Pharmaceutical composition containing trifarotene for use in the treatment of fungal infections

Abstract

The present invention relates to a composition for the treatment of fungal infections of C. albicans and C. non-albicans, such as C. auris, C. glabrata, C. tropicalis, C. krusei and Trichophyton spp., in particular Trichophyton mentagrophytes complex, Tricophyton rubrum.

Description

PHARMACEUTICAL COMPOSITION CONTAINING TRIFAROTENE FOR USE IN THE TREATMENT OF FUNGAL INFECTIONSFIELD OF THE INVENTION

[0001] The present invention relates to a pharmaceutical composition containing trifarotene for the treatment of fungal infections. In particular, the present invention relates to a pharmaceutical composition containing trifarotene for use in the treatment of fungal infections caused by Candida species and in particular on fungal infections caused by C. albicans and by C. non-albicans such as C. auris, C. glabrata, C. tropicalis and C. krusei, and by the genus Trichophyton, in particular Trichophyton mentagrophytes complex and Trichophyton rubrum.STATE OF THE ART

[0002] Over the last 20 years, the incidence of invasive fungal infections (IFIs) has increased significantly. In the global perspective, Candida spp. and Aspergillus spp. are the most frequent fungal aetiological agents, often responsible for invasive fungal infections in severely immunocompromised patients, burdened with a high morbidity and mortality rate. In recent years, fungal infections by environmental mycetes, previously considered non-pathogenic, have also been spreading. A wide range of pathogens belonging to the Mucorales order (e.g. Rhizopus spp), such as hyalohyphomycetes (e.g. Fusarium and Scedosporium spp) or pheohyphomycetes (e.g. Alternaria spp and Cladophialophora bantiana) together with Cryptococcus neoformans, constitute a major cause of infectious morbidity and mortality, with an estimated 600,000 deaths per year.

[0003] Mycoses caused by these organisms, especially the rhino-cerebral forms, are often difficult to treat and require specialist consultations (1 ). The increase of invasive fungal infections (IFIs), but also of non-invasive fungal infections, has been related to the increased prevalence of immunocompromised individuals, especially due to iatrogenic causes, such as oncological chemotherapies, or immunosuppressive therapies in autoimmune diseases, and to the increase of AIDS patients in countries with difficult access to treatment. In addition, improvements in surgical management, the introduction of selective immunosuppressive drugs, and the increase in the number of organ, solid or bone marrow transplant recipients experiencing neutropenia are further risk factors for the development of fungal infections. Similarly, the increased use of intravenous devices, prolonged stay in intensive care units and the administration of antibacterial therapies that alter the normal human microbiota are further predisposing factors for the occurrence of invasive fungal infections.

[0004] The GAFFI (Global Action Fund for Fungal Infections) has estimated that at least 150 patients die every hour worldwide from invasive fungal infections (IFIs), equal to 1 ,350,000 per year. Some neutropenic oncology patients need to undergo antifungal therapy in 1 / 3 of the cases when they do not respond to broad-spectrum antibiotic therapies because they are predisposed to developing fungal infections (2).

[0005] Among invasive fungal infections, invasive pulmonary aspergillosis (IPA) and systemic infections sustained by fungi belonging to the Candida genus are those most frequently encountered in the clinical setting. In particular, IPA represents the most common fungal infection among the invasive ones in the world, which is difficult to diagnose early. The responsible fungus, Aspergillus fumigatus, a ubiquitous fungus widespread in the environment, can cause a broad spectrum of diseases such as bronchopneumonia, allergic diseases, sinusitis, aspergilloma and pneumonia. The lung represents the hotbed from which the infection can spread to many other organs in susceptible patients, turning into a rapidly progressive systemic disease with often fatal outcome. During the recent international congress of the European Respiratory Diseases Society, it was stated that the aspergillosis problem is underestimated, despite the fact that there are about 300000 cases per year and 240000 in Europe alone. This is a major threat to the health of patients at risk, thus, with a mortality rate, if left untreated, of over 90%. However, even when invasive pulmonary aspergillosis (IPA) is treated, the mortality rate is 50% in immunocompromised patients. Guidelines for the treatment of IPA call for the use of antifungal drugs of the azole class. However, cases of resistance to the drugs used to occur.

[0006] Furthermore, during the pandemic triggered by Sars-CoV-2, there was an exponential increase in Mucormycosis infections, infections caused by various fungal organisms belonging to the Mucorales order, which are mainly widespread in India. This suggests that infectious diseases that are considered rare, may instead emerge especially in immunocompromised individuals, leading to a worsened prognosis.

[0007] In addition to aspergillosis, an increased occurrence of fungal infections caused by fungi belonging to the Candida genus has been observed in recent years, especially in immunocompromised patients.

[0008] Infections caused by microorganisms of the Candida genus can manifest themselves in various ways, for example, as localised diseases of the skin or nails, diseases affecting the mucous surfaces of the mouth, such as thrush, or of the vagina, or even as life-threatening systemic candidiases that spread through the blood stream,involving many organs such as the central nervous system, heart, kidneys, liver, bones, muscles, joints, spleen, and eyes.

[0009] The species most frequently responsible for candidiasis include C. albicans, C. tropicalis, Candida kefyr, C. glabrata, C. krusei and Candida parapsilosis (3).

[0010] The treatment of such candidiasis is currently complicated by the fact that some strains, particularly C. glabrata and C. krusei, have recently developed resistance to commonly used antifungal drugs. These drugs are usually azole-based. In particular, C. auris is proving to be a real global health threat, as this fungus is responsible for the most frequent nosocomial infections acquired by individuals weakened by other diseases, undergoing surgery or immunocompromised, and is proving resistant to the most common antifungal drugs currently available, such as azoles, echinocandins and amphotericin B.

[0011] In recent years, in addition to the growing number of infections caused by species of the Candida genus, there has been a significant increase in fungal infections sustained by dermatophytes, in particular Trichophyton spp.. These infections affect a heterogeneous population, affecting paediatric subjects as well as adults and the elderly, with a particularly high prevalence in individuals exposed to conditions conducive to fungal proliferation. The age groups most frequently affected include children and adolescents, who are often exposed in community settings such as schools and sports facilities, where inter-human transmission is facilitated. In adults, the increase is particularly evident in individuals who engage in sporting activities, frequent hot and humid environments or use occlusive footwear for long periods, as well as in individuals with predisposing conditions such as immunosuppression, diabetes mellitus, vascular dysfunction, alterations in the skin microbiota or microtrauma of the skin and adnexa. A further relevant factor in the epidemiology of these infections is the zoonotic species of Trichophyton, which can be transmitted from animals to humans, particularly from domestic and farm animals; this mode contributes increasingly to the spread of dermatophytoses, especially in children and those in close contact with animals. The combination of these factors, coupled with the circulation of more virulent or sometimes resistant dermatophyte strains to conventional therapies, makes dermatophyte infections an expanding clinical problem, with a significant impact on the skin and skin adnexa, including body hair, head hair and nails. A further element contributing to the therapeutic difficulty of nail infections from Trychophyton spp. is their marked propensity to form biofilms within and beneath the nail plate. This compact aggregate of hyphae, fungal cellsand extracellular matrix, commonly referred to as a “dermatophytoma” constitutes a physical and biological barrier that hinders the penetration of conventional antifungal drugs. The presence of biofilm in the nail bed significantly reduces the effectiveness of topical treatments and, in some cases, also limits the response to systemic therapies, making eradication of the infection slower and more complex. This phenomenon is therefore a significant pathogenetic and clinical factor, associated with more recalcitrant forms of onychomycosis and a greater likelihood of recurrence. Resistance to traditional drugs greatly complicates the treatment of such infections, compromising the final outcome of the therapies used. The level of resistance of some fungal species is of particular concern as it severely limits the treatment options available to patients with both invasive and superficial infections.

[0012] Moreover, the drugs currently available for the treatment of fungal infections have high toxicity that risks further compromising the general health of patients using them, especially in the presence of a compromised clinical picture and / or comorbidities. These problems are even more critical when long-term treatment with currently used antifungal drugs is required.

[0013] The development of drug-resistant strains of fungi, the high toxicity of traditional antifungal drugs, especially in the case of prolonged and massive use, has increasingly reinforced the need to find new and more effective therapeutic strategies for fungal infections.

[0014] In particular, there is a strong need to identify new compounds with antifungal activity. There is also a particular need to identify new antifungal compounds that are both effective in treating fungal infections and selective so as to reduce undesirable effects for the treated patient.

[0015] There is also a particular need to identify antifungal compounds that are both effective in treating fungal infections and have reduced cytotoxicity in order to avoid side effects for the patient.

[0016] There is also a particular need to identify antifungal compounds that are both effective in treating fungal infections and less likely to generate drug resistance.

[0017] There is also a particular need to identify compositions that can effectively treat fungal infections caused by C. krusei, C. tropicalis, C. auris, C. glabrata.

[0018] There is also a particular need to identify compositions that can effectively treat fungal infections caused by Trichophyton spp., in particular Trichophyton mentagrophytes complex and Trichophyton rubrum.

[0019] There is therefore a need to identify new compositions for use in the treatment of fungal infections that are effective and possibly safe for patients, and preferably easy to access and clinically manage.

[0020] Of the several hundred known species belonging to the Candida genus, only a few are capable of causing human infections, and these species differ significantly, however, in both virulence and adaptation strategies. These Candida species are located at divergent points in a continuous spectrum of types of interactions between the host species and the microorganism. It appears, in fact, that the various species of Candida have developed distinct biochemical, metabolic and physiological adaptations over the course of evolution, which they use to adapt to commensal niches and achieve full virulence. The different species of Candida therefore differ from each other both in structure and genetic characteristics, but above all in different virulence factors, such as: isotype switching, drug resistance, biofilm formation and quorum sensing.

[0021] In fact, each species of Candida has peculiar features both genotypically and phenotypically and virulence characteristics that differ from the other species and that characterise and distinguish one species of Candida from another.

[0022] These differentiation factors between species of Candida include various hostrecognition biomolecules, e.g. adhesin receptors, i.e. proteins on the surface of cells that recognise fungal pathogens, morphogenesis, i.e. the reversible transition between unicellular yeast cells and filamentous growth forms. In particular, Candida species are in yeast form at room temperature, when exposed to temperatures of 37°C or higher they become a filamentous fungus and produce structures called hyphae, which can lead to infection in humans. Further differences can be found in the aspartic proteases secreted by the fungi of the different Candida species and phospholipases, i.e. molecules that destroy human tissue and allow Candida species to cause systemic or local infections in humans. By destroying tissue with these proteases, hyphae make their way into the skin or mucous membranes and can reach blood vessels, causing systemic infections.

[0023] Among the different virulence factors possessed by the various Candida species, “phenotypic switching”, i.e. the transition from yeast to filamentous form, is accompanied by changes in antigen expression, colony morphology and tissue affinities in all Candida species.

