Injectable compositions of triterpenoid antifungals encapsulated in liposomes

Abstract

To provide a composition and a method of treatment for use in a method for treating fungal infections.SOLUTION: Injectable compositions of single bilayer vesicles encapsulating enfumafungin-derived triterpenoid antifungal compounds can be administered intravenously to treat or prevent fungal infections, while having reduced local injection site reactions (ISRs). The enfumafungin-derived triterpenoid antifungal compounds are inhibitors of (1,3)-β-D-glucan synthesis and are useful in the treatment or prevention of fungal infections, such as systemic fungal infections, including those caused by Candida and Aspergillus species.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to a composition containing liposomal vesicles encapsulating a triterpenoid antifungal drug compound derived from amphomycin. More specifically, the present invention relates to a liposomal bilayer vesicle encapsulating an amphomycin-derived triterpenoid which is an inhibitor of (1,3)-β-D-glucan synthesis (or its pharmaceutically acceptable salt or hydrate), and it can be used for treating and / or preventing systemic fungal infections. The composition of the present invention has good local tolerance characteristics and is well suitable for administration by injection.

Background Art

[0002] Fungal infections are a major medical problem and most commonly appear as invasive fungal diseases (such as candidiasis, invasive aspergillosis), localized fungal infections (such as pleural empyema and abscesses localized in the abdomen, brain, lung, etc.) and mucocutaneous infections (such as oral, esophageal and vulvovaginal candidiasis). The type and extent of the infection depend on the pathogenic factors of the fungal pathogen, the host's defense and the anatomical regions involved.

[0003] Severe systemic fungal infections are more common in patients with reduced immunity, such as patients receiving chemotherapy for treating malignant tumors, or patients receiving immunosuppressants for treating chronic inflammatory conditions, or patients suffering from acquired or genetic disorders causing immunodeficiency. Despite currently available antifungal therapies, systemic fungal infections have a mortality rate of up to 50% depending on the pathogen and the underlying condition of the patient. ​​​​​​​​​​​​​​​

[0004] Enhumafungin is a living plant of the common juniper (Juniperus communis). In the fermentation of Hormonema spp. on leaves It is a hemiacetal triterpene glycoside produced by the synthesis of glycerol (U.S. Pat. No. 5,756,472). ; Pelaez et al., Systematic and Applied Microbiology, 23:333-343, 2000; Schwartz et al., JACS, 122:4882-4886, 2000; Schw artz, RE, Expert Opinion on Therapeuti c Patents, 11(11):1761-1772, 2001). Ngin is one of several triterpene glycosides that have in vitro antifungal activity. The mechanism of action of enfumafungin and other antifungal triterpenoid glycosides is ( The specific action of 1,3)-β-D-glucan synthase on fungal cell wall glucan synthesis It was confirmed that the inhibitory effect was on inhibition of cytochrome P450 synthesis (Onishi et al., Antimicrob Bial Agents and Chemotherapy, 44:368-377 , 2000; Pelaez et al., Systematic and Ap plied Microbiology, 23:333-343, 2000). 1, 3-β-D-glucan synthase is present in many types of pathogenic fungi and is therefore a widespread enzyme. It remains an attractive target for antifungal activity as it offers a broad antifungal spectrum. In addition, there is no equivalent of 1,3-β-D-glucan synthase in mammals. Based on its mechanism, it has little or no toxicity.

[0005] Various enfumafungin derivatives have been disclosed, for example, in International Patent Publication No. 200 7 / 126900 and International Patent Publication No. 2007 / 127012.

[0006] SCY-078 is a representative compound of the enfumafungin derivatives described herein, and is active against various Candida species, including drug-resistant strains, and various Aspergillus species. SCY-078 is a triterpenoid derivative substituted with a carboxylic acid and an alkylamino ether, represented by the following formula:

Chemical formula

[0007]

[0008] Without intending to be bound by theory: due to the amino functional group and the carboxylic acid functional group, SCY-078 is an amphiphilic molecule and exhibits behavior similar to that of a surfactant, such as micelle formation, binding to surface and plasma proteins.

[0009] Intravenous administration of SCY-078 is accompanied by local injection site reactions such as pain, irritation, inflammation and phlebitis (hereinafter referred to as "ISR"), which have prevented its administration via peripheral veins. The underlying mechanism of the observed ISR is not clearly understood, and without intending to be bound by theory: the observed ISR is due to the local direct interaction between SCY-078 and the vascular endothelium at the injection site when SCY-078 is introduced into the bloodstream via the vein. ​​​​​​​​​​​may be caused by.

Prior Art Documents

Patent Documents

[0010]

Patent Document 1

Patent Document 2

Patent Document 3

Non-Patent Documents

[0011]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

Summary of the Invention

Problems to be Solved by the Invention

[0012] In the present technical field, SCY-078 and another enfumafungin derivative antifungal drug are It is possible to administer intravenously via a peripheral vein with fewer and / or less severe ISRs, and it is required to improve the adaptability and convenience for caregivers and patients. This is required.

Means for Solving the Problem

[0013] The present invention provides an injectable composition of a triterpenoid compound derived from enfumafungin encapsulated in unilamellar vesicles or liposomes hydrated in an aqueous phase, which is suitable for intravenous administration via a peripheral vein. The vesicles may be composed of phospholipids and cholesterol. The pH of the aqueous phase is preferably from about 5.0 to about 7.0. Monosaccharide solutions and disaccharide solutions are examples of such aqueous phases. The injectable composition preferably contains SCY-078 or a salt thereof as an enfumafungin derivative. The injectable composition can be intravenously administered for treating and / or preventing fungal infections such as systemic fungal infections (which include systemic fungal infections caused by various Candida species or various Aspergillus species). By using the injectable composition of the present invention, local injection site reactions (such as pain, irritation, inflammation, and phlebitis) after intravenous administration can be reduced in frequency and / or degree compared to such reactions after intravenous administration of a non-liposomal composition containing the same active ingredient. The present invention further provides a method for treating or preventing a fungal infection in a patient, where

[0014]

[0015]

[0016] ​, the method involves intravenously administering an injectable composition containing unilamellar vesicles encapsulating a triterpenoid compound derived from emfumafungin. This is achieved by intravenously administering an injectable composition containing unilamellar vesicles.

[0017] The present invention further provides a method for producing a composition containing unilamellar liposomes encapsulating a triterpenoid compound derived from emfumafungin, which is suitable for injection and intravenous administration. This is achieved by intravenously administering an injectable composition containing unilamellar vesicles encapsulating a triterpenoid compound derived from emfumafungin. The present invention further provides a method for producing a composition containing unilamellar liposomes encapsulating a triterpenoid compound derived from emfumafungin, which is suitable for injection and intravenous administration.

Mode for Carrying Out the Invention

[0018] Phospholipids are amphiphilic molecules containing a hydrophilic ionizable head or "polar head" and a hydrophobic tail consisting of a long-chain fatty acid. When hydrated, multiple phospholipid molecules (optionally together with another molecule such as cholesterol) can form liposomes, which are closed bilayer concentric vesicles that enclose water or another aqueous medium. The fatty acid tails of the phospholipid molecules form part of the interior of the bilayer of the liposome. The polar heads on one surface of the membrane face the aqueous interior of the liposome, and the polar heads on the other surface of the membrane face the aqueous external environment. When hydrated, multiple phospholipid molecules (optionally together with another molecule such as cholesterol) can form liposomes, which are closed bilayer concentric vesicles that enclose water or another aqueous medium. When hydrated, multiple phospholipid molecules (optionally together with another molecule such as cholesterol) can form liposomes, which are closed bilayer concentric vesicles that enclose water or another aqueous medium. The fatty acid tails of the phospholipid molecules form part of the interior of the bilayer of the liposome. The fatty acid tails of the phospholipid molecules form part of the interior of the bilayer of the liposome. The polar heads on one surface of the membrane face the aqueous interior of the liposome, and the polar heads on the other surface of the membrane face the aqueous external environment. The polar heads on one surface of the membrane face the aqueous interior of the liposome, and the polar heads on the other surface of the membrane face the aqueous external environment.

[0019] Depending on their physicochemical properties, drugs can be incorporated into either the aqueous interior or the lipid bilayer of liposomes, where hydrophobic molecules mainly associate with the lipid bilayer and / or hydrophilic molecules are incorporated (e.g., dissolved or dispersed) into the aqueous medium inside the liposome. Depending on their physicochemical properties, drugs can be incorporated into either the aqueous interior or the lipid bilayer of liposomes, where hydrophobic molecules mainly associate with the lipid bilayer and / or hydrophilic molecules are incorporated (e.g., dissolved or dispersed) into the aqueous medium inside the liposome. Depending on their physicochemical properties, drugs can be incorporated into either the aqueous interior or the lipid bilayer of liposomes, where hydrophobic molecules mainly associate with the lipid bilayer and / or hydrophilic molecules are incorporated (e.g., dissolved or dispersed) into the aqueous medium inside the liposome. Depending on their physicochemical properties, drugs can be incorporated into either the aqueous interior or the lipid bilayer of liposomes, where hydrophobic molecules mainly associate with the lipid bilayer and / or hydrophilic molecules are incorporated (e.g., dissolved or dispersed) into the aqueous medium inside the liposome.

