Antifungal prodrugs

Abstract

The present invention relates to antifungal prodrugs comprising an antifungal moiety linked to a trigger moiety by a self-immolative spacer. The trigger moiety is selected from glycosyl residues and oligosaccharides, stabilizes the self-immolative spacer, and is cleavable by a pathogen hydrolase, preferably an extracellular glycosidase (EC 3.2.1). When the trigger moiety is cleaved by the pathogen hydrolase, the self-immolative spacer spontaneously decomposes, releasing the antifungal moiety. The present invention also relates to pharmaceutical compositions comprising the prodrugs and their use in treating infectious diseases.

Description

[Technical Field]

[0001] FIELD OF THE INVENTION The present invention relates to the treatment of infectious diseases, particularly fungal and parasitic diseases. [Background technology]

[0002] Background of the Invention Currently, over 300 species of opportunistic fungi capable of infecting humans and animals have been identified. Nearly 1.7 billion people worldwide suffer from diseases caused by opportunistic fungal species. Immunocompromised individuals are particularly vulnerable to severe infections known as invasive fungal infections (IFIs). Invasive fungal infections refer to systemic proliferation of opportunistic fungi in a host organism, resulting in compromised survival. IFIs can affect various organ systems, including the lungs, meninges, sinuses, and / or bone dissemination. The incidence of IFIs is increasing, primarily due to an increase in immunocompromised patients, including those with neutropenia, HIV, chronic immunosuppression, indwelling prosthetics, burns, diabetes, and those taking broad-spectrum antibiotics. Notably, IFIs contribute to substantial morbidity and mortality in immunocompromised patients. Despite current treatments, IFIs are fatal in more than half of infected patients. The main pathogens involved in IFI belong to the genera Candida, Aspergillus, and Cryptococcus. Currently, there are three main types of antifungal agents used to treat invasive fungal infections (IFI): polyene antifungals, such as amphotericin B; azole antifungals, such as fluconazole and voriconazole; and echinocandins, such as caspofungin.

[0003] Among these compounds, amphotericin B is the most commonly used antifungal treatment due to its broad spectrum of action and low incidence of drug resistance. Amphotericin B has been shown to be effective in treating fungal infections, such as candidiasis, aspergillosis, and cryptococcosis, as well as severe tropical mycoses, such as blastomycosis and coccidioidomycosis. Amphotericin B is also effective against protozoan infections, such as leishmaniasis. Amphotericin B was isolated from the culture medium of Streptomyces nodosus in 1953. Like other antifungal polyenes, amphotericin B acts by binding to ergosterol, a sterol found in the cell membranes of fungi and protozoa, where it depolarizes the membrane, forming pores and causing cell death. Unfortunately, despite its broad spectrum and low incidence of resistance, its use in human therapy remains limited to the management of severe, life-threatening infections, particularly in immunocompromised patients, due to a narrow therapeutic window. Indeed, amphotericin B frequently causes adverse effects, the most serious of which is nephrotoxicity. Nephrotoxicity encompasses the well-known renal tubular injury and can even lead to acute renal failure. Amphotericin B-induced nephrotoxicity is not fully understood and is certainly multifactorial. This may involve amphotericin B's high affinity for cholesterol, which is highly expressed on lipoprotein receptors, potentially resulting in increased exposure of kidney cells to the drug. Several strategies have been developed to improve the solubility of amphotericin B (AmB) and / or mitigate adverse side effects.

[0004] The first formulations for intravenous injection were based on complexing amphotericin B with sodium deoxycholate to improve solubility. Subsequently, several liposomal or lipid complex formulations were developed to improve the solubility and tolerance of amphotericin B. - A lipid complex (ABLC) was jointly developed by Enzon Pharmaceuticals and Cephalon and marketed under the trade name Abelcet®. Abelcet® consists of AmB complexed with two phospholipids: 1-α-dimyristoylphosphatidylcholine (DMPC) and 1-α-dimyristoylphosphatidylglycerol (DMPG). - Colloidal dispersion (ABCD) was developed by Three River Pharmaceuticals Laboratories and sold under the name Amphocil® or Amphotec®. It was a complex of AmB and cholesteryl sulfate to form a colloidal dispersion. This drug was discontinued in 2011. - A liposomal formulation (L-AmB) was developed by Gilead and Astellas Pharma under the trade name Ambisome®. Ambisome® consists of a unilamellar bilayer liposome made of phosphatidylcholine, cholesterol, and distearoylphosphatidylglycerol, with AmB inserted into the membrane.

[0005] These formulations have been shown to be less nephrotoxic and cause fewer infusion-related reactions than the original formulations. However, these formulations are very expensive to manufacture, limiting their availability in low-income countries. In addition, they have other significant drawbacks. Abelcet® exhibits high clearance and a lower Cmax than the original drug, while Ambisome® has limited distribution in the kidney and lung.

[0006] Chemical modifications to improve AmB solubility have also been investigated. For example, Sedlak et al. (Bioorganic & Medicinal Chemistry Letters, 2001, 11, 2833-2835) described the synthesis of AmB-polyethylene glycol (PEG) conjugates. These conjugates were shown to have improved aqueous solubility, but in vitro bioassays showed significantly reduced antifungal activity compared to free AmB (Tan et al., 2016, PLOS ONE | DOI:10.1371 / journal.pone.0152112). Conjugates with other structures, such as B-calix[4]arene (Paquet et al., Bioconj.Chem, 2006, 1460-3) and AmB derivatives with double alkylation of the amino group (WO2007096137), have also been described. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] W02007096137 [Patent Document 2] WO2010019203 [Patent Document 3] WO2016079536 [Non-patent literature]

[0008] [Non-Patent Document 1] Sedlak et al. (Bioorganic & Medicinal Chemistry Letters, 2001, 11, 2833-2835) [Non-patent document 2] Tan et al., 2016, PLOS ONE|DOI:10.1371 / journal.pone.0152112 [Non-patent document 3] Paquet et al., Bioconj.Chem, 2006, 1460-3 [Non-patent document 4] Schmidt et al., Angew. Chem. Int, 2015, 54, 7492-7509 [Non-patent document 5] Gopin et al., Bioorg. Med. Chem. 2004, 12, 1853-1858 [Non-patent document 6] Gopin et al., Angew. Chem. Int. Ed. 2003, 42, 327-332 [Non-Patent Document 7] Remington: The Science and Practice of Pharmacy (Lippincott Williams & Wilkins; Twenty first Edition, 2005) [Non-patent document 8] Handbook of Pharmaceuticals Excipients, American Pharmaceutical Association (Pharmaceutical Press; 6th revised edition, 2009) [Non-Patent Document 9] Le Pape et al., Acta Parasitologica 2002, 47, 79-81 [Non-Patent Document 10] Le Pape et al., 2019, Int J Infect Dis. 2019 Apr;81:85-90 Summary of the Invention [Problem to be solved by the invention]

[0009] However, there remains a need for new derivatives of antifungal drugs that have improved solubility, improved distribution, better targeting to the site of infection, and / or fewer side effects than antifungal drugs currently on the market. [Means for solving the problem]

[0010] SUMMARY OF THE INVENTION The present invention relates to a compound of formula (A): [ka] [In the formula, - AFD refers to antifungal drugs, - SIS refers to a self-immolative spacer covalently attached to the AFD and TM; - TM refers to a trigger moiety selected from glycosyl residues and oligosaccharides, said TM stabilizing the SIS and being cleavable by a pathogen hydrolase, preferably an extracellular glycosidase (EC 3.2.1); Here, when TM is cleaved by pathogen hydrolases, SIS spontaneously degrades to release AFD], an antifungal prodrug.

[0011] In some embodiments, the antifungal prodrug of Formula (A) is - TM is selected from the group consisting of hexosamine, N-acetylhexosamine, neuraminic acid, sialic acid and their oligosaccharides containing 2 to 50, preferably 2 to 10, glycosyl residues; and / or - AFD is selected from the group consisting of azole antifungals, polyene antifungals, echinocandins, orotomides, and enfumafungin aglycon derivatives.

[0012] In some embodiments, the TM is selected from the group consisting of glucosamine, galactosamine, mannosamine, neuraminic acid, N-acetylglucosamine, N-acetylgalactosamine, sialic acid, N-acetylmannosamine, and chitin. For example, the TM is N-acetylglucosamine or N-acetylgalactosamine.

[0013] In some other embodiments, the AFD is selected from the group consisting of amphotericin B, nystatin, natamycin, caspofungin, micafungin, anidulafungin, rezafungin, Boli Conazole (v oriconazole, ketoconazole, itraconazole, fluconazole, ibrexafungerp, olorofim and derivatives thereof.

[0014] In a preferred embodiment, the AFD is caspofungin, Boli Conazole or amphotericin B, more preferably amphotericin B.

[0015] In some embodiments of the prodrugs of the present invention, SIS is selected from self-immolative spacers that spontaneously decompose with an electronic cascade or cyclization. For example, SIS is represented by formula (Ia1), (Ib1), (Ic1), or (Id1): [ka] [In the formula, - X is O, S, -O(C=O)-NH-, O(C=O)O-, -O(P=O)O-, -O(P=S)O-, NR, where R is H or C1-C3 alkyl, preferably CH3; - R1 is H, halogen, such as F, Br, or Cl, -NO2, C1-C3 alkyl, -CF3-NHR, -OR, -C(=O)OR, -S02R, where R is H or C1-C3 alkyl, preferably CH3 or a targeting moiety, and - R3 is H or a targeting moiety, and - R1 and R3 may also comprise or consist of a moiety that is not a targeting moiety.

[0016] Preferably, R3 is H, R1 is H, halogen, -NO2, -CF3-OR, -C(=O)OR, -SO2R, and R is H or C1-C3 alkyl, preferably CH3.

[0017] In certain embodiments, the antifungal prodrug of Formula (A) has the formula (A2): [ka] [In the formula, - TM is a glycosyl residue selected from the group consisting of glucosamine, galactosamine, N-acetylglucosamine, N-acetylgalactosamine, mannosamine, neuraminic acid, and sialic acid; - AFD includes amphotericin B, nystatin, natamycin, caspofungin, micafungin, anidulafungin, and rezafungin. Boli conazole, ketoconazole, itraconazole, fluconazole and derivatives thereof, preferably amphotericin B, caspofungin and Boli and pharmaceutically acceptable salts thereof.

