WO2024246911A1

WO2024246911A1 - Cannabinoid derivatives and uses thereof in the treatment of fungal infections

Info

Publication number: WO2024246911A1
Application number: PCT/IL2024/050540
Authority: WO
Inventors: Natalya M KOGAN; Aviva Breuer; Francesca Levi-Schaffer; Ilan ZAFFRAN; Prince OFORI; Raphael Mechoulam
Original assignee: Yissum Research Development Co of Hebrew University of Jerusalem
Current assignee: Yissum Research Development Co of Hebrew University of Jerusalem
Priority date: 2023-06-01
Filing date: 2024-05-30
Publication date: 2024-12-05
Anticipated expiration: 2025-12-01

Abstract

The present invention provides cannabinoid derivatives, compositions comprising them and uses thereof in a method of treating fungal infections.

Description

CANNABINOID DERIVATIVES AND USES THEREOF IN THE TREATMENT OF FUNGAL INFECTIONS

BACKGROUND OF THE INVENTION

[001] Antimicrobial and antifungal resistance are serious public health concerns globally. The burden and consequences of antifungal resistance have not been fully recognized. In the past few decades, incidences of fungal infections have increased dramatically due to impaired immune systems, complex surgical procedures, and extensive antibiotic treatment. [002] Infections due to Candida range from cutaneous to severe invasive infections. Candida albicans, one of the leading opportunistic fungal pathogens, causes high mortality rate especially in immunocompromised and high-risk surgical patients. C. albicans is responsible for more than 50% of human candidiasis, including two major types of infections, superficial infections (nonlethal), such as oral or vaginal candidiasis; and systemic infections (~40% mortality). Despite this prevalence, therapeutic options are limited. Currently, there are three main classes of antifungal drugs used to treat severe C. albicans infections. They include the azoles, the echinocandins, and the polyenes. Resistance to first- and second-line antifungal s, such as echinocandins and fluconazole (FLU), is a major problem in C. albicans infections.

[003] Antifungals are very important for the treatment of fungal infections particularly in immunocompromised and organ transplant patients. The development of new class of antifungals with different mechanism of action and antifungal spectrum is desperately needed.

[004] Current antifungals that are used to treat systemic infections only target membrane and cell wall integrity. Azoles and echinocandins are fungistatic and moderately fungicidal respectively. This often poses a problem for successful treatment, given that individuals with serious fungal infections are usually severely immunocompromised. Current antifungal treatments lead to altered liver function tests and have limitations with respect to their spectra of activity and toxicities.

[005] Limitations of one or more of the currently used antifungal drugs include high recurrence of infection, high mortality rates, poor oral bioavailability, limitations in spectrum of activity, and liver toxicity. Therefore, there are three thresholds must be met in order to develop a novel antifungal for clinical use. The first threshold is to have a unique chemical composition. The second threshold is having a similar or better spectrum of activity as compared to amphotericin B and fluconazole. The third threshold is having a similar or better toxicity profile compared to amphotericin B and fluconazole.

[006] There is a long term need to develop effective treatment for fungal infections, in which currently available antifungals are failing with respect to effectiveness, toxicity and resistance.

SUMMARY OF THE INVENTION

[007] The invention thus provides a compound (also referred to herein as abnCBD) having the general formula (I):

Wherein

R1 is selected from H, straight or branched C3 - C10 alkyl, straight or branched

C3 - C10 alkenyl, straight or branched C3 - C10 alkynyl; optionally substituted with at least one phenyl, hydroxyphenyl, hydroxy, carboxy, halogen, cyano and any combinations thereof;

Ri l l is selected from H, straight or branched Cl - C10 alkyl, straight or branched Cl - C10 alkenyl, straight or branched Cl - C10 alkynyl; optionally substituted with at least one phenyl, hydroxyphenyl, hydroxy, carboxy, halogen, cyano and any combinations thereof; provided that at least one of R1 and Ri l l is different than H;

R2 and R3 are each independently selected from, H, straight or branched Cl - C10 alkyl, straight or branched Cl - C10 acyl; optionally substituted by morpholino;

R4 is a cyclic terpene having one or two double bond and optionally substituted by at least one of straight or branched Cl - C10 alkyl, straight or branched C2 - C10 alkenyl, straight or branched C2 - C10 alkynyl, hydroxy, carboxy; halogen, cyano and any combinations thereof.

[008] In some embodiments, R1 is a straight or branched C3 - C10 alkyl. In some embodiments, R1 is a straight or branched C3 - C10 alkenyl. In some embodiments, R1 is selected from a straight or branched C3 - C10 alkyl and a straight or branched C3 - C10 alkenyl. In some embodiments, R1 is substituted with at least one phenyl, hydroxy phenyl. In some embodiments, R1 is substituted with at least one phenyl. In some embodiments, R1 is substituted with at least one hydroxy phenyl.

[009] In some embodiments, Ri l l is a straight or branched Cl - C10 alkyl. In some embodiments, R111 is a straight or branched Cl - C10 alkenyl. In some embodiments, Ri l l is selected from a straight or branched Cl - C10 alkyl and a straight or branched Cl - C10 alkenyl. In some embodiments, R111 is substituted with at least one phenyl, hydroxyphenyl. In some embodiments, Ri l l is substituted with at least one phenyl. In some embodiments, R111 is substituted with at least one hydroxyphenyl.

[0010] In some embodiments, R1 is a C3 - C10 alkyl and R111 is a straight or branched Cl - C10 alkyl. [0011] In other embodiments, R2 and R3 both H. In some embodiments, R2 and R3 each independently straight or branched Cl-C5alkyl. In some embodiments, R2 and R3 each independently straight or branched Cl - C10 acyl. In some embodiments, R2 and R3 are each selected from H and straight or branched Cl-C5alkyl. In some embodiments, R2 and R3 are each selected from H and Ac. In some embodiments, R2 and R3 are each selected from straight or branched Cl-C5alkyl and Ac.

[0012] In further embodiments, R4 is a cyclic terpene having one double bond. In some embodiments, R4 is a cyclic terpene being a bicyclic terpene. In some embodiments, R4 is a moiety of general formula (II):

Wherein - is a single bond, double bond or null; provided that one or two of - is a double bond; and provided that no more than two - are optionally null;

Wherein one of R5 - R10 is a connection to general formula (I); and

Wherein each of R5 - R10 is selected from: H, straight or branched Cl - C10 alkyl, straight or branched C2 - C10 alkenyl, straight or branched C2 - C10 alkynyl; each optionally substituted by at least one hydroxy, carboxy, halogen, cyano; or two of R5 - R10 together with the carbon atoms they are connected to form a C3 - C8 ring.

[0013] In other embodiments, R4 is a moiety of general formula (III):

Wherein

is a single bond, double bond or null; provided that one or two of is a double bond; and provided that no more than two -

are optionally null; and

Wherein is a connection to general formula (I); and

Wherein each of R11 -R14 is selected from: H, straight or branched Cl - C10 alkyl, straight or branched C2 - C10 alkenyl, straight or branched C2 - C10 alkynyl; each optionally substituted by at least one hydroxy, halogen, cyano; or two of R11 - R14 together with the carbon atoms they are connected to form a C3 - C8 ring.

[0014] In some embodiments, R4 is selected from:

Wherein is a connection to general formula (I); and

Wherein each of R15 -R25 is selected from: H, straight or branched Cl - C10 alkyl, straight or branched C2 - C10 alkenyl, straight or branched C2 - C10 alkynyl; each optionally substituted by at least one hydroxy, carboxy, halogen, cyano; or two of R15 - R25 together with the carbon atoms they are connected to form a C3 - C8 ring.

[0015] In some embodiments, R4 is a moiety of general formula (IV). In some embodiments, R4 is a moiety of general formula (V). In some embodiments, R4 is a moiety of general formula (VI). In some embodiments, R4 is a moiety of general formula (V).