[0024] Moreover, each Candida species has different susceptibility to antifungals, so that, at the clinical level, each Candida infection is unique and requires specific therapy based on susceptibility testing, and no certain response can be predicted in themanagement of such infections. Each species of Candida in particular may only be sensitive to some of the existing antifungals, so to combat infections by each Candida species, a precise therapeutic strategy must be devised, both in terms of the compounds used and the methods of administration.

[0025] Candida is a saprophytic fungus normally present in several districts of the body (including the vagina and the gastrointestinal tract) in a latent form (as blastospora) and is usually controlled by the immune system and the local bacterial flora (the lactobacilli). Several types of Candida have been isolated, which are usually distinguished into C. albicans and C. non-albicans. C. albicans is usually identifiable in about 80% of cases, other species, grouped and identified as C. non-albicans, such as C. glabrata, C. parapsilosis, C. krusei and C. tropicalis, responsible for about the remaining 20% of candidiasis cases. In recent years we are witnessing an increase in infections caused by C. non-albicans species resistant to common treatments.

[0026] These species of C. non-albicans, which used to cause infections only in immunocompromised individuals (such as HIV-positive individuals or diabetics), now also affect apparently healthy individuals, and their presence should be suspected especially in the presence of an infection with less pronounced symptoms but a greater tendency to become chronic.

[0027] It is well known that the treatment of Candida infections must be appropriate and differentiated according to the type of Candida underlying the infection: in particular, while C. albicans responds well and quickly to common “azole” antimycotics, C. nonAlbicans strains have proven resistant to conventional treatments. On the other hand, as mentioned, such "non-albicans" species are becoming increasingly widespread and can cause severe infections especially in clinically compromised patients.

[0028] In particular, C. albicans differs from non-albicans species such as C. krusei, C. glabrata, C. parapsilosis and C. auris in that it is the only species capable of growing in hyphal form during virulence. Isotype switching from the yeast to the hyphal form plays a key role in the pathogenesis of C. albicans. Hyphae, in fact, not only promote the penetration of the fungus into the tissues and blood vessels of the host, causing systemic infections, but in the hyphal form C. albicans also expresses the gene ECE1, absent, however, in yeast cells, which codes for a toxin known as “candidalysin”. This toxin exerts a cytolytic action against epithelial cells, thus destroying the epithelial barrier and further promoting the spread of Candida at the tissue level. Candidalysin also induces the recruitment of neutrophils, and activates the TH17 lymphocyte response, triggeringimmune-mediated tissue damage. Fungal hyphae also contribute to the production of biofilms as they act as a support, promoting the adhesion of the Candida yeast cells. Biofilms can be produced on both biotic and abiotic surfaces (catheters, valve prostheses, joints) and are more structured in C. albicans than biofilms produced by non-albicans species. C. non-albicans species, on the other hand, can grow in pseudohyphal form during virulence but are not capable of producing true hyphae, with the exception of C. krusei, which can grow in hyphal form under certain conditions, similar to C. albicans. C. glabrata, on the other hand, exists in nature, both as a commensal and as a pathogenic fungus, only in the yeast form.

[0029] Therefore, C. albicans, represents the most virulent species even though, unlike the non-albicans species, C. albicans has retained sensitivity to drugs traditionally used to treat both superficial and systemic candidiasis.

[0030] In contrast, C. glabrata, C. parapsilosis and C. auris have developed resistance to these drugs, while C. krusei has innate resistance to fluconazole.

[0031] The emerging species C. auris has acquired resistance to azoles and also to echinocandins, another class of antifungal drugs commonly used for systemic infections. The phenomenon of multi-drug resistance makes the management of such infections extremely difficult. Furthermore, C. auris produces biofilms that have been shown to be resistant to common disinfectants even on the skin of apparently healthy individuals. This encourages the spread of C. auris infections both by direct contact between different individuals and by contact with contaminated devices. Therefore, while the treatment of such infections is extremely difficult, it is of paramount importance to find treatment methods that can be effective in treating them.

[0032] Therefore, there remains a need to identify molecules or compounds that may be effective in treating fungal infections sustained by C. albicans strains.

[0033] Therefore, there remains a need to identify molecules or compounds that may be effective in treating fungal infections sustained by C. non-albicans strains.

[0034] The need therefore remains to identify molecules or compounds that may be effective in treating fungal infections sustained by C. krusei, C. tropicalis, C. auris, C. glabrata.

[0035] In particular, there is a need to identify molecules or compounds that have a non-negative impact on the patient's general health and whose use does not generate undesirable side effects.

[0036] There is a need to identify molecules or compounds that do not induce drug resistance.

[0037] There is therefore a need to identify molecules or compounds of natural origin that can be used to develop new antifungal therapeutic strategies.

[0038] A study conducted by researchers from the Department of Haematology at the University of Rome "La Sapienza" entitled Infectious complications in patients with acute promyelocytic leukaemia treated with the AIDA regimen published on 15 May 2003 in the journal Leukemia showed that patients with promyelocytic leukaemia treated with all-trans retinoic acid (ATRA) had a lower risk of developing fungal infections from C. albicans, and Aspergillus spp. Based on this observation, experiments in vitro and subsequently in vivo were conducted to test the use of ATRA for the prevention of fungal infections. Tests were conducted in vitro in a preclinical model of invasive pulmonary aspergillosis, which showed that ATRA exerts a fungistatic action in vitro against Aspergillus fumigatus and C. albicans, blocking the germination and replication of these fungi. These in vitro experiments have also shown that ATRA exerts a synergistic action in combination with conventional antifungals such as Amphotericin B or Posaconazole in counteracting the germination of aspergillar conidia.

[0039] All-trans retinoic acid (ATRA) is a derivative of vitamin A, of the retinoid class, and is its active metabolite. Retinoids are chemical compounds derived from vitamin A and chemically related to it. Retinoids, through their related nuclear receptors, exert powerful effects on cell growth, differentiation and apoptosis and hold significant promise for cancer therapy and chemoprevention. Retinoids are divided into four groups according to their molecular structure and receptor selectivity. The first generation group comprises isomers and naturally occurring compounds such as retinol, retinal, tretinoin (ATRA), isotretinoin and al itretinoin. The second-generation group comprises synthetic analogues formulated for oral and non-topical administration, such as etretinate and its metabolite acitretin. The third-generation group comprises retinoidal derivatives of benzoic acid such as adapalene, bexarotene and tazarotene. Finally, the fourth-generation group comprises topical retinoids with selectivity towards the RAR receptor located in the epidermis. Differentiation therapy with ATRA has marked a major breakthrough and has become the drug of first choice in the treatment of acute promyelocytic leukaemia (APL). Conversions of 13-cis-retinoic acid and 9-cis-retinoic acid to all-trans-retinoic acid are very rapid. Currently, two distinct families of retinoid-responsive nuclear receptors have beenidentified and characterised: retinoic acid receptors (RAR) and retinoid receptors (RXR), each of which comprises three isoforms, a, [3 and y.

[0040] Although ATRA was shown to be an active compound in the treatment of C. albicans, no activity was shown against fungi of the Candida non- albicans species. On the other hand, strains of C. non-albicans species differ from C. albicans genetically and also in their aetiology and mode of dissemination in the host organism.

[0041] Moreover, apart from ATRA, the other known first, second and third generation retinoids have not proved active in the treatment of fungal infections. On the other hand, the different molecular structure and kinetics of retinoids of the various generations do not allow one to assume activity against fungal infections.

[0042] However, the inventors have highlighted for the first time that trifarotene has an unexpected antifungal effect. Furthermore, the inventors have highlighted that trifarotene exerts its antifungal action against C. non-albicans species.

[0043] Trifarotene is a compound belonging to the retinoid class but with a completely different chemical structure to that of ATRA.

[0044] Below are the ATRA and trifarotene molecules for immediate comparison:ENE

[0045] The differences between ATRA and TRIFAROTENE both from a molecular point of view and in terms of interaction with other molecules did not allow an antifungal effect to be assumed for trifarotene.

[0046] Trifarotene is a fourth-generation retinoid, selective for gamma receptors. Unlike all other retinoids, trifarotene is defined as such not by its chemical structure, i.e. a triaryl, but by the same intranuclear retinoid target, RAR receptors (alpha, beta and gamma). However, trifarotene has some peculiarities, also in its action with RAR receptors since trifarotene acts selectively on gamma RAR and has no action on RXR receptors, unlike other retinoids.SUMMARY OF THE INVENTION

[0047] An object of the present invention is to provide a molecule or composition having antifungal activity.

[0048] An object of the present invention is to provide a molecule or composition having antifungal activity against C. albicans.

[0049] An object of the present invention is to provide a molecule or composition having antifungal activity against C. non-albicans species, such as C. auris, C. glabrata, C. tropicalis and C. krusei.

[0050] A further object of the invention is to provide a pharmaceutical composition for the treatment of fungal infections in particular fungal infections by C. albicans and / or by C. non-albicans, such as for example C. auris, C. glabrata, C. tropicalis and C. krusei and of the genus Trichophyton spp, in particular Trichophyton mentagrophytes complex and Tricophyton rubrum.

[0051] According to the invention, a pharmaceutical composition comprising trifarotene is provided for the treatment of fungal infections.

[0052] A composition suitable for treating human or animal fungal infections is provided.

[0053] According to the invention, a pharmaceutical composition comprising trifarotene is provided for the treatment of fungal infections sustained by one or more of the following species: C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei and of the genus Trichophyton spp., in particular Trichophyton mentagrophytes complex or Trichophyton rubrum.

[0054] According to the invention, a pharmaceutical composition comprising trifarotene is provided for the treatment of cutaneous or mucocutaneous fungal infections sustained by one or more of the following species: C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei.

[0055] According to the invention a pharmaceutical composition comprising trifarotene is provided for the treatment of cutaneous fungal infections of skin adnexa caused by dermatophytes of the genus Trichophyton spp., in particular Trichophyton mentagrophytes complex or Tricophyton rubrum.

[0056] In a further aspect of the invention, a pharmaceutical composition is provided for the treatment of fungal infections comprising at least one compound suitable for binding the HSP90 protein.

[0057] In a further aspect of the invention a pharmaceutical composition is provided for the treatment of fungal infections comprising at least one compound suitable forbinding the HSP90 protein of a fungus, wherein said fungal infections are fungal infections sustained by one or more of the following species: C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei and Trichophyton spp., in particular Trichophyton mentagrophytes complex or Trichophyton rubrum.

[0058] Preferably, said compound suitable for binding the HSP90 protein of a fungus is selected from the retinoid group. Preferably, said compound suitable for binding the HSP90 protein of a fungus is selected from the group of first generation retinoids, for example retinol, retinal, tretinoin (Retin-A), isotretinoin and alitretinoin; of the second- generation retinoids, e.g. etretinate and its metabolite acitretin; of the third-generation retinoids, e.g. tazarotene, bexarotene and adapalene; of the fourth-generation retinoids, which include trifarotene and seletinoid G. To a greater extent preferred said compound suitable for binding the HSP90 protein of a fungus is selected from the group of fourth generation retinoids, and in particular trifarotene and seletinoid G.

