EA045936B1 - INJECTABLE COMPOSITIONS WITH ANTI-FUNGAL ACTIVITY, AND METHODS FOR THEIR OBTAINING AND APPLICATION - Google Patents

Description

Field of invention

The present invention relates to compositions containing liposomal vesicles in which a triterpenoid antifungal compound, an enfumafungin derivative, is encapsulated. In particular, the invention relates to compositions containing vesicles with a phospholipid bilayer in which a triterpenoid derivative of enfumafungin (or a pharmaceutically acceptable salt or hydrate thereof) is encapsulated, which is an inhibitor of (1,3)-β-O-glucan synthesis, which can be used for the treatment and/or prevention of systemic fungal infections. The compositions of the present invention are well tolerated when applied topically and are well suited for administration by injection.

Level of invention

Fungal infections are a major problem in healthcare and most often manifest as invasive fungal disease (eg, candidemia, invasive aspergillosis), localized fungal infections (eg, pleural empyema and abscess localized to the abdomen, brain, lung, etc.) , and in the form of infections of the skin and mucous membranes (such as candidiasis of the oral cavity, esophagus and vulvovaginal localization). The type and extent of infection depend on the virulence factors of the fungal pathogen, the level of host defense, and the anatomical sites involved.

Severe systemic fungal infections are more common in immunocompromised patients, such as patients undergoing chemotherapy for malignancies, or receiving immunomodulatory agents to treat chronic inflammatory conditions, or patients with immunodeficiency syndromes, either acquired or genetic. Despite the availability of modern antifungal therapy, systemic fungal infections are associated with mortality rates of up to 50%, depending on the pathogen and the patient's underlying condition.

Enfumafungin is a hemiacetal triterpene glycoside that is produced by fermented Hormonema spp. associated with living leaves of Juniperus communis (U.S. Patent No. 5,756,472; Pelaez et al., Systematic and Applied Microbiology, 23:333-343, 2000; Schwartz et al., JACS, 722:4882-4886, 2000; Schwartz, R.E., Expert Opinion on Therapeutic Patents, 11(11):1761-1772, 2001). Enfumafungin belongs to a series of triterpene glycosides with antifungal activity in vitro. The mechanism of antifungal action of enfumafungin and other antifungal triterpenoid glycosides has been found to be inhibition of glucan synthesis in the fungal cell wall due to their specific action on (1,3)-β-O-glucan synthase (Onishi et al., Antimicrobial Agents and Chemotherapy, 44:368-377, 2000; Pelaez et al., Systematic and Applied Microbiology, 23:333-343, 2000). For antifungal action, (1,3)-β-O-glucan synthase is an attractive target because it is present in many pathogenic fungi and therefore a broad spectrum of antifungal activity can be obtained. Additionally, due to the absence of a (1,3)-β-O-glucan synthase analog in mammals, the enfumafungin derivatives of the invention have little or no mechanism-related toxicity. The enfumafungin-derived triterpenoid compounds used according to the present invention are active against fungal isolates of Candida spp., including isolates that are resistant to azoles or other glucan synthase inhibitors (for example, lipopeptide substances such as echinocandins), indicating a difference in biological and molecular target enfumafungin derivatives from the molecular target of other glucan synthase inhibitors.

Various enfumafungin derivatives have been disclosed in a number of international patent publications, for example, WO 2007/126900 and WO 2007/127012.

Representative of the enfumafungin derivatives of the invention is the compound SCY-078, which has activity against a number of Candida and Aspergillus species, including drug-resistant strains. SCY-078 is a carboxylic acid-alkylamino ester substituted triterpenoid derivative having the following formula:

Without being bound by a specific theory, due to the presence of amino and carbonic functional groups

- 1 045936 acid, SCY-078 is an amphiphilic molecule and exhibits surfactant-like properties, including the formation of micelles and binding to surface and plasma proteins.

When SCY-078 was administered intravenously, injection site reactions (hereinafter referred to as ISR), including pain, irritation, inflammation and phlebitis, were observed, precluding administration through peripheral veins. Although the underlying mechanism of the observed ISRs is not entirely clear, without reference to a specific theory, the occurrence of ISRs can be explained by local direct interaction of SCY-078 with the vascular endothelium at the injection site when SCY-078 is administered intravenously into the bloodstream.

There is a need in the art for the ability to administer SCY-078 and other antifungal enfumafungin derivatives intravenously via a peripheral vein while reducing local injection site reactions and/or less severe ISRs, and increasing variability and convenience for healthcare providers and patients.

The essence of the invention

The present invention provides injectable compositions having antifungal activity, comprising an aqueous phase and one or more monolamellar vesicles, each containing a phospholipid and cholesterol, and each of which encapsulates a compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof

which is (1S,4aR,6aS,7R,8R,10aR,10bR,12aR,14R,15R)15-[[2-amino-2,3,3trimethylbutyl]oxy]-8-[(^)-1, 2-dimethylpropyl]-14-[5-(4-pyridinyl)-III-1,2,4-triazol-1-yl]-1,6,6a,7,8,9,10,10a,10b,11 ,12,12a-dodecahydro-1,6a,8,10a-tetramethyl-4H-1,4a-propano-2H-phenanthro[1,2-c]pyran-7carboxylic acid;

where one or more monolamellar vesicles are hydrated in the aqueous phase. The pH of the aqueous phase is preferably from about 5.0 to about 7.0. Examples of the aqueous phase are a monosaccharide solution and a disaccharide solution.

The present invention also provides injectable compositions having antifungal activity, comprising an aqueous phase and one or more monolamellar vesicles, each containing a phospholipid and cholesterol, and each of which encapsulates a compound of formula (IIa) or a pharmaceutically acceptable salt or hydrate thereof

which is (1S,4aR,6aS,7R,8R,10aR,10bR,12aR,14R,15R)-15-[[(2R)-2-amino-2,3,3trimethylbutyl]oxy]-8-[( ^)-1,2-dimethylpropyl]-14-[5-(4-pyridinyl)-III-1,2,4-triazol-1-yl]-1,6,6a,7, 8,9,10, 10a,10b,11,12,12a-dodecahydro-1,6a,8,10a-tetramethyl-4H-1,4a-propano-2H-phenanthro[1,2-c]pyran-7carboxylic acid,

- 2 045936 where one or more monolamellar vesicles are hydrated in the aqueous phase.

Injectable compositions can be administered intravenously for the treatment and/or prevention of fungal infections, such as systemic fungal infections, including those caused by Candida or Aspergillus species. The use of the injectable compositions of the present invention is characterized in that local reactions at the injection site after intravenous administration (including pain, irritation, inflammation and phlebitis) can be reduced in number and/or severity compared to similar reactions after intravenous administration of a non-liposomal composition containing the same active substance.

The present invention also provides methods for treating a fungal infection in a subject in need thereof by intravenous administration of said injectable compositions.

The present invention further provides methods for preparing compositions containing one or more monolamellar vesicles, each of which encapsulates a compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof.