[0016] In some embodiments, a compound of the invention is selected from:

• (1'R,2'R)-5'-methyl-6-pentyl-2'-(prop-l-en-2-yl)-r,2',3',4'-tetrahydro-[l,r- biphenyl]-2,4-diol (Compound 1, NK-1-25);

• (1'R,2'R)-5'-methyl-6-pentyl-2'-(prop-l-en-2-yl)-r,2',3',4'-tetrahydro-[l,r- biphenyl]-2,4-diyl diacetate (Compound 2);

• (rS,2'S)-2'-isopropyl-5'-methyl-6-pentyl-l',2',3',4'-tetrahydro-[l,r-biphenyl]- 2,4-diol (Compound 3, NK1-69);

• (1'R,2'R)-2-methoxy-5'-methyl-6-pentyl-2'-(prop-l-en-2-yl)-r, 2', 3', d'- tetrahydro-fl, l'-biphenyl]-4-ol (Compound 4);

• (1'R,2'R)-4-methoxy-5'-methyl-6-pentyl-2'-(prop-l-en-2-yl)-l', 2', 3', d'- tetrahydro-fl, l'-biphenyl]-2-ol (Compound 5);

• (1'R,2R)-2'-methoxy-5-methyl-6'-pentyl-2 -(prop- l-en-2-yl)-l, 2,3, 4-tetrahydro- l,l'-biphenyl (Compound 6);

• (l'R,2'R)-2'-(3-hydroxyprop-l-en-2-yl)-5'-methyl-6-pentyl-r,2',3',4'-tetrahydro- [l,l'-biphenyl]-2,4-diyl diacetate (Compound 7, TE1-4);

• (l'R,2'R)-2'-(3-hydroxyprop-l-en-2-yl)-5'-methyl-6-pentyl-r,2',3',4'-tetrahydro-

[l,l'-biphenyl]-2,4-diol (Compound 8, Tl-5);

[0017] The invention further provides a composition comprising at least one compound as disclosed herein above and below. In some embodiments, said composition further comprises at least one further anti-fungal agent. In some embodiments, said composition further comprises at least one further pharmaceutical active agent.

[0018] The invention further provides a compound as disclosed herein above and below for use in the treatment of a medical condition, a disease or a disorder associated with a fungal infection.

[0019] The invention further provides a compound as disclosed herein above and below for use in the treatment of a medical condition, a disease or a disorder associated with a resistant fungal infection.

[0020] In some embodiments, said medical condition, disease or disorder associated with fungal infection is caused by at least one fungi selected from Histoplasma (e.g. H. capsulatum), Coccidioides, Blastomyces, Paracoccidioides, Cryptococcus (e.g. C. neoformans), Aspergillus (e.g. A. fumigatus, A. niger, A. nidulans, A. terreus, A. sydowi, A. flavatus, and A. glaucus), Zygomycetes (e.g. Basidiobolus, Conidiobolus, Rhizopus, Mucor, Absidia, Mortierella, Cunninghamella, and Saksenaea), Candida (e.g. C. albicans, C. tropicalis, C. parapsilosis, C. stellatoidea, C. krusei, C. parakrusei, C. lusitaniae, C. pseudotropicalis, C. guilliermondi and C. glabrata), Cryptosporidium parvum, Sporothrix schenckii, Piedraia hortae, Trichosporon beigelii, Malassezia furfur, Phialophora verrucosa, Fonsecae pedrosoi, Madurella mycetomatis and Pneumocystis carinii and any combinations thereof.

[0021] The present embodiments further encompass any enantiomers, prodrugs, solvates, hydrates and/or pharmaceutically acceptable salts of the compounds described herein. [0022] As used herein, the term "enantiomer" refers to a stereoisomer of a compound that is superposable with respect to its counterpart only by a complete inversion/reflection (mirror image) of each other. Enantiomers are said to have “handedness” since they refer to each other like the right and left hand. Enantiomers have identical chemical and physical properties except when present in an environment which by itself has handedness, such as all living systems.

[0023] The term “prodrug” refers to an agent, which is converted into the active compound (the active parent drug) in vivo. Prodrugs are typically useful for facilitating the administration of the parent drug. They may, for instance, be bioavailable by oral administration whereas the parent drug is not. A prodrug may also have improved solubility as compared with the parent drug in pharmaceutical compositions. Prodrugs are also often used to achieve a sustained release of the active compound in vivo. An example, without limitation, of a prodrug would be a compound as described herein, having one or more carboxylic acid moieties, which is administered as an ester (the “prodrug”). Such a prodrug is hydrolyzed in vivo, to thereby provide the free compound (the parent drug). The selected ester may affect both the solubility characteristics and the hydrolysis rate of the prodrug.

[0024] The term “solvate” refers to a complex of variable stoichiometry (e.g., di-, tri-, tetra- , penta-, hexa-, and so on), which is formed by a solute (the compound of the present invention) 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.

[0025] The term “hydrate” refers to a solvate, as defined hereinabove, where the solvent is water.

[0026] The phrase "pharmaceutically acceptable salt" refers to a charged species of the parent compound and its counter ion, which is typically used to modify the solubility characteristics of the parent compound and/or to reduce any significant irritation to an organism by the parent compound, while not abrogating the biological activity and properties of the administered compound. An example, without limitation, of a pharmaceutically acceptable salt would be a carboxylate anion and a cation such as, but not limited to, ammonium, sodium, potassium and the like.

[0027] As is well known in the art, the phrase “acid addition salt” describes a complex of two ionizable moieties, a base and an acid, which, when interacted in a particular stoichiometric proportion and under suitable conditions, form a salt that comprises one or more cations of the base moiety and one or more anions of the acid moiety. As used herein, the phrase "acid addition salt" refers to such a complex, in which the base moiety in amine, such that the salt comprises a cationic form of the amine (ammonium) and an anionic form of an acid.

[0028] Depending on the stochiometric proportions between the base and the acid in the salt complex, as is detailed hereinbelow, the acid additions salts can be either mono addition salts or poly addition salts.

[0029] The phrase “mono addition salt”, as used herein, refers to a salt complex in which the stochiometric ratio between the acid anion and amine cation is 1: 1, such that the acid addition salt includes one molar equivalent of the acid per one molar equivalent of the conjugate.

[0030] The phrase “poly addition salt”, as used herein, refers to a salt complex in which the stochiometric ratio between the acid anion and the amine cation is greater than 1:1 and is, for example, 2: 1, 3 : 1, 4: 1 and so on, such that the acid addition salt 15 includes two or more molar equivalents of the acid per one molar equivalent of the conjugate. [0031] The stoichiometric proportions between the base and the acid of the salt complex, according to some embodiments of the present invention, ranges from 6:1 to 1:6 base:acid equivalents, from 4:1 to 1:4 base:acid equivalents, from 3:1 to 1 :3 base:acid equivalents or from 1: 1 to 1 :3 base:acid equivalents.

