Azapyridinophane compound for use as an antifungal

Abstract

The invention relates to a compound of Formula (I) or a pharmaceutically acceptable solvate or salt thereof, for use against pathogenic fungi, particularly pathogenic moulds, and in particular against A. fumigatus, and preferably Pytren-2Q.

Description

[0001] AN AZAPYRIDINOPHAN COMPOUND FOR USE AS AN ANTIFUNGAL

[0002] Technical Sector

[0003] The present invention is in the field of antifungal or antimycotic agents.

[0004] State of the art

[0005] Currently, four families of compounds are available for the treatment of systemic fungal infections:

[0006] - Azoles, including fluconazole, itraconazole, voriconazole, posaconazole, and isavuconazonium. These drugs are administered orally.

[0007] - Echinocandins, such as anidulafungin, caspofungin, micafungin, and rezafungin. These are drugs administered intravenously.

[0008] - Pyrimidines, such as flucytosine.

[0009] - Polyenes, particularly amphotericin B in its liposomal formulation.

[0010] All of these drugs exhibit adverse effects ranging from mild to severe. None of the currently used drugs are structurally related to the compounds provided by the present invention.

[0011] The discovery of amphotericin B in the 1950s marked the first significant advance in this field. This compound, which belongs to the polyene group, showed efficacy against a wide range of pathogenic fungi, but its high toxicity limited its use. From that point on, the search for safer and more targeted treatments accelerated.

[0012] The use of pyrimidines in the treatment of fungal infections was introduced in the 1960s with the development of flucytosine, initially synthesized as an antineoplastic agent. However, its effectiveness against some fungi led to its use as an antifungal. Flucytosine acts by interfering with the synthesis of fungal DNA and RNA, thus inhibiting fungal growth. It is primarily used in combination with amphotericin B to treat severe fungal infections, such as cryptococcosis and systemic candidiasis, as it readily develops resistance on its own. This advance was significant in the fight against systemic fungal infections, although its use has been limited compared to other antifungals due to its toxicity and the rapid emergence of resistance. In the 1970s, azoles were introduced, a class of compounds that interfere with the synthesis of ergosterol, a key component of the fungal cell membrane.Drugs like miconazole and ketoconazole were among the first in this group, followed by more advanced versions such as fluconazole and itraconazole in the 1990s, which offered greater safety and a broader spectrum of activity. Echinocandins represent a more recent advance in the treatment of serious fungal infections. These drugs work by inhibiting the synthesis of the fungal cell wall, making them highly effective against fungi such as Candida and Aspergillus, with less toxicity than other drugs.

[0013] The emergence of resistance to these drugs is currently driving research into new compounds and therapeutic strategies, such as drug combinations and the development of antifungals with novel mechanisms of action.

[0014] The mechanism by which synthetic polyamines can alter polyamine biosynthesis in fungi and thus be used as antifungals has been described. However, research has primarily focused on linear polyamines, mainly diamines, and their application in plant treatment.

[0015] In 2022, the WHO established a list of 19 fungi (yeasts and molds) for which the development of new drugs is a priority due to their high risk of morbidity and mortality. The limited number of antifungal compounds is explained by the difficulty of developing them, due to the close biomolecular similarity between fungi and human cells, which restricts the number of potential biological targets.

[0016] The synthesis of the compound PyTren-2Q and the use of its metal complexes as mimetics of the enzyme superoxide dismutase have been disclosed in EP2492270A2. This same compound, as well as PyTrenEl-2Py, also appears in patent ES2414291 B2, which protects its use as an antiparasitic, preferably against the parasites Trypanosoma cruzi and Leishmania spp.

[0017] None of the aforementioned patents refer to the antifungal activity of these compounds, and none of the previously presented results suggest they might have such activity. It is important to note that kinetoplastid parasites and fungi belong to different kingdoms (protista and fungi, respectively) and therefore have significant differences in their cellular structure and metabolism. Because of this, it is not obvious that a compound active against one would also be active against the other, although there are examples of drugs (for example, amphotericin B) that exhibit this dual activity.