[0020] Liposomes can be monolayer (i.e., having one bilayer of phospholipids surrounding an aqueous core) or multilamellar (i.e., containing a number of concentric aqueous-filled bilayer vesicles). Liposomes can be monolayer (i.e., having one bilayer of phospholipids surrounding an aqueous core) or multilamellar (i.e., containing a number of concentric aqueous-filled bilayer vesicles). can be in the form of multilamellar vesicles (“MLV”). MLV can have a wide range of heterogeneous size distributions and generally, a reproducible manufacturing process is not easy. MLV has large particle sizes and is not desirable for intravenous administration. Additionally, MLV is difficult to sterilize by filtration . Injectable compositions for intravenous administration preferably have relatively small unilamellar liposomes .

[0021] Liposomes have been used to deliver drugs to target tissues and reduce systemic toxicity but their usefulness in reducing ISR that occurs locally at the site of injection or infusion has not been clearly understood. Additionally, prior to the present invention, it was not known whether it was possible to incorporate triterpenoids derived from enfumafungin into liposomes . Furthermore, particularly with respect to SCY - 078; this drug shows a high degree of protein binding and it was not known whether release or leakage of the encapsulated drug would occur upon intravenous administration of SCY - 078 into the bloodstream ; an effective liposomal composition would need to 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 injection . Using the present invention, it is possible to intravenously administer a composition of phospholipid - cholesterol vesicles encapsulating a triterpenoid antifungal drug (e.g., SCY - 078 or its pharmaceutically acceptable salts) derived from enfumafungin, and the vesicles can remain intact in the bloodstream until they are taken up by macrophages in organs of the reticuloendothelial system (e.g., liver, spleen, etc.). Additionally, signs of ISR are at extremely low concentrations of free (non - encapsulated) SCY ​​​​​​ -078 may occur, but by using the present invention, the availability of the free drug at the injection site is locally minimized, and the severity and frequency of ISR during intravenous administration of SCY-078 can be reduced. The present invention provides an approach to reducing the ISR caused by intravenous administration while providing the beneficial effects of an antifungal agent. SCY-078 (or a pharmaceutically acceptable salt thereof) or another enfumafungin-derived triterpenoid antifungal drug is encapsulated in a phospholipid (``PL'')-based system such as liposomes. By using the present invention, the free (uncapsulated) drug is minimized and the liposome composition is made suitable for commercial manufacture by encapsulating SCY-078 into liposomes with high efficiency. Furthermore, the physicochemical properties of the liposomes described herein are stable, which contributes to the storage of the composition.

[0022] The present invention provides an aqueous phase; and one or more unilamellar vesicles, where each unilamellar vesicle contains a phospholipid and cholesterol, and each contains a compound of formula (I) [Chemical formula]

[0023] wherein X is O or H, H; R e is C(O)NR f R g or a 6-membered heteroaryl group containing one or two nitrogen atoms where the heteroaryl group may be monosubstituted at a ring carbon with fluoro or chloro, or may be monosubstituted at a ring nitrogen with oxygen is good); R f , R g , R 6 and R 7 are each independently hydrogen or C1-C3 alkyl ; R 8 is C1-C4 alkyl, C3-C4 cycloalkyl or C4-C5 cycloalkyl-alkyl; R 9 is methyl or ethyl; and, R 8 and R 9 may together form a 6-membered saturated ring containing one oxygen atom) encapsulating a compound represented by formula (I) or a pharmaceutically acceptable salt or hydrate thereof] ; containing (wherein the one or more unilamellar vesicles are hydrated in the aqueous phase) an injectable composition is provided.

[0024] The pH of the aqueous phase is preferably from about 5.0 to about 7.0. Monosaccharide solutions and disaccharide solutions are examples of the aqueous phase. The injectable composition can be administered intravenously for treating and / or preventing systemic fungal infections (which include systemic fungal infections caused by various Candida species or various Aspergillus species). (Candida species) or various Aspergillus species (Aspergillu s species)

[0025] The present invention further provides a method for treating and / or preventing a fungal infection, wherein the method comprises administering intravenously an injectable composition containing one or more unilamellar vesicles (wherein each unilamellar vesicle contains phospholipid and cholesterol, and each encapsulates a compound represented by formula (I) or a pharmaceutically acceptable salt or hydrate thereof). method comprises administering intravenously an injectable composition containing one or more unilamellar vesicles (wherein each unilamellar vesicle contains phospholipid and cholesterol, and each encapsulates a compound represented by formula (I) or a pharmaceutically acceptable salt or hydrate thereof). and each encapsulates a compound represented by formula (I) or a pharmaceutically acceptable salt or hydrate thereof), and wherein the injectable composition contains)​​ By intravenous administration. Furthermore, the present invention provides an injection for treating or preventing fungal infections In the preparation of a pharmaceutically injectable agent, the use of a vesicle encapsulating a compound represented by formula (I) or a pharmaceutically acceptable salt or Hydrate thereof is also provided.

[0026] The present invention further provides An aqueous phase; and One or more unilamellar vesicles [wherein each of the unilamellar vesicles contains phospholipid and cholesterol, and each contains a compound represented by formula (Ia) —

Chemical formula

[0027] 〔wherein the substituents are as given in formula (I)〕 Or a pharmaceutically acceptable salt or hydrate thereof encapsulated] ; Containing (wherein the one or more unilamellar vesicles are hydrated in the aqueous phase) an injectable Composition is also provided.

[0028] The pH of the aqueous phase is preferably from about 5.0 to about 7.0. Monosaccharide solutions and disaccharide solutions are Examples of the aqueous phase. The injectable composition can be administered intravenously for treating and / or preventing fungal infections such as systemic fungal infections (which include systemic fungal infections caused by various Candida species or various Aspergillus species) (Candida species) or various Aspergillus species (Aspergillu s species). Infections can be treated and / or prevented.

[0029] The present invention further provides a method for treating and / or preventing fungal infections, wherein the method comprises one or more unilamellar vesicles (wherein each of the unilamellar vesicles contains phospholipid and cholesterol — comprising a rule, and each of which encapsulates a compound represented by formula (Ia) or a pharmaceutically acceptable salt or hydrate thereof). The injectable composition is administered intravenously. Further, the present invention provides the use of a vesicle encapsulating a compound represented by formula (Ia) or a pharmaceutically acceptable salt or hydrate thereof in the preparation of an injectable agent for treating or preventing fungal infections.

[0030] In Embodiment 1: X is H, H, and the other substituents are as given in formula (I).

[0031] In Embodiment 2: R e is pyridyl or pyrimidinyl (wherein the pyridyl or pyrimidinyl may be monosubstituted with fluoro or chloro at a ring carbon or may be monosubstituted with oxygen at a ring nitrogen), and the other substituents are as in Embodiment 1 or as given in formula (I).

[0032] In Embodiment 3: R e is 4-pyridyl, and the other substituents are as in Embodiment 1 or as given in formula (I).

[0033] In Embodiment 4: R e is C(O)NH2 or C(O)NH(C1-C3 alkyl) and the other substituents are as in Embodiment 1 or as given in formula (I).

[0034] In Embodiment 5: R 8 is C1-C4 alkyl, and R 9 is methyl ​​​​; and the other substituents are as given in Embodiment 1, Embodiment 2, Embodiment 3 or Embodiment 4 or as given in formula (I).

[0035] In Embodiment 6: R 8 is t-butyl, R 9 is methyl; and the other substituents are as given in Embodiment 1, Embodiment 2, Embodiment 3 or Embodiment 4 or formula (I) as given.

[0036] In Embodiment 7: R 6 and R 7 are each independently hydrogen or methyl, and , the other substituents are as given in Embodiment 1, Embodiment 2, Embodiment 3, Embodiment 4, Embodiment 5 or Embodiment 6 or formula (I).

[0037] In Embodiment 1': X is H, H, and the other substituents are as given in formula (Ia) as given.

[0038] In Embodiment 2': R e is pyridyl or pyrimidinyl (wherein the pyridyl or pyr imidinyl may be monosubstituted with fluoro or chloro at the ring carbon, or monosubstituted with oxygen at the ring nitrogen), and the other substituents are as given in Embodiment 1' or formula (Ia).

[0039] In Embodiment 3': R e is 4-pyridyl, and the other substituents are as given in Embodiment 1' or formula (Ia).

[0040] In Embodiment 4': R eis C(O)NH2 or C(O)NH(C1-C3 alkyl ), and the other substituents are as given in Embodiment 1' or Formula (Ia) .

[0041] In Embodiment 5': R 8 is C1-C4 alkyl, and R 9 is methyl ; and the other substituents are as given in Embodiment 1', Embodiment 2', Embodiment 3' or Embodiment 4' or Formula (Ia).