[0032] The acid addition salts of a chemical conjugate according to the present invention are therefore complexes formed between one or more amino groups of the compound and one or more equivalents of an acid. The acid addition salts may therefore include a variety of organic and inorganic acids, such as, but not limited to, halogen acids such as hydrochloric acid which affords an hydrochloric acid addition salt (as well as salts of bromide and iodide), acetic acid which affords an acetic acid addition salt, ascorbic acid which affords an ascorbic acid addition salt, benzoic acid which affords a benzoic acid addition salt (benzoate), benzenesulfonic acid which affords a benzenesulfonic acid addition salt, camphor sulfonic acid which affords a camphorsulfonic acid addition salt, naphthyl sulfonic acid which affords a naphthyl sulfonic acid addition salt, toluenel sulfonic acid (p-toluenesulfonic acid) which affords a toluenesulfonic acid addition salt (tosylate), trifluoroacetic acid which affords a trifluoroacetic acid addition salt, citric acid which affords a citric acid addition salt, maleic acid which affords a maleic acid addition salt (maleate), methanesulfonic acid which affords a methanesulfonic acid (mesylate or methanesulfonate) addition salt, naphthalenesulfonic acid which affords a napsylate addition salt, oxalic acid which affords an oxalic acid addition salt, phosphoric acid which affords a phosphoric acid addition salt, succinic acid which affords a succinic acid addition salt (succinate), sulfuric acid which affords a sulfuric acid addition salt and tartaric acid which affords a tartaric acid addition salt. Each of these acid addition salts can be either a mono acid addition slat or a poly acid addition salt, as these terms are defined hereinabove. [0033] As used herein, the term "alkyl" describes an aliphatic hydrocarbon including straight chain and branched chain groups. According to some embodiments, the alkyl group has 1 to 10 carbon atoms, according to other embodiments, 1 to 5 carbon atoms, according to yet other embodiments, 6 to 10 carbon atoms, and according to still other embodiments, 4 to 6 carbon atoms. Whenever a numerical range; e.g., “1-10”, is stated herein, it implies that the group, in this case the alkyl group, may contain 1 carbon atom, 2 carbon atoms, 3 carbon atoms, etc., up to and including 10 carbon atoms. The alkyl can be substituted or unsubstituted. When substituted, the substituent can be, for example, an alkyl, an alkenyl, an alkynyl, a cycloalkyl, an aryl, a heteroaryl, a halogen (halo), a hydroxy, a carboxy, an oxo, an alkoxy, an aryloxy, a thiohydroxy, a thioalkoxy, a thioaryl oxy, ahaloalkyl, an amine, a carbonyl, a carboxyl, an amide, a thioamide, a cyano and a carbamate, as these terms are defined herein.

[0034] The term "alkenyl" describes an unsaturated alkyl, as defined herein, having at least two carbon atoms and at least one carbon-carbon double bond. The alkenyl may be unsubstituted or substituted by one or more substituents, as described hereinabove for alkyl. [0035] The term "alkynyl", as defined herein, is an unsaturated alkyl having at least two carbon atoms and at least one carbon-carbon triple bond. The alkynyl may be unsubstituted or substituted by one or more substituents, as described hereinabove for alkyl.

[0036] As used herein, the terms "halo", "halogen" and "halide", which are referred to herein interchangeably, describe an atom of a fluorine, chlorine, bromine or iodine, also referred to herein as fluoride, chloride, bromide and iodide.

[0037] The term “hydroxy” or “hydroxyl” , as used herein interchangeably, refers to an -OH group.

[0038] The term “cyano", as used herein, refers to a -C=N group. [0039] The term “acyl”, as used herein, refers to a -C(=0)R group, wherein R is a straight or branched Cl - C10 alkyl.

[0040] The term “carboxy” as used herein, refers to a -C(=O)R group, wherein R is H or

OH group.

[0041] The term “cyclic terpene” as used herein, refers to a monocyclic and bicyclic terpene hydrocarbon, comprising at least one unsaturated bond, optionally substituted as disclosed herein above and below.

[0042] As described and demonstrated in the Examples section that follows, the compounds according to embodiments of the present invention have been tested for their antiinflammatory activity and were indeed found to be highly potent candidates for antiinflammatory agents and drugs.

[0043] Thus, according to another aspect of the present invention, there is provided a method of treating a medical condition, a disease or a disorder associated with fungal infection, which is affected by administering to a subject in need thereof a therapeutically effective amount of one or more of the compounds as presented herein, as well enantiomers, hydrates, solvates, prodrugs or any pharmaceutically acceptable salts thereof, as defined hereinabove.

[0044] According to some embodiments of the present invention, there is provided a method of treating a medical condition, a disease or a disorder associated with fungal infection, which is effected by administering to a subject in need thereof a therapeutically effective amount of one or more of any one of the compounds presented in Table I which is presented in the Examples section that follows hereinbelow, as well enantiomers, hydrates, solvates, prodrugs or any pharmaceutically acceptable salts thereof, as defined hereinabove. [0045] As used herein, the terms “treating" and “treatment"" includes abrogating, substantially inhibiting, slowing or reversing the progression of a condition, substantially ameliorating clinical or aesthetical symptoms of a condition or substantially preventing the appearance of clinical or aesthetical symptoms of a condition.

[0046] Accordingly, another aspect of the present invention provides a use of one or more of the compounds as presented herein, as well enantiomers, hydrates, solvates, prodrugs or any pharmaceutically acceptable salts thereof, as defined hereinabove, in the preparation of a medicament.

[0047] According to some embodiments of the present invention, the medicament is for the treatment of a medical condition, a disease or a disorder associated with fungal infection.

[0048] According to some embodiments of the present invention, there is provided a use of one or more of any one of the compounds presented in the Examples section that follows hereinbelow, as well as any enantiomers, hydrates, solvates, prodrugs or any pharmaceutically acceptable salts thereof, as defined hereinabove, in the preparation of a medicament for the treatment of a medical condition, a disease or a disorder associated with fungal infection.

[0049] In yet another embodiment the invention provides for the use of a compound of the invention for the inhibition, prevention or eradication of the growth and/or proliferation of fungi, optionally in combination with one or more known anti-fungal agents. In one embodiment, the invention provides a method of inhibiting fungal growth by contacting a fungus with an effective amount of one or more compounds of the invention optionally in combination with one or more other anti-fungal agents. Representative examples of fungi that may be inhibited with compounds of the invention include, but are not limited to, Histoplasma (e.g. H. capsulatum), Coccidioides, Blastomyces, Paracoccidioides, Cryptococcus (e.g. C. neoformans), Aspergillus (e.g. A. fumigatus, A. niger, A. nidulans, A. terreus, A. sydowi, A. flavatus, and A. glaucus), Zygomycetes (e.g. Basidiobolus, Conidiobolus, Rhizopus, Mucor, Absidia, Mortierella, Cunninghamella, and Saksenaea),

Candida (e.g. C. albicans, C. tropicalis, C. parapsilosis, C. stellatoidea, C. krusei, C. parakrusei, C. lusitaniae, C. pseudotropicalis, C. guilliermondi and C. glabrata), Cryptosporidium parvum, Sporothrix schenckii, Piedraia hortae, Trichosporon beigelii, Malassezia furfur, Phialophora verrucosa, Fonsecae pedrosoi, Madurella mycetomatis and Pneumocystis carinii. In a specific embodiment, the compounds according to the invention have anti-fungal activity against yeasts. Examples of yeasts that are susceptible to the anti-fungal effects of these compounds include but are not limited to Candida sp. and Cryptococcus sp. In one embodiment, the compounds according to the invention exhibit anti-fungal activity including activity against Cryptococcus neoformans. In another embodiment, the compounds according to the invention exhibit anti-fungal activity including activity against Candida albicans. In still another embodiment, the compounds according to the invention exhibit anti-fungal activity including activity against Candida glabrata, Candida krusei or Candida parapsilosis.

[0050] In another embodiment, the compounds according to the invention have anti-fungal activity that includes activity against filamentous fungi. Examples of filamentous fungi include, but are not limited to Aspergillus sp., Fusarium sp., and Rhizopus sp. In one embodiment, the compounds according to the invention exhibit anti-fungal activity including activity against Aspergillus sp. In another embodiment, the compounds according to the invention exhibit anti-fungal activity including activity against Aspergillus fumigatus. [0051] In another embodiment, the compounds of the invention exhibit anti-fungal activity against a broad spectrum of fungi and are suitable for use as broad-spectrum anti-fungal agents. One embodiment of the invention thus provides for the use of compounds of the invention as broad spectrum anti-fungal agents. Another embodiment of the invention provides for the use of compounds of the invention as broad spectrum anti-fungal agents.

[0052] In a specific embodiment, the compounds of the invention are capable of inhibiting the growth of or killing dermatophytes. Examples of dermatophytes include, but are not limited to Epidermophyton sp., Microsporum sp., and Trichophyton sp.