[0018] In the article Inclán, M.; Torres Hernández, N.; Martínez Serra, A.; Torrijos Jabón, G.; Blasco, S.; Andreu, C.; del Olmo, MI; Jávega, B.; O'Connor, J.-E.; García-España, E. Antimicrobial Properties of New Polyamines Conjugated with Oxygen-Containing Aromatic Functional Groups. Molecules 2023, 28, 7678. with DOI 10.3390 / molecules28227678, the activity of structurally similar, but different, compounds against a series of bacteria and yeasts was studied, without finding significant activity against them.

[0019] US-6191273 refers to the synthesis of pyridinophane libraries and mentions an assay against Candida albicans, but no activity data (MIC) for any compound is shown, although it notes that the compounds were tested in duplicate to determine the minimum inhibitory activity concentration.

[0020] Other documents such as W02017030728A1 or the article “Synthesis and activities of novel mononuclear or dinuclear cyclon complexes bearing azole pendants as antibacterial and antifungal agents”; European Journal of Medicinal Chemistry; 12 September 2014; Shuo Li Jia-Xuan Chen Fang-Zhen Li, disclose the use of polyazacyclophane-type macrocyclic derivatives against pathogenic fungi, including A. fumigatus, but these compounds are structurally different.

[0021] The compounds of the present invention, in addition to exhibiting activity against fungi (yeasts and molds), display an optimal pharmacological profile: they have low toxicity, are water-soluble, have low molecular weight, and their synthesis is easily scalable. This makes them excellent candidates for the development of new orally administered therapeutic agents for the treatment of invasive mycoses and paves the way for the development of an entire new family of antifungal agents based on macrocyclic polyamines.

[0022] The compounds whose use is the subject of the present invention belong to a chemical family or class called scorpion-type azacyclophanes. They belong to the family of macrocyclic polyamines, also called pyridinophanes, which exhibit remarkable antifungal activity against yeasts and molds. Among these are some of the species included in the QMS priority pathogen list. Particularly interesting is the fact that the compounds are active against both the wild-type and azole-resistant strains of A. fumigatus, a mold included in the QMS list of the four most dangerous pathogens (Critical Group).

[0023] Fungal infections are difficult to treat because of the similarity between fungal and human cells, which has complicated the search for compounds that are effective against fungi while also being safe for humans.

[0024] The present invention represents an important advance, since it is not a simple structural modification of the classes of compounds already used, nor is it a combination therapy.

[0025] Detailed description of the invention

[0026] The present invention relates to a compound of Formula (I),

[0027]

[0028] which is a pyridinophane where Ri is selected from one of the following groups:

[0029]

[0030] or a pharmaceutically acceptable solvate or salt thereof, for use against pathogenic fungi.

[0031] The diagram above, which describes the Ri options, shows the position where the group is attached to the pyridinophane skeleton. This representation should be considered equivalent to the one shown below, which is how the options of formula (I) appear in Spanish priority application P202530332:

[0032]

[0033] The pyridinophane skeleton shown in formula (I) is called in this memory “pytren” (compound with R1 = H), and is named according to IUPAC (6-(2-aminoethyl)-3,6,9-triaza-1-(2,6)-pyridinoddodecaphan).

[0034] Depending on the specific implementation, the pharmaceutically acceptable salt is selected from hydrochloride, hydrobromide, sulfate, nitrate, phosphate salts, carboxylic acid salts, sulfonic acid salts, fatty acid salts, or amino acid salts.

