-atrum, Verticillium dahliae). In some embodiments, the fungus comprises Penicillium digitatum and / or Geotrichum candidum;

[0089] In some embodiments, non-limiting examples of yeast are selected from Cryptococcus neoformans, Candida albicans, Candida tropicalis, Candida stellatoidea, Candida glabrata, Candida krusei, Candida parapsilosis, Candida guilliermondii, Candida viswanathii, Candida lusitaniae, and Rhodotorula mucilaginosa, or any combination thereof.

[0090] The term "biofilm," as used herein, refers to an aggregate of living cells that are attached to each other and / or fixed onto a surface as a colony. The cells are often embedded within an autocrine matrix of extracellular polymeric substances (EPS), a sticky polymeric mixture of nucleic acids, proteins, and polysaccharides, also known as "slime."

[0091] In the context of embodiments of the present invention, the living cells that form a biofilm may be cells of unicellular microorganisms (prokaryotes, archaea, bacteria, eukaryotes, protists, fungi, algae, euglena, protozoa, dinoflagellates, apicomplexa, trypanosoma, amoebae, etc.) or cells of multicellular organisms in which case the biofilm may be considered a subform of a colony of cells or tissue (as is the case for unicellular organisms).

[0092] In the context of the present embodiment, the cells are cells of microbial origin, and the biofilms are biofilms of microorganisms, such as bacteria and fungi. The cells of microorganisms growing in biofilms can be physiologically distinct from the cells of the "planktonic form" of the same organism, which in contrast are single cells that can float or swim in liquid media. Biofilms can go through several life cycle steps, including initial attachment, reversible attachment, one or more stages of mutation, and dispersal.

[0093] The term "anti-biofilm" refers to the ability of a material to impede the formation of a biofilm of bacterial, fungal, and / or other cells on the surface of the material and / or to affect a reduction in the rate of formation of bacterial, fungal, and / or other cells.

[0094] As used herein, the term "prevent" in the context of an antimicrobial agent or preservative refers to reducing the rate of microbial cell growth to essentially zero or to at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% (including any range therebetween) of the occurrence of microorganisms in a comparable situation lacking the presence of the preservative of the present invention or an article containing the same. Alternatively, preventing refers to a reduction to at least 15%, 10%, or 5% of the occurrence of microbial cells in a comparable situation (e.g., an aqueous composition) lacking the presence of the preservative.

[0095] As used herein, the term "reduce" in the context of preservative activity or effectiveness indicates that the growth rate (and / or microbial load expressed in CFU or CFU / ml) of microorganisms (including their spores, cells, or biofilms) is substantially reduced compared to a similar article (or composition) lacking the preservative of the present invention. In some embodiments, the term "essentially reduced" includes a reduction in CFU / ml of at least 2-fold, at least 5-fold, at least 10-fold, at least 50-fold, at least 100-fold, at least 1000-fold, at least 10.000-fold, at least 100.000-fold, at least 1000.000-fold, between 10-1000.000-fold, between 1000-10.000.000-fold, between 1000-1000.000-fold, between 10.000-1000.000-fold, including any range therebetween, as compared to the CFU / ml content (also referred to herein as the initial microbial load) of a similar composition lacking the preservative of the present invention. In some embodiments, the preservative substantially prevents microbial infestation of the article (or composition). In some embodiments, articles (or compositions) comprising the preservative compositions of the present invention maintain their sterility after exposing them to non-sterile conditions (e.g., ambient atmosphere containing one or more microorganisms) for a period of between 1 day and 2 months, between 1 day and 1 month, between 1 and 20 days, between 1 and 60 days, between 1 and 50 days, between 10 and 60 days, between 1 and 10 days, between 10 and 50 days, between 10 and 40 days, between 10 and 30 days (including any range therebetween).

[0096] In another aspect, a method of treating or preventing a disease or disorder associated with a fungal burden in a subject is provided, comprising administering to the subject an effective amount of a pharmaceutical composition of the invention.

[0097] According to another aspect, a method of reducing and / or inhibiting fungal activity in a subject is provided, comprising administering to a subject an effective amount of a pharmaceutical composition of the present invention. In some embodiments, the effective amount (or dose) of the composition comprises an amount sufficient to reduce or ameliorate fungal burden in a subject. In some embodiments, the effective amount (or dose) of the composition is an amount sufficient to prevent or treat a disease or disorder associated with a fungal infection in a subject, where treatment includes reducing at least one symptom thereof, reducing the severity thereof, or inhibiting the progression thereof. In some embodiments, treating a fungal infection includes preventing or treating a disease or disorder associated with fungal activity in a subject, The objective of the present invention is to provide a method for treating a disease, comprising the steps of: preventing, attenuating, and / or inhibiting a disorder or disease state.

[0098] In another aspect, there is provided a method of reducing or preventing the fungal load of an edible substance (e.g., a food product), comprising contacting the food product with a fungicidally effective amount of a fungicidal composition of the invention. In some embodiments, an effective amount of a fungicidal composition of the invention is determined based on the pathogen load (e.g., CFU, CFU / cm) on or in the food product. 3 ) by at least 10-fold, at least 30-fold, at least 50-fold, at least 60-fold, at least 65-fold, at least 70-fold, at least 100-fold, at least 200-fold, at least 400-fold, at least 800-fold, at least 1000-fold, at least 10,000-fold, at least 100,000-fold, or at least 1,000,000-fold compared to a similar food lacking the preservative (also referred to herein as a "control").

[0099] In another aspect of the invention, a food product is provided that comprises the phosphorylcholine-tuftsin conjugate of the invention. In some embodiments, the food product comprises an effective amount (e.g., a fungicidally effective amount) of the phosphorylcholine-tuftsin conjugate of the invention. In some embodiments, the effective amount is sufficient to inhibit or reduce fungal growth and / or fungal load for at least 4 days, at least 5 days, at least 6 days, at least 7 days (including any range or value therebetween) compared to a control. In some embodiments, the effective amount is sufficient to extend the shelf life of the food product compared to a control. In some embodiments, the extension is at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 10 days, at least 20 days, at least 30 days (including any range or value therebetween).

[0100] In some embodiments, the phosphorylcholine-tuftsin conjugate comprises a phosphorylcholine moiety or a derivative thereof and a tuftsin or a derivative thereof covalently linked via a spacer. In some embodiments, the spacer comprises at least two amino acids. In some embodiments, the phosphorylcholine moiety or a derivative thereof is covalently linked to the spacer via a diazo group.

[0101] The term "phosphorylcholine (PC) conjugate," as used herein, refers to a phosphorylcholine moiety or a derivative thereof linked, optionally via a spacer, to tuftsin (T).

[0102] As used herein, the term "tuftsin" refers to a tetra-peptide (threonine-lysine-proline-arginine, TKPR; SEQ ID NO: 1). Tuftsin may be chemically synthesized. Tuftsin is known for its in vitro and in vivo phagocytosis stimulating activity and enhanced antigen presentation capacity of macrophages. In some embodiments, tuftsin may be considered an immunomodulatory molecule.

[0103] The term "derivative of phosphorylcholine" as used herein refers to any compound based on phosphorylcholine. The term "derivative of tuftsin" as used herein refers to any polypeptide based on TKPR. In some embodiments, the derivative retains the immunomodulatory effect of phosphorylcholine and / or tuftsin. In some embodiments, the derivative is a derivative that includes phosphorylcholine. In some embodiments, the derivative is a derivative that includes TKPR. A derivative is not simply a fragment of a polypeptide, nor one in which amino acids have been replaced or removed (analogue), but rather it may have further modifications made to the polypeptide, such as post-translational modifications.

[0104] In some embodiments, the derivative of phosphorylcholine is selected from 4-amino-phenyl-phosphocholine, 4-diazonio-phenyl-phosphorylcholine, 4-nitro-phenyl-phosphocholine, and 12-(3-iodophenyl)dodecyl-phosphocholine, among others. Each possibility is a separate embodiment of the invention.

[0105] The terms "tuftsin derivative," "TD," and "carrier moiety derived from tuftsin" are interchangeable and refer to tuftsin (TKPR, SEQ ID NO: 1) bound to at least two additional independently selected amino acids. Unnatural amino acids, preferably uncharged and non-polar unnatural amino acids, such as β-alanine-6-aminohexanoic acid and 5-aminopentanoic acid, may also be included in the tuftsin derivative. In some embodiments, the tuftsin derivative is threonine-lysine-proline-arginine-glycine-tyrosine (TKPRGY, SEQ ID NO: 2).

[0106] The term "moiety" as used herein refers to a part of a molecule that is missing one or more atoms compared to the corresponding molecule. The term "moiety" as used herein further relates to a part of a molecule that may contain either an entire functional group or a part of a functional group as a substructure. The term "moiety" further refers to a part of a molecule that exhibits a particular set of chemical and / or pharmacological characteristics that are similar to the corresponding molecule.

[0107] The terms "linked" or "attached" as used herein refer to a bond between at least two molecules or moieties such that they are a single molecule. In some embodiments, the bond is a chemical bond. In some embodiments, the bond is a covalent bond. In accordance with the principles of the present invention, the natural amino acids and the unnatural amino acids contained in the tuftsin derivatives are adjacently bonded to each other, while at least one phosphorylcholine derivative is directly or indirectly bound to at least one tuftsin derivative via a spacer. In some embodiments, at least one phosphorylcholine or derivative thereof is linked to the N-terminus of at least one tuftsin or derivative thereof. In some embodiments, at least one phosphorylcholine or derivative thereof is linked to the C-terminus of at least one tuftsin or derivative thereof.

[0108] The term "spacer" as used herein refers to a connecting or otherwise bridging element between a tuftsin derivative and a PC derivative, usually linked thereto by chemical or biological means. Non-limiting examples of spacers include amino acids, peptides, polypeptides, proteins, carbohydrates, and polymers, among others. Each possibility is a separate embodiment of the invention. In some embodiments, the spacer is at least two amino acids. In some embodiments, the spacer is glycine-tyrosine. In some embodiments, the spacer is attached to the C-terminus of TKPR. In some embodiments, the spacer is attached to the N-terminus of TKPR.

[0109] In some embodiments, the phosphorylcholine-tuftsin conjugates described above comprise one phosphorylcholine derivative bound to one tuftsin derivative. In certain embodiments, the phosphorylcholine-tuftsin conjugates described above comprise multiple phosphorylcholine derivatives bound to multiple tuftsin derivatives. In certain embodiments, the phosphorylcholine-tuftsin conjugates described above comprise multiple tuftsin derivatives bound to one phosphorylcholine derivative. In certain embodiments, the phosphorylcholine-tuftsin conjugates described above comprise multiple phosphorylcholine derivatives bound to one tuftsin derivative.