[0053] One embodiment of the invention provides for the use of the anti-fungal compounds of the invention in the treatment of subjects having an infection with a species of Candida, Cryptococcus, or Aspergillus, or having a disease or disorder related to infection with a species of Candida, Cryptococcus, or Aspergillus. Examples of diseases and disorders related to Candida infection include, but are not limited to, candidiasis, vaginitis, monilia, thrush, skin rash, diaper rash, nail bed infections and esophagitis. Examples of diseases and disorders related to Cryptococcus infection include, but are not limited to, cryptococcosis, meningitis, hepatitis, osteomyelitis, prostatitis, pyelonephritis and peritonitis. Examples of diseases and disorders related to Aspergillus infection include, but are not limited to, aspergillosis, chronic lung irritation, hypersensitivity pneumonia, allergic bronchopulmonary aspergillosis, aspergilloma, tracheobronchitis, acute necrotising Aspergillus pneumonia, chronic necrotising Aspergillus pneumonia and granulomatous aspergillosis. In accordance with a further embodiment of the invention, one or more compounds of the invention may be used in combination with one or more known antifungal agents in combination or synergistic therapy for the treatment of fungal infection, or disorders or diseases associated therewith. The compounds of the invention can be administered before, during or after treatment with the known anti-fungal agent(s). Such combination therapy is known in the art and selection of the appropriate anti-fungal agent(s) to be administered with the compounds of the invention is readily discernible by one of skill in the art. For example, for the treatment of fungal infections and fungus-related diseases, known anti-fungal compounds include, but are not limited to, amphotericin B and the structurally related compounds nystatin and pimaricin; flucytosine; azole derivatives such as ketoconazole, clotrimazole, miconazole, econazole, butoconazole, oxiconazole, sul conazole, terconazole, fluconazole and itraconazole; allylamines-thiocarbamates, such as tolnaftate and naftifme, and griseofulvin. In yet another embodiment the invention also contemplates the use of compounds of the invention as the active ingredient in anti-fungal compositions for non-therapeutic uses including, for example, anti-fungal cleansers, polishes, paints, sprays, soaps, and detergents. The compounds of the invention can also be included as an anti-fungal agent in cosmetic, personal care, household and industrial products, for example, to improve shelf-life by inhibiting the growth of fungi within the products. The compounds may be formulated for application to surfaces to inhibit the growth of a fungal species thereon, for example, surfaces such as countertops, desks, chairs, laboratory benches, tables, floors, sinks, showers, toilets, bathtubs, bed stands, tools or equipment, doorknobs and windows. Alternatively, the compounds may be formulated for laundry applications, for example, for washing clothes, towels, sheets and other bed linen, washcloths or other cleaning articles. The anti-fungal cleansers, polishes, paints, sprays, soaps, and detergents comprising the compounds of the invention can optionally contain suitable solvent(s), carrier(s), thickeners, pigments, fragrances, deodorizers, emulsifiers, surfactants, wetting agents, waxes, or oils, as required for the formulation of such products as is known in the art. The cleansers, polishes, paints, sprays, soaps, and detergents comprising the compounds of the invention are useful in institutions, such as in hospital settings, for the prevention of nosocomial infections, as well as in home settings. In one embodiment, the invention provides a formulation containing one or more compounds of the invention for external use as a pharmaceutically acceptable skin cleanser. In yet another embodiment of the invention, the compounds of the invention can be used in agriculture. In the latter case they are formulated in the form of an agricultural formulation.

[0054] In addition, in one embodiment, the invention contemplates the use of compounds of the invention in formulations to inhibit the growth of fungal species in food preparations. In another embodiment, the invention contemplates the use of compounds of the invention in formulations to sterilize surgical and other medical equipment and implantable devices, including prosthetic joints. The compounds can also be formulated for use in the in-situ sterilization of indwelling invasive devices such as intravenous lines and catheters, which are often foci of infection. As such the compounds of the invention can be used for the coating of surfaces of devices or implants to prevent biofilm associated growth of fungi on the latter.

[0055] In another embodiment, the invention contemplates the use of the compounds of the invention as the active ingredient in personal care items, such as soaps, deodorants, shampoos, mouthwashes, toothpastes, and the like. Many compositions used in personal care applications are susceptible to fungal growth and it is thus desirable to incorporate into these compositions an effective anti-fungal agent. The anti-fungal agent may be incorporated into the personal care formulation using techniques known in the art. Thus, the anti-fungal agent may be added to the personal care formulation as a solution, emulsion or dispersion in a suitable liquid medium. Alternatively, the anti-fungal agent may be added, undiluted, to the personal care formulation or may be added with a suitable solid carrier or diluent. The antifungal agent may be added to the pre-formed personal care formulation or may be added during the formation of the personal care formulation, either separately or premixed with one of the other components of the formulation.

[0056] In another embodiment, the invention contemplates the use of the compounds of the invention to treat fungal infections in a patient and/or preventing fungal infections in a patient, comprising identifying a patient in need of treatment and administering a therapeutically effective amount of at least one abn-cannabinoid derivive or a pharmaceutically acceptable salt thereof. In some embodiments, the patient is immunocompromi sed.

[0057] In some embodiments, the fungal infection is caused by one or more fungi selected from the group consisting of Epidermophyton floccosum, Trichophyton rubrum, Trichophyton mentagrophytes, the Microsporum genera, the Trychophyton genera, Candida auris, Candida albicans, Candida lusitaniae, Candida kruseii, Candida glabrata, Candida parapsilosis, Candida tropicalis, Candida guilliermondii, Cryptococcus neoformans, Trichophyton tonsurans, Microsporum canis, Epidermophyton floccosum, Histoplasma capsulatum, blastomyces, Cryptoccus neoformans, Pneumocystis jiroveci, Cocidioides immitis, Aspergillus fumigatus, Aspergillus niger, Penicillium genera, and Cladosporium genera.

[0058] In some embodiments, the fungal infection is resistant to one or more antifungal agents selected from the group consisting of terbinafine, amphotericin B, candicidin, fllipin, hamycin, nystatin, rimocidin, bifonazole, butoconazole, clotrimazole, econazole, fenticonazole, isoconazole, ketoconazole, luliconazole, miconazole, omoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole, albaconazole, fluconazole, isavuconazole, itraconazole, psoaconazole, ravuconazole, terconazole, voriconazole, abafungin, amorolfin, butenafine, naftifine, anidulafungin, caspofungin, micafungin, ciclopirox, flucytosine, haloprogin, griseofulvin, and tolnaftate.

[0059] In some embodiments, the fungal infection is a skin infection. In some embodiments, the skin infection is Athlete's Foot. In some embodiments, the fungal infection is caused by one or more fungi selected from from the group consisting of Epiderm ophyton floccosum, Trichophyton rubrum, or Trichophyton mentagrophytes. In some embodiments, the skin infection is ringworm. In some embodiments, the skin infection is ringworm caused by one or more fungi from the Microsporum or Trichophyton genera. In some embodiments, the skin infection is Tinea curis (jock itch). In some embodiments, the skin infection is caused by one or more fungi from the group consisting of Trichphyton rubrum, Candida albicans, Trichophyton mentagrophytes, and Epidermophyton floccosum.

[0060] In some embodiments, the fungal infection is fungal meningitis. In some embodiments, the fungal infection is a lung infection. In some embodiments, the lung infection is pneumonia. In some embodiments, the patient has a chronic lung disease. In some embodiments, the chronic lung disease is associated with cystic fibrosis. In some embodiments, the lung infection is caused by one or more fungi from the group consisting of Candida albicans, Aspergillus funigatus, Histoplasma capsulatum, blastomyces, Cryptoccus neoformans, Pneumocystis jiroveci, and Cocidioides immitis. In some embodiments, the infection is an eye infection.