[0035] According to particular embodiments, the compound is selected from pytren-2Py (6-[4-(2-pyridyl)-3-azabutyl]-3,6,9-triaza-1-(2,6)-pyridinedodecafane), pytren-3Py (6-[4-(3-pyridyl)-3-azabutyl]-3,6,9-triaza-1-(2,6)-pyridinedodecafane), pytren-4Py (6-[4-(4-pyridyl)-3-azabutyl]-3,6,9-triaza-1-(2,6)-pyridinedodecafane), pytren-2Q (6-[4-(2-quinolyl)-3-azabutyl]-3,6,9-triaza-1-(2,6)-pyridinedodecafane), pytren-4Q (6-[4-(4-quinolyl)-3-azabutyl]-3,6,9-triaza-1-(2,6)-pyridinedodecaphane), pytrenEL-2py (6-[8-(2-pyridyl)-3,7-diazaoctyl]-3,6,9-triaza-1-(2,6)-pyridinedodecaphane), pytrenEL-3py (6-[8-(3-pyridyl)-3,7-diazaoctyl]-3,6,9-triaza-1-(2,6)-pyridinedodecaphane), pytrenEL-4py (6-[8-(4-pyridyl)-3,7-diazaoctyl]-3,6,9-triaza-1-(2,6)-pyridinecycle decaphane), pytren-N (6-[4-(1-naphthyl)-3-azabutyl]-3,6,9-triaza-1-(2,6)-pyridinecyclodecaphane), pytren-A (6-[4-(9-antrii)-3-azabutyl]- 3.6.9-triaza-1-(2,6)-pyridinecyclodecaphane), pytren-Flu (6-[4-(2-fluoryl)-3-azabutyl]-3,6,9-triaza-1-(2,6)-pyridinecyclodecaphane), pytren-6IZ (6-[4-(6-indazolyl)-3-azabutyl]-3,6,9-triaza-1-(2,6)-pyridinecyclodecaphane), pytren-2QI (6-[4-(8-iodoquinolin-2-yl)-3-azabutyl]-3,6,9-triaza-1-(2,6)-pyridinecyclodecaphane), pytren-2QCI (6-[4-(8-chloroquinolin-2-yl)-3-azabutyl]-3, 6, 9-triaza- 1-(2, and pytren.

[0036] “EL” means “elongated”.

[0037] Compounds in which Ri is a quinolinyl radical, and more preferably 2-quinolinyl, have potent antifungal activity, especially against pathogenic fungi.

[0038] According to particular realizations, the minimum inhibitory concentration of compounds of formula (I) is between 0.12-8 µg / ml (MIC90) and 0.12->64 pg / ml (CMhoo).

[0039] According to particular realizations, the fungus is a yeast and the minimum inhibitory concentration (MIC100) of the compounds of formula (I) is between 0.12-8 µg / ml.

[0040] According to particular realizations, the pathogenic fungus is a pathogenic mold.

[0041] According to particular realizations, the fungus is a pathogenic fungus, and the inhibitory concentration (MIC90 or MIC100) of the compounds of formula (I) is between 0.25-8 pg / ml (MIC90) and 0.25-64 pg / ml (MIChoo).

[0042] We consider CMI100 and CMI90 to be those concentrations (expressed in µg of compound per ml of medium used in the test) capable of inhibiting all or at least 90% of the growth of the fungus, respectively, compared to the untreated control.

[0043] For particular realizations, the user is selected among Candida albicans, Pichia kudriavzevii, Candida tropicalis, Candida parapsilosis, Nakaseomyces glabratus, Meyerozyma guilliermondii, Candida dubliniensis, Clavispora lusitanea, Cryptococcus neoformans, Cryptococcus cats, Trichosporon spp; Aspergillus fumigatus ss, Aspergillus niger, Aspergillus flavus, Aspergillus terreus, Rizhopus arrhizus, Mucor circinelloides, Paecilomyces variotii, Fonsecaea pedrosoi, Fusarium solani, Trichophyton mentagrophytes, Trichophyton rubrum, Scedosporium apiospermum y Lomentospora prolificans, Trichophyton sp, Microsporum sp, Epidermophyton sp, Blastomyces sp, Histoplasma sp, Sporothrix sp, Saksenaea sp, Cladophialophora sp, Absidia sp, Coccidioides sp, Paracoccidioides sp, Emmonsia sp, Talaromyces sp.

[0044] According to preferred realizations, the tree is of the genus Aspergillus, and preferably is A. fumigatus.

[0045] According to particular embodiments of the invention, the use of the compound of formula (I) against pathogenic fungi is a use as a medicament.