[0059] The invention provides a composition effective in the treatment of fungal infections, particularly effective in the treatment of fungal infections caused by mycetes that have demonstrated resistance to traditional drugs. This makes it possible to treat with the composition of the invention fungal infections for which known drugs have proved ineffective.

[0060] Moreover, thanks to the invention, a composition is provided that has extremely limited or no side effects on the patient’s health. Thus, the invention provides a composition that can be used to treat fungal infections of patients with a delicate or compromised general clinical picture without risking further worsening the patient’s clinical condition. The inventors found such a composition to be extremely effective in treating fungal infections caused by C. non-albicans species and in particular by one or more of C. auris, C. tropicalis, C. glabrata and C. krusei and in treating infections from Trichophyton spp.

[0061] To date, there is no commercially available formulation or composition based on trifarotene alone or in combination with traditional antifungals.

[0062] The inventors found that trifarotene has a unique mechanism of action different from all molecules used as antifungals. Trifarotene interacts with two key proteins in the life cycle of Candida spp., heat shock protein (Hsp) 90 and 14a-demethylase. While 14a- demethylase is the target of azoles, drugs used in common clinical practice for fungal infections, trifarotene is the only molecule capable of binding HSP90, potentially being used in all those cases of infections sustained by azole-resistant mycetes.

[0063] Moreover, to date there is no known resistance on the HSP90 protein, allowing trifarotene the first molecule in its class to function as an antifungal. In addition, trifarotene has an extremely different chemical structure from other retinoids, and is even chemically classified as a triaryl.

[0064] Experiments conducted in vitro have shown that trifarotene has an antifungal effect, not only fungistatic but also fungicidal, against C. albicans and C. non-albicans species and against Trichophyton spp, in particular Trichophyton mentagrophytes complex, or Tricophyton rubrum.

[0065] Furthermore, the composition of the invention can be used effectively to treat fungal infections caused by a plurality of mycetes. Indeed, as discussed above, trifarotene has been shown to be effective for various fungal populations. Whereas with the known compositions in the case of infections caused by a plurality of mycetes it was necessary to use different antifungal agents, the composition of the invention is effective towards various species of Candida, which have different structural and biochemical characteristics, and the antifungal effect has also been shown on Trichophyton spp., in particular Trichophyton mentagrophytes complex or Tricophyton rubrum. This further reduces possible side effects for the patient without compromising the effectiveness of the treatment, in fact guaranteeing good treatment effectiveness.

[0066] According to the invention topical formulations for topical use comprising a composition containing trifarotene are provided for the treatment of fungal infections, in particular for the treatment of fungal infections sustained by one or more of the following species: C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei.

[0067] According to the invention, formulations for topical use are also provided comprising a composition containing trifarotene for the treatment of fungal infections of the skin and skin adnexa caused by dermatophytes of the genus Trichophyton spp., in particular Trichophyton mentagrophytes complex or Tricophyton rubrum.

[0068] According to the invention compositions are provided for oral use comprising a composition containing trifarotene for the treatment of infections sustained by one or more of the following species C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei or of the genus Trichophyton spp, in particular Trichophyton mentagrophytes complex or Tricophyton rubrum.

[0069] According to the invention compositions are provided for intravenous or intramuscular administration comprising a composition containing trifarotene for thetreatment of infections sustained by one or more of the following species C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei.

[0070] Preferably, a composition containing trifarotene is provided for the treatment of mucosal infections sustained by one or more of the following species C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei.

[0071] According to the invention, a pharmaceutical composition comprising trifarotene is provided for the treatment of infections sustained by one or more of the following species: C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei wherein said fungal infections are fungal infections of the mucous membranes.

[0072] According to the invention, a pharmaceutical composition comprising trifarotene is provided for the treatment of infections sustained by one or more of the following species: C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei wherein said fungal infections are fungal infections of the vaginal, nasal, oral, anal mucosa.

[0073] According to the invention, a composition containing trifarotene is provided for the treatment of fungal infections of C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei in immunocompromised patients and in particular in patients with one or more of acute myeloid leukaemia (AML), human stem cell transplantation (HSCT) and graft versus host disease (GvHD).

[0074] According to the invention, a composition containing trifarotene is provided for the treatment of fungal infections of C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei, or from the genus Trichophyton spp., specifically Trichophyton mentagrophytes complex or Trichophyton rubrum, wherein said fungal infections are epidermal fungal infections.

[0075] According to the invention, a composition containing trifarotene is provided for the treatment of fungal infections of C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei wherein said fungal infections are pulmonary fungal infections.

[0076] According to the invention, a composition containing trifarotene is provided for the treatment of fungal infections of C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei wherein said fungal infections are fungal infections of the nails.

[0077] According to the invention a composition containing trifarotene is provided for the treatment of fungal infections of Trichophyton spp., in particular Trichophyton mentagrophytes complex or Trichophyton rubrum.

[0078] According to the invention, a composition containing trifarotene is provided for the treatment of fungal infections of Trichophyton spp., in particular Trichophytonmentagrophytes complex or Trichophyton rubrum wherein said fungal infections are fungal infections of the nails.According to the invention, a composition containing trifarotene is provided for the treatment of fungal infections of Trichophyton spp., in particular Trichophyton mentagrophytes complex or Trichophyton rubrum, wherein said infections are infections of the skin or skin adnexa, in particular of the nails.

[0079] According to the invention a composition containing trifarotene is provided for the treatment of fungal infections of Trichophyton spp., in particular Trichophyton mentagrophytes complex or Trichophyton rubrum, wherein said infections are infections of the skin or skin adnexa wherein said composition further comprises an additional fungal agent, said additional fungal agent preferably being terbinafine.

[0080] According to the invention, a composition is provided for use in the treatment of fungal infections of Trichophyton spp., in particular Trichophyton mentagrophytes complex or Trichophyton rubrum, comprising trifarotene and terbinafine, wherein said composition comprises terbinafine between about 1 mg / L and about 0.06 mg / L and trifarotene between about 1 mM and about 0.25 mM.

[0081] Skin adnexa are specialised structures of the skin such as hair, nails, sebaceous glands and sweat glands, which develop from the epidermis and perform vital protection, thermoregulation and sensitivity functions. These structures are mainly found in the dermis and are composed of keratin, protecting the body and regulating hydration and temperature.

[0082] The composition of the invention containing trifarotene is suitable for use in the treatment of fungal infections in humans or animals. Preferably the formulation for topical use is in the form of an ointment, topical cream, topical gel, liniment, paste, film, solution, hydrogel, liposome, transferable vesicle, cream, lotion, dermal patch, transdermal patch, transdermal spray.

[0083] Preferably, the pharmaceutical composition comprises trifarotene in a percentage between about 0.001 wt% and about 10wt%, preferably between about 0.005wt% and about 5wt%, more preferably between about 0.01 wt% and about 1wt%.

[0084] Preferably, the pharmaceutical composition comprises trifarotene in a percentage between 0.1 and 2 mM, preferably between 0.25 mM and 1.5 mM, more preferably between 0.5 mM and 1 mM.

[0085] In some preferred versions, the pharmaceutical composition also comprises an additional antifungal agent.

[0086] Preferably at least one additional antifungal agent is selected from the group consisting of azoles such as fluconazole, voriconazole, posaconazole, itraconazole, ketoconazole, isavuconazole, thioconazole, opelconazole, miconazole, from polyenes such as amphotericin B, nystatin or liposomal and lipid forms, from purine or pyrimidine nucleotide inhibitors such as flucytosine, from polyoxins such as nikkomycins, preferably nikkomycin Z or other chitin inhibitors, from echinocandins such as micafungin and anidulafungin or combinations thereof, from alylamines, preferably terbinafine.

[0087] Preferably the composition comprises the at least one additional fungal agent in a percentage between about 0.001 wt% and about 10wt%, preferably between about 0.005wt% and about 5wt%, more preferably between about 0.01 wt% and about 1wt%.

[0088] In a preferred version, said composition is formulated for the administration of an amount of trifarotene between 5 and 100 mg / m2 / day, preferably between 10-95 mg / m2 / day, more preferably between 20-70 mg / m2 / day, preferably between 30-60 mg / m2 / day, preferably between 50-60 mg / m2 / day. Preferably this composition is administered over a period of 1 to 20 days. Administration can be carried out 1 to 3 times a day.

[0089] In one version, the composition is configured so that an amount of trifarotene between 10 mg / m2 / day and 30 mg / m2 / day is administered over a period of 7 to 15 days for between 1 and 3 administrations per day.

[0090] Each dose of the composition may comprise 10-50mg of trifarotene.

[0091] In a preferred version the composition for oral administration contains an amount of trifarotene between 1 and 100 mg, preferably between 5 and 95 mg, preferably between 10 and 80 mg, preferably between 15 and 75 mg, preferably between 20 and 70 mg, preferably between 25 and 65 mg, preferably between 30 and 60 mg, preferably between 35 and 55 mg, preferably between 40 and 50 mg.

[0092] Preferably the composition is administered orally twice a day.

[0093] Preferably the composition is administered orally twice a day for up to 90 days.

[0094] Preferably the composition for oral administration contains 10-30 mg of trifarotene and is intended to be administered over a period of 30 to 60 days.

[0095] In one version, the composition is configured in such a way that an amount of trifarotene between 25 and 50 mg / day is administered over a period of 7 to 28 days, preferably between 7 and 15 days for between 1 and 3 administrations per day.

[0096] In some preferred versions, the composition further comprises at least one compound selected from at least one excipient, at least one carrier, at least one adjuvantsuch as glycerin, gelatin, propylene glycol, butylene glycol, panthenol, sorbitol, urea, hyaluronic acid, glycolic acid, lactic acid, sodium pyrrolidine carboxylic acid, cholesterol, squalene, linoleic acid, stearic acid, oleic acid, fatty alcohols, white / petrolatum soft paraffin, beeswax, mineral oil, dimethicone, lanolin, carnauba wax, cetyl alcohol, caprylic / capric triglyceride.

[0097] In some preferred embodiments, the composition is inserted into a medical device such as, for example, an intrauterine device (IUD), vaginal ring, mucoadhesive microdisk.

[0098] In some preferred embodiments, the composition is in the form of an ointment, pessary, extra-amniotic infusion composition, intravesical infusion, nasal spray, ear drops, ointment, insufflation composition.

[0099] According to an embodiment of the present invention, the composition is used for topical use, the composition will therefore preferably be in the form of: cream, ointment, pomade, lotion, tonic, gel, paste, foam, hydrophilic gel, emulsion, suspension or liquid.

[0100] Depending on the type of formulation, the composition may further comprise one or more excipients and / or carriers.