Detailed Description of the Invention

Phospholipids are amphiphilic molecules having a hydrophilic ionizable head or polar head and a hydrophobic tail containing long-chain fatty acids. When hydrated, several phospholipid molecules (optionally with other molecules such as cholesterol) can form liposomes, which are closed bilayered concentric vesicles capable of entrapping water or other aqueous media. The fatty acid tails of phospholipid molecules form part of the inner layer of the liposome's bilayer membrane. The polar heads on one surface of the membrane are oriented towards the aqueous internal environment of the liposome, and the polar heads on the other surface of the membrane are oriented towards the aqueous external environment.

Drugs, depending on their physicochemical properties, can be included in the liposome either inside the aqueous environment or in the lipid bilayer, with hydrophobic molecules mainly associated with the lipid bilayer, and/or hydrophilic molecules included in the aqueous environment inside the liposome ( e.g. dissolved or dispersed).

Liposomes can be monolamellar, that is, they can have a single bilayer of phospholipids surrounding an aqueous center, or multilamellar, that is, they can contain several concentric bilayer vesicles filled with aqueous content. The size distribution of multilamellar vesicles (MLVs) can be broad and heterogeneous, and the method of producing such vesicles is not reproducible. MLV is not recommended for intravenous administration due to its large particle size. Additionally, MLV is difficult to sterilize by filtration. For injectable compositions intended for intravenous administration, monolamellar liposomes that are relatively small in size are desirable.

Liposomes have previously been used to deliver drugs to target tissues and reduce systemic toxicity, but there has been little understanding of their utility in reducing ISRs occurring locally at the site of injection or infusion. In addition, prior to the present invention, there was no information on the possibility of including enfumafungin-derived triterpenoids in liposomes. Moreover, specifically with respect to SCY-078: this drug exhibits high protein binding, and it was not known whether intravenous administration of SCY-078 would release or leak the encapsulated drug into the bloodstream; An effective liposomal composition must have liposomes that remain intact in the bloodstream for a sufficient time to minimize release or leakage of the encapsulated drug at the site of infusion. According to the present invention, a composition of phospholipid cholesterol vesicles encapsulated with antifungal triterpenoids derived from enfumafungin, such as SCY-078 or an acceptable salt thereof, can be administered intravenously, and the vesicles in the bloodstream can remain intact until they are taken up by macrophages in the organs of the reticuloendothelial system ( RES), such as the liver, spleen and others. In addition, ISRs can occur at very low concentrations of free (unencapsulated) SCY-078, and the present invention can reduce the severity and frequency of ISRs when SCY-078 is administered intravenously by minimizing the local presence of free drug at the injection site. The present invention contemplates the encapsulation of SCY-078 (or a pharmaceutically acceptable salt thereof) or another enfumafungin-derived triterpenoid antifungal agent in a phospholipid (PL)-based system, such as liposomes, as an approach to reduce ISR resulting from intravenous administration. while providing the benefits of an antifungal agent. In the present invention, the drug SCY-078 can be encapsulated in liposomes with high efficiency, in order to minimize the amount of free (unencapsulated) drug substance and make the liposomal composition suitable for commercial production. Additionally, according to the invention, the physicochemical characteristics of the liposomes are stable, which facilitates the storage of the composition.

The present invention provides an injectable composition having antifungal

- 3 045936 activity containing an aqueous phase and one or more monolamellar vesicles, each of which contains a phospholipid and cholesterol, and each of which is encapsulated a compound of formula (II) (in the present invention called SCY-078), or a pharmaceutically acceptable salt thereof or hydrate

(P) which is (1S,4aR,6aS,7R,8R,10aR,10bR,12aR,14R,15R)-15-[[2-amino-2,3,3trimethylbutyl]oxy]-8-[(^ )-1,2-dimethylpropyl]-14-[5-(4-pyridinyl)-III-1,2,4-triazol-1-yl]-1,6,6a,7, 8,9,10,10a ,10b,11,12,12a-dodesahydro-1,6a,8,10a-tetramethyl-4/7-1,4a-propano-2N-phenanthro[1,2-c]pyran-7carboxylic acid, where one or more monolamellar vesicles are hydrated in the aqueous phase.

The pH of the aqueous phase is preferably from about 5.0 to about 7.0. Examples of the aqueous phase are a monosaccharide solution and a disaccharide solution. The injectable composition can be administered intravenously for the treatment and/or prevention of fungal infections, such as systemic fungal infections, including those caused by Candida or Aspergillus species.

The invention also provides methods for treating and/or preventing fungal infection by intravenous administration of an injectable composition containing one or more monolamellar vesicles, each of which contains a phospholipid and cholesterol, and each of which encapsulates a compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof . Additionally, the invention provides the use of vesicles encapsulated with a compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof, for the preparation of an injectable medicament for the treatment or prevention of fungal infection.

In other preferred embodiments, the present invention provides an injectable composition comprising an aqueous phase and one or more monolamellar vesicles, each containing a phospholipid and cholesterol, and each of which encapsulates a compound of formula (IIa) or a pharmaceutically acceptable salt or hydrate thereof

(Pa) which is (1S,4aR,6aS,7R,8R,10aR,10bR,12aR,14R,15R)-15-[[(2R)-2-amino-2,3,3trimethylbutyl]oxy[(^ )-1,2-dimethylpropyl]-14-[5-(4-pyridinyl)-III-1,2,4-triazol-1-yl]-1,6,6a,7,8,9, 10,10a ,10b,11,12,12a-dodecahydro-1,6a,8,10a-tetramethyl-4H-1,4a-propano-2H-phenanthro[1,2-c]pyran-7carboxylic acid, where one or more monolamellar vesicles hydrated in the aqueous phase.

The pH of the aqueous phase is preferably from about 5.0 to about 7.0. Examples of the aqueous phase are a monosaccharide solution and a disaccharide solution. The injectable composition can be administered intravenously for the treatment and/or prevention of fungal infections, such as systemic fungal infections, including those caused by Candida or Aspergillus species.

The invention also provides methods for treating and/or preventing fungal infection

- 4 045936 for intravenous administration of an injectable composition containing one or more monolamellar vesicles, each of which contains a phospholipid and cholesterol, and each of which encapsulates a compound of formula (IIa) or a pharmaceutically acceptable salt or hydrate thereof. Additionally, the invention relates to the use of vesicles encapsulated with a compound of formula (IIa) or a pharmaceutically acceptable salt or hydrate thereof, for the preparation of an injectable medicament for the treatment or prevention of fungal infection.

In preferred embodiments, the phosphate salt of a compound of formula (II) or (IIa) is used or administered according to the present invention.

In preferred embodiments, the citrate salt of a compound of formula (II) or (IIa) is used or administered according to the present invention.

The present invention also provides the use of an injectable composition containing: one or more monolamellar vesicles in which a compound of formula (II) or (IIa), or a pharmaceutically acceptable salt or hydrate thereof, is encapsulated; and a pharmaceutically acceptable carrier, adjuvant or carrier in a subject for treating or preventing a fungal infection.

In the above embodiments, the specified substitutions in the description of the compounds are included only if the presence of the substituents ensures the stability of the compounds as defined.