[0110] In some embodiments, said phosphorylcholine-tuftsin conjugate comprises at least one phosphorylcholine or derivative thereof and at least one tuftsin or derivative thereof separated by a spacer.

[0111] In some embodiments, the phosphorylcholine-tuftsin conjugate has Formula 1: [ka] It is expressed by: In some embodiments, the phosphorylcholine-tuftsin conjugate is in the form of a salt containing a counterion.In some embodiments, the counterion is a pharma- ceutically acceptable ion.In some embodiments, the counterion is any of chloride, acetate, and trifluoroacetate, or any combination thereof.

[0112] Pharmaceutical Compositions In some embodiments, a pharmaceutical composition is provided that includes one or more phosphorylcholine-tuftsin conjugates of the invention (e.g., conjugates of formulas 1-5) and a pharma- ceutically acceptable carrier. In some embodiments, the pharmaceutical composition includes a therapeutically effective amount of one or more phosphorylcholine-tuftsin conjugates of the invention. In some embodiments, the pharmaceutical composition includes a fungicidally effective amount of one or more phosphorylcholine-tuftsin conjugates of the invention.

[0113] In some embodiments, the pharmaceutical composition comprises a preservative effective amount, as disclosed herein above.

[0114] In some embodiments, the storage effective amount is a concentration of one or more phosphorylcholine-tuftsin conjugates of the invention in the pharmaceutical composition of at least 1 nM, at least 10 nM, at least 100 nM, at least 500 nM, at least 1 μM, at least 2 μM, at least 5 μM, at least 10 μM, at least 15 μM, at least 20 μM, at least 30 μM, at least 40 μM, at least 50 μM, at least 75 μM, at least 100 μM, at least 150 μM, at least 200 μM, at least 400 μM, at least 500 μM, at least 600 μM, at least 1,000 μM, or any range or value therebetween. Each possibility represents a separate embodiment of the present invention.

[0115] In some embodiments, a preservative effective amount comprises a concentration of one or more phosphorylcholine-tuftsin conjugates of the present invention (e.g., a conjugate of Formula 2) within the pharmaceutical composition of at least 1 ppm, at least 5 ppm, at least 10 ppm, at least 100 ppm, at least 500 ppm, at least 1000 ppm, or any range or value therebetween.

[0116] In some embodiments, a preservative effective amount comprises a concentration of one or more phosphorylcholine-tuftsin conjugates of the present invention (e.g., conjugates of Formula 1) within the pharmaceutical composition of at least 1 ppm, at least 5 ppm, at least 10 ppm, at least 100 ppm, at least 500 ppm, at least 1000 ppm, or any range or value therebetween.

[0117] In some embodiments, the concentration of one or more phosphorylcholine-tuftsin conjugates of the present invention in the pharmaceutical composition is between 5 ppm and 10 (w / w)%, between 5 ppm and 99 (w / w)%, between 5 ppm and 90 (w / w)%, between 5 ppm and 95 (w / w)%, between 5 ppm and 50 (w / w)%, between 5 ppm and 60 (w / w)%, between 5 ppm and 80 (w / w)%, between 5 ppm and 5 (w / w)%, between 5 ppm and 2 (w / w)%, between 0.5 and 10 (w / w)%, between 0.5 and 90 (w / w)%, between 0.5 and 50 (w / w)%, between 5 ppm and 0.5 (w / w)%, including any range therebetween.

[0118] In some embodiments, the concentration of phosphorylcholine-tuftsin conjugate of formula 2 or formula 2A in the pharmaceutical or fungicidal composition of the invention is between 5 ppm and 10 (w / w)%, between 5 ppm and 50 (w / w)%, between 5 ppm and 20 (w / w)%, between 5 ppm and 30 (w / w)%, between 5 ppm and 40 (w / w)%, between 5 ppm and 2 (w / w)%, between 5 ppm and about 1 (w / w)%, between 10 ppm and 50 (w / w)%, between 10 ppm and 20 (w / w)%, between 10 ppm and 30 (w / w)%, between 10 ppm and 10 (w / w)%, between 10 ppm and 2 (w / w)%, between 10 ppm and about 1 (w / w)%, between 20 ppm and 50 (w / w)%. , between 20ppm and 20(w / w)%, between 20ppm and 30(w / w)%, between 20ppm and 10(w / w)%, between 20ppm and 2(w / w)%, between 20ppm and about 1(w / w)%, between 5ppm and 99(w / w)%, between 5ppm and 90(w / w)%, between 5ppm and 95(w / w)%, between 5ppm and 50(w / w)% 50(w / w)%、5ppm-60(w / w)%、5ppm-80(w / w)%、5ppm-5(w / w)%、5ppm-2(w / w)%、0.5-10(w / w)%、0.5-90(w / w)%、0.5-50(w / w)%、5ppm-0.5(w / w)%(including any range therebetween). In some embodiments, the concentration of phosphorylcholine-tuftsin conjugate of Formula 2 or Formula 2A in the pharmaceutical or fungicidal composition of the invention is at least a fungicidally effective concentration as disclosed herein (e.g., at least about 10 ppm or at least about 20 ppm, which is a concentration that experimentally induced substantial inhibition of yeast activity in aqueous medium as disclosed in the Examples section).

[0119] In some embodiments, the concentration of phosphorylcholine-tuftsin conjugate of formula 1 in the pharmaceutical or fungicidal composition of the invention is between 5 ppm and 10 (w / w)%, between 5 ppm and 50 (w / w)%, between 5 ppm and 20 (w / w)%, between 5 ppm and 30 (w / w)%, between 5 ppm and 40 (w / w)%, between 5 ppm and 2 (w / w)%, between 5 ppm and about 1 (w / w)%, between 10 ppm and 50 (w / w)%, between 10 ppm and 20 (w / w)%, between 10 ppm and 30 (w / w)%, between 10 ppm and 10 (w / w)%, between 10 ppm and 2 (w / w)%, between 10 ppm and about 1 (w / w)%, between 20 ppm and 50 (w / w)%, between 20 ppm and 50 (w / w)%, between 20 ppm and 50 (w / w)%, between 20 ppm and 50 (w / w), ... Between ppm and 20(w / w)%, Between 20ppm and 30(w / w)%, Between 20ppm and 10(w / w)%, Between 20ppm and 2(w / w)%, Between 20ppm and about 1(w / w)%, Between 5ppm and 99(w / w)%, Between 5ppm and 90(w / w)%, Between 5ppm and 95(w / w)%, Between 5ppm and 50(w / w)% , between 5 ppm and 60 (w / w)%, between 5 ppm and 80 (w / w)%, between 5 ppm and 5 (w / w)%, between 5 ppm and 2 (w / w)%, between 0.5 and 10 (w / w)%, between 0.5 and 90 (w / w)%, between 0.5 and 50 (w / w)%, between 5 ppm and 0.5 (w / w)% (including any range therebetween). In some embodiments, the concentration of phosphorylcholine-tuftsin conjugate of formula 1 in the pharmaceutical or fungicidal composition of the invention is at least a fungicidally effective concentration as disclosed herein (at least about 10 ppm or at least about 20 ppm, which is a concentration that experimentally induced substantial inhibition of yeast activity in aqueous medium as disclosed in the Examples section).

[0120] In some embodiments, the combined concentration of the phosphorylcholine-tuftsin conjugate of Formula 1 and the phosphorylcholine-tuftsin conjugate of Formula 2 (or Formula 2A) in the pharmaceutical or fungicidal composition of the invention is between 10 ppm and 50 (w / w)%, between 10 ppm and 20 (w / w)%, between 10 ppm and 30 (w / w)%, between 10 ppm and 10 (w / w)%, between 10 ppm and 2 (w / w)%, between 10 ppm and about 1 (w / w)%, between 20 ppm and 50 (w / w)%, between 20 ppm and 20 (w / w)%, between 20 ppm and 30 (w / w)%, between 20 ppm and 1 ... (w / w)%, between 20 ppm and 2 (w / w)%, between 20 ppm and about 1 (w / w)%, between 10 ppm and 99 (w / w)%, between 10 ppm and 90 (w / w)%, between 10 ppm and 95 (w / w)%, between 10 ppm and 50 (w / w)%, between 10 ppm and 60 (w / w)%, between 10 ppm and 80 (w / w)%, between 10 ppm and 5 (w / w)%, between 10 ppm and 2 (w / w)%, between 0.5 and 10 (w / w)%, between 0.5 and 90 (w / w)%, between 0.5 and 50 (w / w)%, between 10 ppm and 0.5 (w / w)% (including any range therebetween).

[0121] In some embodiments, the concentration of phosphorylcholine-tuftsin conjugate of formula 1 in the pharmaceutical or fungicidal composition of the invention is 0.5-10 (w / w)%, 0.5-50 (w / w)%, 0.5-5 (w / w)%, 0.5 to about 2 (w / w)%, 0.5 to about 1 (w / w)%, 10 ppm to 0.5 (w / w)% (including any range therebetween) and the concentration of phosphorylcholine-tuftsin conjugate of formula 2 or 2A in the pharmaceutical or fungicidal composition of the invention is between 10 ppm to 50 (w / w)%, between 10 ppm to 20 (w / w)%, between 10 ppm to 30 (w / w)%, between 10 ppm to 10 (w / w)%, between 10 ppm to 2 (w / w)%, between 10 ppm to about 1 (w / w)%, 20 Between ppm and 50(w / w)%, Between 20ppm and 20(w / w)%, Between 20ppm and 30(w / w)%, Between 20ppm and 10(w / w)%, Between 20ppm and 2(w / w)%, Between 20ppm and about 1(w / w)%, Between 10ppm and 99(w / w)%, Between 10ppm and 90(w / w)%, Between 10ppm and 95(w / w)%, Between 10ppm and 50 (w / w)%, between 10 ppm and 60 (w / w)%, between 10 ppm and 80 (w / w)%, between 10 ppm and 5 (w / w)%, between 10 ppm and 2 (w / w)%, between 0.5 and 10 (w / w)%, between 0.5 and 90 (w / w)%, between 0.5 and 50 (w / w)%, between 10 ppm and 0.5 (w / w)% (including any range therebetween).