[0101] For lipophilic ointments, excipients can be selected from: solid, semi-solid and / or liquid paraffins, vegetable oils, waxes, liquid silicones, which generally have occlusive or protective properties. In some versions, the composition further comprises surfactants such as sorbitan esters, lanolin alcohols, fatty alcohols, fatty acid sulphates, fatty acid esters and / or polysorbates to make an ointment that emulsifies in water.

[0102] In some versions, said composition is in the form of a hydrophilic ointment. In this case, the composition further comprises excipients such as mixtures of polyethylene glycols.

[0103] In some versions, said composition is in cream form. In this case, as the cream is a multiphase preparation consisting of a lipophilic and a hydrophilic phase, emulsifiers are added. If the composition is a hydrophobic cream, the addable emulsifiers are preferably selected from wool alcohols, sorbitan esters and / or monoglycerides. If the composition is a hydrophilic cream, the emulsifiers will be sodium soaps, polysorbates and / or sulphates of fatty alcohols.

[0104] In some versions, the composition is a hydrophilic gel. In this case, this composition further comprises hydrophilic macromolecules such as sodium alginate, agar-agar, gums, gelatin, starch, cellulose derivatives and / or carboxy-vinyl polymers.

[0105] In some versions, the composition is a hydrophobic gel. In this case, the composition further preferably comprises excipients such as micronised silica, hydrogenated castor oil, stearyl ammonium hectorite, zinc stearates, calcium stearates and / or aluminium stearates.

[0106] In other versions, the composition further comprises other excipients selected from moisturising substances such as collagen, elastin, various protein hydrolysates, nucleic acid hydrolysates, natural moisturising factor; or absorption promoters such as DMSO, fatty acids, urea-azone and / or menthol.

[0107] Preferred features of the present invention are the subject of dependent claims.

[0108] According to preferred aspects of the invention, formulations are provided for topical use comprising or consisting of the composition according to any one of the embodiments described herein in the form of an ointment, topical cream, topical gel, liniment, paste, film, solution, hydrogel, liposome, transferable vesicle, cream, lotion, dermal patch, transdermal patch, transdermal spray.

[0109] According to preferred aspects of the invention, compositions for preventing or treating mucosal infections are provided according to any one of the embodiments described herein in the form of an ointment, pessary, vaginal ring, intrauterine device (IUD), extra-amniotic infusion, intravesical infusion, nasal spray, ear drops, eye drops, mouthwash, ointment, insufflation, mucoadhesive microdisk.

[0110] The composition of the invention can be inserted into a medical device such as a vaginal ring or intrauterine device (IUD).

[0111] It is an object of the present invention to provide a medical device containing a composition containing trifarotene for local and / or controlled release of the composition into a body district for use in the treatment of fungal infections of C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei and Trichophyton spp., in particular Trichophyton mentagrophytes complex or Trichophyton rubrum. Such a medical device is an insertable or implantable medical device selected from devices for gynaecological, urological, rectal, gastrointestinal, cutaneous, intranasal, otological, ophthalmic administration and combinations thereof.

[0112] It is an object of the present invention to provide a medical device such as a vaginal ring or intrauterine device (IUD) containing a composition containing trifarotene, preferably for use in the treatment of fungal infections of C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei.DETAILED DESCRIPTION OF THE INVENTIONDefinitions

[0113] In the context of the present invention, mycoses or fungal infections are understood to be any form of infection by mycetes in humans or animals.

[0114] In the context of the present invention, mycoses or fungal infections from Candida are understood to be any form of infection by mycetes in humans or animals sustained by C. albicans and / or C.non-albicans.

[0115] In the context of the present invention, the singular forms “a” and “an” are intended to designate both the singular and the plural, unless expressly indicated to designate only the singular.

[0116] Furthermore, as used herein, “and / or” refers to and comprises all possible combinations of one or more of the associated listed elements, as well as the lack of combinations when interpreted alternatively (“or”). Thus, fungal infection caused by C. albicans and / or C. non-albicans species; C. albicans infections; or C. non-albicans infections, Trichophyton spp. infections, in particular Trichophyton mentagrophytes complex and Trichophyton rubrum.

[0117] The terms “administering”, “administration” or “administer” as used herein refer to (1 ) providing, dispensing, dosing and / or prescribing, e.g. by or under the direction of a health care professional or his authorised agent, and (2) insertion, application, intake or consumption, e.g. by a health care professional or the subject. For example, the administration may include, without limitation, topical routes of administration (e.g., gel, ointment, cream, aerosol, etc.) and may be formulated, alone or together, in suitable unit dosage formulations containing pharmaceutically acceptable non-toxic conventional carriers, adjuvants, excipients and carriers appropriate for each route of administration. The invention is not limited by the route of administration, formulation or dosage schedule

[0118] The term “treatment” as used herein includes the alleviation, mitigation or improvement of fungal infections caused by C. albicans or non-albicans or of one or more symptoms and signs thereof, irrespective of whether the infection is considered resolved and whether or not all symptoms and signs are resolved. The terms also include reducing or preventing the progression of the disease, or one or more signs and symptoms thereof, by blocking or preventing an underlying mechanism of infection, and achieving any therapeutic and / or prophylactic benefit.

[0119] The term “carrier” refers to a diluent, adjuvant, excipient or carrier with which the therapeutic agent is administered.

[0120] FiguresIn the graphs in Figures 1 -8 the following indications are adopted:CTRL indicates control, i.e. a full well without solvent or compound;TRIFA indicates the histograms for the wells with different concentrations of Trifarotene: 1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM;AmB indicates the histograms for the wells with different concentrations of Amphociterin: 2 pg / mL, 1 pg / mL, 0.5 pg / mL, 0.25 pg / mL, 0.12 pg / mL;DMSO indicates the histograms for the wells with different concentrations of DMSO (without additional compound) 1 %v / v, 0.5%v / v, 0.25%v / v, 0.06 %v / v.

[0121] Figure 1. Figure 1 shows the inhibitory effect of trifarotene on the biofilm biomass of C. krusei. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). Amphotericin B (AmB) was used as a positive control drug. The following AmB concentrations were used: 2 pg / mL, 0.5 pg / mL, 0.25 pg / mL, and 0.12 pg / mL. Biofilm growth was assessed as total biomass (A) by staining with crystal violet (CV) and quantified by measuring optical density (OD) using a spectrophotometer at 595 nm. The results are the mean of the O.D. values ± SD of three independent experiments, each conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001.

[0122] Figure 2. Figure 2 shows the inhibitory effect of trifarotene on the metabolic activity of the C. krusei biofilm. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). AmB was used as a positive control drug. The metabolic activity of the biofilm was assessed by XTT tetrazolium salt reduction assay and quantified by measuring the optical density (O.D.) using a spectrophotometer at 490 nm. The results are the mean of the O.D. values ± SD of three independent experiments, each conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001 .

[0123] Figure 3. Figure 3 shows the inhibitory effect of trifarotene on the biofilm biomass of C. tropicalis. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). AmB was used as a positive control drug. Biofilm growth was assessed as total biomass (A) by staining with crystal violet (CV) and quantified by measuring optical density (OD) using a spectrophotometer at 595 nm. The results are the mean of the O.D. values ± SD of three independent experiments, each conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001.

[0124] Figure 4. Figure 4 shows the inhibitory effect of trifarotene on the metabolic activity of C. tropicalis biofilm. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). AmB was used as a positive control drug. The metabolic activity of the biofilm was assessed by XTT tetrazolium salt reduction assay and quantified by measuring the optical density (O.D.) using a spectrophotometer at 490 nm. The results are the mean of the O.D. values ± SD of three independent experiments, each conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001 .

[0125] Figure 5. Figure 5 shows the inhibitory effect of trifarotene on the biofilm biomass of C. auris. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). AmB was used as a positive control drug. Biofilm growth was assessed as total biomass (A) by staining with crystal violet (CV) and quantified by measuring optical density (OD) using a spectrophotometer at 595 nm. The results are the mean of the O.D. values ± SD of three independent experiments conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001.

[0126] Figure 6. Figure 6 shows the inhibitory effect of trifarotene on the metabolic activity of C. auris biofilm. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). AmB was used as a positive control drug. The metabolic activity of the biofilm was assessed by XTT tetrazolium salt reduction assay and quantified by measuring the optical density (O.D.) using a spectrophotometer at 490 nm. The results are the mean of the O.D. values ± SD of three independent experiments conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001.

[0127] Figure 7. This is an image showing the inhibitory effect of trifarotene on the biofilm biomass of C. glabrata. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). AmB was used as a positive control drug. Biofilm growth was assessed as total biomass (A) by staining with crystal violet (CV) and quantified by measuring optical density (OD) using a spectrophotometer at 595 nm. The results are the mean of the O.D. values ± SD of three independent experiments conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001 .

[0128] Figure 8. Figure 8 shows the inhibitory effect of trifarotene on the metabolic activity of C. glabrata biofilm. The Candida cells were cultured for 24 hours in the absenceor presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). AmB was used as a positive control drug. The metabolic activity of the biofilm was assessed by XTT tetrazolium salt reduction assay and quantified by measuring the optical density (O.D.) using a spectrophotometer at 490 nm. The results are the mean of the O.D. values ± SD of three independent experiments conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001.

[0129] Figure 9. Figure 9 shows the effects of trifarotene on the growth of C. albicans. Yeast cells of C. albicans were cultured for 24 hours in the absence or presence of various concentrations of trifarotene (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). AmB was used as a positive control drug. The results are presented as the mean ± SD of three independent experiments, conducted in triplicate, and expressed as the percentage of growth inhibition compared to the control group. Student’s t-test, * p <0.05; ** p <0.01 ; *** p <0.001. In this figure, the graph on the left shows the histograms for the wells with different concentrations of Trifarotene: 1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM and the graph on the right shows the histograms for the wells with different concentrations of Amphociterin: 2.2 pM, 1.1 pM, 0.55 pM, 0.27pM, 0.14 pM.

[0130] Figure 10. Figure 10 shows the effects of trifarotene on phenotypic switching from yeast to hyphal form of C. albicans. Yeast cells of C. albicans were cultured in the absence or presence of various concentrations of trifarotene (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). The phenotypic transition from yeast to filamentous morphotype was analysed under the microscope after 3 hours (Figure A) and 24 hours (Figure B) of incubation. A representative experiment of three is shown, as corresponding results were obtained in the three experiments.