The compounds of formulas (II) and (IIa) and their pharmaceutically acceptable salts and/or hydrate forms have antimicrobial (e.g., antifungal) activity against yeasts and other fungi, including one or more of the following: Acremonium, Absidia (e.g., Absidiacorymbifera), Alternaria, Aspergillus (e.g. Aspergillus clavatus, Aspergillus flavus, Aspergillus fumigatus, Aspergillus nidulans, Aspergillus niger, Aspergillus terreus and Aspergillus versicolor), Bipolaris, Blastomyces (e.g. Blastomycesdermatitidis), Blastoschizomyces (e.g. Blastoschizomycescapitatus), Candida (e.g. Cand ida albicans , Candida glabrata, Candida guilliermondii, Candida kefyr, Candida krusei, Candida lusitaniae, Candida parapsilosis, Candida pseudotropicalis, Candida stellatoidea, Candida tropicalis, Candida utilis, Candida lipolytica, Candida famata and Candida rugosa), Cladosporium (e.g. Cladosporium carrionii and Clados poriumtrichloides), Coccidioides (eg Coccidioidesimmitis), Cryptococcus (eg Cryptococcus neoformans), Curvularia, Cunninghamella (eg Cunninghamellaelegans), Dermatophyte, Exophiala (eg Exophialadermatitidis and Exophialaspinifera), Epidermophyton (eg Epidermophytonfloccosum), Fonsecaea (eg Fonsecaeapedrosoi), Fusarium (eg Fusariumsolani), Geotrichum (eg Geotrichumcandidum and Geotrichumclavatum), Histoplasma (eg Histoplasmacapsulatum var. capsulatum), Malassezia (e.g. Malassezia furfur), Microsporum (e.g. Microsporumcanis and Microsporumgypseum), Mucor, Paracoccidioides (e.g. Paracoccidioides brasiliensis), Penicillium (e.g. Penicilliummarneffei), Phialophora, Pityrosporumovale, Pneumocystis (e.g. Pneumocystis carinii), Pseudallescheria ( e.g. Pseudallescheriaboydii), Rhizopus (e.g. Rhizopusmicrosporus var. rhizopodiformis and Rhizopusoryzae), Saccharomyces (e.g. Saccharomyces cerevisiae), Scedosporium (e.g. Scedosporiumapiosperum), Scopulariopsis, Sporothmix (e.g. Sporothmixschenckii), Trichoderma, Trichophyton (e.g. Trichophyton chophytonmentagrophytes and Trichophytonrubrum ), and Trichosporon (eg Trichosporonasahii, Trichosporonbeigelii and Trichosporoncutaneum). These compounds are not only useful against organisms that cause systemic pathogenic fungal infections in humans, but are also useful against organisms that cause superficial fungal infections such as Trichoderma spp. and other Candida spp.

The compounds of formulas (II) and (IIa) and their pharmaceutically acceptable salts and/or hydrate forms have an antifungal effect and for this reason are suitable for the treatment and/or prevention of one or more various fungal infections of the external integument, skin, mucous membranes, subcutaneous tissue and systemic infections, including fungal infections of the vulva, vagina, skin, eyes, hair, nails, oral mucosa, gastrointestinal tract, bronchi, lungs, pleura, peritoneum, endocardium, brain, meninges, urinary organs, area vagina, oral cavity, kidneys, heart, external auditory canals, bone, nasal cavity, paranasal cavity, spleen, liver, subcutaneous tissue, lymphatic ducts, joints, muscles, tendons, interstitial plasma cells in the lungs, blood, etc.

The compounds of formulas (II) and (IIa) and their pharmaceutically acceptable salts and/or hydrate forms are useful for the prevention and treatment of one or more various infectious diseases, such as vulvovaginal candidiasis (VVC), dermatophytosis (for example, trichophytosis, ringworm or herpes zoster ), paronychia, lichen versicolor, erythrasma, diaper rash, fungal diaper dermatitis, vulvitis candidiasis, balanitis candidiasis, otitis externa, candidiasis (cutaneous and mucocutaneous), chronic mucocandidiasis (for example, thrush and vaginal candidiasis), cryptococcosis, geotrichosis, trichosporosis, aspergillosis, penicilliosis, fusarium, zygomycosis, sporotrichosis, chromomycosis, coccidioidomycosis, histoplasmosis, blastomycosis, paracoccidioidomycosis, Pseudallescheria infections, mycetoma, fungal keratitis, otomycosis, pneumocystosis, fungal abscess, fungal pleural empyema and fungemia. Compounds of formulas (II) and (IIa) and their pharmaceutically acceptable salts and/or

- 5 045936 hydrated forms can also be used as prophylactic agents to prevent systemic and local fungal infections. Prophylactic use may be considered, for example as part of a selective intestinal decontamination regimen to prevent infection in immunocompromised patients (eg, patients with AIDS, patients receiving anticancer treatment, or transplant patients). It may also be desirable to prevent fungal overgrowth during antibiotic treatment in certain pathological syndromes or iatrogenic conditions.

The compounds of formulas (II) and (IIa) and their pharmaceutically acceptable salts and/or hydrate forms can be prepared according to the synthetic methods disclosed in US Pat. No. 8,188,085, the contents of which are incorporated herein by reference in their entirety.

As used in the present invention, or denotes alternatives, which may be combined if necessary.

Unless otherwise expressly stated, all ranges given herein are inclusive.

A stable compound is a compound that can be prepared and isolated, and the structure and properties of which remain, or can remain or remain substantially unchanged, for a period of time sufficient to permit use of the compound for the purposes described in the present invention (for example, administration to a subject for therapeutic or prophylactic purposes). Also included within the concept of such a compound are stable complexes of the compound, such as a stable hydrate.

As a result of the choice of substituents and substituent groups, some of the compounds of formulas (II) and (IIa) may have asymmetric centers and may occur as mixtures of stereoisomers or as individual diastereomers or enantiomers. Unless otherwise indicated, all isomeric forms of these compounds (and their pharmaceutically acceptable salts and/or hydrated forms), both isolated and in mixtures, are included within the scope of the present invention. Also included within the scope of the present invention are tautomeric forms of these compounds (and their pharmaceutically acceptable salts and/or hydrates).

If any variable appears more than once in any component or in formulas (II) or (IIa), its definition in each case is independent of its definition in each other case. Also, combinations of substituents and/or variable elements are permissible only if stable compounds are formed in the presence of such combinations.

The compounds of formulas (II) and (IIa) and their pharmaceutically acceptable salts and/or hydrate forms are also useful in preparing and performing screening assays for antifungal compounds. For example, the compounds are useful for isolating mutants, which are excellent screening tools for identifying additional antifungal compounds.