[0122] In some embodiments, the concentration of phosphorylcholine-tuftsin conjugate of formula 2 or 2A in the pharmaceutical or fungicidal composition of the invention is 0.5-10 (w / w)%, 0.5-50 (w / w)%, 0.5-5 (w / w)%, 0.5 to about 2 (w / w)%, 0.5 to about 1 (w / w)%, 10 ppm to 0.5 (w / w)% (including any range therebetween), and the concentration of phosphorylcholine-tuftsin conjugate of formula 1 in the pharmaceutical or fungicidal composition of the invention is between 10 ppm to 50 (w / w)%, between 10 ppm to 20 (w / w)%, between 10 ppm to 30 (w / w)%, between 10 ppm to 10 (w / w)%, between 10 ppm to 2 (w / w)%, between 10 ppm to about 1 (w / w)%, 20 Between ppm and 50(w / w)%, Between 20ppm and 20(w / w)%, Between 20ppm and 30(w / w)%, Between 20ppm and 10(w / w)%, Between 20ppm and 2(w / w)%, Between 20ppm and about 1(w / w)%, Between 10ppm and 99(w / w)%, Between 10ppm and 90(w / w)%, Between 10ppm and 95(w / w)%, Between 10ppm and 50 (w / w)%, between 10 ppm and 60 (w / w)%, between 10 ppm and 80 (w / w)%, between 10 ppm and 5 (w / w)%, between 10 ppm and 2 (w / w)%, between 0.5 and 10 (w / w)%, between 0.5 and 90 (w / w)%, between 0.5 and 50 (w / w)%, between 10 ppm and 0.5 (w / w)% (including any range therebetween).

[0123] In some embodiments, the pharmaceutical compositions comprise a phosphorylcholine-tuftsin conjugate of formula 2, including any salts or any functional derivatives thereof. In some embodiments, the pharmaceutical compositions comprise a phosphorylcholine-tuftsin conjugate of formula 2, and further comprise at least a trace amount of any one of the phosphorylcholine-tuftsin conjugates of formulas 3-5, including any salts or any functional derivatives thereof. In some embodiments, the w / w concentration of phosphorylcholine-tuftsin conjugate of formula 2 in the pharmaceutical composition is between about 1 ppm and about 10%, between about 5 ppm and about 10%, between about 5 ppm and about 0.5%, between about 5 ppm and about 1%, between about 5 ppm and about 2%, between about 10 ppm and about 10%, between about 10 ppm and about 200 ppm, between about 10 ppm and about 0.5%, between about 10 ppm and about 0.1%, between about 10 ppm and about 2%, between about 10 ppm and about 1%, between about 10 ppm and about 500 ppm, between about 10 ppm and about 2000 ppm, between about 10 ppm and about 1000 ppm, between about 10 ppm and about 100 ppm, between about 10 ppm and about 50 ppm, between about 5 ppm and about 50 ppm, between about 100 ppm and about 500 ppm, between about 100 ppm and about 1000 ppm, between about 1000 ppm and about 0.1%, between about 100 ppm and about 0.1%, between about 100 ppm and about 1%, between about 1000 ppm and about 1%, between about 0.1 and about 10 (w / w)% (including any range therebetween).

[0124] In some embodiments, the pharmaceutical composition comprises a pharma- ceutical active agent and a carrier, wherein the pharma- ceutical active agent consists essentially of one or more of the phosphorylcholine-tuftsin conjugates of the present invention, phosphorylcholine-tuftsin conjugates as the only pharma- ceutical active ingredient. In some embodiments, the pharmaceutical composition is substantially devoid of any additional pharma- ceutical active ingredient. In some embodiments, the pharma- ceutical active agent consists essentially of the phosphorylcholine-tuftsin conjugate of formula 2, and optionally one or more functional derivatives thereof. In some embodiments, the pharma- ceutical active agent consists essentially of the phosphorylcholine-tuftsin conjugate of formula 2, and optionally one or more functional derivatives thereof. In some embodiments, the pharma- ceutical active agent consists essentially of a phosphorylcholine-tuftsin conjugate of formula 2 and a phosphorylcholine-tuftsin conjugate of formula 1, wherein the w / w ratio between the phosphorylcholine-tuftsin conjugate of formula 1 and the phosphorylcholine-tuftsin conjugate of formula 2 is between 10:1 and 10.000:1, between 10:1 and 1000:1, between 100:1 and 10.000:1, between 200:1 and 10.000:1, between 200:1 and 1000:1 (including any range therebetween).

[0125] In some embodiments, the pharmaceutical composition comprises a phosphorylcholine-tuftsin conjugate of formula 2, including any salt or any functional derivative thereof, and further comprises a phosphorylcholine-tuftsin conjugate of formula 1. In some embodiments, the w / w concentration of the phosphorylcholine-tuftsin conjugate of formula 1 in the pharmaceutical composition is between 0.01-10%, between 0.05-10%, between 0.1-10%, between 0.1-1%, between 0.1-0.5%, between 0.5-1%, between 0.5-1.5%, between 0.5-2%, between about 0.8 to about 1%, between 0.5-10%, between 0.5-5%, between 1-10% (including any range therebetween).

[0126] In some embodiments, the w / w concentration of phosphorylcholine-tuftsin conjugate of formula 1 in the pharmaceutical composition is between 0.1-10%, 0.5-2%, 0.1-about 1%, 0.1-about 2% (including any range therebetween), and the w / w concentration of phosphorylcholine-tuftsin conjugate of formula 2 in the pharmaceutical composition is between about 5 ppm and about 10%, between about 5 ppm and about 0.5%, between about 5 ppm and about 1%, between about 5 ppm and about 2%, between about 10 ppm and about 10%, between about 10 ppm and about 200 ppm, between about 10 ppm and about 0.5%, between about 10 ppm and about 0.1%, between about 10 ppm and about 0.2%, between about 10 ppm and about 0.3%, between about 10 ppm and about 0.4%, between about 10 ppm and about 0.5 ... between about pm and about 2%, between about 10 ppm and about 1%, between about 10 ppm and about 500 ppm, between about 10 ppm and about 2000 ppm, between about 10 ppm and about 1000 ppm, between about 10 ppm and about 100 ppm, between about 10 ppm and about 50 ppm, between about 5 ppm and about 50 ppm, between about 100 ppm and about 500 ppm, between about 100 ppm and about 1000 ppm, between about 1000 ppm and about 0.1%, between about 100 ppm and about 0.1%, between about 100 ppm and about 1%, between about 1000 ppm and about 1%, between about 0.1 and about 10 (w / w)% (including any range therebetween).

[0127] In some embodiments, the w / w concentration of the phosphorylcholine-tuftsin conjugate of formula 1 in the pharmaceutical composition is between 0.1 and about 2% (including any range therebetween), and the w / w concentration of the phosphorylcholine-tuftsin conjugate of formula 2 in the pharmaceutical composition is between about 5 ppm and about 2%.

[0128] In some embodiments, the w / w concentration of the phosphorylcholine-tuftsin conjugate of formula 1 in the pharmaceutical composition is between 0.1 and about 2% (including any range therebetween), and the w / w concentration of the phosphorylcholine-tuftsin conjugate of formula 2 in the pharmaceutical composition is between about 5 ppm and about 200 ppm.

[0129] In some embodiments, the pharmaceutical composition comprises a preservative effective amount of at least one phosphorylcholine-tuftsin conjugate of any one of formulas 1 to 5. In some embodiments, the pharmaceutical composition comprises a preservative effective amount of a phosphorylcholine-tuftsin conjugate of formula 1, or a preservative effective amount of a phosphorylcholine-tuftsin conjugate of formula 2.

[0130] In some embodiments, the pharmaceutical composition is an ophthalmic composition. In some embodiments, the pharmaceutical composition is a liquid, a flowable composition, a semi-solid as a semi-liquid, a foam, or a gel.

[0131] In one aspect of the invention, there is a pharmaceutical composition comprising: (i) one or more phosphorylcholine-tuftsin conjugates as disclosed hereinabove; (ii) between 0.1-5 weight / weight (w / w)% of a viscosity increasing agent; and (iii) between 0.1-5 w / w% of a mucoadhesive polymer. In some embodiments, the pharmaceutical composition is a liquid composition. In some embodiments, the pharmaceutical composition is formulated for administration to the eye. In some embodiments, the terms "ophthalmic composition" and "pharmaceutical composition" are used interchangeably herein.

[0132] In some embodiments, the ophthalmic composition or pharmaceutical composition is liquid at a temperature between -5 and 95°C. In some embodiments, the ophthalmic composition is liquid under physiological conditions (e.g., between 20 and 40°C, pH between 2 and 8, etc.). In some embodiments, the ophthalmic composition is an aqueous composition. In some embodiments, the ophthalmic composition is a solution (e.g., an aqueous solution). In some embodiments, the components of the ophthalmic composition are substantially soluble in the solution. In some embodiments, the ophthalmic composition is substantially devoid of any particulate matter. In some embodiments, the ophthalmic composition is substantially devoid of particles having a particle size greater than 200 nm, greater than 300 nm, greater than 400 nm, greater than 500 nm, greater than 600 nm, greater than 700 nm, greater than 800 nm, greater than 900 nm, greater than 1 μm (including any range or value therebetween). In some embodiments, the ophthalmic composition is a clear solution. In some embodiments, the ophthalmic composition is an opaque solution.

[0133] In some embodiments, the ophthalmic composition is characterized by a transmittance of greater than 80%, greater than 85%, greater than 90%, greater than 95%, greater than 97%, greater than 99%, where the transmittance is measured at a wavelength in the range of 200 nM to 1000 nM. One of ordinary skill in the art will appreciate that transmittance is an indication of the optical clarity of an ophthalmic composition (e.g., in the form of a solution).

[0134] In some embodiments, the ophthalmic composition is a colored solution, hi some embodiments, the colored solution is characterized by a slight yellow to red color.

[0135] In some embodiments, the ophthalmic composition comprises a solvent. In some embodiments, the solvent is an aqueous solvent. In some embodiments, the ophthalmic composition comprises between 50-99(w / w)%, between 50-60(w / w)%, between 60-70(w / w)%, between 70-80(w / w)%, between 80-85(w / w)%, between 85-90(w / w)%, between 90-95(w / w)%, between 90-92(w / w)%, between 92-95(w / w)%, between 95-97(w / w)%, between 97-99(w / w)% (including any range or value therebetween) of the solvent.

[0136] In some embodiments, the w / w concentration of the thickening agent in the pharmaceutical composition is between 0.1-5%, between 0.1-0.2%, between 0.2-0.3%, between 0.3-0.5%, between 0.4-0.6%, between 0.5-0.7%, between 0.7-0.8%, between 0.8-1%, between 1-2%, between 2-3%, between 3-5%, including any range or value therebetween. In some embodiments, the w / w concentration of the thickening agent in the pharmaceutical composition is between 0.4-0.6%.