[0018] Examples of antifungal prodrugs of the present invention are: [ka] or a pharmaceutically acceptable salt thereof.

[0019] The invention also relates to the use of an antifungal prodrug as defined above for the treatment or prevention of an infectious disease. The infectious disease may be caused by a pathogen belonging to the genera Candida, Aspergillus, Cryptococcus, Mucorales, Fusarium, Scedosporium, Lomentospora, Blastomyces, Mucorales, or species of the genera Leishmania, Trypanosoma, or Plasmodium. The antifungal prodrug is particularly useful for treating or preventing invasive fungal diseases in immunocompromised subjects.

[0020] The present invention also relates to a pharmaceutical composition comprising an antifungal prodrug as defined above and a pharmaceutically acceptable excipient.

[0021] The present invention also relates to a method of treating or preventing an infectious disease in a subject, the method comprising the step of administering, preferably by oral or intravenous route, an effective amount of an antifungal prodrug as defined herein.

[0022] The present invention further relates to the use of an antifungal product as defined herein in the preparation of a pharmaceutical composition for the treatment or prevention of infectious diseases, preferably for oral or intravenous administration. drawing [Brief explanation of the drawings]

[0023] [Figure 1A] 1 shows an AmB prodrug of the present invention. This compound provides proof of concept for the present invention and was evaluated in the Examples section of this application. [Figure 1B] FIG. 1 shows the mechanism of release of AmB from the prodrug, which involves hydrolysis of the targeting moiety by fungal hydrolases followed by spontaneous decomposition of the self-immolative spacer. [Figure 2] FIG. 1 shows the synthetic route and reaction conditions used to prepare AmB prodrugs. [Figure 3] Figure 1 shows the kinetics of release of AFD from the AmB prodrug upon incubation with β-N-acetylhexosaminidase, as well as the kinetics of formation of intermediate 3 and residue 5 shown in Figure 1B. Of note is that the AmB prodrug is stable in aqueous medium at 37°C (in the absence of enzyme). [Figure 4a] FIG. 1 shows survival for different groups of animals in a mouse model infected with budding conidia of C. albicans: (i) treated with Fungizone®, (ii) treated with Ambisome®, (iii) treated with an AmB prodrug of the invention (compound of FIG. 1A, referred to herein as GOG), and (iv) administered vehicle (control). [Figure 4B]FIG. 1 shows the fungal charge in the kidney measured after euthanasia for different groups of animals in a mouse model infected with budding conidia of C. albicans: (i) treated with Fungizone®, (ii) treated with Ambisome®, (iii) treated with an AmB prodrug of the invention (compound of FIG. 1A, herein referred to as GOG), and (iv) administered vehicle (control). [Figure 5A] FIG. 1 shows survival curves for Galleria mellonella moths (G. mellonella) treated with amphotericin B (AmB), an AmB prodrug of the invention (compound of FIG. 1A), and a control. [Figure 5B] FIG. 1 shows survival curves of Galleria mellonella infected with Cryptococcus neoformans treated with amphotericin B (AmB), an AmB prodrug of the invention (compound of FIG. 1A), and a control. DETAILED DESCRIPTION OF THE INVENTION

[0024] Detailed Description of the Invention The present inventors have conceived a new prodrug of amphotericin B that has improved solubility, biodistribution, tolerance, and better targeting of the site of infection compared to amphotericin B. This new prodrug is based on a vectorization platform that allows increasing the solubility, masking the toxicity of the antifungal drug, and facilitating the release of the active drug at a very precise site of infection.

[0025] This vectorization platform is based on a trigger moiety linked to an antifungal drug by a self-immolative group. The trigger moiety is selected to stabilize the self-immolative group and to be selectively recognized and cleaved by hydrolytic enzymes spontaneously secreted by pathogens at the site of infection. Following cleavage of the trigger moiety, the self-immolative group spontaneously rearranges, leading to the release of the active antifungal drug.

[0026] Therefore, the vectorization platform we have conceived takes advantage of the fact that pathogens, e.g., fungi, spontaneously secrete hydrolases at the site of infection. By selecting a trigger moiety specific for the hydrolases secreted by the pathogen, we can limit the release of antifungal drugs at the site of fungal infection, preventing damage to patient cells and limiting side effects.

[0027] We have provided a proof-of-concept for an innovative vectorization platform using AmB. We conceived the AmB prodrug, shown in Figure 1A, in which the vectorization platform is linked to the amino group of mycosamine and contains N-acetylglucosamine as the trigger moiety and 4-hydroxy-3-nitrobenzyl alcohol as the self-immolative group.

[0028] As illustrated in the Examples section, the present inventors have demonstrated that AmB is rapidly released from the prodrug by the action of β-N-acetylhexosaminidase.

[0029] As illustrated in Figure 1B and shown in Figure 2, the hydrolase efficiently cleaves the trigger moiety, i.e., the N-acetylglucosamine group, releasing intermediate 3, which spontaneously rearranges to release the active drug AmB.

[0030] Remarkably, the prodrug was shown to be stable in aqueous buffer at 37° C. without undergoing significant hydrolysis.

[0031] Next, we evaluated the antifungal activity of the prodrug against different fungal cell types, namely, blastospores and filamentous yeasts, as well as promastigotes and intracellular amastigotes of Leishmania. Remarkably, the prodrug was shown to be as effective as AmB against these different cell types, confirming that AmB is efficiently released from the prodrug by the action of pathogen hydrolases.

[0032] Furthermore, the prodrug does not show any significant toxicity to HELA cells, in contrast to AmB, which has an IC50 of about 23 μM. Thus, the prodrug is not metabolized by human cells and does not have any significant toxicity with respect to human cells.

[0033] The inventors have also shown that the AmB prodrugs of the present invention are at least as effective as Fungizone® (AmB) and Ambisome® (AmB in a liposomal formulation) in treating fungal infection in a mouse model infected with budding conidia of C. albicans. Notably, the group treated with the AmB prodrugs of the present invention showed significantly improved survival and a significant reduction in kidney fungal load compared to the control group.

[0034] The inventors also studied the effects of AmB and AmB prodrugs of the invention on the Galleria mellonella model, a larval model that allows for the assessment of the efficacy and intrinsic toxicity of active drugs. The inventors showed that the AmB prodrugs of the invention are significantly less toxic than AmB, confirming the data obtained in human cell lines.

[0035] Furthermore, AmB prodrugs were shown to be as effective as AmB against Cryptococcus neoformans and Cryptococcus gattii infections in the Galleria mellonella model.

[0036] Taken together, these results strongly support the fact that the vectorization platform conceived by the inventors does not impair the antifungal activity of drugs, increases their solubility, and promotes their specific release near the site of infection, while preventing adverse side effects caused by the large diffusion of antifungal agents, thus increasing the therapeutic window of antifungal drugs.

[0037] Without being bound by any theory, the inventors believe that this vectorization platform used to vectorize AmB may also be effective for vectorization of other antifungal drugs, such as echinocandins.

[0038] Thus, the present invention provides a compound of formula (A): [ka] [In the formula, - AFD refers to antifungal drugs, - SIS refers to a self-immolative spacer covalently attached to the AFD and TM; - TM refers to a trigger moiety that stabilizes the SIS and can be cleaved by pathogen hydrolases; wherein, upon cleavage of the TM, the SIS spontaneously decomposes to release the AFD. Thus, the active AFD is released from the prodrug of the present invention via a two-step process involving (i) enzymatic hydrolysis of the covalent bond between the TM and the SIS, and (ii) spontaneous decomposition of the SIS.

[0039] - Antifungal drugs (AFD) As used herein, an antifungal drug (AFD) refers to any drug that has fungicidal or fungistatic activity against at least one pathogenic fungal species. In some embodiments, the antifungal drug is active against at least one pathogenic fungus belonging to the genera Candida, Aspergillus, and Cryptococcus. In certain embodiments, the antifungal drug has broad-spectrum activity, meaning that it exhibits antifungal activity against multiple fungal species.

[0040] Antifungal drugs are usually administered in a dose of 2 000 g.mol -1 Less than 1 500 g.mol, preferably -1 has a molecular weight of less than

[0041] Antifungal agents include, but are not limited to, azole antifungals, polyene antifungals, echinocandins, orotomide, and enfumafungin aglycone derivatives.

[0042] As used herein, an azole antifungal agent refers to an antifungal compound containing at least one five-membered heterocyclic moiety containing a nitrogen atom and at least one other non-carbon atom (i.e., nitrogen, sulfur, or oxygen) as part of the ring. Preferred heterocycles are triazoles and imidazoles. Azole antifungals can act by inhibiting lanosterol 14α-demethylase, thereby blocking the conversion of lanosterol to ergosterol. Azole antifungals include, but are not limited to, ketoconazole, itraconazole, fluconazole, efinaconazole, albaconazole, voriconazole, ravuconazole, and posaconazole.

[0043] If present, the azole antifungal agent may be linked to a self-immolative spacer (SIS), for example, via its hydroxyl group.

[0044] As used herein, polyene antifungal agents (also referred to herein as polyene antibiotics or polyene antifungal agents) refer to antifungal drugs comprising a macrocycle containing a highly hydroxylated region opposite a region containing multiple conjugated double bonds (the polyene moiety). Polyene antifungal macrocycles generally possess aminoglycosides, e.g., D-mycosamine.

[0045] Polyene antifungals generally act as ionophores: they bind to ergosterol, a major component of fungal cell membranes, forming pores in the membrane, allowing K+ to leak out, acidifying the membrane and leading to fungal death.

[0046] Polyene antifungal agents include, but are not limited to, amphotericin A and B, nystatin, as well as natamycin, rezafungin, rimocidin, filipin, hamycin, and perimycin.

[0047] When the antifungal drug (AFD) is a polyene antifungal drug containing an aminoglycoside group, the AFD is preferably linked to the self-immolative spacer (SIS) via the amino group of the aminoglycoside. Alternatively, the AFD can be linked to the SIS via one of the hydroxyl groups present on the macrocycle.