[0042] In Embodiment 6': R 8 is t-butyl, R 9 is methyl; and the other substituents are as given in Embodiment 1', Embodiment 2', Embodiment 3' or Embodiment 4' or Formula (Ia).

[0043] In Embodiment 7': R 6 and R 7 are each independently hydrogen or methyl, and also, the other substituents are as given in Embodiment 1', Embodiment 2', Embodiment 3', Embodiment 4', Embodiment 5' or Embodiment 6' or Formula (Ia).

[0044] In a preferred embodiment, the present invention is an aqueous phase; and one or more unilamellar vesicles [wherein each unilamellar vesicle contains phospholipid and cholesterol and each contains a compound represented by Formula (II)

Chemical formula

[0045] represented by (1S,4aR,6aS,7R,8R,10aR,10bR ,(12aR,14R,15R)-15-[[2-Amino-2,3,3-trimethylbutyl oxy]-8-[(1R)-1,2-dimethylpropyl]-14-[5-(4-pyrid nyl)-1H-1,2,4-triazol-1-yl]-1,6,6a,7,8,9,1 0,10a,10b,11,12,12a-dodecahydro-1,6a,8,10a-tetra methyl-4H-1,4a-propano-2H-phenanthro[1,2-c]pyran-7- carboxylic acid (hereinafter referred to as SCY-078 in this specification) or a pharmaceutically acceptable salt or hydrate thereof encapsulated]; An injectable composition comprising (wherein the one or more unilamellar vesicles are hydrated in the aqueous phase) is provided.

[0046] The pH of the aqueous phase is preferably from about 5.0 to about 7.0. Solutions of monosaccharides and disaccharides are examples of the aqueous phase. The injectable composition can be administered intravenously for treating and / or preventing systemic fungal infections (which include systemic fungal infections caused by various Candida species or various Aspergillus species). infections).

[0047] The present invention further provides a method for treating and / or preventing a fungal infection, wherein the method comprises intravenously administering the injectable composition comprising one or more unilamellar vesicles (wherein each unilamellar vesicle contains phospholipid and cholesterol and each encapsulates a compound of formula (II) or a pharmaceutically acceptable salt or hydrate thereof). Further, the present invention relates to an injection for treating or preventing a fungal infection. Use of the vesicles encapsulating a compound represented by formula (II) or a pharmaceutically acceptable salt or hydrate thereof in the preparation of an injectable agent is also provided.

[0048] In another preferred embodiment, the present invention provides an aqueous phase; and one or more unilamellar vesicles [wherein each unilamellar vesicle contains a phospholipid and cholesterol and each contains a compound represented by formula (IIa)

Chemical formula

[0049] (which is (1S,4aR,6aS,7R,8R,10aR,10bR ,12aR,14R,15R)-15-[[(2R)-2-amino-2,3,3-trimethyl butyl]oxy]-8-[(1R)-1,2-dimethylpropyl]-14-[5-( 4-pyridinyl)-1H-1,2,4-triazol-1-yl]-1,6,6a,7, 8,9,10,10a,10b,11,12,12a-dodecahydro-1,6a,8,1 0a-tetramethyl-4H-1,4a-propano-2H-phenanthro[1,2-c]py ran-7-carboxylic acid) or a pharmaceutically acceptable salt or hydrate thereof encapsulated]; and (wherein the one or more unilamellar vesicles are hydrated in the aqueous phase) an injectable composition is provided. The pH of the aqueous phase is preferably from about 5.0 to about 7.0. Monosaccharide solutions and disaccharide solutions are

[0050] examples of the aqueous phase. The injectable composition is for systemic fungal infections (which are various Candida species ​(Candida species) or various species of Aspergillus (Aspergillu s species), including systemic fungal infections caused by such fungal infections, can be administered intravenously for the treatment and / or prevention thereof.

[0051] The present invention further provides a method for treating and / or preventing a fungal infection, wherein the method comprises intravenously administering an injectable composition comprising one or more unilamellar vesicles, wherein each of the unilamellar vesicles contains phospholipids and cholesterol, and each encapsulates a compound represented by formula (IIa) or a pharmaceutically acceptable salt or hydrate thereof. Further, the present invention provides the use of such vesicles encapsulating a compound represented by formula (IIa) or a pharmaceutically acceptable salt or hydrate thereof in the preparation of an injectable agent for treating or preventing a fungal infection. In a preferred embodiment, the phosphate salt of the compound represented by formula (I), formula (Ia), formula (II) or formula (IIa) is used or administered as described herein. In a preferred embodiment, the citrate salt of the compound represented by formula (I), formula (Ia), formula (II) or formula (IIa) is used or administered as described herein. The present invention further provides, for treating or preventing a fungal infection in a subject,

[0052] one or more unilamellar vesicles, wherein each of the unilamellar vesicles encapsulates a compound represented by formula (I), formula (Ia), formula (II) or formula (IIa) or a pharmaceutically acceptable salt or hydrate thereof.

[0053] In a preferred embodiment, the citrate salt of the compound represented by formula (I), formula (Ia), formula (II) or formula (IIa) is used or administered as described herein. In a preferred embodiment, the citrate salt of the compound represented by formula (I), formula (Ia), formula (II) or formula (IIa) is used or administered as described herein.

[0054] The present invention further provides, for treating or preventing a fungal infection in a patient (subject), for one or more unilamellar vesicles, wherein each of the unilamellar vesicles encapsulates a compound represented by formula (I), formula (Ia), formula (II) or formula (IIa) or a pharmaceutically acceptable salt or hydrate thereof. cyclized); and, a pharmaceutically acceptable carrier, adjuvant or vehicle; To provide the use of an injectable composition comprising.

[0055] In the description of the compounds in the embodiments described above, the indicated substitutions are included only within the range in which the substituents result in stable compounds that are compatible with the definition.

[0056] Compounds represented by formula (I), formula (Ia), formula (II) and formula (IIa) and their pharmaceutically acceptable salts and / or hydrate forms include one or more of the following yeasts and other fungi exhibit antibacterial (e.g., antifungal) activity: Acremonium, Absidia (e.g., Absidia corymbifera), Alternaria, Aspergillus (e.g., Aspergillus clavatus, Aspergillus flavus, Aspergillus fumigatus, Aspergillus nidulans, Aspergillus niger, Aspergillus terreus, and Aspergillus versicolor), Bipolaris, Blastomyces (e.g., Blastomyces dermatitidis) ium), Absidia (e.g., Absidia corymbifera), Alternaria bsidia corymbifera)), Alternaria ), Aspergillus (e.g., Aspergillus clavatus s), Aspergillus flavus ergillus flavus), Aspergillus fumigatus us fumigatus), Aspergillus nidulans nidulans), Aspergillus niger , Aspergillus terreus, and, Asper ergillus versicolor)), Bipolaris laris, Blastomyces (e.g., Blastomyces dermatitidis ​is)), Blastoschizomyces (e.g., Blas toschizomyces capitatus), Candida (e.g., Candida albicans, Cand ida albicans), Candida glabrata, Candida guilliermondii, Cand ida glabrata), Candida guilliermondii, Candida kefyr, Cand ida guilliermondii), Candida kefyr, Candida krusei, Cand ida krusei), Candida lusitaniae, Candida parapsilosis, Cand ida lusitaniae), Candida parapsilosis, Candida pseudotropicalis, Cand ida pseudotropicalis), Candida stellatoidea, Candida tropica lis), Candida utilis, Candida lipolytica, Candida famata, and C andida rugosa), Cladosporium (e.g., Cladosporium carrionii, and C ladosporium trichloides), Coccidioides (e.g., Coccidioides immitis), Cryptococcus (e.g., Cryptococcus neoformans), Cryptococcus (e.g., Cryptococcus neoformans), Cryptococcus (e.g., Cryptococcus neoformans), Cladosporium (e.g., Cladosporium carrionii, and C ladosporium trichloides), Coccidioides (e.g., Coccidioides immitis), Cryptococcus (e.g., Cryptococcus neoformans), Cryptococcus (e.g., Cryptococcus neoformans), Cryptococcus (e.g., Cryptococcus neoformans), Cryptococcus neoformans, Curvularia Cunninghamella (e.g., Cunninghamella elegans), Dermatophyte, Exophiala ( e.g., Exophiala dermatitidis and Exophiala spinifera), Epidermophyton (e.g., Epidermophyton floccosum), Fonsecaea (e.g., Fonsecaea pedrosoi), Fusarium (e.g., Fusarium solani), Geotrichum (e.g., Geotrichum candidum and Geotrichum clavatum), Histoplasma (e.g., Histoplasma capsulatum var . capsulatum), Malassezia (e.g., Malassezia furfur), Microsporum (e.g., Microsporum canis ), and ), anis), and Microsporum gypse um), Mucor, Paracoccidi oides) (for example, Paracocci dioides brasiliensis), Penicilli um) (for example, Penicillium marnef fei), Phialophora, Pityrosporum ovale , Pneumocyst is) (for example, Pneumocystis carin ii), Pseudallescheria (for example, Pseudallescheria boydii) , Rhizopus (for example, Rhizopus microsporus var. rhizop odiformis), and Rhizopus oryzae ), Saccharomyces (for example, Saccharomyces cere visiae), Scedosporium (for example, Scedosporium apiosperum ), Scopular iopsis, Sporothrix (for example, Sporothrix schenckii), Trichoderma Aspergillus, Trichophyton (e.g., Tri chophyton mentagrophy tes), and Trichophyton rubrum ), and Trichosporon (e.g., Trichosporon · asahii, Trichosporon beigelii (Trichosporon beigelii), and Trichosporon cutaneum (Trichosporon cutaneum)). The compound is not only useful only against organisms that cause systemic human pathogenic fungal infections, but also useful against organisms that cause any superficial fungal infections such as Trichoderma spp. and other Candida spp. ichoderma spp.) and other Candida spp. The compound is useful against organisms that cause any superficial fungal infections. The compound is effective against various Candida species and various Aspergillus species.