[0137] As used herein, "thickening agent" refers to any substance that increases the viscosity of a solution administered to the eye. In some embodiments, the thickening agent increases the viscosity of an aqueous solution.

[0138] Non-limiting examples of thickening agents include, but are not limited to, hydroxypropyl methylcellulose (HPMC), hydroxymethyl cellulose, hydroxyethyl cellulose, cellulose, polyalcohols polyvinylpyrrolidone, polyethylene glycol, polypropylene glycol, polyacrylates, poly(2-hydroxyethyl methacrylate), and polyvinyl alcohol, or any combination thereof.

[0139] Additional thickening agents, such as poloxamers, galactomannans, hydrolyzed galactomannans, glucomannans, hydrolyzed glucomannans, guar gum, xanthan gum, nanocrystalline cellulose, cyclodextrins, poly(cyclodextrins), dextran, dextrins, starch, heparin, β-glucans, mannans, and levans or any combination thereof, are well known in the art.

[0140] In some embodiments, the pharmaceutical composition comprises a mucoadhesive polymer. In some embodiments, the w / w concentration of the mucoadhesive polymer in the pharmaceutical composition is between 0.1-5%, between 0.1-0.2%, between 0.2-0.3%, between 0.3-0.5%, between 0.4-0.6%, between 0.5-0.7%, between 0.7-0.8%, between 0.8-1%, between 1-2%, between 2-3%, between 3-5% (including any range or value therebetween). In some embodiments, the w / w concentration of the mucoadhesive polymer in the pharmaceutical composition is between 1-5%.

[0141] In some embodiments, the mucoadhesive polymer is an ionic mucoadhesive polymer, hi some embodiments, the mucoadhesive polymer is a cationic polymer, an anionic polymer, or both.

[0142] Non-limiting examples of mucoadhesive polymers include, but are not limited to, alginic acid, chitosan, hyaluronic acid, carboxymethylcellulose, pectin, fucoidan, dermatan sulfate, chondroitin sulfate, polycarbophil-cysteine, polyoxyethylene glycol esters, heparan sulfate, keratan sulfate, and gelatin (including any salt, any polymer, or any combination thereof).

[0143] In some embodiments, the mucoadhesive polymer is alginic acid (or alginate), and / or a salt thereof.

[0144] In some embodiments, the w / w ratio between the mucoadhesive polymer and the viscosity increasing agent in the ophthalmic composition is between 1:1 and 10:1, between 1:1 and 2:1, between 2:1 and 4:1, between 2:1 and 3:1, between 3:1 and 5:1, between 3:1 and 4:1, between 5:1 and 7:1, between 7:1 and 10:1 (including any range or value therebetween).

[0145] In some embodiments, the w / w ratio between the mucoadhesive polymer and the phosphorylcholine-tuftsin conjugate in the ophthalmic composition is between 5:1 and 1:5, between 5:1 and 3:1, between 3:1 and 2:1, between 2:1 and 1:1, between 1:1 and 1:2, between 1:2 and 1:4, between 1:4 and 1:5 (including any range or value therebetween).

[0146] In some embodiments, the ophthalmic composition comprises up to 0.1-10% (w / w) of an osmolality adjusting agent.

[0147] 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 in the ophthalmic composition of the present invention is reduced compared to the label declaration of the preservative (i.e., the effective concentration of the particular preservative as defined by the relevant regulatory agency).

[0148] In some embodiments, the effective amount of preservative in the ophthalmic compositions of the present invention is reduced by at least 10%, at least 50%, at least 70%, at least 2-fold, at least 5-fold, at least 10-fold, at least 50-fold, at least 100-fold, at least 500-fold, or between about 2-fold to about 20-fold, between about 2-fold to about 15-fold, between about 2-fold to about 10-fold, between about 5-fold to about 20-fold, between about 5-fold to about 15-fold (including any range or value therebetween), where reduced is a reduction compared to the labeling of the preservative. In some embodiments, the ophthalmic compositions comprise a reduced amount of preservative, where reduced is a reduction compared to the effective amount of the preservative in a similar composition lacking the phosphorylcholine-tuftsin conjugate.

[0149] In some embodiments, an effective amount of a preservative is <51> The decision is based on a PET study with or any other PET study.

[0150] In some embodiments, the ophthalmic composition comprises between 0.001-1% (w / w) of a preservative, including any range therebetween. In some embodiments, the ophthalmic composition comprises at least 0.001% (w / w) of a preservative, and less than the label labeling, where less than the label labeling includes a concentration of preservative that is reduced by at least 10%, at least 50%, at least 70%, at least 2-fold, at least 5-fold, at least 10-fold, at least 50-fold, at least 100-fold, at least 500-fold, or between about 2-fold to about 20-fold, between about 2-fold to about 15-fold, between about 2-fold to about 10-fold, between about 5-fold to about 20-fold, between about 5-fold to about 15-fold, including any range or value therebetween.

[0151] In some embodiments, the w / w concentration of the preservative in the ophthalmic composition is between 0.001-1%, between 0.001-0.1%, between 0.001-0.01%, between 0.001-0.02%, between 0.001-0.018%, between 0.001-0.015%, between 0.002-0.018%, between 0.002-0.015%, between 0.003-0.018%, between 0.003-0.015%, between 0.05-0.1%, between 0.1-0.2%, between 0.2-0.3%, between 0.3-0.4%, between 0.4-0.5%, between 0.5-0.7%, between 0.7-1%, including any range or value therebetween. In some embodiments, the preservative is suitable for use in an ophthalmic composition. In some embodiments, the preservative is or comprises a quaternary ammonium cation, such as benzalkonium, including any salt thereof, e.g., benzalkonium chloride. In some embodiments, the quaternary ammonium cation is a pharma- ceutically acceptable compound. In some embodiments, the quaternary ammonium cation comprises a pharma-ceutically acceptable counter anion.

[0152] Surprisingly, the inventors have discovered a synergistic preservative activity of phosphorylcholine-tuftsin conjugate (specifically, the phosphorylcholine-tuftsin conjugate of Formula 1) and BAK that results in a significant reduction in the concentration of BAK in the compositions of the present invention required for sufficient preservative activity compared to the labeled claim of BAK in ophthalmic formulations (0.02 (w / w)%).

[0153] In some embodiments, the quaternary ammonium cation has the formula NR 4 + wherein each R is independently selected from an alkyl group, an aryl group, and an alkyl-aryl group, each of which optionally has one or more substituents, each of which independently is -NO 2 , -CN, -OH, OXO, IMINO, -CONH 2 ,-CONR' 2 ,-CNNR' 2 ,-CSNR' 2 , -CONH-OH, -CONH-NH2 , -NHCOR', -NHCSR', -NHCNR', -NC(=O)OR', -NC(=O)NR', ​​-NC(=S)OR', -NC(=S)NR', ​​-SO 2 R', -SOR', -SR', -SO 2 OR', -SO 2 N(R') 2 , -NHNR' 2 , -NNR', C 1 -C 6 Haloalkyl, optionally substituted C 1 -C 6 Alkyl, -NH 2 , -NR' 2 -NH(C 1 -C 6 alkyl), -N(C 1 -C 6 Alkyl) 2 , C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkoxy, Hydroxy (C 1 -C 6 Alkyl), Hydroxy (C 1 -C 6 Alkoxy), Alkoxy(C 1 -C 6 Alkyl), alkoxy (C 1 -C 6 Alkoxy), C 1 -C 6 Alkyl NR' 2 , C 1 -C 6 Alkyl SR', -CONH(C 1 -C 6 alkyl), -CON(C 1 -C 6 Alkyl) 2 , -CO 2 H, -CO 2 R', -OCOR, -OCOR', -OC(=O)OR', -OC(=O)NR', ​​-OC(=S)OR', -OC(=S)NR', ​​or combinations thereof; each R' independently represents hydrogen or an optionally substituted C 1 -C10 Alkyl, optionally substituted C 3 -C 10 Cycloalkyl, optionally substituted C 3 -C 10 Heterocyclyl, optionally substituted heteroaryl, optionally substituted aryl, hydroxy, amino, -NH 2 , -NR' 2 -NH(C 1 -C 6 alkyl), -N(C 1 -C 6 Alkyl) 2 , C 1 -C 6 Alkoxy, C 1 -C 6 Haloalkoxy, Hydroxy (C 1 -C 6 Alkyl), Hydroxy (C 1 -C 6 Alkoxy), Alkoxy(C 1 -C 6 Alkyl), alkoxy (C 1 -C 6 Alkoxy), C 1 -C 6 Alkyl NR' 2 , C 1 -C 6 alkylSR', or combinations thereof.

[0154] As used herein, the term "alkyl" describes an aliphatic hydrocarbon, including straight and branched chain groups. An alkyl group has 1-50 carbon atoms, 1-30 carbon atoms, or between 1-20 carbon atoms. Whenever a number range is described herein, such as "1-30," it is implied that the group, in this case the alkyl group, may contain 1, 2 carbon atoms, 3 carbon atoms, 4 carbon atoms, 5 carbon atoms, 6 carbon atoms, 7 carbon atoms, 10 carbon atoms, 15 carbon atoms, 20 carbon atoms, etc., up to 30 carbon atoms. In some embodiments, the alkyl group is a short alkyl containing between 1-10 or between 1-6 carbon atoms (including any range therebetween). In some embodiments, the alkyl group is a long alkyl having at least 10 carbon atoms or between 10-30 carbon atoms (including any range therebetween). The alkyl may be substituted or unsubstituted as defined herein.

[0155] The term "alkyl," as used herein, also includes saturated or unsaturated hydrocarbons, and thus this term further includes alkenyl and alkynyl.

[0156] In some embodiments, the quaternary ammonium is represented by the formula [ka] wherein each R is independently as disclosed hereinabove and the wavy bond represents the point of attachment to the polymer backbone. Exemplary polymeric quaternary ammonium cations include, but are not limited to, polyquaterniums (e.g., polyquaternium-2, polyquaternium-80, polyquaternium-11, polyquaternium-52, polyquaternium-17, etc.).

[0157] In some embodiments, the quaternary ammonium cation and / or salt thereof is selected from, but not limited to, benzyldimethyldodecylammonium chloride, didecyldimethylammonium chloride, dodecyltrimethylammonium chloride, hexadecyltrimethylammonium chloride, sodium cocoamidopropyl PG-dimonium chloride phosphate (Cola Lipid C), laurdimonium hydroxypropyl decylglucoside chloride (SugaQuat L-1010), berberine, berberubine, Berberis vulgaris root extract, isopropylbenzylbutylnorberberine iodide, palmatine, jatrorrhizine, coptisine, ungeremine, epiberberine, pseudoberberine, stepharanine, sinapine, and any combination thereof.