[0048] As used herein, echinocandins refer to macrocyclic lipopeptide antifungals that act by inhibiting the enzyme (1→3)-β-D-glucan synthase, thereby disrupting the integrity of fungal cell walls. Echinocandins typically contain a lipophilic tail attached to a peptidic macrocycle. Echinocandins include, but are not limited to, caspofungin, micafungin, anidulafungin, rezafungin, echinocandin B (also known as CD 101, CAS No. 1396640-59-7), pneumocandin B0, biafungin, and aminocandins. When AFD is an echinocandin, it can be linked to SIS via one of its free hydroxyl or amino groups, preferably, if present, via one of its primary amino groups.

[0049] As an alternative to echinocandins, enfumafungin aglycone derivatives, such as ibrexafungelp (also known as SCV 078 and MK 3118), can be used. Like echinocandins, these compounds are inhibitors of fungal beta-1,3-D-glucan synthase. ibrexafungelp is a new antifungal drug under development (Phase III clinical trials are underway). Its CAS number is 1207753-03-04. Other enfumafungin aglycone derivatives of interest are disclosed in patent application WO2010019203.

[0050] As used herein, orotomide refers to a new class of antifungal agents that contain a pyrrole moiety and act by halting pyrimidine biosynthesis in fungal cells. Orotomid causes reversible inhibition of dihydroorotate dehydrogenase (DHODH), which in turn blocks mycelial growth.

[0051] Olotomides of interest are described, for example, in patent application WO2016079536. A preferred orotomide is olofim (CAS number 1928707-56-5), which is currently in Phase III clinical trials.

[0052] As used herein, "derivative" refers to any AFD that contains one or several types of chemical modifications while retaining its antifungal activity.

[0053] In some embodiments, the AFD is amphotericin B, nystatin, natamycin, caspofungin, micafungin, anidulafungin, rezafungin, Boli The active ingredient is selected from the group consisting of conazole, ketoconazole, itraconazole, fluconazole, ibrexafungelp, olorofim and derivatives thereof.

[0054] - Self-Destructing Spacer (SIS) A self-immolative spacer (SIS) (also referred to herein as a self-immolative group) is a chemical group that links an antifungal drug (AFD) and a trigger moiety (TM) and spontaneously decomposes once the TM is cleaved.

[0055] Indeed, upon release of the trigger moiety (TM), i.e., cleavage of the covalent bond between the TM and the SIS by the action of pathogen hydrolases, the SIS undergoes spontaneous structural rearrangement, releasing the active antifungal drug at the site of infection.

[0056] The SIS is selected to increase the solubility of the AFD and / or limit steric hindrance around the TM, allowing recognition of the TM by the hydrolases of interest. The SIS is also selected to rapidly degrade once the TM is cleaved by the fungal hydrolases, thereby releasing the AFD.

[0057] The SIS may be a bifunctional spacer or a trifunctional spacer. When a trifunctional SIS is used, the SIS carries a further entity, such as an additional AFD moiety, a moiety for increasing solubility, such as a PEG moiety, or a targeting moiety, as defined further below.

[0058] Self-immolative groups are well known in the art and have been widely studied. See Schmidt et al., Angew. Chem. Int, 2015, 54, 7492-7509, which is a review of self-immolative spacers, the contents of which are incorporated by reference. As explained by Schmidt et al., the spontaneous decomposition of self-immolative groups is primarily driven by two types of processes: (i) an electronic cascade that can lead to the formation of quinones or azaquinones, and (ii) cyclization that can lead to imidazolidinone, oxazolidinone, or 1,3-oxathiolan-2-one ring structures.

[0059] In some embodiments, the self-immolative spacer relies on an electronic cascade for decomposition and comprises an aromatic structure bearing an O-, N-, or S-group.

[0060] In certain embodiments, the SIS has formula (Ia), (Ib), (Ic) or (Id): [ka] [In the formula, - X is O, S, -O(C=O)-NH-, O(C=O)O-, -O(P=O)O-, -O(P=S)O-, NR, where R is H or C1-C3 alkyl, preferably CH3; - R1 is H, halogen, for example F, Br, or Cl, -NO2, C1-C3 alkyl, -CF3-NHR, -OR, -C(=O)OR, -S02R, where R is H or C1-C3 alkyl, preferably CH3 or a targeting moiety, and - R3 is H or a targeting moiety].

[0061] R1 may be in the para, meta, or ortho position of the X group. Preferably, R1 is in the ortho or para position. Preferably, R1 and R3 are not both targeting moieties.

[0062] As used herein, a "targeting moiety" refers to any group that enables delivery of a prodrug to a specific organ, tissue, or cell of a subject, or to a specific pathogen. The targeting moiety can be of any type. Typically, the targeting moiety is capable of specifically binding to a target component expressed by the targeted organ, tissue, cell, or pathogen.

[0063] For example, the targeting moiety may be selected from antibodies, antibody fragments or derivatives, such as Fab, Fab', and ScFv, aptamers, spiegelmers, peptide aptamers, and ligands or substrates of the target component of interest, e.g., having a molecular weight of 1000 g.mol -1 The target component may be any type of small chemical molecule, peptide, sugar, hormone, oligosaccharide, protein, or receptor or receptor fragment that can bind to the target component.

[0064] The targeted component may be, for example, a membrane protein, e.g., a membrane receptor, a component of the membrane or cell wall, etc. In certain embodiments, the targeted component is a component of the cell wall of a pathogen, or a component present on the surface of a pathogen, e.g., Asl3 (agglutinin-like protein 3), HWP1 (hyphal protein 1), beta-D-glucan, or an external fragment of HSP90 (heat shock protein 90).

[0065] Thus, the target may be Asl3, HWP1, HSP90, or beta-D-glucan. In certain embodiments, the targeting moiety comprises a spacer that limits steric hindrance and / or increases solubility while allowing covalent binding to the core structure of the SIS. For example, the spacer may be hydrophilic, such as a PEG-based spacer.

[0066] In certain embodiments, the SIS has the formula (Ia1), (Ib1), (Ic1) or (Ia1): [ka] wherein X, R1 and R3 are as defined above.

[0067] In certain embodiments, the SIS comprises or consists of formula (Ib2): [ka] wherein R1 is as defined above. In a preferred embodiment, R1 is selected from the group consisting of H, halogen, e.g., F, Br, or Cl, —NO2, —CF3, —C(═O)OR, and —S02R, and R is H or C1-C3 alkyl, preferably CH3. R1 may be in the ortho or para position, and is preferably in the ortho position.

[0068] In some other embodiments, the self-immolative spacer relies on a cyclization mechanism and comprises an alkyl chain and / or an aromatic moiety. For example, the self-immolative spacer may be represented by formula (Ie), (If), (Ig), (Ih), or (Ii): [ka] [In the formula, X1 is CH2, O, S, NR, where R is H or C1-C3 alkyl, preferably CH3; Y1 is CH2, O, NH or a single bond; R2 is H, halogen, for example F, Br, or Cl, —NO2, C1-C3 alkyl, —CF3, —NHR, —OR, —C(═O)OR, -SO2R, where R is H or C1-C3 alkyl, preferably CH3, or a targeting moiety; and - n is an integer from 1 to 5, preferably 1 or 2].

[0069] As described above, the prodrug may include a targeting moiety, typically carried by a self-immolative spacer. The self-immolative spacer may be a trifunctional linker that connects the TM, AFD, and targeting moiety together. Such self-immolative spacers, also called chemical adapters, are described, for example, in Gopin et al., Bioorg. Med. Chem. 2004, 12, 1853-1858 and Gopin et al., Angew. Chem. Int. Ed. 2003, 42, 327-332.

[0070] For example, the SIS may be represented by the formula (Ij) or (Ik): [ka] wherein TargM is a targeting moiety as defined above.

[0071] In certain embodiments, the SIS has the formula (Ib3): [ka] wherein R1 is as defined above and R3 is H or a targeting moiety (TargM).

[0072] Preferably, R1 is selected from the group consisting of H, halogen, such as F, Br, or Cl, —NO2, —CF3, —C(═O)OR, and —S02R, where R is H or C1-C3 alkyl, preferably CH3.

[0073] Thus, the prodrugs of the present invention have the formula (A1): [ka] wherein R1 is as defined above and R3 is H or a targeting moiety, preferably H.

[0074] In certain embodiments, R3 is H and R1 is NO2. Thus, the prodrug has the formula (A2): [ka] This is shown in Figure 1.

[0075] - Trigger part(TM) As used herein, a trigger moiety (TM) refers to a chemical group that stabilizes the SIS, i.e., prevents its spontaneous decomposition, thereby acting as a protecting group. In the context of the present invention, the TM is selected so that it is selectively cleaved by the action of an enzyme expressed by the pathogen of interest. The enzyme of interest is a pathogen hydrolase, e.g., a fungal hydrolase, which is secreted into the extracellular environment and can catalyze the release of a glycosyl moiety from a substrate of interest.

[0076] For example, pathogen hydrolases are extracellular glycosidases (EC 3.2.1) that can catalyze the hydrolysis of O-, N-, or S-glycosides. The hydrolases of interest are beta-N-acetylhexosaminidase (EC 3.2.1.52), beta-N-acetylgalactosaminidase (EC 3.2.1.53), chitinase (EC 3.2.1.14), beta-glucosidase (EC 3.2.1.21), alpha-D-mannosidase (EC 3.2.1.24), beta-D-mannosidase (EC 3.2.1.25), chitobiase (EC 3.2.1.29), beta-D-acetylglucosaminidase (EC 3.2.1.30), exo-alpha-sialidase (EC 3.2.1.18), endo-alpha-sialidase (EC 3.2.1.129), exo-1,4-β-D-glucosaminidase (EC 3.2.1.165), including but not limited to:

[0077] Thus, the trigger moiety is typically a glycosyl group. In some embodiments of the invention, the trigger moiety is selected from hexosamine and N-acetylhexosamine, preferably N-acetylhexosamine. The trigger moiety may also be selected from nine-carbon sugars, such as neuraminic acid and sialic acid, such as N-acetylneuraminic acid.

[0078] The trigger moiety may also be selected from hexosamine or N-acetylhexosamine, e.g., chitin-based, and / or neuraminic acid- or sialic acid-based oligosaccharides. Typically, the oligosaccharide may contain 2 to 50 glycosyl residues, e.g., 2 to 10 glycosyl residues.

[0079] As used herein, hexosamine refers to a hexose in which one of the hydroxyl groups has been replaced by an amino group. Hexosamines include, but are not limited to, fructosamine, galactosamine, glucosamine, and mannosamine.