[0131] Figure 11. Figure 11 shows the inhibitory effect of trifarotene on the biofilm biomass of C. albicans. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). AmB was used as a positive control drug. Biofilm growth was assessed as total biomass (A) by staining with crystal violet (CV) and quantified by measuring optical density (OD) using a spectrophotometer at 595 nm. The results are the mean of the O.D. values ± SD of three independent experiments conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001 . In this figure, the graph on the left shows the histograms for the wells with different concentrations of Trifarotene: 1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM and the graph on the right shows the histograms for the wells with different concentrations of Amphociterin: 2.2 pM, 1.1 pM, 0.55 pM, 0.27pM, 0.14 pM;

[0132] Figure 12. Figure 12 shows the inhibitory effect of trifarotene on the metabolic activity of C. albicans biofilm. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). AmB was used as a positive control drug. The metabolic activity of the biofilm was assessed by XTT tetrazolium salt reduction assay and quantified by measuring the optical density (O.D.) using a spectrophotometer at 490 nm. The results are the mean of the O.D. values ± SD of three independent experiments conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001 . In this figure, the graph on the left shows the histograms for the wells with different concentrations of Trifarotene: 1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM and the graph on the right shows the histograms for the wells with different concentrations of Amphociterin: 2.2 pM, 1.1 pM, 0.55 pM, 0.27 pM, 0.14 pM;

[0133] Figure 13. Figure 13 shows the assessment of the viability of C. albicans by double staining with Calcofluor White (CW) and propidium iodide (PI); planktonic cells of C. albicans were incubated in the absence or presence of various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM) of trifarotene. After 24 hours of incubation, Candida cells were double-stained with CW and PI. Fluorescent images of the labelled cells show: in the left column, the blue fluorescence represents the CW-stained cell wall; in the middle column, the red fluorescence represents the Pl-stained nucleic acids of the dead cells; in the right column, the merge of images shows the double-labelled cells, stained with CW and PI (blue and red). All images were obtained at 100x magnification, using a fluorescence microscope. The results, represented graphically with histograms, show the percentage of dead cells. At least 10 fields per slide were counted. Chi-square test; * p <0.05, ** p <0.01 ; ***p<0.001.

[0134] Figure 14. Figure 14 shows the effect of trifarotene on C. auris biofilm formation. Cultures of Candida were incubated for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). The phenotypic transition from yeast to hyphal morphotype was analysed by light microscopy after staining with crystal violet. AmB was used as a positive control drug. The images were acquired by an optical microscope (Olympus, Carl Zeiss, UK) with 40* magnification objectives. A representative experiment of three is shown. The column on the left shows images of the wells with different concentrations of Trifarotene: 1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM; the middle column shows the images for the wells with different concentrations of Amphociterin: 2 pg / mL, 1 pg / mL, 0.5 pg / mL, 0.25 pg / mL, 0.12 pg / mL; the column on the right shows images of the wells with different concentrations of DMSO(without any additional compound) 1 %v / v, 0.5%v / v, 0.25%v / v, 0.06 %v / v, at the top a control image (CTRL) is shown, i.e. full well without solvent or compound.

[0135] Figure 15. Figure 15 shows the effect of trifarotene on C. tropicalis biofilm formation. Cultures of Candida were incubated for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). The phenotypic transition from yeast to hyphal morphotype was analysed by light microscopy after staining with crystal violet. AmB was used as a positive control drug. The images were acquired by an optical microscope (Olympus, Carl Zeiss, UK) with 40* magnification objectives. A representative experiment of three is shown. The column on the left shows images of the wells with different concentrations of Trifarotene: 1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM; the middle column shows the images for the wells with different concentrations of Amphociterin: 2 pg / mL, 1 pg / mL, 0.5 pg / mL, 0.25 pg / mL, 0.12 pg / mL; the column on the right shows images of the wells with different concentrations of DMSO (without any additional compound) 1 %v / v, 0.5%v / v, 0.25%v / v, 0.06 %v / v, at the top a control image (CTRL) is shown, i.e. full well without solvent or compound.

[0136] Figure 16. Figure 16 shows the effect of trifarotene on C. krusei biofilm formation. Cultures of Candida were incubated for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). The phenotypic transition from yeast to hyphal morphotype was analysed by light microscopy after staining with crystal violet. AmB was used as a positive control drug. The images were acquired by an optical microscope (Olympus, Carl Zeiss, UK) with 40* magnification objectives. A representative experiment of three is shown. The column on the left shows images of the wells with different concentrations of Trifarotene: 1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM; the middle column shows the images for the wells with different concentrations of Amphociterin: 2 pg / mL, 1 pg / mL, 0.5 pg / mL, 0.25 pg / mL, 0.12 pg / mL; the column on the right shows images of the wells with different concentrations of DMSO (without any additional compound) 1 %v / v, 0.5%v / v, 0.25%v / v, 0.06 %v / v, at the top a control image (CTRL) is shown, i.e. full well without solvent or compound.

[0137] Figure 17. Figure 17 shows the effect of trifarotene on C. glabrata biofilm formation. Cultures of Candida were incubated for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM). The phenotypic transition from yeast to hyphal morphotype was analysed by light microscopy after staining with crystal violet. AmB was used as a positive control drug. The images were acquired by an optical microscope (Olympus, Carl Zeiss, UK) with 40* magnificationobjectives. A representative experiment of three is shown. The column on the left shows images of the wells with different concentrations of Trifarotene: 1 mM, 0.5 mM, 0.25 mM, 0.12 mM, 0.06 mM; the column on the right shows images of the wells with different Amphotericin concentrations: 2 pg / mL, 1 pg / mL, 0.5 pg / mL, 0.25pg / mL, 0.12 pg / mL; control image (CTRL) is shown above, i.e. full well without solvent or compound.

[0138] Figure 18. Figure 18 shows the effect of trifarotene on Trichophyton mentagrophytes complex biofilm formation. Cultures of Trichophyton mentagrophytes complex were incubated for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.75 mm, 0.5 mM, 0.25 mM, 0.12 mM). Biofilm inhibition by trifarotene was analysed by light microscopy, evaluating the biomass after staining with crystal violet. The images were acquired using an optical microscope (Olympus, Carl Zeiss, UK) with 40* magnification objectives. A representative experiment of three is shown. The column on the left shows images of the wells with different concentrations of Trifarotene: 1 mM, 0.75 mm, 0.5 mM, 0.25 mM, 0.12 mM; the column on the right shows images of the wells with different concentrations of DMSO, the diluent used as a negative control: 1v / v, 0.75 v / v, 0.5 v / v, 0.25 v / v; control image (CTRL) is shown above, i.e. full well without solvent or compound.

[0139] Figure 19. Figure 19 shows the effect of terbinafine alone and in combination with trifarotene on biofilm formation by Trichophyton mentagrophytes complex. Terbinafine is an antifungal drug used as a first-line treatment in infections of the skin and skin adnexa caused by fungi of the genus Trichophyton spp. In the experiments conducted, terbinafine was used at various concentrations, ranging from 1 mg / L to 0.06 mg / L, alone (1 st column on the left) and in combination with trifarotene at various concentrations, to assess a potential synergistic effect between the two drugs.

[0140] According to the present invention, a composition comprising trifarotene is provided for use in the treatment of fungal infections.

[0141] In one version, the composition of the invention is intended for use in the treatment of fungal infections caused by C. albicans species.

[0142] In a preferred embodiment, the composition of the invention is intended for use in the treatment of fungal infections caused by C. non-albicans species.

[0143] Preferably, the composition of the invention is intended to be used for the treatment of fungal infections caused by one or more of C. krusei, C. tropicalis, C. auris, C. glabrata and Trichophyton spp., in particular Trichophyton mentagrophytes complex, Trichophyton rubrum.

[0144] Trifarotene is a fourth-generation retinoid, selective for gamma receptors, whose formula is given below.

[0145] Unlike all other retinoids, trifarotene is defined as such not by its chemical structure, i.e. a triaryl, but by the same intranuclear retinoid target, RARs. However, it also has some peculiarities here, as it selectively acts on the gamma RAR and has no action on RXR receptors, unlike the other retinoids. Randomised trials evaluating the tolerability, safety and efficacy of trifarotene in congenital ichthyosis and acne showed excellent results and mild side effects, leading to FDA approval of trifarotene for the treatment of lamellar ichthyosis in 2014 and acne vulgaris in October 2019. Trifarotene is a selective RAR-y receptor agonist, with lower activity on RAR-[3 and RAR-a (16- and 65-fold, respectively) and no activity on RXR receptors, which are activated by the other molecules of the retinoid class. Binding of trifarotene on RAR-y results in dimerisation of the receptor, leading to retinoid-specific RARE binding. Downstream alterations in gene expression are the main way in which trifarotene exerts its anti-inflammatory, comedolytic and depigmenting actions.

[0146] Trifarotene influences three different pathways, identified by a large-scale gene expression analysis: skin hydration: trifarotene induces skin peptidyl arginine deaminase 1 and aquaporin-3 channels and thus influences skin barrier functions; cell adhesion: trifarotene weakens haemidesmosomes, reducing intercellular adhesion. The lower cohesion between the keratinocytes explains its comedolytic properties; proteolysis: trifarotene subregulates matrix metalloproteinases (MMPs), which act as proteolytic enzymes on elastin and collagen, thus improving skin structure.

[0147] Ex vivo pharmacokinetic models on trifarotene demonstrated the high stability of the compound, with a half-life >24 hours. Despite this, it is rapidly metabolised by hepatic microsomal enzymes, with a half-life of minutes, with respect to tazarotenic acid, which has a 10-fold stability in hepatic microsomes. This is an indicator of a favourable safety profile. Trifarotene metabolism is mainly catalysed by cytochrome (CY) P2C9, CYP3A4, CYP2C8 and, to a lesser extent, CYP2B6. Systemic exposures in mice, afterboth topical and oral administration, were up to 1642 times higher than those observed in humans at the maximum recommended human dose, and these systemic concentrations did not result in observed carcinogenicity. Trifarotene does not appear to pose any risk of carcinogenesis when used at standard doses.

[0148] Current studies confirm that trifarotene cream 50 pg / g (commercial formulation) is well tolerated systemically and safe when applied under maximised conditions in adults and paediatric acne patients, including patients with severe acne. Considering that trifarotene belongs to the retinoid class and is also intended for use in women of childbearing age, further studies are needed to rule out any potential teratogenic effects. Currently, clinical pharmacological data show that trifarotene cream50 pg / g generates low systemic absorption when applied daily under conditions of maximum use. Furthermore, due to its RAR-y selectivity, it could be hypothesised that trifarotene is safer than other retinoids in pregnancy as the placenta has a lower expression of RAR-y (4.739 ± 0.399), resulting in less absorption of the drug than other topical retinoids.

[0149] Advantageously, the inventors have demonstrated that trifarotene, unexpectedly, has an anti-fungal effect.

[0150] Advantageously, the inventors have demonstrated that trifarotene, unexpectedly, has an antifungal effect on infections caused by C. albicans.

[0151] Advantageously, the inventors have demonstrated that trifarotene, unexpectedly, is effective in treating fungal infections from C. non-albicans fungus species. The inventors have demonstrated that trifarotene is effective in the treatment of fungal species such as C. auris, C. krusei, C. tropicalis and C. glabrata which have, on the contrary, proved resistant to conventional therapies commonly used in clinical practice. These species have been shown to be intrinsically resistant to fluconazole, or to other azoles that have been shown to be effective in treating C. albicans infections. These C. non-albicans species are resistant to the drugs currently commonly used in the treatment of C. albicans infections.