[0158] Additional preservatives are well known in the art. Non-limiting examples of preservatives include, but are not limited to, chlorobutanol, sodium perborate, and stabilized oxychloro complexes (SOC), or any combination thereof.

[0159] In some embodiments, the preservatives disclosed herein are the only preservatives in the ophthalmic compositions of the present invention. In some embodiments, the preservative in the ophthalmic composition is benzalkonium (including any salt thereof and its derivatives).

[0160] In some embodiments, the term "derivative" refers to stereoisomers (e.g., enantiomers and / or diastereomers) and / or structural isomers (e.g., alkylated, aminated, hydroxylated, carboxylated derivatives), esters, tautomers, and / or prodrugs or precursors of the disclosed compounds above. In some embodiments, the term "derivative" refers to structural derivatives having similar (or enhanced) antimicrobial activity.

[0161] In some embodiments, the ophthalmic composition of the present invention comprises less than 0.02% (w / w), less than 0.019% (w / w), or less than 0.018% (w / w) of BAK or any derivative of BAK. In some embodiments, the ophthalmic composition of the present invention comprises between 0.001-0.018%, between 0.001-0.015%, between 0.001-0.01%, between 0.001-0.02%, between 0.001-0.018%, between 0.001-0.015%, between 0.002-0.018%, between 0.002-0.015%, between 0.003-0.018%, or between 0.003-0.015% (w / w) (including any range or value therebetween).

[0162] In another embodiment, the w / w ratio between the phosphorylcholine-tuftsin conjugate and the quaternary ammonium cation in the pharmaceutical composition (also referred to herein as a "synergistically effective ratio") is between about 1000:1 and about 1:1, between about 500:1 and 1:1, between about 700:1 and 1:1, between about 100:1 and 1:1, between about 50:1 and 1:1, between about 1000:1 and 10:1, between about 100:1 and 10:1, between about 500:1 and 10:1, between about 100:1 and 20:1, between about 50:1 and 10:1, between about 50:1 and 20:1, between about 10:1 and 5:1, between about 10:1 and 1:1 (including any range therebetween).

[0163] In some embodiments, the quaternary ammonium cation is BAK. In some embodiments, the antimicrobially effective amount of BAK in the pharmaceutical composition (i.e., aqueous composition) is less than 0.2 mg / ml, less than 0.18 mg / ml, less than 0.15 mg / ml, between 0.01-0.19 mg / ml, between 0.01-0.18 mg / ml, between 0.01-0.15 mg / ml, between 0.01-0.1 mg / ml, between 0.02-0.18 mg / ml, between 0.02-0.15 mg / ml, between 0.015-0.18 mg / ml, between 0.015-0.15 mg / ml, between 0.03-0.18 mg / ml, between 0.03-0.15 mg / ml (including any range therebetween).

[0164] In some embodiments, the ophthalmic composition comprises or consists essentially of an aqueous solution (e.g., a buffered aqueous solution), between 0.1-5 w / w, between 0.1-2 w / w, between 0.1-3 w / w, between 5 ppm-10 (w / w)% of one or more phosphorylcholine-tuftsin conjugates of the invention, between 0.5-1.5 (w / w)% of a mucoadhesive polymer (e.g., alginate), between 0.3-0.8 (w / w)% of a viscosity increasing agent (e.g., HPMC), about 4 (w / w)% of an osmotic agent (e.g., mannitol), and an antimicrobially effective amount of a preservative (e.g., BAK). Exemplary compositions are described herein below. In some embodiments, the ophthalmic composition comprises or consists essentially of an aqueous solution (e.g., a buffered aqueous solution), between 0.1-2 (w / w) of one or more phosphorylcholine-tuftsin conjugates of the invention (i.e., conjugates of formula 1, further comprising between 5 ppm and 0.1 (w / w)% of a conjugate of formula 2), between 0.5-1.5 (w / w)% of a mucoadhesive polymer (e.g., alginic acid), between 0.3-0.8 (w / w)% of a viscosity increasing agent (e.g., HPMC), about 4 (w / w)% of an osmotic pressure adjusting agent (e.g., mannitol), and between 0.001-0.019 (w / w)% of a preservative (e.g., BAK). Exemplary compositions are described herein below.

[0165] In some embodiments, the ophthalmic composition comprises or consists essentially of an aqueous solution (e.g., a buffered aqueous solution), between 0.5 and about 1 (w / w)% of a conjugate of Formula 1 (further comprising between 5 ppm and 200 ppm of a conjugate of Formula 2), between 0.5 and 1.5 (w / w)% of a mucoadhesive polymer (such as alginic acid), between 0.3 and 0.8 (w / w)% of a viscosity increasing agent (e.g., HPMC), about 4 (w / w)% of an osmotic agent (e.g., mannitol), and between 0.001 and 0.019 (w / w)% of a preservative (e.g., BAK).

[0166] In another aspect of the invention, there is provided an ophthalmic composition comprising or consisting essentially of an aqueous solution (e.g., a buffered aqueous solution), between 0.5-1.5 (w / w)% of a mucoadhesive polymer (e.g., alginic acid), between 0.3-0.8 (w / w)% of a viscosity increasing agent (e.g., HPMC), about 4 (w / w)% of an osmolality adjusting agent (e.g., mannitol), and an effective amount of a preservative of the present invention (i.e., a combination between a quaternary ammonium cation such as BAK and a phosphorylcholine-tuftsin conjugate of formula 1). In some embodiments, the effective amount of the preservative (also used herein as "preservative effective concentration") is reduced compared to the preservative effective concentration of the quaternary ammonium cation in a similar composition lacking the PC phosphorylcholine-tuftsin conjugate, where reduced is as defined herein above.

[0167] An effective amount of preservative in an ophthalmic composition of the present invention corresponds to a concentration of quaternary ammonium cation between 0.01-0.19 mg / ml, between 0.01-0.18 mg / ml, between 0.01-0.15 mg / ml, between 0.01-0.1 mg / ml, between 0.02-0.18 mg / ml, between 0.02-0.15 mg / ml, between 0.015-0.18 mg / ml, between 0.015-0.15 mg / ml, between 0.03-0.18 mg / ml, between 0.03-0.15 mg / ml (including any range therebetween). In some embodiments, an effective amount of preservative in an ophthalmic composition of the present invention further comprises a phosphorylcholine-tuftsin conjugate, wherein the w / w ratio of phosphorylcholine-tuftsin conjugate to quaternary ammonium cation is as described herein. In some embodiments, the term "corresponding to" means that the concentration of phosphorylcholine-tuftsin conjugate may vary when related to the concentration of quaternary ammonium, but the w / w ratio between phosphorylcholine-tuftsin conjugate and quaternary ammonium cation in the ophthalmic composition of the present invention is as described herein above.

[0168] In some embodiments, the ophthalmic composition consists essentially of pharmaceutical grade ingredients. In some embodiments, all ingredients of the ophthalmic composition are pharmaceutical grade compounds. In some embodiments, the ophthalmic composition is sterile. In some embodiments, the effective amount of preservative in the ophthalmic composition of the present invention is sufficient to maintain the sterility of the ophthalmic composition after exposing it to non-sterile conditions (e.g., an atmosphere containing one or more microorganisms), where the maintenance is for a period ranging from 1, between 1 day and 2 months, between 1 day and 1 month, between 1 and 20 days, between 1 and 60 days, between 1 and 50 days, between 10 and 60 days, between 1 and 10 days, between 10 and 50 days, between 10 and 40 days, between 10 and 30 days, including any range therebetween. The term "sterility" as used herein refers specifically to the microbial load of a composition (e.g., an ophthalmic composition of the present invention) or article in accordance with the sterility requirements recorded in the United States Pharmacopoeia or the European Pharmacopoeia, respectively.

[0169] In some embodiments, the pH of the ophthalmic compositions disclosed herein is between 6 and 8, between 6 and 6.5, between 6.5 and 7, between 7 and 7.5, between 7.5 and 8 (including any range therebetween).

[0170] In some embodiments, the ophthalmic composition is characterized by a viscosity of between 30-150 cps, between 50-120 cps, between 1-5 cps, between 5-10 cps, between 10-15 cps, between 15-20 cps, between 20-30 cps, between 30-40 cps, between 40-50 cps, between 50-60 cps, between 60-70 cps, between 70-80 cps, between 80-90 cps, between 90-100 cps, between 100-110 cps, between 110-120 cps (including any range or value therebetween).

[0171] In some embodiments, the ophthalmic composition is characterized by an osmolality of between 200-600 mOsmols / kg, between 200-500 mOsmols / kg, between 200-300 mOsmols / kg, between 300-350 mOsmols / kg, between 350-400 mOsmols / kg, between 400-450 mOsmols / kg, between 450-500 mOsmols / kg, between 500-550 mOsmols / kg, between 550-600 mOsmols / kg (including any range or value therebetween). In some embodiments, the ophthalmic composition is characterized by an osmolality of between 250-450 mOsmols / kg.

[0172] In some embodiments, the pharmaceutical composition comprises phosphorylcholine conjugate and at least one other component, and any product resulting directly or indirectly from the combination, complexation, or aggregation of any two or more components, the dissociation of one or more components, or any other type of reaction or interaction of one or more components.Thus, the term "pharmaceutical composition" as used herein specifically includes any composition made by mixing a pharmacologic effective amount of phosphorylcholine-tuftsin conjugate and one or more pharmacologic acceptable carriers.In some embodiments, the pharmaceutical composition comprises a pharmacologic acceptable carrier, diluent, or excipient.