[0080] In some embodiments, the trigger moiety (TM) is selected from the group consisting of glucosamine, galactosamine, mannosamine, N-acetylglucosamine, N-acetylgalactosamine, N-acetylmannosamine, chitin, neuraminic acid, and sialic acid.

[0081] In additional embodiments, the TM is selected from N-acetylglucosamine, N-acetylgalactosamine, N-acetylmannosamine, and sialic acid moieties. For example, the trigger moiety is N-acetylglucosamine or N-acetylgalactosamine. Such glycosyl residues can be cleaved by fungal beta-N-acetylhexosaminidase (EC 3.2.1.52).

[0082] Examples of compounds according to the invention In some embodiments, the prodrugs of the present invention have the formula (A): [ka] [In the formula, - TM is a glycosyl residue selected from the group consisting of hexosamine, N-acetylhexosamine, neuraminic acid, sialic acid and their oligosaccharides containing from 2 to 50, preferably from 2 to 10, glycosyl residues; - SIS is a self-immolative spacer comprising or consisting of a moiety of formula (Ia), (Ib), (Ic), (Id), (Ij) or (Ik), - AFD is an antifungal agent selected from azole antifungals, polyene antifungals, echinocandins, orotomide, enfumafungin aglycone derivatives and derivatives thereof, or a pharmaceutically acceptable salt thereof.

[0083] In some other embodiments, the prodrug of the present invention is according to formula (A), wherein: - TM is a glycosyl residue selected from the group consisting of glucosamine, galactosamine, mannosamine, N-acetylglucosamine, N-acetylgalactosamine, N-acetylmannosamine residues, neuraminic acid, sialic acid and chitin; SIS is a self-immolative spacer comprising or consisting of a moiety of formula (Ia1), (Ib1), (Ic1), (Id1), (Ib2) or (Ib3), - AFD is an antifungal agent selected from amphotericin B, nystatin, natamycin, caspofungin, micafungin, anidulafungin, rezafungin and their derivatives.

[0084] In some embodiments, the prodrug is according to formula (A), wherein: - TM is a glycosyl residue selected from the group consisting of glucosamine, galactosamine, mannosamine, N-acetylglucosamine, N-acetylgalactosamine, and N-acetylmannosamine residues; - SIS is a self-immolating spacer, - AFD is an antifungal agent selected from amphotericin B, caspofungin, and their derivatives.

[0085] The SIS may comprise or consist of a moiety of formula (Ia), (Ib), (Ic), (Id), (Ie), (If), (Ig), (Ih), (Ij), or (Ik).

[0086] In some other embodiments, the prodrug of the present invention is according to formula (A), wherein: - TM is an N-acetylglucosamine residue and an N-acetylgalactosamine residue, preferably an N-acetylgalactosamine residue; - SIS is a self-immolating spacer, - AFD is an antifungal agent selected from amphotericin B, caspofungin, and their derivatives.

[0087] The SIS may comprise or consist of a moiety of any one of formulae (Ia), (Ib), (Ic), (Id), (Ie), (If), (Ig), (Ih1), (Ih2), (Ij), (Ik), (Ib2) and (Ib3), preferably any one of formulae (Ia1), (Ib1), (Ic1), (Id1), (Ib2) and (Ib3).

[0088] In some preferred embodiments, the AFD is amphotericin B.

[0089] In other embodiments, the prodrug of the present invention has the formula (A1): [ka] [In the formula, - R1 is H, halogen, for example F, Br, or Cl, -NO2, C1-C3 alkyl, -CF3, -NHR, -OR, -C(=O)OR, -S02R, where R is H or C1-C3 alkyl, for example CH3; R3 is H or a targeting moiety, preferably H. - TM is a glycosyl residue selected from the group consisting of glucosamine, galactosamine, mannosamine, N-acetylglucosamine, N-acetylgalactosamine, and N-acetylmannosamine; - AFD refers to polyene antifungals and echinocandins, such as amphotericin B, nystatin, natamycin, caspofungin, micafungin, anidulafungin, rezafungin, Boli An antifungal agent selected from the group consisting of conazole, ibrexafungelp, olorofim and derivatives thereof, or a pharmaceutically acceptable salt thereof.

[0090] In additional embodiments, the prodrug of the present invention has the formula (A1): [In the formula, - R1 is H, halogen, for example F, Br, or Cl, -NO2, -CF3, -NHR, -C(=O)OR, -S02R, and R is H or C1-C3 alkyl, for example CH3; - R3 is H - TM is a glycosyl residue selected from the group consisting of glucosamine, galactosamine, N-acetylglucosamine, and N-acetylgalactosamine; - AFD is a class of antifungal drugs, including polyene antifungals, echinocandins, orotomide, enfumafungin aglycone derivatives, e.g., amphotericin B, nystatin, natamycin, caspofungin, micafungin, anidulafungin, and rezafungin. Boli An antifungal agent selected from the group consisting of conazole, ibrexafungelp, olorofim, and derivatives thereof, or a pharmaceutically acceptable salt thereof.

[0091] In a further embodiment, the prodrug of the present invention has the formula (A2): [ka] [In the formula, - TM is a glycosyl residue selected from the group consisting of glucosamine, galactosamine, N-acetylglucosamine, and N-acetylgalactosamine; - AFD includes polyene antifungals, echinocandins, azole antifungals, orotomide, and enfumafungin aglycone derivatives, e.g., amphotericin B, nystatin, natamycin, caspofungin, micafungin, anidulafungin, rezafungin, Boli or a pharmaceutically acceptable salt thereof.

[0092] In another embodiment, the prodrug of the present invention has the formula (A3): [ka] [wherein R1 is selected from the group consisting of H, -NO2, and -COOMe, and is preferably -NO2; AFD is selected from polyene antifungal drugs, echinocandins, azole antifungal drugs, and derivatives thereof, and is preferably amphotericin B, nystatin, natamycin, caspofungin, micafungin, anidulafungin, rezafungin, Boli conazole and derivatives thereof, or a pharmaceutically acceptable salt thereof.

[0093] The preferred AFD is: Boli Conazole, amphotericin B and caspofungin, more preferably amphotericin B.

[0094] For example, a prodrug of the present invention can be one of the following compounds or a pharmaceutical salt thereof: [ka]

[0095] As used herein, the term "pharmaceutically acceptable salt" refers to a non-toxic salt, which can generally be prepared by contacting the subject prodrug (e.g., an AmB prodrug) with a suitable organic or inorganic acid. For example, the pharmaceutical salt may be, but is not limited to, acetate, benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, bromide, butyrate, carbonate, chloride, citrate, diphosphate, fumarate, iodide, lactate, laurate, malate, maleate, mandelate, mesylate, oleate, oxalate, palmitate, phosphate, propionate, succinate, sulfate, tartrate, and the like.

[0096] The prodrugs of the present invention can be prepared by standard chemical processes. The Examples section describes the synthesis of certain prodrugs of the present invention, which can be adapted to obtain other prodrugs of interest.

[0097] Therapeutic uses and methods of the present invention The present invention also relates to the use of the prodrugs as defined above in the treatment or prevention of infectious diseases. A further object of the present invention is a method for treating or preventing an infectious disease in a subject, comprising administering to the subject an effective amount of a prodrug of the present invention. The present invention also relates to the use of the prodrugs of the present invention for treating or preventing an infectious disease in a subject.

[0098] As used herein, "infectious disease" (also referred to herein as infectious disease) refers to any disease or disorder, and its symptoms, caused by or resulting from contamination of a subject with a pathogen, e.g., a pathogenic bacterium, a fungus, including yeasts and molds, or a protozoan, or a virus. In preferred embodiments, the infectious disease is caused by a pathogenic fungus, e.g., a pathogenic yeast or mold, or by a pathogenic protozoan, more preferably a pathogenic fungus. For example, the infectious disease may be caused by a pathogen belonging to a species of the genera Candida, Aspergillus, Cryptococcus, Mucorales, Fusarium, Scedosporium, Lomentospora, Blastomyces, or Leishmania, Trypanosoma, or Plasmodium.

[0099] Pathogens include, for example, Aspergillus fumigatus, Aspergillus flavus, Candida albicans (including budding conidia of C. albicans), Candida krusei, Candida lusitaniae, Candida parapsilosis, Candida tropicalis, Candida glabrata, Candida auris, Cryptococcus neoformans, Cryptococcus gattii, and Blastomyces dermatitidis.

[0100] An infectious disease may be systemic, involving one or more organs, such as the respiratory or gastrointestinal organ system, or may be localized, i.e., localized to a particular organ or tissue, such as the brain, skin, or oral cavity. The infectious disease may be an infection of a mucous membrane, such as an oral, esophageal, or vaginal infection, or an infection affecting the bone, skin, blood, genitourinary tract, or central nervous system of a subject; this list is not exhaustive.

[0101] Infectious diseases include, but are not limited to, Candida, Aspergillus, Cryptococcus infections, mucormycosis, blastomycosis, fusariosis, leishmaniasis, and the like.

[0102] In some embodiments, the infectious disease may be a nosocomial infection, ie, a hospital-acquired infection, or a community-acquired disease.

[0103] In some embodiments, the infectious disease is an invasive fungal disease (IFD).

[0104] The subjects to be treated with the prodrugs of the present invention are preferably mammals, more preferably humans, and may be of any gender and age, including neonates, infants, children, and the elderly.

[0105] In some embodiments, the subject is immunocompromised. The patient's immunocompromised state may be a primary immunodeficiency (i.e., caused by a congenital or genetic defect) or a secondary immunodeficiency due to, for example, surgery, or immunosuppressive treatments, such as chemotherapy and anti-rejection drugs, cancer, such as leukemia, pathogens, such as the human immunodeficiency virus (HIV) that causes AIDS, and autoimmune diseases. In certain embodiments, the subject may suffer from a disease that predisposes the subject to infectious diseases. For example, the patient may be diabetic.

[0106] In some other embodiments, the patient has undergone or is scheduled to undergo surgery. In such cases, the prodrugs of the present invention can be used to prevent the onset of infectious diseases in subjects who have undergone surgery or are scheduled to undergo surgery. The prodrugs of the present invention can also be used to prevent such infectious diseases in subjects who have been exposed to pathogens. For example, the subject may be a medical worker.