[0152] On the other hand, C. tropicalis and C. glabrata in particular, are species that differ considerably from C. albicans in both morphology and genome and, therefore, have different mechanisms of proliferation but also of interaction with drugs compared to C. albicans.

[0153] C. tropicalis and C. glabrata show increasing rates of resistance not only to azoles but also to other drug classes such as echinocandins. Since these species differnot only in their response to antifungals compared to C. albicans, but also in their genome and the expression of different transmembrane proteins, an antifungal effect of trifarotene against these species was not to be expected. All the more so since trifarotene has a chemically different molecule from ATRA and has different interaction mechanisms from those of ATRA.

[0154] Contrary to expectations, trifarotene proved effective against C. albicans. Contrary to expectations, trifarotene proved effective against C. non-albicans species.

[0155] Knowledge in the field did not allow inference of trifarotene's efficacy in treating fungal infections, and in particular fungal infections caused by C. albicans or C. non- albicans. Due to the above-mentioned differences in both the genetics and dynamics of infection propagation between the different mycetes and the characteristics of trifarotene, it was not possible to imagine or predict the efficacy of trifarotene against infections caused by C. albicans and C. non-albicans. Nor could it be assumed that trifarotene would be effective against such a large number of Candida species and, in particular, against C. albicans and C. non-albicans species, which, as mentioned, are very different from each other. In fact, such mycetes are extremely different from C. albicans and different from each other, despite being classified within the same fungal genus both in aetiology and genetically.

[0156] Trifarotene has been shown to be effective in inhibiting biofilm formation in vitro by Trichophyton spp. and in particular against Trichophyton mentagrophytes complex and / or Trichophyton rubrum.

[0157] The ability of trifarotene to interfere with biofilm formation by Trichophyton mentagrophytes complex and / or Trichophyton rubrum was not expected and is of particular interest, considering that biofilm is often associated with increased persistence of infection and reduced sensitivity to traditional treatments. In addition, the potential ability of trifarotene to synergize with terbinafine lends further importance to its pharmacological profile, suggesting the possibility of developing combination therapeutic approaches capable of enhancing the efficacy of available treatments, especially in more recalcitrant cases or those characterised by suboptimal response to first-line antifungals. Another advantage related to the administration of trifarotene is the zero risk of possible drug interactions with amphotericin, echinocandins, cyclopyroxolamine, and azoles, thus being an adequate and safe drug in combination with the antifungals used in common clinical practice.

[0158] Terbinafine is the antifungal drug currently used in the treatment of many Trichophyton spp. dermatophytosis, in particular also for dermatophytosis caused by Trichophyton mentagrophytes complex and / or Trichophyton rubrum and has for years been considered the standard for efficacy and handling. However, the last decade has seen an increase in reports of reduced susceptibility / resistance, also associated with clinical failures and relapses. A significant proportion of resistance is linked to point mutations in the squalene epoxidase (SQLE) target gene, which reduce drug activity. In parallel, Trichophyton indotineae, frequently associated with terbinafine resistance profiles and infections that are more difficult to eradicate, is emerging and spreading in several countries (including Europe).

[0159] The data obtained show that trifarotene combined with terbinafine can effectively treat mycoses caused or sustained by Trichophyton mentagrophytes complex or Trichophyton rubrum. This effect is maintained even when using lower amounts of trifarotene and terbinafine than those usually considered effective of the two compounds when administered separately. Therefore, the simultaneous use of trifarotene and terbinafine (even at sub-optimal doses) achieves an enhanced effect in the treatment of Trichophyton mentagrophytes complex contrasting biofilm formation compared to treatments with the single compounds.

[0160] In the light of our results, trifarotene shows an inhibiting effect on biofilm formation (reduction of biomass) and, when used in combination with terbinafine at suboptimal doses (0.5, 0.25, 0.125, 0.06mg / l), shows a synergistic effect with terbinafine, enhancing the efficacy of terbinafine and counteracting conditions in which the response to monotherapy is less satisfactory.

[0161] The composition can be administered to a subject in need by conventional methods.

[0162] In an embodiment, said composition is in the form of a preparation for oral administration.

[0163] In an embodiment, said composition is in the form of a preparation for intravenous administration.

[0164] In an embodiment, said composition is in the form of a preparation for intramuscular administration.

[0165] In an embodiment said composition is in the form of a preparation for topical administration, preferably said composition being in the form of a gel, cream, ointment,pomade, lotion, tonic, gel, paste, foam, hydrophilic gel, emulsion, suspension or liquid, solution for nail application, composition for transdermal application.

[0166] For lipophilic ointments, the excipients can be selected from: solid, semi-solid and / or liquid paraffins, vegetable oils, waxes, liquid silicones, which generally have occlusive or protective properties. Surfactants such as sorbitan esters, lanolin alcohols, fatty alcohols, fatty acid sulphates, fatty acid esters and / or polysorbates can also be added to make an ointment that emulsifies in water.

[0167] With regard to the composition of the present invention in the form of a hydrophilic ointment, appropriate excipients to be added will be mixtures of polyethylene glycols.

[0168] With reference to a possible embodiment in which the composition described by the present invention is in cream form, the cream being a multiphase preparation consisting of a lipophilic and a hydrophilic phase, the addition of emulsifiers is essential. In the case of hydrophobic creams, the emulsifiers that can be added are wool alcohols, sorbitan esters and / or monoglycerides, whereas in the case of hydrophilic creams, the emulsifiers will be sodium soaps, polysorbates and / or sulphates of fatty alcohols.

[0169] For compositions formulated as hydrophilic gels, the addition of hydrophilic macromolecules such as sodium alginate, agar-agar, gums, gelatin, starch, cellulose derivatives and / or carboxy-vinyl polymers is essential.

[0170] In the case of hydrophobic gels, includable excipients are: micronised silica, hydrogenated castor oil, stearyl ammonium hectorite, zinc stearates, calcium stearates and / or aluminium stearates.

[0171] Other excipients may be moisturising substances such as collagen, elastin, various protein hydrolysates, nucleic acid hydrolysates, natural moisturising factor; or absorption promoters such as DMSO, fatty acids, urea-azone and / or menthol.

[0172] In an embodiment, said composition is in the form of a preparation for oral liposomal administration.

[0173] In an embodiment, said composition is in the form of an aerosol preparation.

[0174] In other embodiments the composition can be administered in other forms that are equally suitable for implementing the present invention.

[0175] A person skilled in the art may decide to administer the composition by any conventional pharmaceutical form. Reference can be made to Remington’s Pharmaceutical Sciences, latest edition.

[0176] A person skilled in the art will decide the effective timing of administration, depending on the patient’s condition, the level of severity of the pathology, the patient's response, and any other clinical parameters included in the general knowledge of this topic.

[0177] The composition of the invention also contains, together with the active ingredients, at least one pharmaceutically acceptable carrier or excipient.

[0178] Such a carrier may be a macromolecule, such as proteins, polysaccharides, polylactic acids, polyglycolic acids, polymeric amino acids, amino acid copolymers, an antibody, a lipid molecule, an inactive viral particle or any other carrier known in the pharmaceutical industry.

[0179] An extensive discussion of commercially available carriers can be found in Remington's Pharmaceutical Sciences (Mack Pub. Co., N.J. 1991 ).

[0180] The carriers may additionally contain liquids such as water, saline solution, glycerol and ethanol.

[0181] In addition, particularly useful formulation aids may be present, e.g. solubilising agents, dispersing agents, suspending agents and emulsifying agents. These components allow pharmaceutical compositions such as tablets, pills, capsules, dragees, liquids, gels, syrups, slurries, suspensions and the like to be formulated for oral administration.

[0182] All these components are known in the industry and can easily be selected by a person skilled in the art based on their general knowledge of the pharmaceutical industry.

[0183] Once formulated, the composition can be administered directly to the subject. The subjects to be treated can be animals; in particular, they can be human subjects.

[0184] The composition of the invention may be administered by any route, including oral, intravenous, intramuscular, intra-arterial, intramedullary, intrathecal, intraventricular, transdermal or by transcutaneous, subcutaneous, intraperitoneal, intranasal, enteral, topical, sublingual, intravaginal or rectal means of application. A preferred route is the oral route.

[0185] The compositions for oral administration can take the form of liquid solutions or suspensions or powders.

[0186] Most commonly, the compositions are in a unit dosage form to facilitate dosing. The term “unit dosage forms” refers to physically discrete units suitable as unit dosages for humans and other mammals, each unit containing a predetermined amount of activematerial calculated to produce the desired therapeutic effect, in combination with a pharmaceutically acceptable excipient. Typical unit dosage forms comprise premeasured ampoules or syringes for liquid compositions or pills, tablets, capsules or the like in the case of solid compositions. In such compositions, the compounds of the invention are generally a minor component (e.g. from about 0.1 to about 50% by weight or preferably from about 1 to about 40% by weight), the remainder comprising various carriers and auxiliary substances useful in composing the desired dosage form. Treatment may involve single-dose or multiple-dose administration.

[0187] Pharmaceutical compositions according to the present invention may also contain one or more additional active ingredients.

[0188] Average quantities of active compound may vary and should be based on the recommendations and prescription of a qualified physician. The administration regimen, dosage and posology will be decided by the doctor based on his or her experience, the disease to be treated and the patient's condition, and on general knowledge in the field.

[0189] For each compound, the therapeutically effective dose can be estimated initially in cell culture assays or in animal models, usually mice, rats, guinea pigs, dogs or pigs. The animal model can also be used to determine the appropriate concentration range and route of administration. This information can be used to determine useful doses and routes of administration for humans. When calculating the Human Equivalent Dose (HED), it is recommended to use the conversion table provided in the document Guidance for Industry and Reviewers (2002, U.S. Food and Drug Administration, Rockville, Maryland, USA).

[0190] An average dose for administration in humans may be established during clinical trials, as is standard practice in the industry.

[0191] The specific effective dose for a human subject will then depend on the severity of the condition, the subject's general health, age, weight and sex, diet, time and frequency of administration, any combination of drugs and tolerance / response to therapy. This amount can be determined through routine trials and is a matter for the doctor's assessment.

[0192] Depending on the chosen route of administration, the compositions will be in solid or liquid form, suitable for oral or parenteral administration or any other chosen route of administration.

[0193] Methods are provided to treat fungal infection from C. albicans and non albicans species, such as C. krusei, C. tropicalis, C. auris, C. glabrata and infectionssustained by the genus Trichophyton spp., in particular Trichophyton mentagrophytes complex or Trichophyton rubrum. The method comprises, or essentially consists of, administering to the subject a pharmaceutical composition that includes trifarotene or an equivalent thereof.