[0173] As used herein, the term "carrier", "adjuvant", or "excipient" refers to any component of a pharmaceutical composition that is not an active agent. As used herein, the term "pharmaceutical acceptable carrier" refers to any type of non-toxic, inert solid, semi-solid, or liquid filler, diluent, encapsulating material, formulation aid, or simply a sterile aqueous vehicle such as saline. Some examples of substances which can function as pharma- ceutically acceptable carriers are sugars such as lactose, glucose, and sucrose, starches such as corn starch and potato starch, cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt, gelatin, talc; excipients such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn 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; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline, Ringer's solution; ethyl alcohol, phosphate buffer solutions, and other non-toxic compatible substances used in pharmaceutical preparations. Some non-limiting examples of materials that can function as carriers herein include sugar, starch, cellulose and its derivatives, powdered tragacanth, malt, gelatin, talc, stearic acid, magnesium stearate, calcium sulfate, vegetable oils, polyols, alginic acid, pyrogen-free water, isotonic saline, phosphate buffer solution, cocoa butter (suppository base), emulsifiers, and other non-toxic compatible materials used in other pharmaceutical preparations. Wetting agents and lubricants, such as sodium lauryl sulfate, and coloring agents, flavoring agents, excipients, stabilizers, antioxidants, and preservatives may also be present. Any non-toxic, inert, effective carrier may be used to formulate the compositions contemplated herein.In this regard, suitable pharma- ceutically acceptable carriers, excipients, and diluents are well known to those skilled in the art, for example, those described in The Merck Index, Thirteenth Edition, Budavari et al., Eds., Merck & Co., Inc., Rahway, NJ (2001); the CTFA (Cosmetic, Toiletry, and Fragrance Association) International Cosmetic Ingredient Dictionary and Handbook, Tenth Edition (2004); and the "Inactive Ingredient Guide," US Food and Drug Administration (FDA) Center for Drug Evaluation and Research (CDER) Office of Management, the entire contents of which are hereby incorporated by reference. Examples of pharma-ceutically acceptable excipients, carriers, and diluents useful in the present compositions include distilled water, saline, Ringer's solution, dextrose solution, Hank's solution, and DMSO. These additional inactive ingredients, as well as effective formulation and administration techniques, are well known in the art and are described in standard textbooks such as Goodman and Gillman's: The Pharmacological Bases of Therapeutics, 8th Ed., Gilman et al. Eds. Pergamon Press (1990); Remington's Pharmaceutical Sciences, 18th Ed., Mack Publishing Co., Easton, Pa. (1990); and Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott Williams & Wilkins, Philadelphia, Pa., (2005), each of which is incorporated by reference in its entirety.The compositions described herein may also be included in artificially created structures such as liposomes, ISCOMS, sustained release particles, and other vehicles that increase the half-life of peptides or polypeptides. Liposomes include emulsions, foams, micelles, insoluble monolayers, liquid crystals, phospholipid dispersions, lamellar layers, and the like. Liposomes used with the peptides described herein are formed from standard vesicle-forming lipids, which generally include neutral and negatively charged phospholipids and sterols such as cholesterol. The choice of lipid is generally determined by considerations such as liposome size and stability in blood. Various methods are available for preparing liposomes, as reviewed, for example, in Coligan, JE et al, Current Protocols in Protein Science, 1999, John Wiley & Sons, Inc., New York, and see also U.S. Patent Nos. 4,235,871, 4,501,728, 4,837,028, and 5,019,369.

[0174] Carriers may in total constitute from about 0.1% to about 99.99999% (including any range therebetween) of the pharmaceutical compositions presented herein.

[0175] In some embodiments, the pharmaceutical composition is an aqueous solution. In some embodiments, the aqueous solution is a buffered solution. In some embodiments, the aqueous solution comprises a pharma- ceutically acceptable salt. In some embodiments, the buffered solution comprises a phosphate buffer. In some embodiments, the aqueous solution comprises water for injection, which is well known in the art.

[0176] Suitable buffer components or buffers that may be used in the aqueous solution are those conventionally used in pharmaceutical compositions. In some embodiments, the pharmaceutical compositions are formulated for local and / or systemic administration.

[0177] One embodiment of the present invention relates to the phosphorylcholine-tuftsin conjugate of the present invention, which is presented in unit dosage form and prepared by any of the methods well known in the art of pharmacy. In one embodiment, the unit dosage form is in the form of a tablet, capsule, lozenge, wafer, patch, ampoule, vial, or pre-filled syringe. In one embodiment, the pharmaceutical composition is formulated with a compatible pharmaceutical carrier, prepared, placed in an appropriate container, and labeled for the treatment of an indicated condition.

[0178] In addition, in vitro assays can be optionally used to help identify optimal dosage ranges. The precise dosage to be used in the formulation also depends on the route of administration, the nature of the disease or disorder, and should be determined according to the judgment of the practitioner and each patient's circumstances. Effective doses can be extrapolated from dose-response curves derived from in vitro or in vivo animal model test bioassays or systems.

[0179] In one embodiment, the composition of the present invention is administered in the form of a pharmaceutical composition comprising at least one of the active ingredients of the present invention together with a pharma- ceutically acceptable carrier or diluent.In another embodiment, the composition of the present invention can be administered separately or together in any conventional oral, parenteral or transdermal dosage form.

[0180] For topical application, the phosphorylcholine-tuftsin conjugates of the present invention or combinations thereof can be combined with a pharma- ceutically acceptable carrier to deliver an effective dose based on the desired activity. The carrier can be in the form of, for example, but not limited to, an ointment, cream, gel, paste, foam, aerosol, suppository, pad, or gel stick.

[0181] For oral application, the pharmaceutical composition may be in the form of a tablet or capsule, which may contain any of the following ingredients, or compounds of similar nature: binders, such as microcrystalline cellulose, gum tragacanth, or gelatin; excipients, such as starch or lactose; disintegrants, such as alginic acid, Primogel, or corn starch; lubricants, such as magnesium stearate; or glidants, such as colloidal silicon dioxide.When the unit dosage form is a capsule, it may contain, in addition to the above-mentioned types of materials, a liquid carrier, such as fatty oil.In addition, the unit dosage form may contain various other materials that modify the physical form of the dosage unit, such as sugar coating, shellac, or other enteric coating agents.The tablet of the present invention may also be film coated.

[0182] For the purpose of parenteral administration, solutions in sesame or peanut oil or aqueous propylene glycol, as well as sterile aqueous solutions of the corresponding water-soluble salts, can be used. Such aqueous solutions may be suitably buffered if necessary, and the liquid diluent is first rendered isotonic with sufficient saline or glucose. These aqueous solutions are particularly suitable for the purpose of intravenous, intramuscular, subcutaneous, and intraperitoneal injection.

[0183] Compositions also include incorporation of active agents into or on certain formulations of polymeric compounds such as polylactic acid, polyglycolic acid, hydrogels, or onto liposomes, microemulsions, micelles, unilamellar or multilamellar vesicles, erythrocyte ghosts, or spheroplasts. Such compositions affect the physical state, solubility, stability, rate of in vivo release, and rate of in vivo clearance.

[0184] In one embodiment, the present invention provides a combination preparation. In one embodiment, "combination preparation" defines a "kit of parts" in the sense that the combination partners defined above can be administered independently or by using various fixed combinations of distinct amounts of combination partners, i.e. simultaneously, concurrently, separately, or sequentially. In some embodiments, the parts of the kit of parts in this case can be administered, for example, simultaneously or chronologically staggered, i.e. at different time points and at equal or different time intervals, with respect to any part of the kit of parts. In some embodiments, the ratio of the total amounts of the combination partners can be administered in the combination preparation. In one embodiment, the combination preparation can be varied, for example, to address the needs of a subgroup of patients to be treated, or the needs of a single patient, whose different needs may be due to a specific disease, disease severity, age, sex, or weight, as can be easily performed by those skilled in the art.

[0185] In some embodiments, pharma- ceutically acceptable salts include, but are not limited to, alkali metal, alkaline earth metal, and / or ammonium salts, as well as citrate, phosphate, borate, lactate, and the like salts, and mixtures thereof. Conventional organic buffers, such as Good's buffers and the like, may also be used.

[0186] In some embodiments, the pharmaceutical composition comprises a therapeutically effective amount of one or more of the phosphorylcholine-tuftsin conjugates of the present invention. The term "therapeutically effective amount" refers to an amount of conjugate effective for treating a disease or disorder in a mammal. The term "therapeutically effective amount" refers to an amount effective at a dosage or time required to achieve a desired therapeutic or prophylactic result. The exact dosage form and regimen are determined by a physician according to the patient's condition. In some embodiments, the terms "therapeutically effective amount" and "pharmaceutical effective amount" are used interchangeably herein.

[0187] method In another aspect, there is a method of treating or preventing a disease or disorder associated with a fungal infection in a subject, comprising administering to said subject a therapeutically effective amount of a fungicidal composition of the present invention. In some embodiments, the method is for inhibiting, reducing, or decreasing fungal load in a subject in need thereof. In some embodiments, the composition is for use in inhibiting, reducing, or decreasing fungal activity in a subject in need thereof. In some embodiments, the fungal activity is associated with infection by one or more fungal species.

[0188] In some embodiments, the subject suffers from a fungal infection. In some embodiments, the disclosed invention is directed to a fungicide composition for use in inhibiting, reducing or decreasing fungal activity. In some embodiments, the fungal activity comprises any activity selected from the group consisting of proliferation, fungicide resistance, cell-cell communication or quorum sensing, biofilm production, toxin production or secretion, and combinations thereof.

[0189] In some embodiments, the method further comprises a pre-step performed before the step of administering. In some embodiments, the pre-step comprises determining that the subject suffers from a fungal infection. In some embodiments, the determining comprises determining fungal activity. In some embodiments, the determining is performed in a sample from the subject. Fungal activity can be assayed using any common method, non-limiting examples of which include, but are not limited to, spectrophotometry, drug resistance assay using selective substrates, bioluminescence assay, liquid chromatography and mass spectrometry, or other methods that are apparent to those skilled in the art.

[0190] As used herein, the term "treatment" or "treating" of a fungal infection and / or a disease or disorder associated therewith includes alleviating at least one symptom thereof, reducing its severity, or inhibiting its progression. Treatment does not necessarily mean that the disease, disorder, or condition is completely cured. To be an effective treatment, the compositions useful herein need only reduce the severity of the disease, disorder, or condition, reduce the severity of symptoms associated therewith, or provide an improvement in the quality of life of the patient or subject. In some embodiments, treating a fungal infection includes at least one of preventing, attenuating, and / or inhibiting a disease, disorder, or condition associated with fungal activity in a subject.

[0191] The term "subject" as used herein refers to animals, more specifically non-human mammals and human organisms. Non-human animal subjects can also include prenatal forms of animals, such as embryos or fetuses. Non-limiting examples of non-human animals include horses, cows, camels, goats, sheep, dogs, cats, non-human primates, mice, rats, rabbits, hamsters, guinea pigs, and pigs. In one embodiment, the subject is a human. Human subjects can also include fetuses.