[0061] In some embodiments, the fungal infection is nail fungus (e.g., affecting a toenail, fingernail, or analogous structure in a non-human animal). The nail fungus, or onychomycosis, may be caused by dermatophytes, Candida, and nondermatophytic molds. Dermatophytes are the fungi most commonly responsible for onychomycosis in the temperate western countries. While Candida and nondermatophytic molds are more frequently involved in the tropics and subtropics with a hot and humid climate, Trichophyton rubrum is the most common dermatophyte involved in onychomycosis. Other dermatophytes that may be involved are Trichophyton interdigitale, Epidermophyton floccosum, Trichophyton violaceum, Microsporum gypseum, Trichophyton tonsurans, and Trichophyton soudanense. A common outdated name that may still be reported by medical laboratories is Trichophyton mentagrophytes f or Trichophyton interdigitale.

[0062] Other causative pathogens include Candida and nondermatophytic molds, in particular members of the mold genera Scytalidium (name recently changed to Neoscytalidium), Scopulariopsis, and Aspergillus.

[0063] In some embodiments, an antifungal composition compnsing abn-canabinoid derivative is utilized to therapeutically or prophylactically treat or prevent an infection associated with Candida spp. In some embodiments, a treatment composition is configured for application to an open wound, surgical site, catheter (e.g., venous catheter) insertion site, or other such wound. In some embodiments, a treatment composition is configured as a wash, spray, gel, paste, or other formulation suitable for application to an open wound, surgical site, catheter insertion site, or other site of potential fungal infection. Such embodiments may be particularly useful for treating or preventing a Candida infection.

[0064] In some embodiments, an antifungal composition comprising abn-canabinoid derivative is applied to a medical device to prevent fungal colonization of the medical device and/or fungal infections at the medical implant site. Non-limiting examples of medical devices to which the treatment composition may be applied include devices which are implanted into a subject's tissues, deployed at a puncture or wound site, positioned for feeding or withdrawing material from a body cavity, or are otherwise associated with a patient/ subject in such a way that biological compatibility is of concern (e.g., because fungal infection and/or fouling of the device can result).

[0065] In some embodiments, the antifungal composition may be applied to a catheter (e.g., a venous catheter), intravenous needle, intravenous line, oral care device (e.g., dentures, dental implant), intrauterine device (IUD), feeder tube, drain, prosthesis component (e.g., voice prosthesis), peristaltic pump, tympsanostomy tube, tracheotomy tube, endotracheal tube, joint prosthesis, dialysis access graft, or cardiac graft.

[0066] In some embodiments, at least one abnCBD is administered with at least one non- abnCBD therapeutic agent.

[0067] In some embodiments, the non-abnCBD therapeutic agent is selected from the group consisting of an antifungal agent, an antibiotic, a nonsteroidal anti-inflammatory agent, an antiviral agent, an antiretroviral agent, an antipyretic, an antiemetic, an immunomodulator, a chemotherapeutic agent, an antihistamine, an opioid receptor agonist, an anti -cholinergic, and a beta2 -adrenoreceptor agonist.

[0068] In some embodiments, two or more abnCBDs are co-administered. In some embodiments, administration of the abnCBD is selected from the group consisting of topical application (via topical spray, wash, cream, paste, gel), inhalation, intravenous injection, subcutaneous injection, intraperitoneal injection, depot injection, intramuscular injection, transdermal patch, ear drops, and eye drops. In some embodiments, one or more abnCBDs are administered in a pharmaceutically acceptable formulation.

[0069] As used herein, the phrase “therapeutically effective amount” describes an amount of the compound being administered which will relieve to some extent one or more of the symptoms of the condition being treated. [0070] As demonstrated in the examples section that follows, an exemplary therapeutically effective amount of the compounds of the present invention ranges between about 0.1 mg/kg body and about 100 mg/kg body.

[0071] In any of the methods and uses described herein, the cannabinoid derivative compounds of the present embodiments can be utilized either per se or, according to some embodiments, as a part of a pharmaceutical composition that further comprises a pharmaceutically acceptable carrier.

[0072] Thus, according to additional aspects of the present invention, there is provided a pharmaceutical composition, which comprises one or more compounds having general Formula I, as defined hereinabove, and a pharmaceutically acceptable carrier.

[0073] According to some embodiments of the present invention, there is provided a pharmaceutical composition, which comprises one or more of any one of the compounds presented in in the Examples section that follows hereinbelow, and a pharmaceutically acceptable carrier, as well as any enantiomers, hydrates, solvates, prodrugs or any pharmaceutically acceptable salts thereof, as defined hereinabove.

[0074] As used herein a “pharmaceutical composition” refers to a preparation of the compounds presented herein, with other chemical components such as pharmaceutically acceptable and suitable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration of a compound to an organism.

[0075] Hereinafter, the term “pharmaceutically acceptable carrier” refers to a carrier or a diluent that does not cause significant irritation to an organism and does not abrogate the biological activity and properties of the administered compound. Examples, without limitations, of carriers are: propylene glycol, saline, emulsions and mixtures of organic solvents with water, as well as solid (e.g., powdered) and gaseous carriers. [0076] Herein the term “excipient” refers to an inert substance added to a pharmaceutical composition to further facilitate administration of a compound. Examples, without limitation, of excipients include calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols.

[0077] Techniques for formulation and administration of drugs may be found in “Remington’s Pharmaceutical Sciences” Mack Publishing Co., Easton, PA, latest edition, which is incorporated herein by reference.

[0078] Pharmaceutical compositions for use in accordance with the present invention thus may be formulated in conventional manner using one or more pharmaceutically acceptable carriers comprising excipients and auxiliaries, which facilitate processing of the compounds into preparations which can be used pharmaceutically. Proper formulation is dependentupon the route of administration chosen. The dosage may vary depending upon the dosage form employed and the route of administration utilized. The exact formulation, route of administration and dosage can be chosen by the individual physician in view of the patient's condition (see e.g., Fingl et al., 1975, in “The Pharmacological Basis of Therapeutics”, Ch. I p. I). The pharmaceutical composition may be formulated for administration in either one or more of routes depending on whether local or systemic treatment or administration is of choice, and on the area to be treated. Administration may be done orally, by inhalation, or parenterally, for example by intravenous drip or intraperitoneal, subcutaneous, intramuscular or intravenous injection, or topically (including ophtalmically, vaginally, rectally, intranasally).

[0079] Formulations for topical administration may include but are not limited to lotions, ointments, gels, creams, suppositories, drops, liquids, sprays and powders. [0080] Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.

[0081] Compositions for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, sachets, pills, caplets, capsules or tablets. Thickeners, diluents, flavorings, dispersing aids, emulsifiers or binders may be desirable.

[0082] Formulations for parenteral administration may include, but are not limited to, sterile solutions which may also contain buffers, diluents and other suitable additives. Slow release compositions are envisaged for treatment.

[0083] The amount of a composition to be administered will, of course, be dependent on the subject being treated, the severity of the affliction, the manner of administration, the judgment of the prescribing physician, etc.

[0084] Compositions of the present invention may, if desired, be presented in a pack or dispenser device, such as an FDA (the U.S. Food and Drug Administration) approved kit, which may contain one or more unit dosage forms containing the active ingredient. The pack may, for example, comprise metal or plastic foil, such as, but not limited to a blister pack or a pressurized container (for inhalation). The pack or dispenser device may be accompanied by instructions for administration. The pack or dispenser may also be accompanied by a notice associated with the container in a form prescribed by a governmental agency regulating the manufacture, use or sale of pharmaceuticals, which notice is reflective of approval by the agency of the form of the compositions for human or veterinary administration. Such notice, for example, may be of labeling approved by the U.S. Food and Drug Administration for prescription drugs or of an approved product insert.

[0085] Compositions comprising a compound of the invention formulated in a compatible pharmaceutical carrier may also be prepared, placed in an appropriate container, and labeled for treatment of a medical condition, disease or disorder associated with fungal infection, as is detailed hereinabove.

[0086] Thus, according to an embodiment of the present invention, the pharmaceutical composition of the present invention is being packaged in a packaging material and identified in print, in or on the packaging material, for use in the treatment of a medical condition, disease or disorder associated with fungal infection, as is defined hereinabove.