[0107] In certain embodiments, a prodrug of the present invention may be administered to a subject in combination with an additional therapeutic agent. The additional therapeutic compound may be administered simultaneously, separately, or sequentially with the administration of a prodrug of the present invention.

[0108] As used herein, a "therapeutically effective amount" refers to an amount of a prodrug that prevents, eliminates, slows down an infectious disease, or reduces or delays one or more symptoms or disorders caused by or associated with said infectious disease in a subject, preferably a human.

[0109] Effective amounts of the prodrugs of the present invention and pharmaceutical compositions thereof, and more generally, dosing regimens, can be readily determined and adapted by those skilled in the art. Effective doses can be determined using conventional techniques and by observing results obtained under analogous circumstances. The therapeutically effective dose of the prodrugs of the present invention will vary depending on the infectious disease being treated or prevented, the severity of the infectious disease being treated, the route of administration, any associated concomitant therapy, the patient's age, weight, general health, medical history, etc. Typically, the amount of the prodrug administered to a patient can range from about 0.001 mg / day / kg to 100 mg / day / kg of body weight, preferably 0.1 mg / day / kg to 25 mg / day / kg of body weight, and more preferably 0.1 mg / day / kg to 10 mg / day / kg of body weight.

[0110] The prodrugs of the present invention may be administered at least once daily for several consecutive days, weeks, or months until the desired therapeutic effect is achieved.

[0111] Administration of the prodrugs of the present invention may be topical, parenteral, or enteral. Indeed, the prodrugs of the present invention may be administered by any conventional route, including, but not limited to, oral, buccal, sublingual, rectal, intravenous, intramuscular, subcutaneous, intraosseous, cutaneous, transdermal, mucosal, transmucosal, intraarticular, intracardiac, intracerebral, intraperitoneal, intranasal, pulmonary, intraocular, vaginal, or transdermal. Indeed, the route of administration of the prodrugs of the present invention may vary depending on the infectious disease being treated and the organ or tissue of the patient afflicted with the disease.

[0112] In some preferred embodiments, the prodrugs of the present invention are administered by intravenous or oral routes.

[0113] - Pharmaceutical compositions of the present invention In an additional aspect, the present invention relates to a pharmaceutical composition comprising (i) as an active ingredient, a compound of Formula (A), Formula (A1), (A2), or (A3), and a prodrug of any one of the foregoing (or a pharmaceutically acceptable salt or solvate thereof), and (ii) at least one pharmaceutically acceptable excipient.

[0114] The pharmaceutical composition of the present invention comprises: - 0.01% by mass to 90% by mass of the prodrug of the present invention; - 10% to 99.99% by weight of excipients; may include Percentages are expressed relative to the total weight of the composition.

[0115] Preferably, the pharmaceutical composition comprises: - 0.1% by mass to 50% by mass of the prodrug of the present invention; - 50% to 99.9% by weight of excipients; may include:

[0116] Such pharmaceutical compositions are preferably used for the treatment or prevention of infectious diseases caused by fungi, such as Candida, Aspergillus and Cryptococcus species, or protozoa, such as Leishmania.

[0117] Pharmaceutical compositions of the invention can be formulated according to standard methods, such as those described in Remington: The Science and Practice of Pharmacy (Lippincott Williams & Wilkins; Twenty First Edition, 2005).

[0118] Pharmaceutically acceptable excipients that can be used are particularly described in Handbook of Pharmaceuticals Excipients, American Pharmaceutical Association (Pharmaceutical Press; 6th revised edition, 2009). Generally, the pharmaceutical composition of the present invention can be obtained by mixing the prodrug of the present invention with at least one pharmaceutical excipient.

[0119] Suitable excipients include, but are not limited to, solvents such as water or water / ethanol mixtures, fillers, carriers, diluents, binders, anti-caking agents, plasticizers, disintegrants, lubricants, flavors, buffers, stabilizers, colorants, dyes, antioxidants, anti-adherents, softeners, preservatives, surfactants, waxes, emulsifiers, humectants, glidants, etc. Diluents include, but are not limited to, microcrystalline cellulose, starch, modified starch, dibasic calcium phosphate dihydrate, calcium sulfate trihydrate, calcium sulfate dihydrate, calcium carbonate, mono- or disaccharides such as lactose, dextrose, sucrose, mannitol, galactose, and sorbitol, xylitol, and combinations thereof. Binders include, but are not limited to, starches such as potato starch, wheat starch, and corn starch; gums such as tragacanth gum, acacia gum, and gelatin; hydroxypropyl cellulose, hydroxyethyl cellulose, and hydroxypropyl methylcellulose; polyvinylpyrrolidone, copovidone, polyethylene glycol, and combinations thereof. Lubricants include, but are not limited to, fatty acids and their derivatives, such as calcium stearate, glyceryl monostearate, glyceryl palmitostearate, magnesium stearate, zinc stearate, or stearic acid, or polyalkylene glycols, such as PEG. Glidants can be selected from colloidal silica, silicon dioxide, talc, and the like. Disintegrants include, but are not limited to, crospovidone, croscarmellose salts, such as croscarmellose sodium, starch, and derivatives thereof. Surfactants include, but are not limited to, for example, simethicone, triethanolamine, les polysorbate and its derivatives, such as tween® 20 or tween® 40, poloxamer, fatty alcohols, such as lauryl alcohol and cetyl alcohol, and alkyl sulfates, such as sodium dodecyl sulfate (SDS).Examples of emulsifiers include, for example, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, polyethylene glycols and fatty acid esters of sorbitan, or mixtures of these substances.

[0120] It will be appreciated that the excipients combined with the prodrugs of the present invention may vary depending on (i) the physicochemical properties, including stability, of the active prodrug, (ii) the desired pharmacokinetic profile of the active ingredient, (iii) the dosage form, and (iv) the route of administration.

[0121] The pharmaceutical composition may be in any dosage form. For example, the pharmaceutical composition may be a solid oral dosage form, a liquid oral dosage form, a suspension, e.g., for intravenous administration, a dosage form for topical application, e.g., a cream, ointment, gel, etc., a patch, e.g., a transdermal patch, a mucoadhesive patch, or a tablet, particularly a bandage or dressing, a suppository, or an aerosol for intranasal or pulmonary administration. The pharmaceutical composition may provide immediate, controlled, or sustained release of the prodrug of the present invention. Oral solid dosage forms include, but are not limited to, tablets, capsules, pills, and granules. If necessary, the oral solid form may be prepared with coatings and shells, such as enteric coatings or other suitable coatings or shells. Some such coatings and / or shells are well known in the art. Examples of coating compositions that can be used are polymeric substances and waxes. The prodrug may also be used in microencapsulated form, if appropriate, with one or more of the above-mentioned excipients. Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, and elixirs. Liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilizers and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils, polyethylene glycol and fatty acid esters of sorbitan, or mixtures of these substances. If necessary, the compositions may contain adjuvants, such as wetting agents, emulsifiers and suspending agents, sweeteners, flavorings, and / or fragrances. Suspensions may contain suspending agents, such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar, etc.

[0122] Vaginal or rectal suppositories can be prepared by mixing a prodrug of the present invention with a suitable non-irritating excipient or carrier such as cocoa butter, polyethylene glycol, or a suppository wax that is solid at ordinary temperatures but liquid at body temperature.

[0123] The ointments, pastes, creams and gels may contain excipients such as oils, waxes, paraffins, starches, tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonite, silicic acid, talc and zinc oxide, or mixtures thereof. The pharmaceutical composition may be in the form of an aerosol that can be delivered to the lungs by using an inhaler system. For example, the prodrug of the present invention may be adsorbed onto the surface of a nanocarrier or microcarrier. In some embodiments, the pharmaceutical composition of the present invention is an injectable composition, e.g., an injectable composition, e.g., a composition for intramuscular injection, intravenous injection, or infusion.

[0124] Typically, the pharmaceutical composition may be in the form of a liquid composition ready for injection, a concentrated liquid composition that is diluted before injection, or a powder, e.g., a lyophilized powder, that is dissolved or suspended in a suitable vehicle immediately prior to administration to a subject.

[0125] The prodrugs of the present invention may be formulated into liposome compositions, lipid complex compositions using excipients such as phospholipids, cholesterol, similar lipid complexes, or colloidal dispersions such as surfactants and / or lipids, such as those contained in Abelcet® or Ambisome® formulations.

[0126] The present invention also relates to a pharmaceutical kit comprising a prodrug of the present invention or a pharmaceutical composition of the present invention, in combination with means for administration to a subject, such as a reconstitution buffer, and / or means for injection, such as a needle and syringe. The kit may also contain instructions for carrying out the treatment method of the present invention. Further aspects and advantages of the present invention are disclosed in the experimental section below, which should be considered as illustrative and not limiting the scope of this application. [Example]

[0127] All reagents, including enzyme samples, were purchased from various suppliers (Sigma Aldrich®, Fluka®, Alfa Aesar®, Acros®, or TCI Chemical®) and stored according to their detailed specifications. The following solvents and reagents were freshly distilled under argon immediately before use: DCM, MeCN, and EtN over anhydrous calcium hydride; MeOH over sodium, and THF over sodium and benzophenone. DCM was sometimes purified by a solvent purification system (SPS). DMF was purchased anhydrous from Sigma Aldrich®. Where necessary, workup and purification solvents were distilled beforehand on a Buchi R-220-SE rotavapor to remove stabilizers.

[0128] Example 1 Synthesis of the Prodrugs of the Invention The AmB prodrug of Figure 1A was prepared according to the synthetic process described in Figure 2. After optimization of the reaction conditions, the AmB prodrug was obtained in 58% overall yield through six consecutive steps (Figure 2). The synthetic protocol is described below.