[0057] Compounds represented by formula (I), formula (Ia), formula (II) and formula (IIa) and their pharmaceutically acceptable salts and / or hydrate forms are, in view of their antifungal activities, useful for treating and / or preventing one or more of various superficial, cutaneous, mucocutaneous, subcutaneous and systemic fungal infections in the vulva, vagina, skin, eyes, hair, nails, oral mucosa, gastrointestinal tract, bronchi, lungs, pleura, peritoneum, endocardium, brain, meninges , urinary organs, vagina, oral cavity, kidneys, heart, external auditory canal, bones, nasal cavity, paranasal sinuses, spleen, liver, subcutaneous tissue , lymphatic vessels, joints, muscles, tendons, interstitial cells in the lungs and blood, etc. preventing.​​

[0058] Compounds represented by formula (I), formula (Ia), formula (II) and formula (IIa) and their pharmaceutically acceptable salts and / or hydrate forms are useful for preventing and treating vulvovaginal candidiasis (VVC), cutaneous mycoses (e.g., trichophytosis, ringworm infections or tinea infections), paronychia, vitiligo, erythrasma, intertrigo, fungal erythema, candidal vulvitis, candidal balanitis, otitis externa, candidiasis (cutaneous and mucocutaneous) , chronic mucocutaneous candidiasis (e.g., thrush and vaginal candidiasis), cryptococcosis, geotrichum osis, trichosporonosis, aspergillosis, penicilliosis, fusariumosis, zygomycosis, sporotrichosis, chromomycosis, coccidioides osis, histoplasmosis, blastomycosis, paracoccidioides osis, pseudallescheria, mycetoma, filamentous keratitis, otomycosis, pneumocystis, fungal abscess, fungal empyema and fungemia, etc. It is useful for preventing and treating one or more of various infections. Compounds represented by formula (I), formula ( Ia), formula (II) and formula (IIa) and their pharmaceutically acceptable salts and / or hydrate forms can also be used as a prophylactic agent for preventing systemic and local fungal infections. Use as a prophylactic agent can be appropriate, for example, as part of an elective gastrointestinal decontamination dosing regimen for preventing infections in patients with compromised immunity (e.g., AIDS patients, cancer patients undergoing treatment, or transplant patients). During treatment with antibiotics, it may also be desirable to prevent overgrowth of fungi in some disease syndromes or iatrogenic conditions.

[0059] Compounds represented by formula (I), formula (Ia), formula (II) and formula (IIa) and their pharmaceutically acceptable salts and / or hydrate forms are described in U.S. Patent No. 8,188,085 (the contents of this patent are hereby incorporated by reference in their entirety) by the synthetic methods disclosed therein.

[0060] As used herein, the term "alkyl" refers to any straight-chain or branched-chain alkyl group having the number of carbon atoms within the specified range. Thus, for example, " C 1-6 alkyl" (or, "C1-C6 alkyl") refers to all isomers of hexyl alkyl and pentyl alkyl, as well as n-, iso-, sec- and t-butyl, n- and isopropyl, ethyl and methyl. As another example, " 1-4 C alkyl" refers to n-

[0061] cycloalkyl" refers to any cyclic ring of an alkane having the number of carbon atoms within the specified range. Thus, for example, " 3-4 C cycloalkyl" (or, "

[0062] The term "cycloalkyl-alkyl" (or, synonymously, "alkyl-cycloalkyl" ) when used herein refers to a system containing the alkyl moiety described above and also containing the cycloalkyl moiety described above. The bond to " cycloalkyl-alkyl" (or, "alkyl-cycloalkyl") can be to the cycloalkyl moiety or It can be through any of the alkyl moieties. In the "cycloalkyl-alkyl" system the number of carbon atoms specified refers to the total number of carbon atoms in both the alkyl moiety and the cycloalkyl moiety. Examples of C4-C5 cycloalkyl-alkyl include, but are not limited to, methylcyclopropyl, dimethylcyclopropyl, methylcyclobutyl, ethylcyclopropyl, cyclopropylmethyl, cyclopropylethyl, and cyclobutylmethyl, etc.

[0063] The term "halogen" (or "halo") represents fluorine, chlorine, bromine, and iodine (alternatively, also referred to as fluoro, chloro, bromo, and iodo).

[0064] As used herein, the term "or" means alternative, which may be combined where appropriate.

[0065] Unless otherwise expressly stated, all ranges described herein are inclusive. For example, a heterocyclic ring described as containing "1 to 4 heteroatoms" means that the ring may contain 1 atom, 2 atoms, 3 atoms, or 4 heteroatoms. It should also be understood that any range described herein includes all sub-ranges within that range. Thus, for example, a heterocyclic ring described as containing "1 to 4 heteroatoms" is intended to include, as aspects thereof, heterocyclic rings containing 2 to 4 heteroatoms, 3 or 4 heteroatoms, 1 to 3 heteroatoms, 2 or 3 heteroatoms, 1 or 2 heteroatoms, 1 heteroatom, 2 heteroatoms, etc.

[0066] ​​​​ All of the various cycloalkyls and heterocyclic / heteroaryl rings and heterocyclic / heteroaryl ring systems defined herein can be attached to the remainder of the compound with any ring atom (i.e., any carbon atom or any heteroatom), provided that the resulting compound is stable. Suitable 5- or 6-membered heteroaromatic rings include, but are not limited to, pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, and triazolyl.

[0067] A "stable" compound is one that can be prepared and isolated and that remains essentially unchanged or is made to remain unchanged over a period of time sufficient for the use of the compound for the purposes described herein (e.g., therapeutic or prophylactic administration to a subject). When reference is made to a compound, the reference also includes stable complexes of the compound (e.g., stable hydrates).

[0068] As a result of the choice of substituents and substitution patterns, certain of the compounds represented by formula (I), formula (Ia), formula (II), and formula (IIa) can have asymmetric centers and can exist as mixtures of stereoisomers or as individual diastereomers or enantiomers. Unless otherwise indicated, all isomeric forms of these compounds (as well as their pharmaceutically acceptable salts and / or hydrate forms), whether isolated or in admixture, are within the scope of the invention. Also, the tautomeric forms of the compounds shown (as well as their pharmaceutically acceptable salts and / or hydrate forms) ​​​​​​​​​​​​​​​​ is also included within the scope of the present invention.

[0069] If any variable part appears two or more times in any of the components or in formula (I), formula (Ia), formula (II) or formula (IIa ), the definition thereof in each occurrence is independent of its definition in all other occurrences. Also, combinations of substituents and / or variable parts are only permitted if such combinations result in stable compounds.

[0070] The term "substituted" includes mono- and polysubstitution (including multiple substitutions at the same position) with the specified substituents, within the range in which such mono- and polysubstitution is chemically possible. Unless otherwise specified, substitution with the specified substituents is possible on any atom in a ring ( e.g., aryl, cycloalkyl, heteroaryl or heterocyclyl), provided that such ring substitution is chemically possible and results in a stable compound. For example, aryl, cycloalkyl, heteroaryl or heterocyclyl), provided that such ring substitution is chemically possible and results in a stable compound. atom, provided that such ring substitution is chemically possible and results in a stable compound. compound.

[0071] A bond ending with a wavy line is used herein to indicate the attachment point of a substituent or substructure. This usage is illustrated by the following examples: This usage is illustrated by the following examples:

Chemical formula

[0072] The compounds represented by formula (I), formula (Ia), formula (II) and formula (IIa) and their pharmaceutically acceptable salts and / or hydrates are also useful in the preparation and conduct of screening assays for antifungal compounds. For example, the compounds are further antifungal Useful for isolating mutants, which are excellent screening means for confirming compounds It is.