[0046] According to particular embodiments of the invention, the pathogenic fungus is a pathogenic yeast. And according to further particular embodiments of the invention, the fungus is an azole-resistant yeast.

[0047] Pytren-2Q was consistently the most active of the compounds tested with any of the detection methods (diffusion disk or microdilution) and against yeasts and molds.

[0048] The pytren-2Q molecule is small (MW=374.16 g / mol), highly soluble in water (>0.4 g / mL), complies with Lipinski's rules, has good in silico pharmacokinetic properties and showed low toxicity in vitro (vero cells) and in vivo (BALB / c mice).

[0049] Although the antifungal mechanism of action of pytren-2Q has not yet been thoroughly investigated, it could be related, in part, to the metal-chelating properties of pytren-2Q, for the following reasons:

[0050] (i) Metal chelators are being used as antifungal drugs. To function properly, all living organisms, including pathogenic fungi, must regulate their intracellular levels of essential transition metals, such as iron, zinc, manganese, and copper. Effective assimilation and detoxification of these metals are necessary for pathogenic fungi to survive and proliferate within the infected host. In this respect, the mammalian host has developed so-called “nutritional immunity,” which addresses both the essentiality and toxicity of these metals to defend against fungal and bacterial invasion. Preventing pathogenic fungi from acquiring these crucial micronutrients is an effective means of preventing their proliferation.

[0051] This strategy has also been imitated by the topical or systemic use of drugs such as iron chelators like deferasirox and deferiprone, which have demonstrated antifungal efficacy against most human pathogenic fungi.

[0052] (ii) Most scorpion-type macrocycles, including pytren-2Q, are metal chelators.

[0053] (ii) In the metal chelation process, the side chain of the pytren macrocycle folds into the macrocyclic core, forming a structural cage where the metal is trapped more or less tightly, depending on the characteristics of the metal and the nature of the heterocycle. Protonation of the aliphatic amine between the macrocycle and the heterocycle appears to be crucial for achieving this closed conformation.

[0054] (iv) In addition, although being isomers, pytren-4Q is less active than pytren-2Q, a fact may be related to the lower chelating activity of pytren-4Q towards metals (Olmo F, Clares MP, 2014).

[0055] The present invention also relates to a pharmaceutical composition comprising a compound of formula (I) alone or combined with one or more additional active ingredients.

[0056] The pharmaceutical composition is intended in particular for use as an antifungal, and more specifically, against pathogenic fungi.

[0057] According to specific realizations, pathogenic fungi are pathogenic molds.

[0058] According to particular embodiments, the pharmaceutical composition is a semi-solid composition, selected from a cream, a gel or an ointment; a liquid composition, selected from a lotion, an eye drop, a syrup or an injectable solution; or a solid composition, selected from a tablet or a granule.

[0059] In addition to the preparation of oral medications, the compounds will also be used in various formulations for topical, vaginal, and ocular use, such as creams, gels, and eye drops. This latter application could be of great interest, since there is currently only one approved ocular antifungal, natamycin.

[0060] The amount of compound (I) present in the pharmaceutical composition varies depending on the type of preparation. In oral suspensions, concentrations range from 10 mg / ml to 40 mg / ml. In topical formulations (creams, ointments, or eye solutions), concentrations range from 0.5% to 2% (5 mg / g - 20 mg / g), while in shampoos, concentrations range from 1% to 2% (10 mg / g - 20 mg / g). For oral formulations (tablets or capsules), the dose per unit ranges from 50 mg to 500 mg, while in intravenous (injectable) formulations, the concentration ranges from 0.1 mg / ml to 10 mg / ml. For vaginal formulations (suppositories or creams), concentrations range from 1% to 2% (10 mg / g - 20 mg / g). For nail treatments (lacquers or solutions), concentrations include ranges between 5% and 8% (50 mg / g - 80 mg / g).

[0061] The pharmaceutical composition of the invention can be administered orally, topically, vaginally, ocularly, and intravenously.

[0062] Description of the figures

[0063] Fig. 1. Percentage of inhibition relative to the control drug (100%) of the three most active compounds pytren-S, pytrenEL-2Py, and pytren-2Q. Pathogenic yeasts are classified according to their phylogenetic similarities. MA: millions of years.