[0087] According to further embodiments of the any of the methods, uses and compositions presented herein, the compounds of the present invention can be combined with other active ingredients which are commonly used to treat fungal infection-associated diseases and disorders.

[0088] Additional objects, advantages, and novel features of the present invention will become apparent to one ordinarily 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 delineated hereinabove and as claimed in the claims section below finds experimental support in the following examples.

[0089] Additional features and advantages will be set forth in part in the description that follows, and in part will be obvious from the description, or may be learned by practice of the embodiments disclosed herein. The objects and advantages of the embodiments disclosed herein will be realized and attained by means of the elements and combinations particularly pointed out in the appended claims. It is to be understood that both the foregoing brief summary and the following detailed description are exemplary and explanatory only and are not restrictive of the embodiments disclosed herein or as claimed. [0090] Candida species are increasingly resistant to common therapeutic treatments, and in particular to azoles. While resistance to echinocandins is less prevalent, some Candida species, such as C. glabrata and C. parapsilosis can present some intrinsic resistance.

[0091] It should be clear to the skilled practitioner that the dose and dosage regimen will depend mainly on whether abnormal cannabinoid or a combination between abnormal cannabinoid and other drugs, is (are) being administered separately or as a combination (a mixture), the type of fungal infection to treat, the specific application, the patient and the patient's history. The amount must be effective to achieve a fungicidal or fungistatic effect or an amount, which is synergistic particularly when used in a combination. The practitioner will be able to ascertain upon routine experimentation which route of administration and frequency of administration are most effective in any particular case.

[0092] It is understood that the anti-fungal activity of abnormal cannabinoid can be tested using standard techniques known in the art to determine whether the separate compound or the combination is suitable for the use as a specific anti-fungal compound. As used herein a compound of the invention (or compounds of the invention) refers to abnormal cannabinoid alone or the combination of abnormal cannabinoid with other drugs.

[0093] As is known in the art, anti-fungal activity of a compound (or a combination of compounds) may result in the killing or eradication of fungal cells (i.e. fungicidal activity), in the slowing or arrest of the growth or proliferation of fungal cells (i.e. fungistatic activity), and/or in the prevention of fungal cell growth, and/or the prevention of the formation of a fungal biofilm and/or the destruction of an established fungal biofilm. Thus, the compounds of the invention may be fungicidal, fungistatic, and/or may prevent the growth or the biofilm formation of fungal cells. It is understood that compounds of the invention that slow or arrest fungal cell growth may also be useful in combination treatments with other known anti- fungal agents. Exemplary methods of testing the compounds of the invention are provided below and in the examples included herein. These methods can be used to test the anti-fungal activity of provided below and in the examples included herein. These methods can be used to test the anti-fungal activity of provided below and in the examples included herein. These methods can be used to test the anti-fungal activity of abnormal cannabinoid or in combination with other drugs, or in still a further combination with other anti-fungal agents. One skilled in the art will understand that other methods of testing the anti-fungal activity of compounds are known in the art and are also suitable for testing compounds.

[0094] In vitro methods of determining the ability of the compounds of the invention to inhibit the growth of fungal cells are well-known in the art. In general, these methods involve contacting a culture of the cells of interest with various concentrations of the compounds and monitoring the growth of the cell culture relative to an untreated control culture. A second control culture comprising cells contacted with a known anti-fungal agent may also be included in such tests, if desired. For example, the ability of a compound of the invention to inhibit the growth of specific fungal cells can readily be determined by measurement of the minimum inhibitory concentration (MIC) for the compound. The MIC is defined as the lowest concentration that inhibits growth of the specific fungus to a pre-determined extent. For example, a MIC100 value is defined as the lowest concentration that completely inhibits growth of the organism, whereas a MIC90 value is defined as the lowest concentration that inhibits growth by 90% and a MIC50 value is defined as the lowest concentration that inhibits growth by 50%. MIC values are sometimes expressed as ranges, for example, the MIC100 for a compound may be expressed as the concentration at which no growth is observed or as a range between the concentration at which no growth is observed and the concentration of the dilution which immediately follows. Techniques for determining anti- fungal MIC values for candidate compounds include both macrodilution and microdilution methods (see for example Pfaller, M. A. et al (1997), Clin. Infect. Dis. 24:776-84). As is known in the art, different types of fungi may require different testing methods. F or example, suitable reference methods for testing MIC values for candidate compounds in yeasts, include the NCCLS reference method for broth dilution anti-fungal susceptibility testing of yeasts (M27-A2, Vol. 22 No. 15). This method can be used to test MIC values in yeasts such as Candida species, and Cryptococcus neoformans, for example. Alternatively, suitable reference methods for determining MIC values for candidate compounds in filamentous fungi include the NCCLS reference method for broth dilution anti-fungal susceptibility testing of filamentous fungi. In the classical broth microdilution method, the anti-fungal compound is diluted in culture medium in a sterile, covered 96-well microtiter plate. An overnight culture of a single fungal colony is diluted in sterile medium such that, after inoculation, each well in the microtiter plate contains an appropriate number of CFU/ml (typically, approximately 5.103 CFU/ml). Culture medium only (containing no fungal cells) is also included as a negative control for each plate, and known anti-fungal compounds are often included as positive controls. The inoculated microtiter plate is subsequently incubated at an appropriate temperature (for example, 35°C for 16-48 hours). The turbidity of each well is then determined by visual inspection and/or by measuring the absorbance, or optical density (0D), at 595 nm or 600 nm using a microplate reader and is used as an indication of the extent of fungal growth.

[0095] The ability of a compound of the invention to act as an anti-fungal agent can also be tested in vivo using standard techniques. A number of animal models are known in the art that are suitable for testing the activity of anti-fungal compounds and are readily available. [0096] The ability of a compound of the invention to act as an anti-fungal agent can also be tested in vivo using standard techniques. A number of animal models are known in the art that are suitable for testing the activity of anti-fungal compounds and are readily available.

[0097] It is to be understood that although particular embodiments, specific configurations as well as materials and/or molecules, have been discussed herein for cells, animals and humans and methods according to the present invention, various changes or modifications in form and detail may be made without departing from the scope and spirit of this invention. The following examples are provided to better illustrate particular embodiments, and they should not be considered limiting the application. The application is limited only by the claims.

BRIEF DESCRIPTION OF THE DRAWINGS

[0098] The subject matter regarded as the invention is particularly pointed out and distinctly claimed in the concluding portion of the specification. The invention, however, both as to organization and method of operation, together with objects, features, and advantages thereof, may best be understood by reference to the following detailed description when read with the accompanying drawings in which:

[0099] Fig. 1 is the scheme of abnCBD synthesis.

[00100] Figs. 2A-2B show the FACS scanner analyzis of propidium-iodide staining on Candida cells.

[00101] Fig. 3 is an optical density measurement of alive Candida cells in regular and antifungal s-resisrant Candida albicans strains.

[00102] Fig. 4 is an optical density measurement of alive Candida cells in Candida auris, Candida glabrata and Candida parapsilopsis strains. [00103] Fig. 5 shows the colony forming unit (CFU) of C. albicans strain SC5314 from the vagina of mice treated intravaginally with vehicle or NK-1 -25 at low (25mg/kg) and high dose (lOOmg/kg).

[00104] Figs. 6A - 6B shows the H&E staining of vagina of NK 1-25 treated (6 A) and control mice (6B).

[00105] Figs. 7A - 7C show the effect of compounds of the invention (7A: NK1-25, 7B: NK2-28 and 7C: Tl-1) on the growth of C. albicans. Time-Kill assay: growth curve of C. albicans incubated with various concentrations of Abn-CBDs at 30 cC and turbidity was measured at OD 600 nm every 2 hrs for 20 hrs. Data are the means of n=3, performed in triplicates.