[0194] In one or more embodiments, the method includes the topical application of a pharmaceutical composition, which includes trifarotene or an equivalent thereof. In one or more embodiments, the method includes the topical administration of an effective amount of trifarotene or an equivalent thereof to the subject. In one or more embodiments, trifarotene or an equivalent thereof is administered by applying a thin layer sufficient to cover the area to be treated.

[0195] An effective amount can be administered in one or more administrations, applications or dosages. Such dispensing depends on a number of variables, including the length of time for which the individual dosage unit is to be used, the bioavailability of the therapeutic agent, the route of administration, etc. The therapeutic agents of the present disclosure for any particular subject depend on a variety of factors including the activity of the specific compound used, the age, body weight, general health, sex and diet of the subject, time of administration, rate of excretion, combination of drugs, dermal absorption for topical compositions, general systemic absorption for topical and oral compositions, and the seventy of the particular disorder to be treated and the form of administration. Treatment dosages can generally be titrated to optimise safety and efficacy. The dosage can be determined by a doctor and adjusted if necessary to suit the observed treatment effects and to manage and adapt to retinoid dermatitis.

[0196] In one or more embodiments, creams, ointments, foams or lotions comprising from about 1 ug / g to about 100 ug / g of trifarotene or an equivalent thereof are topically administered to the subject, including, but not limited to, from about 5 ug / g to about 95 ug / g, from about 10 ug / g to about 90 ug / g, from about 15 ug / g to about 85 ug / g, from about 20 ug / g to about 80 ug / g, from about 25 ug / g to about 75 ug / g, from about 30 ug / g to about 70 ug / g, from about 35 ug / g to about 65 ug / g, from about 40 ug / g to about 60 ug / g, from about 45 ug / g to about 55 ug / g, from about 46 ug / g to about 54 ug / g, from about 47 ug / g to about 53 ug / g, from about 48 ug / g to about 52 ug / g, or from about 49 ug / g to about 51 ug / g. In some embodiments, the subject is given topically a cream containing from about 1 ug / g to about 100 ug / g of trifarotene or an equivalent thereof. In some embodiments, a cream containing approximately 50 ug / g of trifarotene or anequivalent thereof is administered topically once or several times a day on the affected area of the subject.

[0197] In one or more embodiments, the effective amount of trifarotene or an equivalent thereof varies from about 0.0001 % w / w to about 0.1 % w / w, from about 0.001 % w / w to about 0.5% w / w, from about 0.005% w / w to about 1 % w / w, from about 0.01 % w / w to about 1.5% w / w, or from about 0.1 % w / w to about 10% w / w. In particular embodiments, the effective amount of trifarotene or an equivalent thereof is about 0.0001 % by weight, about 0.005% by weight, about 0.1 % by weight, about 1 % by weight, about 2% by weight, or about 2.5% by weight, or any range comprised and / or between any two of these values and / or as needed depending on the onset of symptoms of fungal infection from C. albicans and non-albicans species. In special embodiments, the effective amount of trifarotene or an equivalent thereof is a fixed dose of 2.5 g (e.g. in cream form).

[0198] In one or more embodiments, trifarotene or an equivalent thereof is administered topically. In one or more embodiments, trifarotene or an equivalent thereof is administered by applying a thin layer sufficient to cover the area to be treated. In one or more embodiments, the dose is administered proximal to the site of the fungal infection. In some embodiments, the subject may choose to avoid applying trifarotene or an equivalent thereof to damaged skin (such as cuts, abrasions) and sunburnt skin. In one or more embodiments, trifarotene or an equivalent thereof is administered topically to the affected area of the subject. In one or more embodiments, trifarotene or an equivalent thereof is administered by applying a thin layer of the pharmaceutical composition containing trifarotene or an equivalent thereof to the affected areas once a day, in the evening, on clean, dry skin.

[0199] In one or more embodiments, trifarotene or an equivalent thereof is administered three times a day, twice a day, once a day, every other day, twice a week, three times a week, four times a week, five times a week, six times a week, once a week, once every fortnight, once every three weeks, once every four weeks, once every five weeks, once every six weeks, once every seven weeks, once every eight weeks, once every nine weeks, once every 10 weeks, once every 11 weeks, once every 12 weeks, twice a year, once a year, or any interval comprised and / or comprised between any two of these values and / or as required depending on the appearance of symptoms of fungal infection. In one or more embodiments, trifarotene or an equivalent thereof is administered once a day.

[0200] Treatments vary in duration, depending on the patient and the severity of the fungal infection. The treatment period can thus last from several days to five years. In one or more embodiments, the duration of the treatment is about 5 days, about 6 days, about 7 days, about 8 days, about 9 days, about 10 days, about 11 days, about 12 days, about 13 days, about 14 days, about 15 days, about 16 days, about 17 days, about 18 days, about 19 days, about 20 days, about 21 days, about 22 days, about 23 days, about 24 days, about 25 days, about 26 days, about 27 days, about 28 days, about 29 days, about 30 days, about one week, about 2 weeks, about 3 weeks, about 4 weeks, about 5 weeks, about 10 weeks, about 20 weeks, about 30 weeks, about 36 weeks, about 40 weeks, about 48 weeks, about 50 weeks, about one year, about two years, about three years, about four years, about five years, or any interval comprised and / or comprised between any two of these values, and / or as needed depending on the appearance of symptoms of fungal infection. In one or more embodiments, the duration of treatment ranges from about 15 days to about 45 days, from about 21 days to about 30 days, from about 12 to about 48 weeks, or from about 24 to about 36 weeks.

[0201] In one or more embodiments, uses of trifarotene, or an equivalent thereof, are envisaged in the production of a medicament for the treatment of fungal infection in a subject. In one or more embodiments, a pharmaceutical composition comprising an effective amount of trifarotene or an equivalent thereof and a pharmaceutically acceptable carrier is provided for use in a method of treating fungal infection in a subject.

[0202] The invention will now be further illustrated by the following examples.EXAMPLESEXPERIMENTAL TESTS

[0203] Studies in vitro were conducted to evaluate the antifungal effect of trifarotene at different concentrations against opportunistic pathogenic mycetes such as C. albicans and C. non-albicans species, in particular C. auris, C. krusei, C. glabrata, and C. tropicalis, according to standard protocols.

[0204] The results showed that trifarotene exerted fungistatic activity at a concentration of 0.5 mM against all C. non-albicans species tested. This effect was maintained up to seven days, the last evaluated time point.

[0205] Strong fungicidal activity of trifarotene was demonstrated at a concentration of 1 mM on C. albicans and the C. non-albicans species tested, i.e. C. auris, C. krusei, C. glabrata, and C. tropicalis.MATERIALS AND METHODS

[0206] For Candida non albicans species, clinical strains of C. tropicalis, C. krusei, C. glabrata were isolated. For C. auris the reference strain C. auris CDC B11903 was used. For C. albicans the reference strain ATCC 2091 was used. A clinical isolate was used for Trichophyton spp.

[0207] The same experimental tests were performed on all strains of Candida used in the experiment, so in the following generic reference will be made to Candida species and it is understood that this definition includes, within the scope of this discussion, strains of C. tropicalis, C. krusei, C. glabrata and C. auris.

[0208] Strains of each Candida species were grown on Sabouraud dextrose agar plates (Difco Laboratories, Detroit, Ml, USA), fortified with chloramphenicol, and incubated at 30°C for each Candida species. After 24 hours of incubation, the Candida cells were harvested by washing with sterile saline solution or distilled water with 0.05% (v / v) Tween 20. The Candida cells were counted using a Burker chamber, and diluted to the desired seeding density in RPMI 1640 medium with 10% fetal bovine serum (FCS) (catalogue 9014-81 -7 Sigma-Aldrich, Milan, Italy). The strain of Trichophyton was cultivated on DTM agar plates (Dermatophyte Test Medium; teknopharma). After 2 weeks, the microconidia were collected by washing with sterile saline or distilled water with 0.05% (v / v) Tween 20. The conidia were counted using a Burker chamber, and diluted to the desired seeding density in RPMI 1640 medium with 10% fetal bovine serum (FCS) (catalogue 9014-81 -7 Sigma-Aldrich, Milan, Italy).Antimicrobial agents

[0209] Trifarotene was supplied in freeze-dried form by the company Sigma-Aldrich, Milan, Italy (catalogue no. AMBH93D58E72; Merck Life Science, Milan, Italy). The powder was dissolved in 50% dimethylsulfoxide (DMSO; Sigma-Aldrich, Milan, Italy) and diluted with the appropriate RPMI 1640 culture medium at a final DMSO concentration of 2.5% (v / v).

[0210] RPMI 1640 containing 2.5% DMSO was used for each experimental point in all tests. The following concentrations of trifarotene were used: 1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM. 3 mg of trifarotene was dissolved in 1 mL of DMSO to obtain a solution containing 10 mM trifarotene, called the stock solution, (solution A). From this stock solution, dilutions are made to obtain the desired concentration. From solution A, five further solutions were prepared with different dilution degrees in order to obtain solutions with different concentrations of trifarotene. In particular, the following solutionswere prepared: solution B: trifarotene 1 mM; solution C: trifarotene 0.5 mM; solution D: trifarotene 0.25 mM; solution E: trifarotene 0.12 mM; solution F: trifarotene 0.06 mM. Specifically, solution B (= 1 mM) was obtained from solution A by taking 100 microlitres of solution A and adding 900 microlitres of RPMI 1640 (which is the culture medium in which the Candida species used for the experiments grow). From solution B, solution C was prepared by performing a 1 :1 dilution, i.e. 500 microlitres of solution B was taken and added to 500 microlitres of RPMI, resulting in solution C (0.5 mM). The same process was used to prepare solutions D, E and F.Compositions containing trifarotene and having different final concentrations of trifarotene were prepared. Each concentration of trifarotene (1 mM, 0.5 mM, 0.25 mM, 0.12 mM and 0.06 mM) was added in a volume of 100 pL / well. Each test was repeated in triplicate for each concentration of trifarotene.For Amphotericin B, the following concentrations were used: 2 pg / mL, 1 pg / mL, 0.5 pg / mL, 0.25 pg / mL, and 0.12 pg / mL. Positive control wells were prepared for each strain of Candida by plating 2x105cells of Candida in 200 pL of culture medium (RPMI), and negative control wells by plating 200 pL of culture medium / well only or 100 pL of culture medium plus 100 pL of each solution prepared for the different concentrations of trifarotene or AmB in the absence of Candida. For terbinafine, the following concentrations were used: 1 mg / L, 0.5 mg / L, 0.25 mg / L, 0.12 mg / L and 0.06 mg / L. For the Trichophyton strain, positive control wells were prepared by plating 2x105conidia of Trichophyton in 200 pL of culture medium (RPMI), and negative control wells by plating 200 pL of culture medium / well only or 100 pL of culture medium plus 100 pL of each solution prepared for the different concentrations of terbinafine in the absence of Trichophyton conidia.Each of the experiments was conducted including positive control wells and negative control wells; three wells were prepared for each experiment, i.e. for each concentration of trifarotene.Cell growth rate of C. albicans

[0211] To evaluate the antifungal potential of trifarotene on the growth of C. albicans, 2x105cells of each Candida strain were cultured in 96-well plates (Thermo Scientific™ Nunc™ MicroWell™ 96-Well, Nunclon Delta-Treated, Flat -Microplate bottom) in 100 pL of RPM1 1640 medium in 10% FCS (no. catalogue 9014-81 -7; Sigma-Aldrich, Milan, Italy) in the absence or presence of trifarotene or AmB (Amphotericin B).