The compounds of formulas (II) and (IIa) can be administered in the form of pharmaceutically acceptable salts or hydrates, depending on the situation. However, other salts may be useful for preparing the compounds or pharmaceutically acceptable salts thereof. For example, if the compounds contain a basic amino group, they can be easily isolated as trifluoroacetic acid salts (eg after HPLC purification). The conversion of trifluoroacetic acid salts to other salts, including pharmaceutically acceptable salts, can be achieved using certain standard methods known in the art. For example, a suitable ion exchange resin can be used to obtain the desired salt. Alternatively, conversion of the trifluoroacetic acid salt to the free amine starting material can be accomplished by standard methods known in the art (eg, neutralization with a suitable inorganic base such as NaHCO ₃ ). Other desired amine salts can be prepared in the usual manner by reacting the free base with a suitable organic or inorganic acid. By way of example, pharmaceutically acceptable quaternary ammonium salts include the following salts: hydrochloride, sulfate, phosphate, carbonate, acetate, tartrate, citrate, malate, succinate, lactate, stearate, fumarate, hippurate, maleate, gluconate, ascorbate, adipate, gluceptate, glutamate , glucoronate, propionate, benzoate, mesylate, tosylate, oleate, lactobionate, lauryl sulfate, besylate, caprylate, isethionate, gentisate, malonate, napsylate, edisylate, pamoate, xinafoate, napadisylate, hydrobromide, nitrate, oxalate, cinnamate, mandelate, undecylenate and camsylate . Many of the compounds of formulas (II) and (IIa) contain an acidic carboxylic acid functionality, in which case suitable pharmaceutically acceptable salts thereof may include alkali metal salts, for example sodium or potassium salts; alkaline earth metal salts, for example calcium or magnesium salts; and salts formed with suitable organic ligands, for example, quaternary ammonium salts.

The present invention includes the use of prodrugs having formulas (II) and (IIa). Typically, such prodrugs are functional derivatives of compounds that are readily convertible in vivo into the desired compound. Thus, in the methods of treatment of the present invention, the term administration covers the treatment of the various described conditions with a specifically disclosed compound or with a compound that is converted to the specified compound in vivo after administration to a patient. Conventional methods for selecting and preparing suitable derivatives of prodrugs are described, for example, in the manual Design of Prodrugs, ed. H. Bundgaard, Elsevier, 1985, which is incorporated herein by reference in its entirety. Metabolites of compounds of formulas (II) and

- 6 045936 (IIa) include active substances produced when these compounds are introduced into a biological environment.

The term administration and its variations (eg, administering a compound) means delivering a compound or a prodrug of that compound into the body of a subject in need of treatment. If a compound of formula (II) and (IIa) or a pharmaceutically acceptable salt thereof, or a hydrate or prodrug thereof is administered in combination with a second active agent (for example, another antifungal and/or antibacterial agent useful for the treatment of fungal and/or bacterial infections) , the term administration and its variations include simultaneous and sequential administration of the compound (or a salt, hydrate or prodrug thereof) and another active substance.

As used herein, the term composition refers to a product containing the specified ingredients, as well as any product that directly or indirectly results from the combination of the specified ingredients.

The term pharmaceutically acceptable means that the ingredients of the pharmaceutical composition must be compatible with each other and not cause harm to the recipient.

As used herein, subject (alternatively referred to herein as patient) refers to an animal, preferably a mammal, most preferably a human, who is being treated, observed or experimented.

As used herein, an effective amount means the amount of an active ingredient or pharmaceutical substance that produces in a tissue, system, animal or human the biological or clinical response that the researcher, veterinarian, physician or other clinician seeks to achieve. In one embodiment, the effective amount may be a therapeutically effective amount that reduces the symptoms of the disease or condition being treated. In another embodiment, the effective amount may be a prophylactically effective amount to prevent symptoms of a preventable disease or condition, or to reduce the likelihood of its occurrence. The term may also refer to an effective inhibitory amount of an enfumafungin derivative that is sufficient to inhibit (1,3)-β-O-glucan synthase and thereby produce the desired response.

The terms treat, treatment, and variations thereof generally refer to treatment that, when administered, results in the disappearance or improvement of one or more signs or symptoms associated with a fungal infection, or results in the destruction of the fungi causing the infection, or any combination of these results.

For the prevention or treatment of fungal infection, a compound of formula (II) or (IIa) (optionally in the form of a salt or hydrate) can be administered by conventional methods available for use in combination with pharmaceutical agents.

For the purpose of preventing or treating fungal infections occurring in conditions or anatomical areas having an acidic pH, a compound of formula (II) or (IIa) (optionally in the form of a salt or hydrate) can be administered alone as a stand-alone therapeutic agent, or together with one or more other antifungal agents (sequentially or simultaneously) as a combination of therapeutic agents.

For the purpose of preventing or treating a fungal infection, a compound of formula (II) or (IIa) (optionally in the form of a salt or hydrate) can be administered together with a pharmaceutical carrier selected based on the preferred route of administration and standard pharmaceutical practice.

For example, the compounds of formulas (II) and (IIa) and pharmaceutical forms of their salts and/or hydrates can be administered by one or more of the following routes: orally, parenterally (including subcutaneous injection, intravenous, intramuscular, intralesional injection or infusion), by inhalation (eg, by nasal or buccal inhalation spray, aerosol metered dose inhaler and dry powder inhaler), via nebulizer, ocular, topical, transdermal or rectal, in unit dose form of a pharmaceutical composition containing an effective amount of the compound and conventional non-toxic pharmaceutically acceptable carriers, adjuvants and fillers. Liquid preparations suitable for oral administration (for example, suspensions, syrups, elixirs and the like) can be prepared in accordance with techniques known in the art, and conventional media such as water, glycols, oils, alcohols and the like can be used. . Solid preparations suitable for oral administration (for example, powders, pills, capsules and tablets) can be prepared according to techniques known in the art, and solid excipients such as starches, sugars, kaolin, lubricants, binders can be used. substances, disintegrants and the like. Parenteral compositions can be prepared according to techniques known in the art, typically using sterile water as the carrier and optionally other ingredients, such as a solubility aid. Injectable solutions can be prepared according to methods known in the art, wherein the carrier comprises saline solution, glucose solution, or a solution containing a mixture of saline solution and glucose.

- 7 045936

Additional descriptions of methods suitable for use in preparing pharmaceutical compositions and ingredients useful for use in said compositions are provided in Remington's Pharmaceutical Sciences, ^20th edition, ed. AR Gennaro, Mack Publishing Co., 2000.

The compounds of formulas (II) and (IIa) and pharmaceutically acceptable forms of their salts and/or hydrates can be administered, for example, orally or intravenously, in doses ranging from, for example, 0.001 to 1000 mg/kg body weight of a mammal (e.g., human) per day, as a single dose or in divided doses. An example dosage range is 0.01 to 500 mg/kg body weight per day orally or intravenously in a single dose or in divided doses. Another example dosage range is from 0.1 to 50 mg/kg body weight per day orally or intravenously in single or divided doses. Compositions for oral administration may be presented in the form of tablets or capsules containing, for example, from 1.0 to 1000 mg of active ingredient, in particular 1, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200 , 250, 300, 400, 500, 600, 750 and 1000 mg of the active ingredient to adjust the dose according to the symptoms of the patient being treated. The specific dosage level and frequency of dosing for any given patient may vary and depend on a variety of factors, including the potency of the particular compound being used, the metabolic stability and duration of action of that compound, the route and timing of administration, the rate of elimination, the drug combination, the severity of the particular condition, and age. , body weight, general health, gender and diet of the patient and the type of host being treated.