[0064] Fig. 2. Percentage of inhibition relative to the control drug (100%) of pytren-S, pytrenEL-2Py, and pytren-2Q. Pathogenic molds are classified according to their phylogenetic similarities. MA: millions of years.

[0065] EXAMPLES

[0066] The present invention is further illustrated by the following examples, which cannot be considered as a limitation of its scope.

[0067] Reagents

[0068] The ligands pytren-2Q (L12) and pytren-4Q (L13) were synthesized as previously described [Clares, MP; 2011, 47, 5988]. Forbol 12-myristate 13-acetate (PMA), LPS (from Escherichia coli O111: B4), L-ascorbic acid (L-ASC), and resveratrol (RSV) were supplied by Sigma-Aldrich (St. Louis, MO, USA). Strains, isolates, cultures, and chemical reagents

[0069] C. parapsilosis (Ashford) Langeron et Talice (ATCC 22019), A. fumigatus Fresenius (ATCC MYA-3626), A. flavas Link (ATCC 204304) and P. variotii Byssochlamys spectabilis (Udagawa et Suzuki) Houbraken et Samson (ATCC 22319).

[0070] They analyzed a total of 57 clinical isolates (29 yeasts and 28 molds). Candida albicans (5), Pichia kudriavzevii (formerly Candida krusei) (4), Candida tropicalis (4), Candida parapsilosis (3), Nakaseomyces glabratus (formerly Candida glabrata (3), Meyerozyma guilliermondii (formerly Candida guilliermondii) (3), Candida dubliniensis (1), Clavispora lusitanea (1), Cryptococcus neoformans (2), Cryptococcus gattii (2), Trichosporon spp (1); Aspergillus fumigatus ss (A. fumigatus sensu stricto) (5), Aspergillus niger (4), Aspergillus flavus (3), Aspergillus terreus (3), Rizhopus arrhizus (formerly Rizhopus oryzae) (2), Mucor circinelloides (2), Paecilomyces variotii (1), Fonsecaea pedrosoi (1), Fusarium solani (2), Trichophyton mentagrophytes (1), Trichophyton rubrum (1), Scedosporium apiospermum (2), Lomentospora prolificans (formerly Scedosporium prolificans (1).The isolates were provided by the University of Chile (Chile) and the Faculty of Medicine of Reus (Spain), and the test panel included one strain of A. fumigatus and one of C. glabrata resistant to azoles. C. parapsilosis (Ashford) Langeron et Talice (ATCC 22019), A. fumigatus Fresenius (ATCC MYA-3626), A. flavus Link (ATCC 204304), and Byssochlamys spectabilis variotii (Udagawa et Suzuki) Houbraken et Samson (ATCC 22319) were used as control strains.

[0071] Antifungal susceptibility testing.

[0072] The disk diffusion test method was performed using unsupplemented Mueller-Hinton agar and 6-mm diameter paper disks containing 25 pg of the corresponding compounds. Control disks were loaded with 25 pg of fluconazole, 1 pg of voriconazole, or 20 pg of amphotericin B (CLSI m51-P, 10).

[0073] Broth microdilution methods specific to yeasts or molds were performed according to Clinical and Laboratory Standards Institute guidelines for yeasts (10) (CLSI 2-27 a3 08) and for molds (11) (m-38 a2 08). Two-dilution microdilution plates were prepared, with final drug concentrations (compound (I) or the reference drug used as a control) ranging from 64 to 0.12 pg / ml for all compounds tested. Amphotericin B and voriconazole were used as reference drugs. MICs were the lowest drug concentrations that visually achieved >90% or 100% inhibition of fungal growth compared to the control well.

[0074] Drug interactions were assessed using the checkerboard method and the fractional inhibitory concentration index (FICI) was used to rank drug interactions (Dannaoui E, Lortholary O., 2004).