[0073] The compounds represented by formula (I), formula (Ia), formula (II) and formula (IIa) can, where appropriate be administered in the form of "pharmaceutically acceptable salts" or hydrates. However others, other salts may be useful in the preparation of the compounds or their pharmaceutically acceptable salts For example, when the compound contains a basic amine group, they can, conveniently, be isolated as tri fluoroacetate (e.g., by HPLC purification). Converting the trifluoro acetate to another salt (which includes pharmaceutically acceptable salts) can be achieved by many standard methods known in the art. For example, the desired salt can be produced using an appropriate ion exchange resin. Alternatively, converting the trifluoro acetate to the free amine of the parent compound can be achieved by standard methods known in the art such as neutralization with an appropriate inorganic base such as NaHCO3. Then, by reacting the free base with an appropriate organic acid or inorganic acid, another desired amine salt can be prepared in a conventional manner. Representative pharmaceutically acceptable quaternary ammonium salts can include the following: hydrochloride, sulfate, phosphate , carbonate, acetate, tartrate, citrate, malate, succinate, lactate, stearate , fumarate, hippurate, maleate, gluconate, ascorbate, adipate , glycerophosphate, glutamate, glucuronate (glucoronate e), propionate, benzoate, mesylate, tosylate, oleate, lactobionate ​Oxalates, lauryl sulfates, besylates, caprylates, isethionates, gentisates , malonates, napsylates, edisylates, pamoates, xinafoates, napadisylates , hydrobromides, nitrates, oxalates, cinnamates, mandelates, undecylenates, and cancilates. Many of the compounds represented by formula (I), formula (Ia), formula (II) and formula (IIa) have an acidic carboxylic acid moiety, in which case suitable pharmaceutically acceptable salts can include alkali metal salts (e.g., sodium or potassium salts); alkaline earth metal salts (e.g., calcium or magnesium salts); and salts formed using suitable organic ligands (e.g., quaternary ammonium salts). The present invention includes within its scope the use of prodrugs represented by formula (I), formula (Ia), formula (II) and formula (IIa). Generally, such prodrugs are functional derivatives of the compounds, which are readily convertible in vivo to the required compounds. Thus in the treatment methods of the present invention, the term “administering” includes treating the various conditions described with the specifically disclosed compounds, or treating with compounds that are convertible in vivo to the compounds identified after administration to a patient. Conventional methods for the selection and preparation of suitable prodrug

[0074] derivatives are described, for example, in ““Design of P rodrugs,” ed. H. Bundgaard, Elsevier, 19 85” (which is incorporated herein by reference in its entirety). Metabolites of the compounds represented by formula (I), formula (Ia), formula (II) and formula (IIa) include the active chemical species produced when the compounds are introduced into a biological environment. (I), formula (Ia), formula (II) and formula (IIa) include the active chemical species produced when the compounds are introduced into a biological environment. rodrugs,” ed. H. Bundgaard, Elsevier, 19 85” (which is incorporated herein by reference in its entirety). Metabolites of the compounds represented by formula (I), formula (Ia), formula (II) and formula (IIa) include the active chemical species produced when the compounds are introduced into a biological environment. The metabolites of the compounds represented by formula (I), formula (Ia), formula (II) and formula (IIa) include the active chemical species produced when the compounds are introduced into a biological environment.

[0075] The term “administer” and variations thereof (e.g., “administering” a compound) mean providing a particular compound or a particular prodrug of that compound to a subject in need of treatment. When a compound represented by Formula (I), Formula (Ia), Formula (II), or Formula (IIa), or a pharmaceutically acceptable salt thereof, or a hydrate or prodrug thereof is administered in combination with a second active agent (e.g., another antifungal / antibacterial agent useful for treating a fungal / bacterial infection), “administer” and variations thereof are each understood to encompass co-administering and sequentially administering the compound (or a salt, hydrate, or prodrug thereof) and the other active agent. When used herein, the term “composition” is intended to encompass a composition containing the specified ingredients, and any product directly or indirectly resulting from combining the specified ingredients. “Pharmaceutically acceptable” means that the components of the pharmaceutical composition must be compatible with each other and not harmful to the recipient. The term “subject” (alternatively, also referred to herein as “patient”) represents an animal (preferably a mammal, most preferably a human) that is the subject of treatment, observation, or experiment when used herein. The term “effective amount” when used herein means an amount that is sufficient for a researcher, veterinarian, physician, or other

[0076]

[0077]

[0078]

[0079] ​​​​​​​​​​​​The amount of an active ingredient or a medicament that elicits a biological or medical response in a tissue, system, animal, or human as required by a clinician. In one embodiment, an "effective amount" can be a "therapeutically effective amount" that alleviates the symptoms of a disease or condition in a subject. In another embodiment, an "effective amount" can be a "prophylactically effective amount" for preventing the symptoms of a disease or condition to be prevented or reducing the likelihood of onset. This term can also further mean an inhibitory effective amount of the enfamafungin derivative that inhibits (1,3)-β-D-glucan synthase and thereby is sufficient to elicit the required response. References to "treating," "treatment," "treat," and variations thereof generally mean a treatment that, after administration, results in the alleviation or improvement of one or more signs or symptoms associated with a fungal infection, or a treatment that results in the eradication of the fungus causing the infection, or any combination of these results. For the purpose of preventing or treating a fungal infection, a compound represented by formula (I), formula (Ia), formula (II), or formula (IIa) (optionally in the form of a salt or hydrate) can be administered by conventional methods available for use in combination with a medicament. For the purpose of preventing or treating a fungal infection occurring in a state or anatomical region having an acidic pH, a compound represented by formula (I), formula (Ia), formula (II), or formula (IIa) (optionally in the form of a salt or hydrate) can be administered alone as an individual therapeutic agent or together with one or more other antifungal agents as a combination of therapeutic agents (sequentially or simultaneously).

[0080] References to "treating," "treatment," "treat," and variations thereof generally mean a treatment that, after administration, results in the alleviation or improvement of one or more signs or symptoms associated with a fungal infection, or a treatment that results in the eradication of the fungus causing the infection, or any combination of these results.

[0081] For the purpose of preventing or treating a fungal infection, a compound represented by formula (I), formula (Ia), formula (II), or formula (IIa) (optionally in the form of a salt or hydrate) can be administered by conventional methods available for use in combination with a medicament.

[0082] For the purpose of preventing or treating a fungal infection occurring in a state or anatomical region having an acidic pH, a compound represented by formula (I), formula (Ia), formula (II), or formula (IIa) (optionally in the form of a salt or hydrate) can be administered alone as an individual therapeutic agent or together with one or more other antifungal agents as a combination of therapeutic agents (sequentially or simultaneously). ​​​​​​​​​​​) can be administered.

[0083] For the purpose of preventing or treating fungal infections, a compound represented by formula (I), formula (Ia), formula (II) or formula (I Ia) (optionally in the form of a salt or hydrate) can be administered together with a pharmaceutically acceptable carrier selected based on the selected administration route and standard pharmaceutical practice.

[0084] For example, the compounds represented by formula (I), formula (Ia), formula (II) and formula (IIa) and their pharmaceutically acceptable salts and / or hydrate forms can be administered by one or more of the following routes: in unit dosage form of a pharmaceutical composition containing an effective amount of the compound and a conventional non-toxic pharmaceutically acceptable carrier, adjuvant and vehicle, orally , parenterally (which includes subcutaneous injection, intravenous, intramuscular, intralesional injection or infusion techniques ), by inhalation (e.g., nasal or oral inhalation sprays, aerosols from metered-dose inhalers and dry powder inhalers), by nebulizer, ophthalmically, topically, transdermally, or rectally . Liquid preparations suitable for oral administration (such as suspensions, syrups and elixirs, etc.) can be prepared according to techniques known in the art, and conventional media such as water, glycols, oils and alcohols can be used. Solid preparations suitable for oral administration (such as 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 and disintegrants can be used. Parenteral compositions can be prepared according to techniques known in the art, and typically, a sterile carrier is used as . Solid preparations suitable for oral administration (such as 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 and disintegrants can be used. Parenteral compositions can be prepared according to techniques known in the art, and typically, a sterile carrier is used and is typically a sterile Water is used and, optionally, another component such as a solubilizing agent is used. Injectable solutions can be prepared according to methods known in the art, where the carrier contains a physiological saline solution, a glucose solution, or a solution containing a mixture of physiological saline and glucose .

[0085] Further explanation of the methods suitable for use in the preparation of pharmaceutical compositions and the components suitable for use in such compositions is provided in " " Remington’s Pharmace utical Sciences , 20 th edition, edited by A. R. Gennaro, Mack Publishing Co., 200 0".

[0086] The compounds represented by formula (I), formula (Ia), formula (II) and formula (IIa) and their pharmaceutically acceptable salts and / or hydrate forms can be administered, for example, orally or intravenously, for example , per day, per kg of body weight of a mammal (e.g., a human), within a dosage range of 0.001 - 1000 m g, in a single dose or divided doses. One example of the dosage range is, in a single dose or divided doses, orally or intravenously, per day, per kg of body weight , 0.01 - 500 mg. Another example of the dosage range is, in a single dose or divided doses, orally or intravenously, per day, per kg of body weight 0.1 - 50 mg. For oral administration , the composition may, for example, contain 1.0 - 1000 milligrams of the active ingredient, in particular 1, 5, 10, 15, 20, 25, 50, 75, 100, 150, 200, 250, 300, 400, 500, 600, 75 500, 600, 75 It can be provided in the form of tablets or capsules containing 0 and 1000 milligrams of the active ingredient. The specific dosage level and frequency of administration for any particular patient can vary, and they will depend on various factors including the activity of the specific compound being used, the metabolic stability and duration of action of that compound, age, weight, general health, sex, diet, method and time of administration, rate of excretion, drug combinations, the severity of the particular condition, and the host being treated, etc.