[00106] Fig. 8 shows the disruptive effect of compounds of the invention on the membrane of C. albicans. The relative proportion of Pl-positive yeast cells normalized to the control group treated with medium (considered as zero). Data are the means ± SD, n=2. [00107] Fig. 9 shows the effect of compounds of the invention on the inhibition of the formation of C. albicans biofilm. Anti-biofilm formation activity of Abn-CBDs. The Graph shows the percentage of viable C. albicans biofilm after incubation with various concentrations of Abn-CBDs in a biofilm-inducing medium at 37 c-C for 24 hrs. The MTT assay was used to determine the percentage of viable biofilm cells with reference to cell treated with medium. Data are the means ± SD, n=4.

[00108] Fig. 10 shows the eradication of C. albicans biofilm by compounds of the invention. Disruption of mature biofilm by Abn-CBDs. The graph shows the percentage of viable C. albicans biofilm after matured C. albicans biofilm (yeasts cells incubated at 37 oC for 24 hrs to form biofilm) were incubated with various concentrations of Abn-CBDs in biofilm at 37 oC for 24 hrs. The MTT assay was employed to determine the percentage of viable biofilm cells with reference to cell treated with medium. Data are the means ± SD, n=4.

[00109] It will be appreciated that for simplicity and clarity of illustration, elements shown in the figures have not necessarily been drawn to scale. For example, the dimensions of some of the elements may be exaggerated relative to other elements for clarity. Further, where considered appropriate, reference numerals may be repeated among the figures to indicate corresponding or analogous elements.

DETAILED DESCRIPTION OF THE PRESENT INVENTION

[00110] In the following detailed description, numerous specific details are set forth in order to provide a thorough understanding of the invention. However, it will be understood by those skilled in the art that the present invention may be practiced without these specific details. In other instances, well-known methods, procedures, and components have not been described in detail so as not to obscure the present invention.

[00111] Material s and methods :

[00112] AU chemical reagents were purchased from Sigma-Aldrich.

[00113] Organic solvents were purchased from Bio-Lab.

[00114] Cannabinoids (such as cannabidiol) were extracted from Cannabis sativa plant as previously described [Gaoni, Y. and Mechoulam, R. (1971) J. Amer. Chem. Soc. 93, 217- 224],

[00115] The synthesis of abnCBD. 1.5 g of olivetol and 1.2 g of para-mentha di enol were dissolved in 50 ml dry DCM, 1g of dry MgSO4 was added under N2 atmosphere. The reaction was cooled to 0°C, and 50 ul of BUetherate were added. The reaction was mixed for 1.5h, washed with 10% w/v NaHCOs solution to neutral, the aqueous phase was extracted three times with DCM and dried over MgSCU. The crude was purified by by silica gel chromatography with 5%— >30% diethyl ether in petroleum ether. CBD was obtained at 45% yield, and abnCBD at 35% yield.

[00116] Infection model: A previously described model of vulvovaginal candidiasis (WC) was utilized. Three days prior infection and on day 4 post-infection (dpi), C57bl/6 mice were injected with 0.5mg/200pl of P-Estradiol subcutaneously to maintain pseudoestrus. On day 0, mice were inoculated intravaginally with 20pL of a suspension of C. albicans at 2.5*106 CFU/ml. NK1-25 was administrated intravaginally daily beginning 1 dpi and continued through 4 dpi at 50 or lOOmg/kg. Control mice were infected but received empty vehicle consisted of DMSO, Tween80, and PBS in a ratio of 1 : 1 : 18 respectively. On day 5, mice were euthanized the fungal burden in the vagina was determined by colony forming unit count (CFU) in the vaginal lavage and the vaginal tissues were collected for histochemistry staining. Fig. 5 shows the colony forming unit (CFU) of C. albicans strain SC5314 from the vagina of mice treated intravaginally with vehicle or NK-1-25 at low (25mg/kg) and high dose (lOOmg/kg). Figs. 6A - 6B shows the H&E staining of vagina of NK1-25 treated (6A) and control mice (6B).

[00117] Preparation of Candida cells: Primary cell culture was prepared by resuspending 3 morphologically isolated colonies from Candida cultures on SDA into a 15 ml Sabouraud Dextrose broth (SDB) medium and then incubated overnight at 30°C, 200 rpm. 300 pl of the primary culture was diluted in a new 13 ml SDB medium and incubated at 30°C, 200 rpm for 2 hours (secondary culture). The secondary culture was washed 4 times with sterile PBS by centrifugation at 3000 rpm at 4°C for 5mins, counted, and resuspended in 10 ml PBS. The Cells were stored on ice for further experiment.

[00118] Effect of Abn-CBDs on the growth of C. albicans: 100 pL containing 10 ⁴

Candida cells from prepared cell culture as described above, was plated in a 96 well plate in the presence of 16 pg/ml, 32 pg/ml, 64 pg/ml, and 128 pg/ml Abn-CBDs or fluconazole (FLC), or 2% dimethyl sulfoxide (DMSO) and incubated at 37 °C for different time points from 0 - to- 20 hours with 2 hours intervals. At each time point, the turbidity of the culture was measured at OD600 in a plate reader.

[00119] Membrane Permeability Assay: The permeability of the C. albicans cell membrane was investigated to identify the possible mechanism of action for our Abnormal Cannabinoid (Abn-CBD) and its derivatives (Abn-CBDs). The propidium Iodide (PI) assay was performed. In this assay, 100 pL of prepared cell culture as described above in sterile SD broth medium will be cultured with Abn-CBDs (32 pg/ml and 64 pg/ml), FLC (as positive control), and DMSO (as solvent control) in a 96 well plate and incubated at 37 °C for 24 hours. Following this, cells will be washed twice with PBS. Next, 10 pl PI will be incubated with the treated cells at 30°C for 10 mins. The percent uptake of PI was analyzed using flow cytometry.

[00120] Effect on Biofilm Formation and mature biofilm disruption: Biofilm formation is one of the virulent factors of Candida and a contributing factor in drug resistance. Here the anti-biofilm effect of Abn-CBDs were tested with FLC as a positive control. Here, 100 pL of prepared cell culture as described above in RPML1640 medium will be dispensed into each well of 96 well plate and incubated with 100 pL of 32 pg/ml Abn-CBDs. 2% DMSO was used as a negative control. The plate was incubated at 37 °C for 24 hours. After this, the biofilm was washed with PBS to remove planktonic cells. The dry biofilm was incubated with 200 pL of 100 mM 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) and the optical density measured at 570 nm using a spectrophotometer. For matured biofilm eradication, cells in their log phase will be incubated with RPML1640 at 37°C for 24 hours and the dry biofilm will be resuspended with 200 pL of 64 pg/ml Abn-CBDs followed by incubation at 37°C for 24 hours. After the removal of the supernatant and planktonic cells as above, MTT staining of dry biofilm will be performed as above.

[00121] RESULTS

[00122] Time-Kill Assay: Kinetic growth curve test (Time-kill assay) was performed to evaluate how Abn-CBDs inhibits fungal growth. Over a period of 20 hours, the effect Abn- CBDs (16 pg/ml, 32 pg/ml, 64 pg/ml, and 128 pg/ml), FLC (positive control) and DMSO (negative control) on C. albicans growth was examined (Figs. 7A - 7C). As shown in Figure 7, Abn-CBDs at 32 pg/ml and FLC completely inhibited fungal growth for the whole 24 hours. However, there is differential inhibition of fungal growth with regards to Abn-CBDs at 16 pg/ml. While Abn-CBD(NKl-25) delays fungal growth until 14 hours where it begins to recover, its derivatives; NK2-28 inhibits fungal growth up to 18 hours and Tl-1 is not different than the negative control which does not inhibit fungal growth.

[00123 ] Membrane Permeability Assay: The di sruption of the fungal cell membrane was investigated to identify the possible mechanism of action for Abn-CBDs. The PI dye can only enter the membrane of dead cells and binds to the nucleus. This florescent binding can be detected using flow cytometry (Figure 8). Based on the results obtained from this study, Abn-CBD (NK1-25) and its derivatives (Tl-1, and TE 1-4) at 64 pg/ml resulted in mean percentages of 33.98, 16.21, 24.50, and 13.04 respectively. This shows that Abn-CBDs can disrupt the membrane of C. albicans.