[0129] compound 6 [ka]

[0130] In a sealed tube, commercially available N-acetyl-D-glucosamine (3.000 g, 13.50 mmol, 1.0 equiv.) was dissolved in HCl gThe reaction mixture was dissolved in freshly prepared acetyl chloride solution saturated with HCl (15 mL, 210.2 mmol, 9.3 equiv.) and cooled to 0 °C. The reaction mixture was warmed and stirred at room temperature for 7 days. Upon completion, the reaction mixture was dissolved in DCM (20 mL) and cooled to 0 °C. The organic layer was carefully washed with saturated aqueous NaHCO (3 × 30 mL) and brine (30 mL). The organic layer was separated, dried over NaSO, filtered, and evaporated under reduced pressure. The crude product was purified by flash column chromatography on silica gel (gradient elution 100:0 to 0:100 DCM / EtOAc) to give compound 6 (3.305 g, 67%) as an air-sensitive white solid. 1 H NMR (300.13 MHz, CDCl3, 298.15 K): δ H 6.19 (d, J 1-2 ) = 3.6 Hz, 1H, H 1 ), 5.79 (d, J 7-2 = 8.7 Hz, 1H, H 7 ), 5.30 (m, 1H, H 3 ), 5.22 (t, J 4-3 , 4-5 = 9.6 Hz, 1H, H 4 ), 4.57-4.50 (m, 1H, H 2 ), 4.32-4.24 (m, 2H, H 5 , H 6a ), 4.14 (m, 1H, H 6b ), 2.11 (s, 3H, H アセチル ), 2.06 (s, 3H, H アセチル ), 2.05 (s, 3H, H アセチル ), 1.99 (s, 3H, H アセチル ) ppm. 13 C NMR (75.48 MHz, CDCl3, 298.15 K): δ C 171.5 (s, C アセチル ), 170.6 (s, C アセチル ), 170.2 (s, C アセチル ), 169.2 (s, C アセチル), 93.6 (s, C 1 ), 70.9 (s, C 5 ), 70.1 (s, C 3 ), 66.9 (s, C 4 ), 61.1 (s, C 6 ), 53.5 (s, C 2 ), 23.1 (s, C アセチル ), 21.5 (s, C アセチル ), 20.7 (s, C アセチル ), 20.6 (s, C アセチル ) ppm. white solid C 14 H 20 ClNO8 Molecular weight: 365.76g.mol -1 R f :0.63(EtOH) Melting point: 122°C Yield: 67%

number

[0131] compound 7 [ka]

[0132] Commercially available compound 4-hydroxy-3-nitrobenzaldehyde (1.372 g, 8.21 mmol, 1.5 equiv.) was dissolved in freshly distilled MeCN (40 mL). Activated molecular sieves 4 Å (1.000 g) and AgO (2.535 g, 10.94 mmol, 2.0 equiv.) were added at room temperature. The reaction mixture was stirred at room temperature for 15 minutes under a positive argon atmosphere. Compound 6 (2.000 g, 5.47 mmol, 1.0 equiv.) was added. The reaction mixture was stirred at room temperature for 18 hours under a positive argon atmosphere, protected from light, and analyzed by TLC (EtOAc, UV). 254nm The reaction mixture was monitored by HPLC (depicted with cerium molybdate). After completion, the reaction mixture was filtered through a pad of Celite, the residue was washed with DCM, and the organic layer was evaporated under reduced pressure. The crude product was purified by flash column chromatography on silica gel (gradient elution 100:0 to 0:100 DCM / EtOAc) to give compound 7 (3.720 g, quantitative) as a white solid. 1 H NMR (300.13 MHz, CDCl3, 298.15 K): δ H 9.97 (s, 1H, H 14 ), 8.29 (s, 1H, H 10 ), 8.05 (dd, J 12-10 = 2.0 Hz, J 12-13 = 8.2 Hz, 1H, H 12 ), 7.49 (d, J 13-12 = 8.2 Hz, 1H, H 13 ), 5.94 (d, J 7-2 = 6.5 Hz, 1H, H 7 ), 5.81 (d, J 1-2 = 7.5 Hz, 1H, H 1 ), 5.70 (t, J 3-2, 3-4 = 9.6 Hz, 1H, H 3 ), 5.13 (t, J 4-3, 4-5 = 9.2 Hz, 1H, H 4 ), 4.34-3.17 (m, 2H, H 5 , H 6a ), 4.00 (m, 1H, H 6b), 3.82 (m, 1H, H 2 ), 2.10 (s, 3H, H アセチル ), 2.07 (s, 3H, H アセチル ), 2.06 (s, 3H, H アセチル ), 1.97 (s, 3H, H アセチル ppm. 13 C NMR (75.48 MHz, CDCl3, 298.15 K): δ C 188.7 (s, C 14 ), 171.3 (s, C アセチル ), 170.6 (s, C アセチル ), 170.4 (s, C アセチル ), 169.6 (s, C アセチル ), 153.8 (s, C 8 ), 141.4 (s, C 9 ), 134.4 (s, C 12 ), 131.5 (s, C 11 ), 126.8 (s, C 10 ), 119.5 (s, C 13 ), 98.6 (s, C 1 ), 72.6 (s, C 5 ), 70.7 (s, C 3 ), 68.5 (s, C 4 ), 62.0 (s, C 6 ), 55.7 (s, C 2 ), 23.4 (s, C アセチル ), 20.8 (repeated, C) アセチル C アセチル C アセチル ,) ppm. White solid C 21 H 24 N2O 12 Molecular weight: 496.43 g·mol -1 R f 0.45 (EtOAc) Melting point: 165℃ Yield: Appropriate amount

number

[0133] compound 8 [ka]

[0134] Compound 7 (4.719 g, 9.51 mmol, 1.0 eq) was dissolved in a mixture of CHCl (74 mL), i-PrOH (21 mL), and silica gel (7.608 g), and the solution was cooled to 0 °C. NaBH (1.079 g, 28.53 mmol, 3.0 eq) was added, and the reaction mixture was stirred at 0 °C for 15 min under a positive atmosphere of argon. The reaction mixture was warmed and stirred at room temperature for 10 h under a positive atmosphere of argon, and TLC (EtOAc, UV) showed 5% CO₂Cl ... 254nm The reaction mixture was monitored by HPLC (denoted by cerium molybdate). Upon completion, the reaction mixture was cooled to 0°C. The organic layer was carefully washed with 1.0 M HCl solution (15 mL) and brine (30 mL). The organic layer was separated, dried over NaSO, filtered, and evaporated under reduced pressure to give compound 8 (5.055 g, quantitative) as a white solid. 1 H NMR (300.13 MHz, CDCl3, 298.15 K): δ H 7.78 (d, J 10-12 = 2.1 Hz, 1H, H 10 ), 7.46 (dd, J 12-10 = 2.1 Hz, J 12-13 = 8.6 Hz, 1H, H 12 ), 7.35 (d, J 13-12 = 8.5 Hz, 1H, H 13), 5.85 (d, J 7-2 = 8.2 Hz, 1H, H 7 ), 5.55 (dd, J 3-2, 3-4 = 9.2, 10.4 Hz, 1H, H 3 ), 5.45 (d, J 1-2 = 8.2 Hz, 1H, H 1 ), 5.12 (t, J 4-3, 4-5 = 9.5 Hz, 1H, H 4 ), 4.71 (s, 2H, H 14 ), 4.27 (dd, J 6a-5 = 5.2 Hz, J 6a-6b = 12.3 Hz, 1H, H 6a ), 4.20 (dd, J 6b-5 = 2.7 Hz, J 6b-6a = 12.2 Hz, 1H, H 6b ), 3.93 (dt, J 2-1, 2-3, 2-7 = 8.1, 10.4 Hz, 1H, H 2 ), 3.86 (m, 1H, H 5 ), 2.67 (s, 1H, H 15 ), 2.09 (s, 3H, H アセチル ), 2.06 (s, 3H, H アセチル ), 2.04 (s, 3H, H アセチル ), 1.98 (s, 3H, H アセチル ) ppm. 13 C NMR (75.48 MHz, CDCl3, 298.15 K): δ C 171.3 (s, C アセチル ), 170.7 (s, C アセチル ), 170.6 (s, C アセチル ), 169 6 (s, C アセチル ), 148.5 (s, C 8 ), 141.8 (s, C 9 ), 137.6 (s, C 11 ), 132.0 (s, C 12 ), 123.0 (s, C 10), 121.5 (s, C 13 ), 100.0 (s, C 1 ), 72.4 (s, C 5 ), 71.5 (s, C 3 ), 68.8 (s, C 4 ), 63.6 (s, C 14 ), 62.1 (s, C 6 ), 55.4 (s, C 2 ), 23.4 (s, C アセチル ), 20.9 (s, C アセチル ), 20.8 (s, C アセチル ), 20.8 (s, C アセチル ) ppm. white solid C 21 H 26 N2O 12 Molecular weight: 498.44g.mol -1 R f :0.28(EtOH) Melting point: 186°C Yield: Moderate

number

[0135] compound 9 [ka]

[0136] Compound 8 (1.317 g, 2.64 mmol, 1.0 equiv.) was dissolved in freshly distilled MeCN (20 mL). EtN (440 μL, 3.17 mmol, 1.2 equiv.) and commercially available N,N'-disuccinimidyl carbonate (743 mg, 2.90 mmol, 1.1 equiv.) were added. The reaction mixture was stirred at room temperature under a positive atmosphere of argon for 24 h and analyzed by TLC (EtOAc, UV). 254nm The reaction mixture was monitored by HPLC (depicted with cerium molybdate). After completion, the organic layer was evaporated under reduced pressure to give crude product 9 as an air-sensitive yellow solid, which was unstable and was used immediately in the next step. For further characterization, one batch of crude product was purified by flash column chromatography on silica gel (gradient elution 100:0 to 0:100 DCM / EtOAc) to give pure compound 9 as an air-sensitive white solid. 1 H NMR (300.13 MHz, CDCl3, 298.15 K): δ H 7.82 (s, 1H, H 10 ), 7.57 (d, J 12-13 = 8.5 Hz, 1H, H 12 ), 7.40 (d, J 13-12 = 8.4 Hz, 1H, H 13 ), 6.58 (d, J 7-2 = 8.2 Hz, 1H, H 7 ), 5.56 (d, J 1-2 = 9.0 Hz, 1H, H 1 ), 5.49 (d, J= 10.1 Hz, 1H, H 3 ), 5.28 (s, 2H, H 14 ), 5.10 (t, J 4-3, 4-5 = 9.5 Hz, 1H, H 4 ), 4.32-4.15 (m, 2H, H 6a , H 6b ), 4.04-3.92 (m, 2H, H 2 , H 5 ), 2.84 (s, 4H, H 17 ), 2.08 (s, 3H, H アセチル), 2.03 (s, 6H, H アセチル ), 1.95 (s, 1H, H アセチル ppm. 13 C NMR (75.48 MHz, CDCl3, 298.15 K): δ C 172.7 (s, C アセチル ), 170.9 (s, C アセチル ), 169.7 (s, C アセチル ), 168.9 (s, C 16 ), 151.5 (s, C 15 ), 150.1 (s, C 8 ), 141.0 (s, C 9 ), 134.2 (s, C 12 ), 129.1 (s, C 11 ), 125.6 (s, C 10 ), 120.3 (s, C 13 ), 99.2 (s, C 1 ), 72.2 (s, C 5 ), 71.3 (s, C 3 ), 70.8 (s, C 14 ), 68.6 (s, C 4 ), 62.0 (s, C 6 ), 55.0 (s, C 2 ), 25.5 (s, C 17 ), 23.0 (s, C アセチル ), 20.9 (s, C アセチル ), 20.8 (s, C アセチル ppm. White solid C 26 H 29 N3O 16 Molecular weight: 639.52 g·mol⁻¹ -1 R f 0.42 (EtOAc) HRMS (ESI) + ):C 26 H 29 N3O 16 Naの[M+Na] +The calculated m / z for is 662.1446, and the observed value is 662.1445 FT-IR (ATR): 1706, 1534, 1369, 1034, 648cm -1