[0075] RESULTS

[0076] Antifungal activity was determined against eight clinical isolates (six yeasts and two molds) as the diameter of the growth inhibition zone measured in mm (Table 1). All pytren derivatives, scorpiande and macrocycle (compounds L7-L21), were active (Table 1).

[0077] Table 1 shows the fungal growth inhibition and molecular structure of the L7-L21 polyamines. Numerical values ​​of the zone diameters in mm within cells are shown for each fungal pathogen. Yeasts: C. albicans (C. A), C. neoformans (C. N), C. tropicalis (C. T), C. parapsilosis (C. P), N. glabratus (formerly C. glabrata) (N. G), P. kudriavzevii (formerly C. krusei) (P. K), and M. guilliermondii (formerly C. guilliermondii) (M. G). Molds: A. fumigatus (A. F), and R. arrhizus (formerly R. oryzae) (R. A). Voriconazole, amphotericin B, and fluconazole were used as control antifungal agents.

[0078] The most active compounds L12 (pytren-2Q), followed by L10 (pytrenEL-2Py), L21 (pytren-S), L7 (pytren) and L15 (pytren-2QI) were selected for an expanded antifungal evaluation against twenty-three pathogenic yeasts and molds.

[0079]

[0080] Table 1. Moderate activity of pytren-2Q against pathogenic yeasts.

[0081] Table 2 shows the activity of pytren, pytren-S, pytrenEL-2Py, pytren-2QI and pytren-2Q against 28 clinical isolates of yeasts using a specific yeast microdilution procedure to calculate their minimum inhibitory concentrations (MICs). (These 28 clinical isolates, including an azole-resistant strain of C. glabrata, belong to 11 different pathogenic yeasts, 7 of which are included in the QMS list of priority fungal pathogens.

[0082]

[0083] Table 2

[0084] Table 2 shows the antifungal activities of selected pytren macrocyclic polyamines against human pathogenic yeasts. The numbers within the boxes represent the minimum inhibitory concentrations (MICs) in g / mL of the selected compounds pytren, pytren-S, pytrenEL-2Py, pytren-2QI, and pytren-2Q against pathogenic yeasts. The yeast strains are classified into critical, high, and moderate priority groups as in the WHO list of pathogenic fungi (WHO, 2022). Voriconazole (VRC) was used as a control antifungal agent.

[0085] The assay showed that all compounds were able to completely inhibit yeast growth. Interestingly, in the case of azole-resistant C. glabrata, it was found to be approximately eight times more sensitive to the compounds (MIC values ​​between 1 and 2 pg / ml) than to voriconazole (MIC 8 pg / ml) (Table 2). As in the disk diffusion assay, the most effective compounds were pytren-2Q, followed by pytrenEL-2Py and pytren-S (Table 2). Pytren2Q showed the closest range of activity to voriconazole, between 19 and 68% of voriconazole's antifungal activity for the WHO priority yeast group included in the assay (Table 2).

[0086] Interestingly, sensitivity and resistance to pytren-S, pytrenEL-2Py, and pytren-2Q were strain-specific, with P. kudriavzevii, C. tropicalis, and C. albicans being the most sensitive fungal pathogens and C. lusitanea, C. dubliniensis, and C. guilliermondii the most resistant (Fig. 1). Phylogenetically close fungal pathogens such as C. albicans and C. dubliniensis showed very different sensitivities to these compounds (Fig. 1).

[0087] Powerful activity of pytren-2Q against pathogenic molds, including WT and the WHO critical priority azole-resistant A. fumigatus.

[0088] The susceptibility of molds to pytren, pytren-S, pytrenEL-2Py, pytren-2QI, and pytren-2Q was further tested in 23 clinical isolates, including an azole-resistant strain of A. fumigatus, using a mold-specific microdilution method (as described above). These isolates belong to 12 species of pathogenic molds, 6 of which are among the WHO priorities (Table 3).

[0089]

[0090] Table 3

[0091] In addition, ATCC strains of A. fumigatus, A. flavas, and P. variotii were included as controls (Table 3).