[0087] The antifungal activity of the compounds can be demonstrated by various assays known in the art, for example, their minimum inhibitory concentration (MIC) against yeast and the minimum effective concentration (MEC) against filamentous fungi and dermatophytes in broth microdilution assays, or by in vivo evaluation of their anti-Candida and anti-Aspergillus activity in mouse or rabbit models. The compounds represented by formula (I) described in the examples of U.S. Patent No. 8,188,085 generally inhibit the growth of Candida spp. in the range of <0.03 - 32 μg / mL, or result in an MEC against Aspergillus fumigatus in the range of <0.03 - 32 μg / mL.

[0088] Unilamellar vesicles or liposomes encapsulating the triterpenoid antifungal drug compounds derived from enfumafungin according to the present invention preferably have an average particle size (vesicle diameter) of about 150 nm or less, more preferably about 100 nm or less, or about 70 - about 80 nm. It has. The vesicles are hydrated in a suitable aqueous phase such as water containing monosaccharides or disaccharides. Any aqueous phase with low ionic strength is suitable for hydrating the vesicles. The aqueous phase is preferably isotonic. The pH of the aqueous phase should be from about 4.0 to about 8.0, preferably from about 5.0 to about 7.0 .

[0089] Methods for producing unilamellar vesicles or liposomes include methods that provide sufficient energy and shear force (e.g., sonication, high-pressure homogenization, microfluidic mixing, etc.) . The unilamellar vesicles described herein are hydrated suspensions containing an antifungal drug derived from enfumafungin (e.g., a compound represented by formula (I), formula (Ia), formula (II) or formula (IIa ), or a pharmaceutically acceptable salt or hydrate thereof), phospholipids and cholesterol . They can be obtained by subjecting them to sonication, microfluidic mixing and / or homogenization. In a preferred embodiment, 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 at a molar concentration of 1:7:2.5 of SCY-0 78:PL:CHOL, and then subjected to sonication, microfluidic mixing, homogenization, and / or another process that promotes the formation of unilamellar vesicles containing phospholipids and cholesterol, thereby encapsulating SCY-078 (or a salt or hydrate thereof) inside the vesicles. The citrate salt of SCY-078 is a preferred drug for encapsulation in the unilamellar vesicles described herein.

[0090] Form the single bilayer vesicles described herein using a single type of phospholipid or use a combination of two or more different types of phospholipids to form a single bilayer vesicle. Suitable phospholipids for use in forming the single bilayer vesicles described herein include, but are not limited to, synthetically derived or naturally occurring phosphatidylcholine ("PC"), phosphatidic acid (" PA"), phosphatidylserine ("PS"), phosphatidylethanolamine ("P E") and phosphatidylglycerol ("PG").

[0091] The phospholipids used in forming the single bilayer vesicles described herein are preferably PC and PG used in combination.

[0092] Suitable PCs include, but are not limited to, synthetically derived dipalmitoylphosphatidyl choline ("DPPC"), distearoylphosphatidylcholine ("DSPC"), dilauroylphosphatidylcholine ("DLPC") and dimyristoylphosphatidyl choline ("DMPC"), as well as PCs from natural sources such as egg PC and soybean PC.

[0093] Suitable PGs include, but are not limited to, synthetically derived dipalmitoylphosphatidyl glycerol ("DPPG"), distearoylphosphatidylglycerol ("D SPG"), dilauroylphosphatidylglycerol ("DLPG") and dimyristoyl phosphatidylglycerol ("DMPG"), as well as PGs from natural sources such as egg PG.

[0094] ​​ Preferably, the phospholipid content of the vesicles encapsulating the enfumafungin derivative according to the present invention is 50 to 80 mol%.

[0095] Cholesterol can stabilize the lipid bilayer membrane and prevent leakage of the encapsulated drug, but an excessive amount of cholesterol may have an adverse effect on the encapsulation efficiency. . Preferably, the cholesterol content of the vesicles encapsulating the enfumafungin derivative according to the present invention is 10 to 30 mol%.

[0096] Preferably, the drug content of the vesicles encapsulating the enfumafungin derivative according to the present invention is 5 to 12 mol%.

[0097] The unilamellar vesicles encapsulating the triterpenoid antifungal drug compound derived from enfumafungin are hydrated in an aqueous phase. Preferably, the aqueous phase contains one or more monosaccharides (e.g., glucose, fructose, and galactose) and / or one or more disaccharides (e.g., sucrose, trehalose, and lactose), which render the injectable composition (unilamellar vesicles encapsulating the drug, hydrated in the aqueous phase) isotonic and can improve the stability of the bilayer membrane. When monosaccharides are used in the aqueous phase, they are preferably present in an amount of about 4 to 6% (w / v) with respect to the injectable composition (vesicles encapsulating the drug, hydrated in the aqueous phase). When disaccharides are used in the aqueous phase, they are preferably present in an amount of about 8 to 10% (w / v) with respect to the injectable composition (vesicles encapsulating the drug, hydrated in the aqueous phase). If they are used, they are preferably present in an amount of about 8 to 10% (w / v) with respect to the injectable composition (vesicles encapsulating the drug, hydrated in the aqueous phase).

[0098] Preferably, the injectable composition (single bilayer vesicles encapsulating the drug, in the aqueous phase) The concentration of encapsulated drug, expressed as mg / mL (hydrated), is 0.01 The concentration is preferably from 0.1 to 50 mg / mL, and more preferably from 0.1 to 5 mg / mL.

[0099] In order to prepare a lipid dispersion containing an antifungal drug derived from enfumafungin, a phospholipid The lipids and cholesterol are dissolved in a C1-C5 alcohol (preferably methanol or ethanol) ), and the solution is dissolved in an organic solvent such as enfumafungin-derived antifungal agent. Alternatively, the antifungal agent is added to a solution of phosphorus in an organic solvent such as ethanol. The lipid and cholesterol solutions can be added directly to the reaction mixture in a suitable reaction vessel, such as a round-bottom flask. In a reaction vessel, the solvent is evaporated (optionally under reduced pressure) to leave a fat containing the antifungal agent. A homogeneous dispersion is obtained. The solvent is removed by another method, for example by using a spray dryer. Advantageously, the lipid fraction containing the antifungal agent derived from enfumafungin may be The dispersion is stable and can be stored and used for any desired period of time. It is convenient in that it can be hydrated at a point and subjected to a separate process to form liposomes. It makes sex possible.

[0100] The lipid dispersion containing the antifungal agent is preferably a water-soluble lipid dispersion containing mono- or disaccharide sugars. The mixture is hydrated in a liquid to form a hydrated suspension. The hydrated suspension is mixed with a high shear mixer. (e.g., at about 10,000 rpm) and the resulting multilamellar vesicles are subjected to high pressure homogenization. (e.g., at about 10,000 to about 30,000 psi and, e.g., at about 25°C to about 70°C) At a processing temperature within the range of ) to form single bilayer vesicles (single layer). The single bilayer Vesicles can be sterilized, for example, by passing them through 0.45 μm and 0.22 μm filters and / or by moist heat sterilization. The filtered suspension of single bilayer Vesicles can be frozen and dehydrated (lyophilized) under high vacuum for long-term storage. A ready-to-use injectable composition is obtained by adding and mixing sterile water to the lyophilized powder.

[0101] The amount of encapsulated antifungal drug in vesicles prepared according to the methods described herein can be confirmed, for example, using a high performance liquid chromatography (HPLC) assay. The size of the vesicles can be confirmed, for example, using a light scattering method such as dynamic light scattering (DLS).

[0102] Single bilayer vesicles encapsulating an antifungal drug derived from enfumafungin were administered to animal models such as rats and rabbits to evaluate the local tolerability (ISR) upon intravenous administration. To evaluate local tolerability, clinical and histopathological evaluations of the injection site tissue were performed. To demonstrate the usefulness of the injectable compositions described herein, the stability of the vesicles was also confirmed under conditions simulating the injection of the vesicles into the bloodstream.

Examples

[0103] The following examples are only useful for explaining the present invention and its implementation. The examples should not be construed as a limitation to the scope or spirit of the present invention.

[0104] Preparation of Liposomes Encapsulating SCY-078 Examples A - I For Examples A - I, a lipid mixture containing DSPC and DSPG, cholesterol, and the enfumafungin derivative S CY - 078 were dissolved in a solvent (ethanol at 70 °C), and water with a solvent:water volume ratio of 1:3 was used. At 65 °C and a flow rate of 10 mL / min, microfluidic mixing (NanoAssemblr TM ) was performed to form unilamellar vesicles or liposomes loaded (encapsulated) with SCY - 078. The solvent and free (unencapsulated) drug in the liposome suspension were removed by tangential flow filtration ("TFF"). In the liposome suspension, the solvent and free (unencapsulated) drug were removed by tangential flow filtration ("TFF").