[0137] compound 10 [ka]

[0138] Crude compound 9 (1.040 g, 1.25 mmol, 5.0 equiv.) was dissolved in anhydrous DMF (5 mL), and the solution was stirred at room temperature for 15 min under a positive atmosphere of argon. Commercially available compound amphotericin B (231 mg, 0.25 mmol, 1.0 equiv.) and EtN (77 μL, 0.55 mmol, 2.2 equiv.) were added. The reaction mixture was stirred at room temperature for 23 h, protected from light, and analyzed by reverse-phase TLC (an organic mixture composed of 15:85 HO / 43:20 MeOH / MeCN, UV). 254nm The reaction mixture was monitored by HPLC (denoted by cerium molybdate). After completion, the reaction mixture was co-evaporated with toluene. The crude product was dissolved in toluene under ultrasound and placed at -18°C. The precipitate was filtered, washed with DCM, and then dried under reduced pressure to give compound 10 (312 mg, 86%) as a brown solid. 1 H NMR (300.13 MHz, 2:1 DMSO-d6 / MeOD, 298.15 K): δ H 7.81 (d, J= 1.7 Hz, 1H, H 9 ), 7.67-7.58 (m, 1H, H 11 ), 7.41 (d,J= 8.7 Hz, 1H, H 12 ), 7.23-7.05 (m, 2H, H 7 , H 7' ), 6.51-5.83 (m, 12H, H 21'' ~H 32'' ), 5.47-5.32 (m, 2H, H 1 , H 33'' ), 5.28-5.13 (m, 2H, H 37'' , H4 ), 5.02 (s, 2H, H 13 ), 4.95 (t, J = 9.5 Hz, 1H, H 5 ), 4.45 - 4.35 (m, 2H, H 1' , H 3 ), 4.35 - 3.92 (m, 9H, H 2 , H 6a , H 6b , H 5' , H 3'' , H 11'' , H 15'' , H 17'' , H 19'' ,), 3.65 - 3.51 (m, 2H, H 2' , H 5'' ), 3.51 - 3.36 (m, 2H, H 3' , H 4' ), 3.26 - 3.12 (m, 2H, H 8'' , H 9'' ), 3.12 - 2.99 (m, 1H, H 35'' ), 2.36 - 2.24 (m, 1H, H 34'' ), 2.24 - 2.06 (m, 2H, H 2'' ), 2.00 (doublet of doublets, 4H, H アセチル , H 16'' ), 1.96 (s, 3H, H アセチル ), 1.91 (s, 3H, H アセチル ), 1.78 (s, 3H, H アセチル ), 1.75 - 1.18 (m, 15H, H 4'' , H 6'' , H 7'' , H 10'' , H 12'' , H 14'' , H 18'' , H 36'' ), 1.16 (d, J = 5.5 Hz, 3H, H 6' ), 1.10 (d, J = 6.3 Hz, 3H, H 38'' ), 1.02 (d, J = 6.2 Hz, 3H, H 40'' ), 0.90 (d, J = 7.1 Hz, 3H, H39'' ) ppm. 13 C NMR (75.48 MHz MHz, 2:1 DMSO-d6 / MeOD, 298.15 K): δ C 171.4 (s, C 1'' ), 170.8 (s, C アセチル ), 170.7 (s, C アセチル ), 170.4 (s, C アセチル ), 169.9 (s, C アセチル ), 156.5 (s, C 14 ), 148.9 (s, C 7 ), 141.2 (s, C 8 ), 124.2 (s, C 9 ), 137.2 (s, C 33'' ), 137.1 (s, C エチレン ), 134.4 (s, C エチレン ), 134.3 (s, C エチレン ), 133.9 (s, C エチレン ), 133.8 (s, C エチレン ), 133.6 (s, C 11 ), 133.2 (m, C 10 , C エチレン ), 133.0 (s, C エチレン ), 132.9 (s, C エチレン ), 132.7 (s, C エチレン ), 132.6 (s, C エチレン ), 132.0 (m, C エチレン , C エチレン ), 129.6 (s, C エチレン ), 118.3 (s, C 12 ), 99.5 (s, C 1 ), 97.9 (s, C 11'' ), 97.5 (s,C 1' ),78.0 (s, C 35'' ), 75.7 (s, C 3 ), 74.9 (s, C 4' ), 74.3 (s, C 19'' ), 72.7 (s, C 4), 72.1 (s, C 5' ), 70.6 (duplicate, C 5'' , C 8'' ,C 9'' , C 11'' ), 69.9 (s, C 2' ), 69.7 (s, C 37'' ), 67.4 (s, C 3'' ), 66.1 (duplicate, C 15'' , C 17'' ), 64.4 (s, C 13 ), 62.1 (s, C 6 ), 57.5 (duplicate, C 3' , C 16'' ), 53.7 (s, C 2 ), 46.8 (s, C 14'' ), 44.7 (duplicate, C 4'' , C 10'' , C 12'' ), 43.2 (s, C 34'' ), 42.4 (s, C 2'' ), 36.1 (s, C 18'' ), 35.6 (duplicate, C 6'' , C 7'' , C 18'' ), 22.6 (s, C アセチル ), 20.6 (s, C アセチル ), 20.4 (s, C アセチル ), 20.4 (s, C アセチル ), 18.8 (s, C 40'' ), 18.3 (s, C 6' ), 17.1 (s, C 38'' ), 12.3 (s, C 39'' ) ppm. this 13 In the NMR assignments for C, some atoms are not assigned. 36 The carbon signal corresponding to is superimposed with the signal of DMSO-d6. brown solid C 69 H 97 N3O 30 Molecular weight: 1448.53g.mol -1 R f : 0.41 (reverse phase 15:85 HO / 43:20 MeOH / MeCN organic mixture) Melting point: 155℃ Yield: 86%

number

[0139] Compound 1 (prodrug) [ka]

[0140] brown solid C 63 H 91 N3O 27 Molecular weight=1322.42.mol -1 R f = 0.37 (reverse-phase 15:85 HO / 43:20 MeOH / MeCN organic mixture) Melting point = 164°C (decomposition) Yield: Moderate HRMS(ESI - ):C 63 H 90 N3O 27 [MH] - The calculated m / z for is 1320.5762, and the observed value is 1320.5820 FT-IR (ATR): 3250, 1559, 1401, 1010cm -1 t 1 / 2 aq,pH7.4,37℃ >24 hours

[0141] Compound 10 (259 mg, 0.18 mmol, 1.0 equiv) was dissolved in a mixture of freshly distilled MeOH (1.8 mL) and THF (720 μL), and the solution was stirred at room temperature for 15 min under a positive atmosphere of argon. KCO (124 mg, 0.90 mmol, 5.0 equiv) was added. The reaction mixture was stirred at room temperature for 22 h, protected from light, and analyzed by reverse-phase TLC (an organic mixture composed of 15:85 HO / 43:20 MeOH / MeCN, UV). 254nm The reaction mixture was monitored by HPLC (depicted with cerium molybdate). Upon completion, sulfonic acid resin was added and the reaction mixture was stirred at room temperature for 15 minutes. The reaction mixture was filtered, the resin was washed thoroughly with MeOH, and the organic layer was evaporated under reduced pressure to give compound 11 (298 mg, quantitative) as an orange solid. If necessary, further purification was performed by preparative liquid chromatography (LC preparative). 1 H NMR (300.13 MHz, 2:1 DMSO-d6 / CD3OD, 298.15 K): δ H 7.82 (br, 1H), 7.76 (d, J= 8.9 Hz, 1H), 7.62 (d, J= 8.9 Hz, 1H), 7.43 (d, J= 8.9 Hz, 1H), 7.29-7.10 (m, 1H), 7.88 (d, J= 8.9 Hz, 1H), 6.53-6.20 (m, 10H), 6.20-6.01 (m, 4H), 5.99-5.84 (m, 2H), 5.49-5.39 (m, 1H), 5.38-5.31 (m, 1H), 5.25-5.14 (m, 2H), 5.12-5.07 (m, 1H), 5.04-4.99 (s, 1H), 4.83-4.73 (m, 3H), 4.68-4.61 (m, 2H), 4.48-4.36 (m, 3H), 4.27-4.17 (m, 2H), 4.11-3.99 (m, 2H), 2.75-2.70 (m, 1H), 2.32-2.24 (m, 2H), 2.19-2.14 (m, 1H), 2.08 (s, 1H), 1.78 (s, 3H, H アセチル), 1.75-1.18 (m, 15H), 1.15 (d, J= 5.5 Hz, 3H), 1.11 (d, J= 6.2 Hz, 3H), 1.03 (d, J= 5.8 Hz, 3H), 0.91 (d, J= 6.9 Hz, 3H) ppm.