The antifungal activity of the compounds can be demonstrated using various assays known in the art, for example, the minimum inhibitory concentration (MIC) assay against yeast and the minimum effective concentration (MEC) assay against filamentous molds and dermatophytes with broth microdilution, or by evaluating the activity of these compounds against Candida and against Aspergillus in vivo in mice or rabbits. The compounds of formula (I) exemplified in US Pat. No. 8,188,885 have been found to generally inhibit the growth of Candida spp. in a concentration range lower than 0.03 to 32 pg/ml, or with MEC values against Aspergillus fumigatus in a range lower than 0.03 to 32 pg/ml.

The single bilayer vesicles or liposomes encapsulating the enfumafungin triterpenoid antifungal compounds of the present invention preferably have an average particle size (vesicle diameter) of about 150 nm or less, more preferably of about 100 nm or less, or of about 70 nm or less. up to 80 nm. The vesicles are hydrated in a suitable aqueous phase, for example water containing a monosaccharide or disaccharide. Any aqueous phase with low ionic strength is suitable for hydration of vesicles. The aqueous phase is preferably isotonic. The pH of the aqueous phase should be from about 4.0 to about 8.0, and preferably from about 5.0 to about 7.0.

Methods for producing single bilayer vesicles or liposomes include methods that provide sufficient energy and shear force to form monolamellar vesicles (such as sonication, high pressure homogenization, microfluidic mixing, etc.). The single bilayer vesicles of the invention can be prepared by sonication, microfluidic mixing and/or homogenization of hydrated suspensions containing an enfumafungin-derived antifungal agent (for example, a compound of formula (II) or (IIa) or a pharmaceutically acceptable salt or hydrate thereof), a phospholipid and cholesterol. In preferred embodiments, the drug SCY-078 (or a pharmaceutically acceptable salt or hydrate thereof), phospholipid (PL) and cholesterol (CHOL) are suspended in an aqueous phase with a pH of about 5.0 to about 7.0 and a molar ratio of SCY-078 :PL:CHOL ratio of 1:7:2.5 and subjected to ultrasonication, microfluidic mixing, homogenization and/or other method, resulting in single bilayer vesicles containing phospholipid and cholesterol encapsulated with SCY-078 (or its salt or hydrate) inside the vesicle. The preferred drug for encapsulation in single bilayer vesicles of the invention is the citrate salt of SCY-078.

To form the single bilayer vesicles of the invention, a single species of phospholipid may be used, or different two or more phospholipids may be used in combination to form the single bilayer vesicles. Phospholipids suitable for use in preparing the single bilayer vesicles of the invention include, but are not limited to, synthetically produced or naturally occurring substances such as phosphatidylcholine (PC), phosphatidic acid (PA), phosphatidylserine (PS), phosphatidylethanolamine (PE) and phosphatidylglycerol (PG).

The phospholipids used to form the single bilayer vesicles of the invention are preferably PC and PG used in combination.

Suitable phosphatidylcholines include, but are not limited to, synthetically derived dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), dilauroylphosphatidylcholine (DLPC), and dimyristoylphosphatidylcholine (DMPC). Naturally occurring phosphatidylcholines are egg PC and soy PC.

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Suitable phosphatidylglycerols include, but are not limited to, synthetically produced dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylglycerol (DSPG), dilauroylphosphatidylglycerol (DLPG) and dimyristoylphosphatidylglycerol (DMPG), and naturally occurring phosphatidylglycerols, such as egg PG.

Preferably, the phospholipid content of the enfumafungin derivative-encapsulated vesicles of the present invention ranges from 50 to 80 mol%.

Cholesterol stabilizes the lipid bilayer and can prevent leakage of the encapsulated drug, but excess cholesterol can negatively affect the effectiveness of encapsulation. The preferred cholesterol content of the enfumafungin derivative encapsulated vesicles of the invention is from 10 to 30 mol%.

The preferred drug content of the enfumafungin derivative encapsulated vesicles of the invention is from 5 to 12 mol%.

Single bilayer vesicles encapsulating a triterpenoid antifungal compound, an enfumafungin derivative, are hydrated in the aqueous phase. Preferably, the aqueous phase contains one or more monosaccharides (such as glucose, fructose and galactose) and/or one or more disaccharides (such as sucrose, trehalose and lactose) that render the injectable compositions (drug-encapsulated single bilayer vesicles and hydrated in the aqueous phase) and can improve bilayer stability. When monosaccharide(s) are used in the aqueous phase, the amount is preferably from about 4 to 6% (w/v) relative to the injectable composition (encapsulated drug vesicles hydrated in the aqueous phase). When disaccharides are used in the aqueous phase, the preferred amount is from about 8 to 10% (w/v) relative to the injectable composition (encapsulated drug vesicles hydrated in the aqueous phase).

Preferably, the concentration of encapsulated drug, expressed in mg per ml of injectable composition (aqueous phase hydrated single bilayer vesicles in which the drug is encapsulated), is from 0.01 to 50 mg/ml and more preferably from 0.1 to 5 mg /ml.

To prepare a lipid dispersion containing an enfumafungin-derived antifungal agent, the phospholipid and cholesterol are dissolved in an organic solvent such as a C1-C ₅ alcohol (preferably methanol or ethanol), and the said solution is added to a solution of the enfumafungin-derived antifungal agent in an organic solvent. , for example, in ethanol. Alternatively, the antifungal agent can be added directly to the phospholipid and cholesterol solution. To obtain a lipid dispersion containing an antifungal agent, the solvent is evaporated (optionally in vacuo) in a suitable reaction vessel, such as a round bottom flask. The solvent can be removed by other means, such as using a spray dryer. Preferably, the lipid dispersion containing the enfumafungin-derived antifungal agent is shelf stable. It is also convenient if such a lipid dispersion can be hydrated at any desired time and subjected to other processing to form liposomes.

The lipid dispersion containing the antifungal agent is hydrated in an aqueous solution, preferably containing a monosaccharide or disaccharide, to obtain a hydrated suspension. The hydrated suspension is mixed using a high shear mixer (eg, about 10,000 rpm), and the resulting multilamellar vesicles are homogenized under high pressure (eg, about 10,000 to about 30,000 psi (about 68. 95 to about 206.8 MPa) and at processing temperatures ranging from, for example, about 25°C to about 70°C) to form single bilayer vesicles (monolamellar vesicles). Such single bilayer vesicles can be sterilized, for example, by filtration through 0.45 and 0.22 μm filters, and/or sterilized by moist heat. The filtered suspension of single bilayer vesicles can be frozen and dehydrated under high vacuum (lyophilized) for long-term storage.

The amount of antifungal agent encapsulated in the vesicles that are obtained in accordance with the methods of the invention can be determined, for example, using high performance liquid chromatography (HPLC). The size of the vesicles can be determined, for example, using light scattering technology, such as dynamic light scattering (DLS).

The local tolerance (ISR) of single bilayer vesicles encapsulating an enfumafungin-derived antifungal agent was assessed when administered intravenously in animal models, including rats and rabbits. To assess local tolerance, clinical and histopathological evaluation of tissue at the infusion site was performed. Also to demonstrate the utility of the injectable compositions of the invention, the stability of the vesicles was determined under conditions simulating the infusion of the vesicles into the bloodstream.

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Examples

The following examples are provided solely to illustrate the invention and its implementation. The examples should not be construed as limiting the scope or spirit of the invention.