[0105] Examples J - P Examples J and K were prepared by the thin - film hydration method. A specific amount of a lipid mixture containing DSPC and DSPG, cholesterol, and SCY - 078 was dissolved in ethanol at 70 °C. In a round - bottom flask as a suitable reaction vessel, the solvent was removed by evaporation under reduced pressure to obtain a lipid dispersion film containing the antifungal drug. In a round - bottom flask as a suitable reaction vessel, the solvent was removed by evaporation under reduced pressure to obtain a lipid dispersion film containing the antifungal drug.

[0106] In Examples L - P, the solvent was removed by spray drying. A specific amount of a lipid mixture containing DSPC and DSPG, cholesterol, and SCY - 078 was dissolved in ethanol at 70 °C, mixed to form a uniform colloidal dispersion. The dispersion was sprayed as a fine mist at a spray pressure of about 70 psi through a 0.7 - mm nozzle at 20 mL / min and dried at about 50 °C. The spray - dried powder was collected and dried under reduced pressure until it reached a constant weight. The dispersion was sprayed as a fine mist at a spray pressure of about 70 psi through a 0.7 - mm nozzle at 20 mL / min and dried at about 50 °C. The spray - dried powder was collected and dried under reduced pressure until it reached a constant weight. ​​​

[0107] In Examples J to P, the antifungal agent obtained by either thin film evaporation or spray drying was hydrated in an aqueous solution containing 7.5% (w / v) sucrose Thereafter, the hydrated suspension was mixed at 10,000 rpm for about 5 minutes using a high shear mixer and the resulting multilamellar vesicles were subjected to high pressure homogenization (Microfluidics TM ) at a pressure in the range of about 10,000 to about 30,000 psi using a treatment temperature in the range of about 25°C to about 70°Cto form unilamellar vesicles.

[0108] The liposomes prepared in Examples M, O and P were subjected to sterilization by filtration through 0.45 μm and 0.2 2 μm filters. The liposomes of Example M were lyophilized The liposomes of Example N were sterilized by moist heat at 121°C for 12 minutes.

[0109] Characterization of Liposomes Encapsulating SCY-078 Formulations having various DSPC:cholesterol:DSPG:SCY -078 molar ratios shown in Tables 1 and 2 were evaluated with respect to unilamellar vesicle formation, particle size distribution, surface charge and the amount of encapsulated SCY-078.

[0110] After formation of the vesicles or liposomes, free (non-encapsulated) SCY-078 was removed by TFF (Examples A to I). When TFF was not used in the manufacturing process non-encapsulated SCY-078 was separated from encapsulated SCY-078 by dialysis using a 20,000 Dalton molecular weight cut-off (MWCO) cellulose ester membrane. CY-078 ​​

[0111] SCY-078 encapsulated in liposomes was separated from unencapsulated SCY-078 by size exclusion chromatography using a Sephadex® G-25 column. The amount of SCY-078 encapsulated in liposomes was confirmed using an HPLC assay. SCY-078 was separated on a C18 column and detected by ultraviolet (UV) spectroscopy at 210 nm. The particle size of the vesicles was confirmed by DLS (Z etasizer Nano, Malvern Instruments). The surface charge (zeta potential) was measured by laser Doppler microelectrophoresis in a disposable electrophoresis cell. As shown in Tables 1 and 2, several formulations with specific molar ratios of DSPC:cholesterol:DS PG:SCY-078 were found to form unilamellar liposome vesicles or liposomes with a capture efficiency of greater than 95%. Surprisingly, even though the process of removing unencapsulated drug was not part of the manufacturing process, free (unencapsulated) drug was found to be essentially undetectable (Examples J - Example O). It was also surprising that the vesicles remained stable after autoclaving, with no change in vesicle size and greater than 95% of SCY-078 retained within the vesicles (Example N).

Table 1

[0112]

Table 2

[0113] Evaluation of Physicochemical Stability of Spray-Dried Lipid Dispersions Containing SCY-078 The stability of the spray-dried intermediate (lipid dispersion containing SCY-078) was evaluated under ambient environmental (25°C / 60% RH) storage conditions and accelerated (40°C / 75% RH) storage conditions. As shown in Table 3, the spray-dried intermediate was stable with respect to the assay and formation of degradation products of SCY-078, and no significant changes or trends were observed when stored under ambient environmental (25°C / 60% RH) storage conditions and accelerated (40°C / 75% RH) storage conditions for up to 3 months. These results indicate that the spray-dried intermediate can be conveniently stored under ambient environmental conditions, for example, before another treatment to form hydration and liposomes, which are commercially desirable attributes.

Table 3

[0114] Evaluation of Physicochemical Stability of Liposomes Encapsulating SCY-078 The stability of an immediately dilutable suspension of liposomes encapsulating SCY-078 was evaluated under refrigerated conditions (2°C to 8°C). As shown in Table 4, there were no changes in the SCY-078 assay and average particle size. Furthermore, there was no leakage during storage of the encapsulated drug, which demonstrates a strong binding between SCY-078 and the lipid bilayer of the liposomes. These results indicate the potential for stability and long-term stability suitable for various commercial applications related to the liposome compositions described herein.

Table 4

[0115] Evaluation of Physiological Stability of Liposomes Encapsulating SCY-078 ​​​​​​​​​​The stability of the vesicles or liposomes encapsulating SCY-078 was determined by incubating the liposomes in fresh 50% bovine serum at 37 °C for 24 h. The liposome fraction was separated from the serum components by size exclusion chromatography using a Sephadex® G-25 column. The collected liposomes and serum fractions were analyzed for SCY-078 content by HPLC. The results shown in Table 5 demonstrate that SCY-078 was not detected in the serum protein fraction, but essentially all of the nominal content of SCY-078 was recovered from the liposome fraction, indicating that, remarkably, under conditions simulating intravenous administration into the bloodstream, SCY-078 remained encapsulated in intact liposomes, considering the high degree of protein binding of SCY-078. In a 14-day intravenous infusion study in rats, animals were injected 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 via an indwelling catheter surgically implanted into the

[0116] Evaluation of Local Tolerance of Liposomes Encapsulating SCY-078 in Animals Intravenous Injection Local Tolerance Test in Sprague-Dawley Rats jugular vein. In this way, evaluation of the reaction at the injection site emphasized histological observation of vascular inflammation. Overall, the incidence and severity of vascular inflammation were increased for SCY-078 administered as a solution compared to vascular inflammation in saline-treated animals; however, for liposome-encapsulated SCY-078, the incidence and severity of vascular inflammation were similar to those in saline-treated animals. The evaluation of the reaction at the injection site emphasized histological observation of vascular inflammation. Overall, the incidence and severity of vascular inflammation were increased for SCY-078 administered as a solution compared to vascular inflammation in saline-treated animals; however, for liposome-encapsulated In animals administered with the formulated SCY-078, vascular inflammation was not observed (Table 6).

[0117] Local Intravenous Irritation Test in New Zealand White Rabbits This study evaluated local irritation of physiological saline control, SCY-078 at a dose of 40 mg / kg / dose in solution and SCY-078 encapsulated in liposomes (10 or 40 mg / kg) by administration twice a day (separated by 6 hours ± 15 minutes). The doses were administered to New Zealand White rabbits via indwelling catheter at a rate of 20 mL / hour for 1 hour of intravenous infusion for 5 consecutive days.

[0118] Twice-daily intravenous infusion of SCY-078 at a dose of 40 mg / kg / dose as a solution formulation over 1 hour resulted in adverse local reactions at the injection site and surrounding areas, manifested as very slight / slight to severe erythema and edema. As a result, after only 1 day of administration all animals in this group were euthanized unexpectedly. In contrast, animals receiving twice-daily intravenous infusion of SCY-078 encapsulated in liposomes at 10 and 40 mg / kg were able to complete the planned 5-day administration, and the formulation was well tolerated.

[0119] The results of the local tolerance test in animals indicate that SCY-078 encapsulated in liposomes is more tolerable than SCY-078 in solution with respect to vascular inflammation and ISR at the injection site and may be suitable for intravenous administration via peripheral veins. [Table 5] [Table 6] ​​​​​​

Claims

1. An injectable composition comprising: an aqueous phase; and one or more unilamellar vesicles, wherein each unilamellar vesicle contains phospholipid and cholesterol, and each encapsulates a compound of formula (I) 【Chemical Formula 1】 wherein: X is O or H, H; R ｅ is C(O)NR ｆ R ｇ , or a 6-membered ring containing one or two nitrogen atoms a heteroaryl group (wherein the heteroaryl group may be mono-substituted with fluoro or chloro at a ring carbon or may be mono-substituted with oxygen at a ring nitrogen); is C₁₋₆-alkyl; R ｆ 、 R ｇ 、 R ６ and R ７ are each independently hydrogen or C １ -C ３ alkyl ； R ８ is C １ -C ４ alkyl, C ３ -C ４ cycloalkyl or C ４ -C ５ cycloalkyl or a pharmaceutically acceptable salt or hydrate thereof; R ９ is methyl or ethyl; and, R ８ and R ９ may together form a 6-membered saturated ring containing one oxygen atom. ; ； wherein the one or more unilamellar vesicles are hydrated in the aqueous phase; the injectable composition.