[0142] Example 2 Evaluation of AmB prodrugs - Materials and Methods Enzymatic release and stability in aqueous solution Commercially available β-N-acetylhexosaminidase EC 3.2.1.52 (22.8 units mg) from jack bean (Canavalia ensiformis) -1 In vitro enzymatic hydrolysis was carried out using 100% ethanol (protein, suspension in 2.5 M ammonium sulfate, pH 7.0) and monitored by analytical LC using a VWR® Cooling Thermal Shake Touch. Prodrug 1 was incubated with the enzyme according to the following table:

[0143] [Table 1]

[0144] Antifungal and antiparasitic activity (in vitro) The half-maximal inhibitory concentration (IC) required to inhibit 50% of cell growth or viability in vitro 50 ) was determined by broth microdilution according to the European Commission's recommendations for antimicrobial susceptibility testing (protocol E.DEF 7.3.1 for Candida species) and the laboratory's certified internal procedure II CiMed (for Leishmania species) (Le Pape et al., Acta Parasitologica 2002, 47, 79-81).

[0145] The different strains, isolates and cell lines used are listed below: - Candida albicans clinical strain (IICI CiMed number CAAL93) - Candida albicans clinical strain (IICI CiMed number CAAL121) - Candida albicans reference strain SC5314 (IICIMed number CAAL146) - Leishmania major reference isolate MHOM / IL / 81 / BNI (IICIMed number LEMA1) - A549 cell reference line, ATCC® CCL-185 (cancer epithelial cells from the lung) - HeLa cell reference line

[0146] Growth or cell viability was performed in flat-bottom microplates and initially assessed by visual reading and always confirmed using a Bio-Rad iMark microplate absorbance reader to measure plate absorbance at 595 nm or a resazurin salt cell viability assay, measuring fluorescence at 590 nm after excitation at 530 nm with a Packard Fluorocount microplate reader BF10000 equipped with a halogen light source. Results are shown as means (± standard error of the mean SEM, if available) calculated from at least two independent experiments, each performed in triplicate, for one or several strains. Values ​​are expressed in μM for each corresponding pathogen.

[0147] result - Evaluation of enzymatic release The release of AmB from prodrug 1 was confirmed by incubating prodrug 1 with the enzyme of interest. Thus, real-time monitoring allowed visualization of the release kinetics in the presence of the enzyme of interest and its good aqueous stability in the absence of this enzyme. The release kinetics are shown in Figure 3.

[0148] - In vitro cell evaluation The prodrugs were tested on various cell types, blastospores or filamentous yeast, to determine antifungal activity, on promastigotes and intracellular amastigotes of Leishmania, to determine antiprotozoal activity, and finally on human cells to detect cytotoxicity. The results are shown in Table 2 below.

[0149] [Table 2]

[0150] Compound 1 exhibited activity against C. albicans or L. major at the same level as amphotericin B. Compound 1 also exhibited activity against Cryptococcus neoformans and Cryptococcus gattii. Notably, compound 1 exhibited low toxicity against HeLa cells. Furthermore, compound 1 showed no toxicity against the lung cell line A549 and human PBMCs (CI 50 >50 μM).

[0151] Example 3 Evaluation of the antifungal activity of AmB prodrugs in vivo Materials and Methods Mice were immunosuppressed by subcutaneous injection of 30 mg / kg prednisolone one day before challenge. On day 0, mice were intravenously infected with budding conidia of C. albicans. One hour after infection, mice were intraperitoneally treated with 1 mg / kg body weight of Fungizone®, Ambisome®, or an AmB prodrug (compound 1, Figure 1A) once daily for three consecutive days. Control groups received sterile distilled water (vehicle). Survival was monitored for 14 days after inoculation. Differences between cohorts were analyzed using the log-rank test.

[0152] On day 14, all mice were euthanized and their kidneys were removed and weighed.

[0153] Tissues were homogenized and serially diluted 10- to 1000-fold in sterile saline, plated on Sabouraud dextrose agar, and incubated for 48 hours to determine the number of CFU. The fungal burden in tissue was expressed as the mean logarithm of CFU / gram of tissue. Differences in mean kidney CFU were compared with vehicle controls using one-way ANOVA with post-hoc Tukey's test. AP values ​​of less than 0.05 were considered statistically significant.

[0154] ·result Figure 1 shows that vehicle-treated mice died before day 5. All treatments (Fungizone® (amphotericin B), Ambisome® (AmB in a liposomal composition), and the AmB prodrug of the present invention (referred to as GOG in Figures 4A and 4B) statistically improved survival (p>0.001). No statistical differences were observed between the treatments used. Regarding kidney fungal burden, mice treated with the AmB prodrug showed a significant reduction in burden compared to the placebo-administered control group (p<0.0079). No statistical differences were measured between the treatments used (p>0.05). In other words, these data indicated that the AmB prodrug of the present invention is at least as effective as the AmB drug.

[0155] Example 4 Evaluation of AmB prodrugs in the Galleria mellonella model Toxicity evaluation in the Galleria mellonella model The larval model is a rapid and practical tool for assessing the intrinsic toxicity of active substances. For further details of the model, see Le Pape et al., 2019, Int J Infect Dis. 2019 Apr;81:85-90. Larvae were incubated with AmB, an AmB prodrug (compound 1), and vehicle. Toxicity results are shown in Figure 5A. 2 mg.kg -1At a dose of 100 mg AmB, AmB was highly toxic, with a 40% survival rate in the treatment group. In comparison, at a comparable dose, its prodrug was much less toxic, with a survival rate of 80%. This statistically significant difference confirmed the in vitro results in human cells and demonstrated reduced toxicity of AmB in the carbamate prodrug form.

[0156] Evaluation of in vivo anti-cryptococcal activity in Galleria mellonella model This model constitutes a screening model and is an excellent alternative to traditional mouse models for evaluating the activity of antifungal molecules. The AmB prodrug of the present invention (compound 1) was evaluated for its antifungal effect against Cryptococcus neoformans and Cr. gattii. Figure 5B shows the survival curves of Galleria mellonella moths infected with Cryptococcus neoformans and treated with amphotericin B or its carbamate N-acetyl-D-glucosamine prodrug. Without treatment, all larvae died after 6 and 5 days, respectively. For Cryptococcus neoformans, amphotericin B at a dose of 2 mg / kg resulted in a 50% survival rate, and its prodrug was also effective, with a 30% survival rate at the same dose.

[0157] For C. gattii, amphotericin B and its prodrug resulted in survival rates of 30% and 20%, respectively.

Claims

1. Formula (A): 【Chemistry 1】 [In the formula, - AFD refers to antifungal drugs, SIS refers to a self-immolative spacer covalently attached to the AFD and the TM and has the formula (Ib1): 【Chemistry 2】 [In the formula, - X is O, S, -O(C=O)-NH-, O(C=O)O-, -O(P=O)O-, -O(P=S)O-, NR, and R is H or C1-C3 alkyl; - R1 is H, halogen, -NO2, C1-C3 alkyl, -CF3-NHR, -OR, -C(=O)OR, -SO2R, where R is H or C1-C3 alkyl; - R 3 is H] containing or consisting of parts of - TM refers to a trigger moiety that stabilizes the SIS and is cleavable by a pathogen hydrolase, said TM being selected from the group consisting of hexosamine, N-acetylhexosamine, neuraminic acid, sialic acid, and their oligosaccharides containing 2 to 50 glycosyl residues; Once the TM is cleaved by the pathogen hydrolase, the SIS spontaneously degrades, releasing the AFD. Antifungal prodrug of.

2. The antifungal prodrug of claim 1, wherein AFD is selected from the group consisting of azole antifungals, polyene antifungals, echinocandins, orotomide, and enfumafungin aglycone derivatives.

3. 2. The antifungal prodrug of claim 1, wherein TM is selected from the group consisting of glucosamine, galactosamine, mannosamine, neuraminic acid, N-acetylglucosamine, N-acetylgalactosamine, sialic acid, N-acetylmannosamine, and chitin.

4. 2. The antifungal prodrug of claim 1, wherein TM is N-acetylglucosamine or N-acetylgalactosamine.

5. 5. The antifungal prodrug of any one of claims 1 to 4, wherein the AFD is selected from the group consisting of amphotericin B, nystatin, natamycin, caspofungin, micafungin, anidulafungin, rezafungin, voriconazole, ketoconazole, itraconazole, fluconazole, ibrexafungelp, olorofim, and derivatives thereof.

6. 6. The antifungal prodrug of any one of claims 1 to 5, wherein AFD is caspofungin, voriconazole, or amphotericin B.

7. SIS is represented by formula (Ib2): 【Transformation 3】 wherein R 1 is as defined in claim 1. containing or consisting of parts of 7. An antifungal prodrug according to any one of claims 1 to 6.

8. R 1 H, halogen, -NO 2 , -CF 3 -OR, -C(=O)OR, -SO 2 R, where R is H or C 1 ~C 3 8. The antifungal prodrug of claim 7, which is alkyl.

9. Formula (A2): 【Chemistry 4】 [In the formula, - TM is a glycosyl residue selected from the group consisting of glucosamine, galactosamine, N-acetylglucosamine, N-acetylgalactosamine, mannosamine, neuraminic acid, and sialic acid; - AFD is an antifungal agent selected from the group consisting of amphotericin B, nystatin, natamycin, caspofungin, micafungin, anidulafungin, rezafungin, voriconazole, ketoconazole, itraconazole, fluconazole, and derivatives thereof.

9. The antifungal prodrug of any one of claims 1 to 8, wherein 10. The antifungal prodrug of claim 9, wherein AFD is amphotericin B.

11. The prodrug is 【Transformation 5】 or a pharmaceutically acceptable salt thereof.

12. 12. A pharmaceutical composition comprising an antifungal prodrug according to any one of claims 1 to 11 for use in the treatment or prevention of an infectious disease.

13. 13. The pharmaceutical composition for use according to claim 12, wherein the infectious disease is caused by a pathogen belonging to the genera Candida, Aspergillus, Cryptococcus, Mucorales, Fusarium, Scedosporium, Lomentospora, Blastomyces, Mucorales, or a species of the genera Leishmania, Trypanosoma, or Plasmodium.

14. 14. A pharmaceutical composition for use according to claim 12 or 13 for the treatment or prevention of invasive fungal disease in an immunocompromised subject.

15. 15. The pharmaceutical composition for use according to any one of claims 12 to 14, wherein the pharmaceutical composition is for oral or intravenous administration.

16. 12. A pharmaceutical composition comprising the antifungal prodrug of any one of claims 1 to 11 and a pharmaceutically acceptable excipient.

17. 12. Use of an antifungal product as defined in any one of claims 1 to 11 in the preparation of a pharmaceutical composition for the treatment or prevention of an infectious disease.

Images