Preparation of liposomes with encapsulated SCY-078.

Examples from A to I.

In Examples A to I: a phospholipid mixture including DSPC and DSPG, and cholesterol and the enfumafungin derivative SCY-078 in different molar ratios, as shown in table. 1, dissolved in a solvent (ethanol at 70°C) and subjected to microfluidic mixing (NanoAssemblr™) with water in a 1:3 volume ratio solvent:water at 65°C at a flow rate of 10 ml/min to form vesicles or liposomes with a single bilayer , carrying SCY-078 (SCY-078-encapsulated). Solvent and free (unencapsulated) material in the liposome suspension were removed by tangential flow filtration (TFF).

Examples from J to P.

Examples J and K used thin film hydration techniques. A phospholipid mixture including DSPC and DSPG, and cholesterol and SCY-078 in amounts according to Table 2 was dissolved in ethanol at 70°C. A round bottom flask was used as a suitable reaction vessel, from which the solvent was removed by evaporation under vacuum to obtain a lipid dispersion film containing the antifungal agent.

In Examples L to P, the solvent was removed by spray drying. Phospholipid mixture including DSPC and DSPG, and cholesterol and SCY-078 in quantities according to table. 2 was dissolved in ethanol at 70°C and mixed to form a homogeneous colloidal dispersion. The dispersion was sprayed as a fine mist at a flow rate of 20 ml/min through a 0.7 mm nozzle at a spray pressure of approximately 70 psi (482.6 kPa) and dried at approximately 50°C. The spray-dried powder was collected and dried under vacuum to constant weight.

In Examples J to P, a lipid dispersion containing an antifungal agent prepared by thin film evaporation or spray drying was hydrated in an aqueous solution containing 7.5% (w/v) sucrose. To form single bilayer vesicles, the hydrated suspension was mixed using a high shear mixer at 10,000 rpm for approximately 5 minutes, and the resulting multilamellar vesicles were subjected to high pressure homogenization (Microfluidics®) at pressures ranging from approximately 10,000 approximately to 30,000 psi (about 68.95 to about 206.8 MPa), using processing temperatures ranging from about 25°C to about 70°C.

The liposomes obtained in examples M, O and P were subjected to sterilization by filtration through a 0.45 μm and 0.22 μm filter. The liposomes in Example M were lyophilized. The liposomes in Example N were sterilized with moist heat at 121°C for 12 minutes.

Characterization of SCY-078-encapsulated liposomes.

Compositions with different molar ratios of DSPC: cholesterol: DSPG: SCY-078 were evaluated, as shown in table. 1 and 2 regarding single bilayer vesicle formation, particle size distribution, surface charge, and amount of SCY-078 encapsulated.

After vesicle or liposome formation, free (unencapsulated) SCY-078 was removed by TFF (Examples A to I). In cases where vesicles or liposomes were prepared without the use of TFF, all unencapsulated SCY-078 preparation was separated from encapsulated SCY-078 by dialysis using a cellulose ether membrane with a molecular weight cutoff (MWCO) of 20,000 Dalton.

Liposome-encapsulated SCY-078 was separated from any non-encapsulated SCY-078 by size exclusion chromatography using a Sephadex® G-25 column. The amount of liposome-encapsulated SCY-078 was determined by HPLC analysis. The SCY-078 preparation was separated on a C18 column and detected using ultraviolet (UV) spectroscopy at 210 nm. Vesicle particle size was determined using DLS (Zetasizer Nano, Malvern Instruments). Surface charge (zeta potential) was determined by laser Doppler microelectrophoresis in a disposable electrophoretic cell.

As shown in table. 1 and 2, in a number of compositions with certain molar ratios of DSPC: cholesterol: DSPG: SCY-078, the formation of liposomal vesicles or liposomes with a single bilayer was detected with an entrapment efficiency of more than 95%. Surprisingly, there was essentially no free (non-encapsulated) drug even though the production process did not include the step of removing any non-encapsulated drug (Examples J-O). It was also surprisingly shown that the vesicles remained stable after moist heat sterilization, with no change in vesicle size and retention of >95% SCY-078 in the vesicles (Example N).

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Table 1

Liposome compositions obtained by microfluidic mixing using NanoAssemblr™ equipment

Molar ratio mol% Example SCY-078: PG : PC:CHOL SCY- 078 FL HOL Average particle size (nm) Zeta potential (charge, mV) % encapsulated SCY078 A 1:1:2.5:1.25 17.4 60.9 21.7 4527.3 -26.31 49.3 IN 1:0.5:5:2.5 11.1 61.1 27.8 4359 -6.43 58.3 WITH 1:1.2: 3:1.5 14.9 62.7 22.4 175.5 -31.59 82.3 D 1:1:5:2.5 10.5 63.2 26.3 173.3 -37.93 81.7 E 1:1.5:3.75:1.87 12.3 64.7 23 89.6 -39.72 98.0 F 1:1.6: 4:2 11.6 65.1 23.3 104.7 -47.05 95.7 G 1:2:5:2.5 9.5 66.7 23.8 124.9 -53.34 96.3 N 1:3:5:2.5 8.7 69.7 21.6 146.0 -59.33 98.3 I 1:2:5: 1.25 10.8 75.7 13.5 122.0 -48.19 97.7

table 2

Liposome compositions obtained by homogenization at high pressure in a microfluidization device

Molar ratio mol% Example SCY-078: PG : PC : CHOL SCY- 078 FL HOL Average particle size (nm) Zeta potential (charge, mV) % encapsulated other SCY-078 J 1:1.5:3.75: 11.87 12.3 64.7 23 81.6 -32.0 99 TO 1:2:5: 2.5 9.5 66.7 23.8 91.2 -47.2 96 L 1:2:5:2.5 9.5 66.7 23.8 95 -54.0 99 M 1:2:5: 2.5 9.5 66.7 23.8 98 - 99 N 1:2:5: 1.25 9.5 66.7 23.8 100 -55.6 >95 ABOUT 1:2:5: 1.25 9.5 66.7 23.8 51.1 -47.3 >99 R 1:2:5: 1.25 9.5 66.7 23.8 78 -74.1 >95

Evaluation of the physicochemical stability of SCY-078-containing lipid dispersion after spray drying.

The stability of the spray-dried intermediate (lipid dispersion containing SCY-078) was assessed under ambient storage (25°C/60% relative humidity (RH)) and accelerated aging (40°C/75% relative humidity). As shown in table. 3, The spray-dried intermediate was stable in SCY-078 and degradation product assays with no significant changes or trends observed for up to 3 months when stored at ambient conditions (25°C/60% RH) and under accelerated storage conditions (40°C/75% RH). These results indicate that the spray-dried intermediate can be conveniently stored, for example under ambient conditions, until

- 11 045936 ratification and other processing to obtain liposomes, which is a commercially desirable property.

Table 3

Stability assessment of spray-dried SCY-078 dispersion

Analysis Initial conditions 3 months 25 °C/60% ov 40 °C/75% OB Appearance (visual) White fine powder White powder White powder Analysis (mg/g) SCY-078 as free base 89.2 92.2 90.7 Decomposition products Individual\(installed and uninstalled) ^A No impurities >0.1% No impurities >0.1% No impurities >0.1% Total No impurities >0.1% No impurities >0.1% No impurities >0.1% Water content (%) 2.3 1.8 3.1

A: Report degradation products >1% area (excluding peaks present at or below the API level).