[00124] Anti-biofilm activity of Abn-CBDs: The ability of fungal to form biofilms, is one of the main reasons for the high rising of drug-resistant in fungal infections. Biofilm is considered one of the virulence factors of fungal. Once Candida form biofilm, the immune system fails to attack them and most of the existing antifungal drugs also fails to disrupt them. The anti-biofilm (anti-biofilm formation and eradication of matured biofilm) activity of Abn-CBDs were evaluated against C. albicans (SC3515) by MTT reduction assay.

[00125] The effect of Abn-CBDs on C. albicans biofilm formation was tested. For this, C. albicans were exposed to 32 pg/ml Abn-CBDs for 24 hours, after which biofilm activity was analyzed using MTT assay. As shown in Figure 9, differential inhibition of the Abn- CBDs was observed. NK1-25 and NK2-28 were able to inhibit C. albicans biofilm formation up to about 90 %, while Tl-5, HUM216, and NK1-69 were able to inhibit up to 50 %. Next, Abn-CBDs was examined to see if it can disrupt preformed biofilm of C. albicans. Abn-CBDs (NK1-25, NK2-28, Tl-5, NK1-69, and Tl-1) at 32 pg/ml decreased mature biofilm by 64 %, 43 %, 50 %, 56 % and 50 % respectively Figure 10.

[00126] It is noteworthy that the Abn-CBDs affect the different stages of C. albicans infection differentially. TE1-4, Tl-5, andNKl-69 could not inhibit the growth of C. albicans at its yeast stage, however, they have been shown to affect biofilm formation and can disrupt C. albicans biofilm.

[00127] While certain features of the invention have been illustrated and described herein, many modifications, substitutions, changes, and equivalents will now occur to those of ordinary skill in the art. It is, therefore, to be understood that the appended claims are intended to cover all such modifications and changes as fall within the true spirit of the invention.

Claims

CLAIMS [00128] What is claimed is:

1. A compound having the general formula (I):

Wherein

R1 is selected from H, straight or branched C3 - C10 alkyl, straight or branched C3 - C10 alkenyl, straight or branched C3 - C10 alkynyl; optionally substituted with at least one phenyl, hydroxyphenyl, hydroxy, carboxy, halogen, cyano and any combinations thereof;

Ri l l is selected from H, straight or branched Cl - C10 alkyl, straight or branched Cl - C10 alkenyl, straight or branched Cl - C10 alkynyl; optionally substituted with at least one phenyl, hydroxyphenyl, hydroxy, carboxy, halogen, cyano and any combinations thereof provided that at least one of R1 and Ri l l is different than H;

2. A compound according to claim 1, wherein R1 is a straight or branched C3 - C10 alkyl.

3. A compound according to claim 1, wherein R1 is a straight or branched C3 - C10 alkenyl.

4. A compound according to claim 1, wherein R1 is selected from a straight or branched C3 - C10 alkyl and a straight or branched C3 - C10 alkenyl.

5. A compound according to any one of claims 1 to 4, wherein R1 is substituted with at least one phenyl, hydroxyphenyl.

6. A compound according to any one of claims 1 to 4, wherein R1 is substituted with at least one phenyl.

7. A compound according to any one of claims 1 to 4, wherein R1 is substituted with at least one hydroxyphenyl.

8. A compound according to claim 1, wherein Ri l l is a straight or branched Cl

- C10 alkyl.

9. A compound according to claim 1, wherein Ri l l is a straight or branched Cl - C10 alkenyl.

10. A compound according to claim 1, wherein Ri l l is selected from a straight or branched Cl - C10 alkyl and a straight or branched Cl - C10 alkenyl.

11. A compound according to any one of claims 1 to 10, wherein Ri l l is substituted with at least one phenyl, hydroxyphenyl.

12. A compound according to any one of claims 1 to 10, wherein Ri l l is substituted with at least one phenyl.

13. A compound according to any one of claims 1 to 10, wherein Ri l l IS substituted with at least one hydroxyphenyl.

14. A compound according to any one of the preceding claims, wherein R1 is a C3 - C10 alkyl and Ri l l is a straight or branched Cl - C10 alkyl.

15. A compound according to any one of claims 1 to 14, wherein R2 and R3 both

16. A compound according to any one of claims 1 to 14, wherein R2 and R3 each independently straight or branched Cl-C5alkyl.

17. A compound according to any one of claims 1 to 14, wherein R2 and R3 each independently straight or branched Cl - C10 acyl.

18. A compound according to any one of claims 1 to 14, wherein R2 and R3 are each selected from H and straight or branched Cl-C5alkyl.

19. A compound according to any one of claims 1 to 14, wherein R2 and R3 are each selected from H and Ac.

20. A compound according to any one of claims 1 to 14, wherein R2 and R3 are each selected from straight or branched Cl-C5alkyl and Ac.

21. A compound according to any one of claims 1 to 20, wherein R4 is a cyclic terpene having one double bond.

22. A compound according to any one of claims 1 to 20, wherein R4 is a cyclic terpene being a bicyclic terpene.

23. A compound according to any one of claims 1 to 20, wherein R4 is a moiety of general formula (II):

Wherein

are optionally null;

Wherein one of R5 - R10 is a connection to general formula (I); and

24. A compound according to any one of claims 1 to 13, wherein R4 is a moiety of general formula (III):

Wherein

are optionally null; and

Wherein connection to general formula (I); and

Wherein each of R11 -R14 is selected from: H, straight or branched Cl - C10 alkyl, straight or branched C2 - C10 alkenyl, straight or branched C2 - C10 alkynyl; each optionally substituted by at least one hydroxy, carboxy, halogen, cyano; or two of R11 - R14 together with the carbon atoms they are connected to form a C3 - C8 ring.

25. A compound according to any one of claims 1 to 17, wherein R4 is selected from:

Wherein

is a connection to general formula (I); and

26. A compound according to any one of claims 1 to 25, wherein R4 is a moiety of general formula (IV).

27. A compound according to any one of claims 1 to 25, wherein R4 is a moiety of general formula (V).

28. A compound according to any one of claims 1 to 25, wherein R4 is a moiety of general formula (VI).

29. A compound according to any one of claims 1 to 25, wherein R4 is a moiety of general formula (V).

30. A compound according to any one of the preceding claims, selected from:

31. A composition comprising at least one compound according to any one of claims 1 to 30.

32. A composition according to claim 31, further comprising at least one further anti-fungal agent.

33. A composition according to claims 31 or 30, further comprising at least one further pharmaceutical active agent.

34. A compound according to any one of claims 1 to 30, for use in the treatment of a medical condition, a disease or a disorder associated with a fungal infection.

35. A compound according to any one of claims 1 to 30, for use in the treatment of a medical condition, a disease or a disorder associated with a resistant fungal infection.

36. The compound for use according to claims 34 or 35, wherein said medical condition, disease or disorder associated with fungal infection is caused by at least one fungi selected from Histoplasma (e.g. H. capsulatum), Coccidioides, Blastomyces, Paracoccidioides, Cryptococcus (e.g. C. neoformans), Aspergillus (e.g. A. fumigatus, A. niger, A. nidulans, A. terreus, A. sydowi, A. flavatus, and A. glaucus), Zygomycetes (e.g. Basidiobolus, Conidiobolus, Rhizopus, Mucor, Absidia, Mortierella, Cunninghamella, and Saksenaea), Candida (e.g. C. albicans, C. tropicalis, C. parapsilosis, C. stellatoidea, C. krusei, C. parakrusei, C. lusitaniae, C. pseudotropicalis, C. guilliermondi and C. glabrata), Cryptosporidium parvum, Sporothrix schenckii, Piedraia hortae, Trichosporon beigelii, Malassezia furfur, Phialophora verrucosa, Fonsecae pedrosoi, Madurella mycetomatis and Pneumocystis carinii and any combinations thereof.