[0192] As used herein, the term "prevention" of a disease, disorder, or condition encompasses delaying, preventing, inhibiting, or inhibiting the onset of the disease, disorder, or condition. As used in accordance with the subject matter described herein, the term "prevention" relates to a prophylactic process in which a subject is exposed to a peptide described herein before the induction or onset of the disease / disorder process. This can be done when an individual has a genetic lineage that indicates a predisposition to the onset of the disease / disorder to be prevented. For example, this can be the case for an individual whose ancestors indicate a predisposition to a particular type of, for example, inflammatory disorder. The term "suppression" is used to describe a condition in which the disease / disorder process has already begun, but no obvious symptoms of the condition have yet been noticed. Thus, an individual's cells may have the disease / disorder, but the outward signs of the disease / disorder have not yet been clinically recognized. In either case, the term prophylaxis can be applied to encompass both prevention and inhibition. Conversely, the term "treatment" refers to the clinical application of an active agent to combat an already existing condition in which clinical symptoms are already noticed in the patient.

[0193] As used herein, the term "pathology" refers to a condition that interferes with or alters the performance of bodily functions. It includes anatomical and physiological deviations from the normal that constitute a dysfunction of the normal state of a living animal or one of its parts.

[0194] As used herein, the term "antifungal activity" refers to the ability to inhibit, prevent, reduce or delay any one of the following: growth, protein production rate, protein production yield, protein synthesis, messenger RNA translation rate, at least one fungal biofilm formation, and / or eradicate live fungal cells or their spores. In some embodiments, antifungal activity is achieved on a surface or in a moist environment. In some embodiments, inhibiting or reducing or delaying the formation of fungal load refers to inhibiting, reducing or delaying the growth of fungi and / or eradicating some or all of the existing population of fungi.

[0195] Methods for determining the reduction in protein synthesis, protein production rate, and yield are common and apparent to one of skill in the art, a non-limiting example of which using a luciferase assay is provided herein below.

[0196] In some embodiments, the inhibition, prevention, delay, or reduction is at least 5%, at least 15%, at least 25%, at least 40%, at least 50%, at least 65%, at least 75%, at least 85%, at least 90%, at least 95%, at least 99%, or any value or range therebetween, as compared to a control. In some embodiments, the alteration is 1-5%, 7-15%, 10-25%, 20-40%, 35-50%, 45-65%, 55-75%, 70-85%, 80-90%, 87-95%, or 92-100% as compared to a control. Each possibility represents a separate embodiment of the present invention.

[0197] In some embodiments, the reduction comprises at least a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99%, or 100% reduction, with each possibility representing a separate embodiment of the present invention.

[0198] As used herein, the terms "administering," "administration," and similar terms refer to any method of delivering a composition containing an active agent to a subject in such a manner as to provide a therapeutic benefit in sound medical practice. In some embodiments, administration is ocular or intraocular administration.

[0199] Those skilled in the art will understand that, especially eye drops, and also ointments to some extent, do not completely reach the site of inflammation. Therefore, to compensate for the mode of administration, the dose needs to be increased or decreased as determined by those skilled in the art. Administration by intraocular injection reaches the site of inflammation more directly, and therefore the administered dose needs to be adjusted again.

[0200] In some embodiments, the amount of composition administered (dosage) will, of course, be dependent on the subject being treated, the medical condition being treated, the severity of the affliction, the manner of administration, the judgment of the prescribing physician, etc.

[0201] In one embodiment, depending on the severity and responsiveness of the condition to be treated, administration may be a single or multiple administrations, where the course of treatment lasts from several days to several weeks, or until a cure is effected or a diminution of the disease state is achieved.

[0202] In some embodiments, the fungicidal composition of the present invention is administered in a therapeutically safe and effective amount. As used herein, the term "safe and effective amount" refers to an amount of the component that is sufficient to obtain a desired therapeutic response commensurate with a reasonable benefit / risk ratio when used in the methods described herein without undue adverse side effects (such as toxicity, irritation, or allergic response). In another embodiment, a therapeutically effective amount of a conjugate, or any derivative or combination thereof, is the amount of the conjugate referred to herein required for the predicted biological effect measurable in vivo. The actual amount administered, as well as the rate and time course of administration, will depend on the nature and severity of the condition being treated. Decisions regarding the prescription of treatment, such as dosage, timing, etc., are within the responsibility of a general practitioner or specialist and will usually take into account the disorder being treated, the condition of the individual patient, the site of delivery, the method of administration, and other factors known to the practitioner. Examples of techniques and protocols can be found in Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott Williams & Wilkins, Philadelphia, Pa., (2005). In some embodiments, the effective amount or dosage formulation can be initially estimated from in vitro assays, hi one embodiment, a dose can be formulated in animal models and such information can be used to more accurately determine useful doses in humans.

[0203] In some embodiments, the daily dose (i.e., the amount of phosphorylcholine-tuftsin conjugate per day) is between 1-2000 μg, between 100-1000 ng, between 200-2000 μg, between 1-10 μg, between 10-50 μg, between 50-100 μg, between 100-200 μg, between 200-300 μg, between 300-400 μg, between 400-500 μg, between 500 and 600 μg, between 600 and 700 μg, between 700 and 800 μg, between 800 and 900 μg, between 900 and 1000 μg, between 1000 and 1100 μg, between 1100 and 1300 μg, between 1300 and 1500 μg, between 1500 and 1800 μg, between 1800 and 2000 μg (including any range or value therebetween).

[0204] In some embodiments, the fungicidal composition is administered 1, 2, 3, 4, 5, 6, 8, 9, 10 times per day (including any range therebetween).

[0205] The pharmaceutical compositions may further comprise additional pharma- ceutical active or inactive agents, such as, but not limited to, antifungals, antioxidants, buffers, bulking agents, surfactants, anti-inflammatory agents, antiviral agents, chemotherapeutic agents, and antihistamines.

[0206] In one embodiment of the present invention, the pharmaceutical compositions described herein above are packaged in packaging material and identified by printing in or on the packaging material for use in treating a disease or disorder as described herein.

[0207] In another embodiment of the invention, the pharmaceutical composition is packaged with packaging material and identified by printing in or on the packaging material for use in monitoring a disease or disorder described herein.

[0208] The product of the invention may be presented in a pack or dispenser device, such as a kit approved by the United States Food and Drug Administration (FDA), which may contain one or more unit dosage forms containing the disclosed compositions, if desired. The pack may, for example, comprise a 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 contained with a notice associated with the container in a form prescribed by a government agency regulating the manufacture, use, or sale of pharmaceuticals, which notice reflects the form of the composition or the agency's approval for human or veterinary administration. Such notice may, for example, be a label approved by the FDA for prescription drugs or an approved product insert.

[0209] In some embodiments, the kit comprises a single dosage form, wherein the dosage form contains a daily dose of the disclosed composition.In some embodiments, the kit comprises multiple dosage forms.In some embodiments, the kit comprises multiple dosage forms, wherein the multiple dosage forms are equivalent to a daily dose of the disclosed composition.

[0210] It will be understood that the compositions of the present invention may be administered in combination with other drugs, including other anti-fungal drugs.

[0211] General As used herein, the term "about" refers to ±10%.

[0212] The terms "comprises," "comprising," "includes," "including," "having," and their cognates mean "including, but not limited to."

[0213] The term "consisting of" means "including and limited to."

[0214] The term "consisting essentially of" means that a composition, method, or structure may include additional ingredients, steps, and / or moieties, provided that the additional ingredients, steps, and / or moieties do not materially alter the basic and novel characteristics of the claimed formulation, method, or structure.

[0215] The word "exemplary" is used herein to mean "serving as an example, instance, or illustration." Any embodiment described as "exemplary" is not necessarily to be construed as preferred or advantageous over other embodiments and / or as excluding the incorporation of features from other embodiments.

[0216] The word "optionally" is used herein to mean "provided in some embodiments and not provided in other embodiments." Any particular embodiment of the invention may include multiple "optional" features unless such features are inconsistent.

[0217] As used herein, the singular forms "a," "an," and "the" include the plural forms unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" can include a plurality of compounds, including mixtures thereof.

[0218] Throughout this application, various embodiments of the invention may be presented in a range format. Descriptions in range format are merely for convenience and brevity and should not be construed as inflexible limitations on the scope of the invention. Thus, descriptions of ranges should be considered to have all possible subranges specifically disclosed, as well as individual numerical values ​​within the range. For example, descriptions of ranges such as 1-6 should be considered to have specifically disclosed subranges such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, as well as individual numerical values ​​within the range, such as 1, 2, 3, 4, 5, and 6. This is true regardless of the breadth of the range.

[0219] Whenever a range of numbers is recited herein, it is meant to include all recited numbers (fractions or integers) within the recited range. The terms "ranging from / between" a first designated number and a second designated number and "ranging from" a first designated number to a second designated number are used interchangeably herein and are meant to include the first and second designated numbers and all fractions and integers therebetween.

[0220] As used herein, the term "method" refers to methods, means, techniques, and procedures for accomplishing a given task, including, but not limited to, methods, means, techniques, and procedures known to or readily developed from known methods, means, techniques, and procedures by practitioners in the fields of chemistry, pharmacology, biology, biochemistry, and medicine.

[0221] It is understood that certain features of the invention that are described for clarity in the context of separate embodiments may also be provided in combination in a single embodiment. Conversely, various features of the invention that are described for convenience in the context of a single embodiment may also be provided separately or in any suitable subcombination. All combinations of the embodiments belonging to the invention are specifically embraced by the invention and are disclosed herein as if each and every combination were individually and expressly disclosed herein. Moreover, all subcombinations of the various embodiments and elements thereof are also specifically embraced by the invention and are disclosed herein as if each and every subcombination were individually and expressly disclosed herein.

[0222] Additional objects, advantages, and novel features of the present invention will become apparent to those skilled in the art upon examination of the following examples, which are not intended to be limiting. Additionally, each of the various embodiments and aspects of the present invention as described hereinabove and as claimed in the claims section below finds experimental support in the following examples.

[0223] Various embodiments and aspects of the present invention as delineated hereinabove and as claimed in the claims section below find experimental support in the following examples.

[0224] Working Example Materials used in the manufacture of exemplary ophthalmic compositions include: TRS (pharmaceutical substance, investigational grade); sodium phosphate, monobasic, dihydrate (buffering agent, USP / NF); sodium phosphate, dibasic, dihydrate (buffering agent, USP / NF); benzalkonium chloride (preservative, USP / NF); sodium alginate (mucoadhesive, Ph Eur); hypromellose (thickening agent, USP / NF); sodium hydroxide and / or hydrochloric acid (pH adjuster, USP / NF); water for injection (solvent, USP / NF); mannitol (osmolality adjuster, USP).

[0225] The drug substance "TRS" is the trifluoroacetate [TFA] salt of a phosphorylcholine-tuftsin conjugate represented by Formula 1 (shown herein). Molecular formula: C43H68N13O13P. Appearance: yellow-orange solid. Solubility: greater than 40 mg / ml in water.