Assessment of physicochemical stability of liposomes encapsulated with SCY-078.

The stability under refrigerated conditions (from 2 to 8°C) of a ready-to-dilute liposomal suspension with encapsulated SCY-078 was assessed. As shown in table. 4, no changes were detected in the SCY-078 analysis or in the average particle size values. Additionally, no leakage of the encapsulated drug was observed during storage, indicating the strong binding of SCY-078 to the lipid bilayer of the liposomes. These results demonstrate stability and potential long-term stability, which is suitable for various commercial applications of the liposomal compositions of the invention.

Table 4

Evaluation of the stability of the injectable form of liposomes with SCY-078 (concentration 4 mg/ml)

Example Conditions storage Storage time Analysis (mg/ml) Average particle size (nm) % encapsulation N from 2°С to 8°С original 3.8 100 >95% N 3 months 3.78 89.6 - ABOUT from 2°С to 8°С original 3.80 51.1 >99% ABOUT 6 months 4.00 53.4 >99%

Assessment of the physiological stability of liposomes encapsulated with SCY-078.

The stability of SCY-078-encapsulated vesicles or liposomes was assessed by incubating the liposomes in fresh 50% bovine serum for 24 h at 37°C. Liposome fractions were separated from serum components using size exclusion chromatography on a Sephadex® G-25 column. The collected liposome and serum fractions were analyzed for SCY-078 content by HPLC. The results presented in table. 5 demonstrate that although SCY-078 was not detected in the whey protein fractions, substantially all of the nominal amount of SCY-078 was recovered from the liposome fractions. Given the high degree of protein binding by SCY-078, these results unexpectedly showed that under conditions simulating intravenous administration into the bloodstream, SCY-078 remained encapsulated in intact liposomes.

Evaluation of local stability of liposomes encapsulated with SCY-078 in animal models.

Local tolerance study of intravenous infusion in Sprague Dawley rats.

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An intravenous infusion study was conducted in rats over a 14-day period. Animals were treated with liposome-encapsulated SCY-078 (10 or 40 mg/kg/day), or SCY-078 in solution (10 or 40 mg/kg/day), or saline control. Animals were administered through indwelling catheters surgically implanted into the vena cava, such that assessment of response at the infusion site was based on histological detection of vascular inflammation. Overall, the incidence and severity of vascular inflammation increased for SCY-078 administered as a solution compared with vascular inflammation in saline-treated animals; however, in animals that were injected with SCY-078 encapsulated in a liposome, vascular inflammation was not observed (Table 6).

A study of local intravenous irritation in New Zealand white rabbits.

This study assessed local irritation during administration of a physiological control, SCY-078 at a dose of 40 mg/kg in solution and liposome-encapsulated SCY-078 (10 or 40 mg/kg), in a twice daily dosing schedule (6 h ±15 min). These doses were administered to New Zealand White rabbits by intravenous infusion over one hour at a rate of 20 ml/h through an indwelling catheter for five consecutive days.

During intravenous infusion of SCY-078 administered as a solution at a dose of 40 mg/kg over 1 hour twice daily, adverse local reactions occurred at the infusion site and surrounding area, manifesting as very mild/mild/to severe swelling and erythema. which resulted in unplanned euthanasia of all animals in this group immediately after the first day of administration. In contrast, animals receiving twice daily intravenous infusions of liposome-encapsulated SCY078 at doses of 10 and 40 mg/kg were able to complete the planned 5-day study with good tolerability of the formulation.

Results from an animal local tolerability study showed that liposome-encapsulated SCY-078 was better tolerated than SCY-078 in solution with regard to vascular inflammation and ISR at the infusion site and may be suitable for intravenous administration via a peripheral vein.

Table 5 Stability of SCY-078 encapsulated liposomes ________after incubation in fetal bovine serum at 37°C_______

Incubation time in serum at 37 °C (hours) Isolated SCY078 in serum (%) Fractions (0-29) Isolated liposomes in serum (%) Factions (0-29) Isolated SCY078 in liposome (%) Fractions (29-80) ¹ 0 hours <LOD <LOD 112 1 hour <LOD <LOD 90 2 hours <LOD <LOD 118 4 hours <LOD <LOD 98 8 ocloc'k <LOD <LOD 91 24 hours <LOD <LOD 96 Average 100.83

¹ Amount of SCY-078 isolated normalized to liposomes isolated. Limit of detection LOD:SCY=0.036 μg/ml; liposomes = 0.017 pg/ml.

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Table 6

Vascular inflammation at the infusion site in Sprague Dawley rats

Group designation Control (saline solution) SCY-078 in solution SCY-078 in liposome Dose (mg/kg/day) 0 10 40 10 40 Infusion site, from skull to catheter tip Total 0/4 2/4 2/4 0/4 0/4 Minimum 0 2 1 0 0 Moderate 0 0 1 0 0 Infusion site, at the tip of the catheter Total 0/4 1/4 0/4 0/4 0/4 Moderate 0 1 0 0 0 Infusion site, from tail to tip of catheter Total 0/4 0/4 0/4 0/4 0/4 Minimum 0 0 0 0 0

Claims (45)