[0092] Table 3 shows the antifungal activities of selected pytren macrocyclic polyamines against human pathogenic molds. The MICs in pg / ml of the selected compounds pytren, pytren-S, pytrenEL-2Py, pytren-2QI, and pytren-2Q are shown in the boxes. The mold strains are classified into the WHO's critical, high, and moderate priority pathogen groups (WHO, 2022). Voriconazole (VRC) or amphotericin B (AMB) were used as controls. Antifungal activity values ​​were calculated as MIC₉₀ (MIC causing 90% inhibition of growth compared to the untreated control) and MIC₁₀₀ (MIC causing 100% inhibition of growth). All compounds were able to stop fungal growth by 100% at the tested concentrations, except for T. mentagrophytes, F. solani, L. prolificans and S. apiospermum in which the compounds did inhibit 90% of their growth at concentrations lower than 8 pg / ml (Table 3).As in the disk diffusion and yeast microdilution assays, the most active compounds were pytren-2Q, pytrenEL-2Py, and pytren-S, while pytren and pytren-2QI had the lowest antifungal activity (Table 3). Pytren-2Q was as effective as voriconazole against WT isolates of A. fumigatus and 32 times more effective against its azole-resistant strain (Table 3).

[0093] By plotting the percentage of activity relative to the controls and classifying the pathogenic molds according to their phylogenetic relationships, we observed that, as with yeasts, sensitivity and resistance to pytren-S, pytrenEL-2Py, and pytren-2Q were strain-specific, and that phylogenetically close fungal pathogens showed different sensitivities, such as 7. mentagrophytes and T. rubrum, or within the Aspergillus / aceae family, A. terreus and A. flavus (Fig. 2). The most sensitive pathogenic fungus was T. rubrum, followed by the two mucoral strains M. circillenoides and R. arrhizus (Fig.

[0094] 2). T. mentagrophytes, a representative of the subclass Hypocreomycetidae, F. solani, L. prolificans and S. apiospermum, while pytren-S, pytrenEL-2Py and pytren-2Q were sometimes even more effective than the control in stopping growth by 90%, did not completely stop fungal proliferation by 100% (Fig. 2).

[0095] REFERENCES

[0096] Martín-Montes A, Clares MP, Martín-Escolano R, Delgado-Pinar E, Marín C, Verdejo B, Martínez-Camarena Á, Molina-Carreño D, García-España E, Sánchez-Moreno M. Heterocyclic Diamines with Leishmanicidal Activity. ACS Infect Dis. 2021 Dec 10;7(12):3168-3181. doi: 10.1021 / acsinfecdis.1c00215. Epub 2021 Nov 4. PMID: 34734686.

[0097] Olmo, F., Marín, C., Clares, MP, Blasco, S., Albelda, MT, Soriano, C., Gutiérrez-Sánchez, R., Arrebola-Vargas, F., García-España, E., and Sánchez-Moreno, M. (2013) Scorpiand-like azamacrocycles prevent the chronic establishment of Trypanosoma cruzi in a murine model. Eur. J. Med. Chem. 70, 189-198.Marín C, Clares MP, Ramírez-Macías I, Blasco S, Olmo F, Soriano C, Verdejo B, Rosales MJ, Gomez-Herrera D, García-España E, Sánchez-Moreno M. In vitro activity of scorpiand-like azamacrocycle derivatives in promastigotes and intracellular amastigotes of Leishmania infantum and Leishmania braziliensis.. Eur J Med Chem. 2013 Apr;62:466-77, doi: 10.1016 / j.ejmech.2013.01.001. Epub 2013 Jan 19. PMID: 23395967.

[0098] Olmo F, Clares MP, Marín C, González J, Inclán M, Soriano C, et al. Synthetic single and double aza-scorpiand macrocycles acting as inhibitors of the antioxidant enzymes iron superoxide dismutase and trypanothione reductase in Trypanosoma cruzi with promising results in a murine model. RSC Adv. 2014:4:65108-20.

[0099] Clares, M. P.; Blasco, S.; Inclán, M.; del Castillo Agudo, L.; Verdejo, B.; Soriano, C.; Doménech, A.; Latorre, J.; García-España, E. Manganese(II) Complexes of Scorpiand- like Azamacrocycles as MnSOD Mimics. Chem. Commun. 2011, 47, 5988.