[0226] Packaging for the Ophthalmic Composition. Single-use media bottle (PETG) with HDPE screw cap. After filling, the closure is sealed with a tamper evident label. Bottles and closures are purchased radiation sterilized and apyrogenic. Storage conditions: -20°C ± 5°C. Long-term stability studies of [TRS] at -20°C ± 5°C and accelerated stability studies at +5°C ± 3°C have recently been initiated for 36 months. Available stability data at -20°C ± 5°C demonstrated that the drug substance is stable for 6 months when stored under these conditions.

[0227] An exemplary pharmaceutical formulation of the invention is as follows: TRS (about 1 (w / w)%), phosphate buffer (10 mM), HPMC (about 0.5 (w / w)%), BAK (about 0.02 (w / w)%), mannitol (about 1.7 (w / w)%).

[0228] Example 1 The phosphorylcholine-Tuftsin conjugate of formula 2A, and those represented by formulas 3A, 4, and 5, have been synthesized by reacting TRS with acetaldehyde, as disclosed hereinbelow.

[0229] Briefly, 50 mg of TRS was dissolved in 2.5 mL of water and 50 equivalents of acetaldehyde were added (at room temperature). The pH was then adjusted to 6.8. The solution was shaken over the weekend and analyzed by analytical HPLC. Various phosphorylcholine-Tuftsin conjugates, such as phosphorylcholine-Tuftsin conjugates of formula 2A and formula 3A, and further phosphorylcholine-Tuftsin conjugates represented by either formula 4 and / or 5, have been detected in the reaction mixture. The reaction mixture was purified by preparative HPLC to isolate phosphorylcholine-Tuftsin conjugate of formula 3A. However, it was not possible to isolate phosphorylcholine-Tuftsin conjugate of formula 2A in the form of a pure compound using the preparative HPLC separation method.

[0230] We then successfully obtained phosphorylcholine-tuftsin conjugate of formula 2A (more than 90% chemical purity) through partial hydrolysis of phosphorylcholine-tuftsin conjugate of formula 3A. Phosphorylcholine-tuftsin conjugate of formula 3A was dissolved in 0.1% aqueous formic acid and the pH was adjusted to 2.7. After 30 minutes, the solution was lyophilized. The crude reaction mixture was purified by preparative HPLC to obtain phosphorylcholine-tuftsin conjugate of formula 2A.

[0231] The chemical structures of the synthesized phosphorylcholine-tuftsin conjugates were confirmed by LC-MS and optionally 1D- and 2D-NMR.

[0232] The phosphorylcholine-Tuftsin conjugate of formula 2A has been found to exhibit robust biological activity and sufficient chemical stability. In contrast, the phosphorylcholine-Tuftsin conjugate of formula 3A undergoes hydrolysis (converts to PC of formula 2A at acidic pH). Either one of the PCs of formula 4 or 5 (further derivatives of PC of formula 2) features reduced chemical stability compared to PC of formula 2.

[0233] Example 2 The present inventors have surprisingly discovered a synergistic preservative activity of the combination of phosphorylcholine-tuftsin conjugate (TRS) of formula 1 and benzalkonium chloride. Briefly, 1.0% (w / w) TRS formulations based on the components of the TRS formulation disclosed below were prepared, containing three different concentrations of benzalkonium chloride. The solutions were then diluted with 100% ethanol, 100% water, and 100% ethanol. <51> The specimens were subjected to PET (Preservative Effectiveness Testing) according to the method described above.

[0234] A TRS formulation containing only 0.003% (w / w) benzalkonium chloride (corresponding to 15% label claim) passed the PET test. Furthermore, a TRS formulation containing an even lower concentration of 0.001% (corresponding to 5% label claim) benzalkonium chloride showed a robust synergistic antibacterial effect. A similar formulation without benzalkonium chloride did not show any significant antibacterial effect, pointing to the synergistic effect of both benzalkonium chloride and TRS in the pharmaceutical composition of the present invention.

[0235] We will conduct similar experiments with additional phosphorylcholine-tuftsin conjugates disclosed herein. Based on the demonstrated fungicidal effects of both phosphorylcholine-tuftsin conjugates of formula 2 and 1 (see Example 3), it is predicted that additional phosphorylcholine-tuftsin conjugates (e.g., conjugates of formula 2 and / or 3) will exhibit similar synergistic antimicrobial effects.

[0236] Example 3 In preliminary studies, inhibition of yeast cell respiration in aqueous suspension was observed as a reduction in fermentation activity in the presence of TRS (phosphorylcholine-tuftsin conjugate (PC) of Formula 1), resulting in reduced gas formation, whereas a control suspension without TRS exhibited normal yeast cell respiration associated with substantial gas formation. Yeast fermentation activity correlated with foam formation within the aqueous suspension.

[0237] Briefly, various amounts of 1% (w / w) TRS solution were added to an aqueous suspension containing yeast (S. cerevisiae) and sufficient sucrose to obtain aqueous compositions containing 4 ppm, 20 ppm, 40 ppm, and 80 ppm TRS. Gas formation (fermentation activity) of aqueous suspensions containing TRS (hereinafter "samples") was quantified by assessing the volume of gas released and compared to a similar aqueous yeast suspension lacking TRS (hereinafter "control"). The results of this experiment are depicted in FIG. 1.

[0238] Surprisingly, it has been observed that samples containing 40 ppm and 80 ppm TRS were almost completely devoid of fermentation activity (no bubbles were observed), whereas the control was characterized by substantial fermentation activity, as inferred from vigorous gas formation therein. As shown in FIG. 1, samples with 20 ppm TRS (20 μg / ml) already showed a significant reduction in fermentation activity compared to the control. It is therefore hypothesized that TRS is effective in inhibiting or reducing fungal load and / or fungal activity.

[0239] Additionally, the phosphorylcholine-tuftsin conjugate of formula 2A, synthesized as disclosed herein, was tested to evaluate its antifungal activity. We performed similar experiments as disclosed above using various concentrations (i.e., 6.7 ppm, 20 ppm, 40 ppm, and 80 ppm) of the phosphorylcholine-tuftsin conjugate of formula 2.

[0240] The results of the above experiment (yeast activity plotted versus the concentration of each PC in suspension) are shown in Figure 2, which demonstrates a significant reduction in yeast activity even at a minimum concentration of approximately 7 ppm of the conjugate of Formula 2. Yeast activity is further reduced with increasing concentrations of the conjugate, almost completely disappearing at 80 ppm of the conjugate of Formula 2.

[0241] Based on this experiment, the inventors speculate that the phosphoryl tuftsin conjugates of the present invention will exhibit robust fungicidal activity against additional fungal / yeast and / or mold species.

[0242] While 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.

[0243] All publications, patents, and patent applications mentioned in this specification are incorporated by reference in their entirety herein to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference herein. Furthermore, the listing or identification of any reference in this application should not be construed as an admission that the reference is available as prior art to the present invention. To the extent section headings are used, they should not be construed as necessarily limiting.

Claims

1. A phosphorylcholine-tuftsin conjugate comprising any functional derivative thereof, a salt thereof, or both, the phosphorylcholine-tuftsin conjugate is 【Chemistry 1】 and the functional derivative thereof is not a phosphorylcholine-tuftsin conjugate of formula 1; Phosphorylcholine-tuftsin conjugate.

2. The phosphorylcholine-tuftsin conjugate of claim 1 characterized by antifungal activity.

3. 3. The phosphorylcholine-tuftsin conjugate of claim 1, wherein the salt is a pharmaceutically acceptable salt and the phosphorylcholine-tuftsin conjugate is characterized by a chemical purity of at least 95%.

4. The functional derivative is 【Chemistry 2】 2. The phosphorylcholine-tuftsin conjugate of claim 1, wherein:

5. A pharmaceutical composition comprising the phosphorylcholine-tuftsin conjugate of claim 1 and a pharmaceutically acceptable carrier.

6. 6. The pharmaceutical composition of claim 5, comprising a fungicidally effective amount of said phosphorylcholine-tuftsin conjugate, wherein said fungicidally effective amount is at least about 5 μg / ml or between about 5 ppm and about 2% (w / w).

7. Formula 1: 【Transformation 3】 6. The pharmaceutical composition of claim 5, further comprising an additional phosphorylcholine-tuftsin conjugate of the formula:

8. 8. The pharmaceutical composition of claim 7, wherein the concentration of the additional phosphorylcholine-tuftsin conjugate in the pharmaceutical composition is between 5 ppm and 30% (w / w).

9. 6. The pharmaceutical composition of claim 5, wherein the pharmaceutically acceptable carrier is an aqueous composition comprising between 0.1 and 5 weight / weight (w / w) % of a viscosity-increasing agent and between 0.1 and 5 (w / w) % of a mucoadhesive polymer.

10. 10. The pharmaceutical composition of claim 9, wherein the mucoadhesive polymer is an ionic mucoadhesive polymer, which may be selected from alginic acid, chitosan, polyacrylate, hyaluronic acid, carboxymethylcellulose, pectin, and gelatin, including any salt, copolymer, or combination thereof; and the viscosity increasing agent may comprise hydroxymethylcellulose, hydroxyethylcellulose, cellulose, hydroxypropylmethylcellulose, polyvinylpyrrolidone, polyethylene glycol, polypropylene glycol, poly(2-hydroxyethyl methacrylate), and polyvinyl alcohol, including any salt, copolymer, or combination thereof.

11. 10. The pharmaceutical composition of claim 9, wherein the aqueous composition further comprises up to 10% (w / w) of an osmolality adjusting agent, the viscosity increasing agent comprises hydroxymethylcellulose, and the mucoadhesive polymer comprises alginic acid, including any salt thereof.

12. The pharmaceutical composition of claim 5 , wherein the pharmaceutical composition is an ophthalmic composition formulated for ocular administration.

13. A fungicidal composition comprising a fungicidally effective amount of one or more phosphorylcholine-tuftsin conjugates, salts thereof, functional derivatives thereof, or any combination thereof, and further comprising a pharmaceutically acceptable carrier.

14. 14. The fungicidal composition of claim 13, wherein the one or more phosphorylcholine-tuftsin conjugates comprise at least one phosphorylcholine moiety or derivative thereof and tuftsin or a derivative thereof covalently linked via a spacer.

15. 14. Use of the fungicidal composition of claim 13 for treating or preventing a disease or disorder associated with a fungal infection in a subject in need thereof.

16. 14. Use of a fungicidal composition according to claim 13 to reduce or inhibit the activity or growth of at least one fungus.

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