2. The injectable composition according to claim 1, wherein the concentration of the encapsulated compound of formula (I) or a pharmaceutically acceptable salt or hydrate thereof in the injectable composition is from about 0.01 to about 50 mg / mL.

3. The injectable composition according to claim 1, wherein the aqueous phase 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, galactose, and combinations thereof.

6. The injectable composition according to claim 3, wherein the pH of the aqueous phase is in the range of about 5.0 to about 7.

0.

7. The injectable composition according to claim 1, wherein the one or more unilamellar vesicles contain phosphatidylcholine, phosphatidic acid, phosphatidylserine, phosphatidylethanolamine, phosphatidylglycerol, or combinations thereof.

8. The injectable composition according to claim 1, wherein the one or more unilamellar vesicles contain phosphatidylcholine and phosphatidylglycerol.

9. The injectable composition according to claim 8, wherein the phosphatidylcholine is selected from dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, egg phosphatidylcholine, soybean phosphatidylcholine, dilauroylphosphatidylcholine, and dimyristoylphosphatidylcholine.

10. The injectable composition according to claim 8, wherein the phosphatidylglycerol is dipalmitoylphosphatidylglycerol, di... Stearoyl phosphatidylglycerol, dilauroyl phosphatidylglycerol and and dimyristoyl phosphatidylglycerol, the injectable composition according to claim 8.

11. The compound represented by formula (I) or a pharmaceutically acceptable salt or hydrate thereof is present in the vesicles in an amount of about 5 to about 12 mol percent, the phospholipid is present in the vesicles in an amount of about 50 to about 80 mol percent, and the cholesterol is present in the vesicles in an amount of about 10 to about 30 mol percent, the injectable composition according to claim 1 。

12. The phospholipid contains phosphatidylglycerol and phosphatidylcholine, and the molar ratio of the compound represented by formula (I) or a pharmaceutically acceptable salt or hydrate thereof to the phosphatidylglycerol, the phosphatidylcholine, and the cholesterol is 1:2:5:2.5, the injectable composition according to claim 1.

13. The average particle size of the one or more unilamellar vesicles is less than about 150 nm, the injectable composition according to claim 1.

14. The average particle size of the one or more unilamellar vesicles is less than about 100 nm, the injectable composition according to claim 13.

15. The average particle size of the one or more unilamellar vesicles is about 70 to about 80 nm, the injectable composition according to claim 14.

16. An injectable composition, an aqueous phase; and, one or more unilamellar vesicles [wherein each of the unilamellar vesicles contains a phospholipid and cholesterol, and each of them contains a compound represented by formula (II) 【Chemical 2】 which is (1S,4aR,6aS,7R,8R,10aR,10bR ,12aR,14R,15R)-15-[[(2-amino-2,3,3-trimethylbutyl oxy]-8-[(1R)-1,2-dimethylpropyl]-14-[5-(4-pyrid nyl)-1H-1,2,4-triazol-1-yl]-1,6,6a,7,8,9,1 0,10a,10b,11,12,12a-dodecahydro-1,6a,8,10a-tetra methyl-4H-1,4a-propan-2H-phenanthro[1,2-c]pyran-7- carboxylic acid) or a pharmaceutically acceptable salt or hydrate thereof encapsulated therein ］； ; wherein the encapsulated compound represented by formula (II) in the injectable composition or the concentration of the pharmaceutically acceptable salt or hydrate thereof is from about 0.01 to about 50 mg / mL and; wherein the phospholipid contains phosphatidylglycerol and phosphatidylcholine and; wherein the molar ratio of the compound represented by formula (II) or a pharmaceutically acceptable salt or hydrate thereof, the phosphatidylglycerol, the phosphatidylcholine, and the cholesterol is 1:2:5:2.5; wherein the aqueous phase contains sugar and has a pH of about 5.0 to about 7.0; and wherein the one or more unilamellar vesicles are hydrated in the aqueous phase; the injectable composition.

17. The injectable composition according to claim 16, wherein the one or more unilamellar vesicles encapsulate a citrate salt of the compound represented by formula (II).

18. A method for treating a fungal infection in a patient in need thereof, the method comprising intravenous administration of the injectable composition according to claim 1.

19. The method according to claim 18, wherein the patient 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 for treating a fungal infection in a patient in need thereof, the method comprising intravenous administration of the injectable composition according to claim 16.

23. A method for producing an injectable composition comprising one or more unilamellar vesicles [wherein each unilamellar vesicle encapsulates a compound represented by formula (I) wherein: X is O or H, H; a heteroaryl group (wherein the heteroaryl group may be mono-substituted with fluoro or chloro at a ring carbon or may be mono-substituted with oxygen at a ring nitrogen); is C₁-C₆-alkyl; or a pharmaceutically acceptable salt or hydrate thereof], comprising: (a) dissolving a phospholipid and cholesterol in an aliphatic alcohol having 1 to 5 carbon atoms to form a first solution; [Chemical Formula 3] (b) dissolving the compound represented by formula (I) or a pharmaceutically acceptable salt or hydrate thereof in the first solution to form a second solution; (c) mixing the second solution. R ｅ is C(O)NR ｆ R ｇ or a 6-membered ring containing one or two nitrogen atoms ​ ​ ​ R ｆ 、R ｇ 、R ６ and R ７ are each independently hydrogen or C １ -C ３ alkyl ； R ８ is C １ -C ４ alkyl, C ３ -C ４ cycloalkyl or C ４ -C ５ cycloalkyl ​ R ９ is methyl or ethyl; and, R ８ and R ９ may together form a 6-membered saturated ring containing one oxygen atom) ​ ​ ​ ​ ​ ​ ​ (d) evaporating the solvent from the second solution to produce a phospholipid-cholesterol dispersion containing the compound represented by formula (I) or a pharmaceutically acceptable salt or hydrate thereof; generating; (e) hydrating the phospholipid-cholesterol dispersion containing the compound represented by formula (I) or a pharmaceutically acceptable salt or hydrate thereof with a sugar solution to produce a hydrated suspension; and (f) forming one or more unilamellar vesicles from the hydrated suspension, wherein each of the unilamellar vesicles contains phospholipid and cholesterol and encapsulates the compound represented by formula (I) or a pharmaceutically acceptable salt or hydrate thereof; The method as described above, which includes Claim 24 The method according to claim 23, wherein the aliphatic alcohol is selected from methanol or ethanol. Claim 25 The method according to claim 23, wherein the sugar solution contains a sugar selected from monosaccharides, disaccharides and combinations thereof. Claim 26 The method according to claim 25, wherein the sugar is selected from sucrose, trehalose, lactose, glucose, fructose and galactose and combinations thereof. Claim 27 About 90% or more of the amount of the compound represented by formula (I) or a pharmaceutically acceptable salt or hydrate thereof present during step (b) is encapsulated within the one or more unilamellar vesicles during step (f), according to the method of claim 23. Claim 28 About 95% or more of the amount of the compound represented by formula (I) or a pharmaceutically acceptable salt or hydrate thereof present during step (b) is encapsulated within the one or more unilamellar vesicles during step (f), according to the method of claim 23. Claim 29 The method according to claim 23, wherein the compound represented by formula (I) or a pharmaceutically acceptable salt or hydrate thereof is present in the vesicles in an amount of about 5 to about 12 mole percent. Claim 30 The phospholipid contains phosphatidylglycerol and phosphatidylcholine, and the molar ratio of the compound represented by formula (I) or a pharmaceutically acceptable salt or hydrate thereof, the phosphatidylglycerol, the phosphatidylcholine and the cholesterol is 1:2:5:2.5, according to the method of claim 23. Claim 31 During step (f), for forming the one or more unilamellar vesicles, sonication, micro- ​ ​ ​ ​ ​ ​ ​ ​ The method according to claim 23, using fluid mixing, homogenization, or a combination thereof.

32. The method according to claim 23, further comprising sterilizing the one or more monolamellar vesicles produced in step (f).

3.

33. The method according to claim 23, further comprising lyophilizing the one or more monolamellar vesicles produced in step (f).

6.

34. The method according to claim 23, wherein the average particle size of the one or more monolamellar vesicles is less than about 150 nm.

9.

35. The method according to claim 34, wherein the average particle size of the one or more monolamellar vesicles is less than about 100 nm.

12.

36. The method according to claim 35, wherein the average particle size of the one or more monolamellar vesicles is about 70 to about 80 nm. 15.

Images