[0100] Dannaoui E, Lortholary O, Dromer F. 2004. In Vitro Evaluation of Double and Triple Combinations of Antifungal Drugs against Aspergillus fumigatus and Aspergillus terreus. Antimicrob Agents Chemother 48:. https: / / doi.org / 10.1128 / aac.48.3.970-978.2004.

Claims

AMENDED CLAIMS received by the International Bureau on September 14, 2026 (14.09.2026)

1. [modified] An azapyridinophane compound of Formula (I) where Ri is selected from one of the following groups: or a pharmaceutically acceptable solvate or salt thereof, for use against pathogenic fungi.

2. A compound according to claim 1 for use against pathogenic fungi, wherein the pharmaceutically acceptable salt is selected from hydrochloride, hydrobromide, sulfate, nitrate, phosphate, a carboxylic acid salt, a sulfonic acid salt, a fatty acid salt, and an amino acid salt.

3. A compound according to claim 1 for use against pathogenic fungi, which is selected from those in which Ri is a 4-quinolinyl or 2-quinolinyl radical, and more preferably, a 2-quinolinyl radical.

4. A compound according to claim 1 for use against pathogenic fungi, which is selected from pytren-2Q, pytrenEL-2Py, pytren-S, pytren and pytren-2QI, preferably pytren-2Q.

5. A compound according to claim 1 for use against pathogenic fungi, having a minimum inhibitory concentration between 0.12-8 μg / ml (MIC90) and 0.12-64 μg / ml (MIC100). [Reivindicación 6] Un compuesto según la reivindicación 1, para su uso como antifúngico contra un hongo patógeno seleccionado entre Candida albicans, Pichia kudriavzevii, Candida tropicalis, Candida parapsilosis, Nakaseomyces glabratus, Meyerozyma guilliermondii, Candida dubliniensis, Clavispora lusitanea, Cryptococcus neoformans, Cryptococcus gattii, Trichosporon spp Aspergillus fumigatus ss, Aspergillus niger, Aspergillus flavus, Aspergillus terreus, Rizhopus arrhizus, Mucor circinelloides, Paecilomyces variotii, Fonsecaea pedrosoi, Fusarium solani, Trichophyton mentagrophytes, Trichophyton rubrum, Scedosporium apiospermum y Lomentospora prolificans, Trichophyton sp, Microsporum sp, Epidermophyton sp, Blastomyces sp, Histoplasma sp, Sporothrix sp, Saksenaea sp, Cladophialophora sp, Absidia sp, Coccidioides sp, Paracoccidioides sp, Emmonsia sp, y Talaromyces sp.

7. A compound according to claim 1, for use against a pathogenic fungus of the genus Aspergillus, and preferably A. fumigatus.

8. A compound according to any one of claims 1 to 7, wherein the use against pathogenic fungi is a use as a medicament.

9. A pharmaceutical composition comprising a compound of formula (I) alone or combined with one or more additional active ingredients for use as an antifungal, preferably against pathogenic fungi.

10. A pharmaceutical composition according to claim 9 for use as an antifungal drug, preferably against pathogenic fungi, selected from a cream, a gel or an ointment; a liquid composition and a solid composition.

11. A pharmaceutical composition according to claim 9 or 10 for use as an antifungal drug, preferably against pathogenic fungi, selected from a composition for oral, topical, vaginal, ocular and intravenous administration.

12. A pharmaceutical composition according to any one of claims 9 to 11 for use [0001]Declaration pursuant to Article 19.1) PCT [0002]With regard to the international application mentioned in the reference, it is hereby stated that the following claims have been amended pursuant to Article 19 of the PCT in view of the International Search Report: [0003]- In claim 1, a compound included in the general formula has been omitted, specifically the compound in which the radical Ri is [0004] [0005]thus limiting the scope of the main claim. [0006]No other claim has been modified from the version originally submitted. [0007]Any modifications to the description that are necessary to adapt it to the amended claims will be made in the national phase.

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