1. An injectable composition having antifungal activity, containing an aqueous phase and one or more monolamellar vesicles, each of which contains a phospholipid and cholesterol and each of which is encapsulated a compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof (P) which is (1S,4aR,6aS,7R,SR,10aR,10bR,12aR,14R,15R)-15-[[2-amino-2,3,3trimethylbutyl]oxy]-8-[(^ )-1,2-dimethylpropyl]-14-[5-(4-pyridinyl)-III-1,2,4-triazol-1-yl]-1,6,6a,7,8, 9,10,10a ,10b,11,12,12a-dodecahydro-1,6a,8,10a-tetramethyl-4H-1,4a-propano-2H-phenanthro[1,2-c]pyran-7carboxylic acid; where one or more monolamellar vesicles are hydrated in the aqueous phase. 2. The injectable composition according to claim 1, wherein the concentration of the encapsulated compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof in the injectable composition is from 0.01 to 50 mg/ml. 3. Injectable composition according to claim 1, in which the aqueous phase additionally contains sugar. 4. The injectable composition according to claim 3, wherein the sugar is selected from monosaccharides, disaccharides and combinations thereof. 5. The injectable composition according to claim 4, wherein the sugar is selected from sucrose, trehalose, lactose, glucose, fructose and galactose and combinations thereof. 6. Injectable composition according to claim 3, in which the pH level of the aqueous phase is in the range from 5.0 to 7.0. 7. The injectable composition according to claim 1, wherein the phospholipid is phosphatidylcholine, phosphatidic acid, phosphatidylserine, phosphatidylethanolamine, phosphatidylglycerol, or combinations thereof. 8. The injectable composition according to claim 1, wherein the phospholipid is phosphatidylcholine and phosphatidylglycerol. - 14 045936 9. The injectable composition according to claim 8, wherein the phosphatidylcholine is selected from dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, egg phosphatidylcholine, soy phosphatidylcholine, dilauroylphosphatidylcholine and dimyristoylphosphatidylcholine. 10. The injectable composition according to claim 8, wherein the phosphatidylglycerol is selected from dipalmitoylphosphatidylglycerol, distearoylphosphatidylglycerol, dilauroylphosphatidylglycerol and dimyristoylphosphatidylglycerol. 11. The injectable composition according to claim 1, wherein the compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof is present in the vesicles in an amount of from 5 to 12 mol.%, the phospholipid is present in the vesicles in an amount of from 50 to 80 mol.%, and cholesterol is present in the vesicles in amounts from 10 to 30 mol.%. 12. The injectable composition according to claim 1, wherein the phospholipid includes phosphatidylglycerol and phosphatidylcholine, and the molar ratio of the compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof to phosphatidylglycerol, to phosphatidylcholine, to cholesterol is 1:2:5:2.5 . 13. The injectable composition according to claim 1, wherein the average particle size of the one or more monolamellar vesicles is less than 150 nm. 14. The injectable composition according to claim 13, wherein the average particle size of the one or more monolamellar vesicles is less than 100 nm. 15. The injectable composition according to claim 14, wherein the average particle size of the one or more monolamellar vesicles is from 70 to 80 nm. 16. The injectable composition according to claim 1, wherein the concentration of the encapsulated compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof in the injectable composition is from 0.01 to 50 mg/ml; phospholipid includes phosphatidylglycerol and phosphatidylcholine; the molar ratio of the compound of formula (II) or its pharmaceutically acceptable salt or hydrate to phosphatidylglycerol, phosphatidylcholine, to cholesterol is 1:2:5:2.5; and the aqueous phase further contains sugar and has a pH level of 5.0 to 7.0. 17. The injectable composition according to claim 16, wherein the citrate salt of the compound of formula (II) is encapsulated in one or more monolamellar vesicles. 18. A method of treating a fungal infection in a subject in need thereof, wherein said method comprises intravenous administration of an injectable composition according to claim 1. 19. The method according to claim 18, wherein the subject is a human. 20. The method according to claim 18, wherein the fungal infection is caused by Candida spp. 21. The method according to claim 18, wherein the fungal infection is caused by Aspergillus spp. 22. A method of treating a fungal infection in a subject in need thereof, wherein said method comprises intravenous administration of an injectable composition according to claim 16. 23. A method for preparing an injectable composition containing one or more monolamellar vesicles, each of which encapsulates a compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof (P) which is (1S,4aR,6aS,7R,8R,10aR,10bR,12aR,14R,15R)-15-[[2-amino-2,3,3trimethylbutyl]oxy]-8-[(^ )-1,2-dimethylpropyl]-14-[5-(4-pyridinyl)-III-1,2,4-triazol-1-yl]-1,6,6a,7,8, 9,10,10a ,10b,11,12,12a-dodecahydro-1,6a,8,10a-tetramethyl-4H-1,4a-propano-2H-phenanthro[1,2-c]pyran-7carboxylic acid; this method includes: a) dissolving the phospholipid and cholesterol in an aliphatic alcohol having from one to five carbon atoms to form a first solution; b) dissolving a compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof in the first solution to form a second solution; c) mixing the second solution; - 15 045936 d) evaporating the solvent from the second solution to obtain a phospholipid-cholesterol dispersion containing a compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof; e) hydrating a phospholipid-cholesterol dispersion containing a compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof with a sugar solution to obtain a hydrated suspension; And f) forming from the hydrated suspension one or more monolamellar vesicles, each containing a phospholipid and cholesterol and in which a compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof is encapsulated. 24. The method according to claim 23, wherein the aliphatic alcohol is selected from methanol or ethanol. 25. The method of claim 23, wherein the sugar solution contains a sugar selected from monosaccharides, disaccharides and combinations thereof. 26. The method of claim 25, wherein the sugar is selected from sucrose, trehalose, lactose, glucose, fructose and galactose and combinations thereof. 27. The method of claim 23, wherein 90% or more of the amount of the compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof present in step (b) is encapsulated in step (f) in one or more monolamellar vesicles. 28. The method of claim 23, wherein 95% or more of the amount of the compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof present in step (b) is encapsulated in step (f) in one or more monolamellar vesicles. 29. The method according to claim 23, wherein the compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof is present in the vesicles in an amount of from 5 to 12 mol%. 30. The method according to claim 23, in which the phospholipid includes phosphatidylglycerol and phosphatidylcholine and in which the molar ratio of the compound of formula (II) or its pharmaceutically acceptable salt or hydrate to phosphatidylglycerol, to phosphatidylcholine, to cholesterol is 1:2:5:2.5 . 31. The method of claim 23, wherein sonication, microfluidic mixing, homogenization, or a combination thereof is used to form one or more monolamellar vesicles in step (f). 32. The method of claim 23, further comprising sterilizing one or more monolamellar vesicles obtained in step (f). 33. The method of claim 23, further comprising lyophilizing one or more monolamellar vesicles obtained in step (f). 34. The method of claim 23, wherein the average particle size of the one or more monolamellar vesicles is less than 150 nm. 35. The method of claim 34, wherein the average particle size of the one or more monolamellar vesicles is less than 100 nm. 36. The method of claim 35, wherein the average particle size of the one or more monolamellar vesicles is from 70 to 80 nm. 37. An injectable composition having antifungal activity, containing an aqueous phase and one or more monolamellar vesicles, each of which contains a phospholipid and cholesterol and each of which encapsulates a compound of formula (IIa) or a pharmaceutically acceptable salt or hydrate thereof (Pa) which is (1S,4aR,6aS,7R,8R,10aR,10bR,12aR,14R,15R)-15-[[(2R)-2-amino-2,3,3trimethylbutyl]oxy]-8 -[(^)-1,2-dimethylpropyl]-14-[5-(4-pyridinyl)-III-1,2,4-triazol-1-yl]-1,6,6a,7,8, 9 ,10,10a,10b,11,12,12a-dodecahydro-1,6a,8,10a-tetramethyl-4H-1,4a-propano-2H-phenanthro[1,2-c]pyran-7carboxylic acid, where one or more monolamellar vesicles are hydrated in the aqueous phase. 38. The injectable composition according to claim 37, wherein the compound of formula (IIa) is encapsulated in one or more monolamellar vesicles. - 16 045936 39. The injectable composition according to claim 37, wherein a pharmaceutically acceptable salt of the compound of formula (IIa) is encapsulated in one or more monolamellar vesicles. 40. The injectable composition according to claim 37, wherein the citrate salt of the compound of formula (IIa) is encapsulated in one or more monolamellar vesicles. 41. A method of treating a fungal infection in a subject in need thereof, wherein said method comprises intravenous administration of an injectable composition according to claim 37. 42. The method according to claim 41, wherein the fungal infection is caused by Candida spp. 43. The method of claim 41, wherein the fungal infection is candidemia. 44. The method according to claim 41, wherein the fungal infection is caused by Aspergillus spp. 45. The method of claim 41, wherein the fungal infection is invasive aspergillosis.