[0212] Compositions containing the following concentrations of trifarotene were prepared: 1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM corresponding to final concentrations of 300 pg / mL, 150 pg / mL, 75 pg / mL, 37.5 pg / mL and 18.75 pg / mL of trifarotene. Each of the above compositions was added in a volume of 100 pL / well, for each experimental point. Each experimental point was repeated in triplicate to validate the results.

[0213] Amphotericin B (AmB) was supplied in freeze-dried form by the company Sigma-Aldrich, Milan, Italy. The powder was dissolved in 50% dimethylsulfoxide (DMSO; Sigma-Aldrich, Milan, Italy) and diluted with the appropriate RPMI 1640 culture medium at a final DMSO concentration of 2.5% (v / v). RPMI 1640 containing 2.5% DMSO was used for each experimental point in all tests. The following amphotericin concentrations were used: 2 pg / mL, 0.5 pg / mL, 0.25 pg / mL, and 0.12 pg / mL.

[0214] For each strain of C. non-albicans (C. tropicalis, C. krusei, C. glabrata and C. auris), positive control wells were prepared by plating 2x105cells of Candida in 200 pL of culture medium, and negative control wells by plating 200 pL of culture medium / well only or 100 pL of culture medium plus 100 pL of each concentration of trifarotene or AmB in the absence of Candida.

[0215] Each of the experiments was conducted by including positive control wells and negative control wells, three positive wells and three control wells for each experimental point.

[0216] The plate was incubated at 30°C for 24 hours. At the end of the incubation period, the cell growth rate was assessed by calculating the optical density using a spectrophotometer at 510 nm.

[0217] The results are presented as the mean OD ± SD of three independent experiments, conducted in triplicate and expressed as the percentage of growth inhibition compared to the untreated control.Hyphal growth inhibition assay

[0218] The impact of trifarotene on C. albicans and hyphal growth was also evaluated. For this purpose, 2x105cells were grown in 96-well plates in 200 pL of 10% FCS RPMI 1640 medium and incubated at 37°C in the absence or presence of different concentrations of trifarotene 1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM corresponding to final concentrations of 300 pg / mL, 150 pg / mL, 75 pg / mL, 37.5 pg / mL and 18.75 pg / mL of trifarotene.

[0219] Experimental tests were conducted after 3 hours and after 24 hours of incubation. Germ tube formation and hyphal growth were assessed by microscopic examination using an optical microscope (Olympus, Carl Zeiss, UK), at 40x magnification, after 3 and 24 hours of incubation. The images were documented with the digital camera provided.Biofilm quantification using Crystal Violet and XTT tests

[0220] In order to evaluate the effects of trifarotene on the biofilm of C. albicans, C. tropicalis, C. krusei, C. auris, C. glabrata, in terms of both biomass and metabolic activity, 2x105Candida cells were cultured in 96-well plates in 200 pL of RPMI 1640 medium at 10% FCS and incubated at 37°C for 24 h in the absence or presence of different concentrations of trifarotene: 1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM corresponding to final concentrations of 300 pg / mL, 150 pg / mL, 75 pg / mL, 37.5 pg / mL and 18.75 pg / mL of trifarotene.

[0221] Crystal violet (CV) staining and the 2,3-Bis-(2-methoxy-4-nitro-5- sulfophophenyl)-2H-tetrazolium-5-carboxanilide (XTT) reduction test were used to assess the biomass and metabolic activity of the biofilm, respectively, by calculating OD by spectrophotometer. Three independent experiments were performed in triplicate and the data were expressed as the arithmetic mean of the absorbance values (OD) ± SD. In all experiments, the OD values correspond to the absorbances of the individual samples from which the ODs of the negative controls (wells not containing cells) were subtracted.Quantification of cell viability of C. albicans

[0222] To assess the impact of trifarotene on the viability of C. albicans, planktonic cells were double-stained with Calcofluor White (CW), which binds to the chitin of the cell wall of fungal cells, irrespective of their metabolic state, and propidium iodide (PI) (Catalogue No. P4170; Sigma Aldrich, Milan, Italy), which penetrates the cell membranes of poorly viable or dead cells. The results are presented graphically with histograms showing the percentage of dead cells stained with PI. At least 10 fields were counted per slide.Quantification of biofilm biomass produced by Trichophyton spp.

[0223] Quantification of biofilm biomass produced by Trichophyton spp. In order to evaluate the effects of trifarotene alone or in combination with terbinafine on Trichophyton biofilm, in terms of biomass, 2x105conidia of Trichophyton were grown in 96-well plates in 200 pL of RPMI 1640 medium at 10% FCS and incubated at 30°C for 24 h in the absence or presence of different concentrations of trifarotene: 1 mM, 0.5 mM, 0.25 mM,0.12mM and 0.06 mM corresponding to final concentrations of 300 pg / mL, 150 pg / mL, 75 pg / mL, 37.5 pg / mL and 18.75 pg / mL of trifarotene, in the presence or absence of terbinafine: 1 mg / L, 0.5 mg / L, 0.25 mg / L, 0.12 mg / L, 0.06 mg / L, alone or in combination with suboptimal doses of trifarotene (0.75 mm, 0.5 mM, 0.25 mM).RESULTSCell growth rate of C. albicans

[0224] The efficacy of trifarotene in inhibiting the growth of C. albicans was studied. Trifarotene from 1.0 to 0.25 mM significantly inhibited (chi-square test, p<0.001.) the growth of Candida, with 1 mM being the most effective concentration, inducing an 88% reduction in fungal growth. As shown in Figure 9, the effects of trifarotene on the growth of C. albicans are evident. Yeast cells of C. albicans were cultured for 24 hours in the absence or presence of various concentrations of trifarotene (1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM trifarotene). AmB was used as a positive control drug. The results are presented as the mean ± SD of three independent experiments, conducted in triplicate, and expressed as the percentage of growth inhibition compared to the control group. Student’s t-test, * p <0.05; ** p <0.01 ; *** p <0.001.Hyphal growth inhibition assay

[0225] The impact of trifarotene on the germination of C. albicans and hyphal growth was also evaluated. For this purpose, 2x105cells were cultured in 96-well plates in 200 pL of RPMI 1640 medium with 10% FCS and incubated at 37°C in the absence or presence of different concentrations of trifarotene at 1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM. As shown in Figure 10, the effects of trifarotene on phenotypic switching from the yeast to the hyphal form of C. albicans are evident. Yeast cells of C. albicans were cultured in the absence or presence of various concentrations of trifarotene (1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM). The phenotypic transition from yeast to filamentous morphotype was analysed under the microscope after 3 and 24 hours of incubation. A representative experiment of three is shown.Biofilm quantification using Crystal Violet and XTT tests

[0226] The impact of trifarotene on the germination of C. albicans, C. tropicalis, C. krusei, C. auris, C. glabrata, in terms of both biomass and metabolic activity, was also evaluated. For this purpose, 2x105Candida cells were cultured in 96-well plates in 200 pL of RPMI 1640 medium with 10% FCS and incubated at 37°C for 24 hours in the absence or presence of different concentrations of trifarotene. The data obtained showthat at concentrations between 1.0 mM and 0.25 mM, trifarotene is able to inhibit biofilm production in C. albicans, C. tropicalis, C. krusei, C. auris, C. glabrata.

[0227] As shown in Figure 1 , the inhibitory effect of trifarotene on the biofilm biomass of C. krusei is evident. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM). Amphotericin B (AmB) was used as a positive control drug. The following AmB concentrations were used: 2 pg / mL, 0.5 pg / mL, 0.25 pg / mL, and 0.12 pg / mL. Biofilm growth was assessed as total biomass (A) by staining with crystal violet (CV) and quantified by measuring optical density (OD) using a spectrophotometer at 595 nm. The results are the mean of the O.D. values ± SD of three independent experiments, each conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001 .

[0228] As shown in Figure 2, the inhibitory effect of trifarotene on the metabolic activity of C. krusei biofilm is evident. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM). AmB was used as a positive control drug. The metabolic activity of the biofilm was assessed by XTT tetrazolium salt reduction assay and quantified by measuring the optical density (O.D.) using a spectrophotometer at 490 nm. The results are the mean of the O.D. values ± SD of three independent experiments, each conducted in triplicate * p <0.05, ** p <0.01 ; *** p <0.001 .

[0229] As shown in Figure 3, the inhibitory effect of trifarotene on the biofilm biomass of C. tropicalis is evident. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM). AmB was used as a positive control drug. Biofilm growth was assessed as total biomass (A) by staining with crystal violet (CV) and quantified by measuring optical density (OD) using a spectrophotometer at 595 nm. The results are the mean of the O.D. values ± SD of three independent experiments, each conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001.

[0230] As shown in Figure 4, the inhibitory effect of trifarotene on the metabolic activity of C. tropicalis biofilm is evident. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM). AmB was used as a positive control drug. The metabolic activity of the biofilm was assessed by XTT tetrazolium salt reduction assay and quantified by measuring the optical density (O.D.) using a spectrophotometer at 490 nm.The results are the mean of the O.D. values ± SD of three independent experiments, each conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001.

[0231] As shown in Figure 5, the inhibitory effect of trifarotene on the biofilm biomass of C. auris is evident. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12 mM and 0.06 mM). AmB was used as a positive control drug. Biofilm growth was assessed as total biomass (A) by staining with crystal violet (CV) and quantified by measuring optical density (OD) using a spectrophotometer at 595 nm. The results are the mean of the O.D. values ± SD of three independent experiments conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001.

[0232] As shown in Figure 6, the inhibitory effect of trifarotene on the metabolic activity of C. auris biofilm is evident. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM). AmB was used as a positive control drug. The metabolic activity of the biofilm was assessed by XTT tetrazolium salt reduction assay and quantified by measuring the optical density (O.D.) using a spectrophotometer at 490 nm. The results are the mean of the O.D. values ± SD of three independent experiments conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001 .

[0233] As shown in Figure 7, the inhibitory effect of trifarotene on the biofilm biomass of C. glabrata is evident. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM). AmB was used as a positive control drug. Biofilm growth was assessed as total biomass (A) by staining with crystal violet (CV) and quantified by measuring optical density (OD) using a spectrophotometer at 595 nm. The results are the mean of the O.D. values ± SD of three independent experiments conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001.

[0234] As shown in Figure 8, the inhibitory effect of trifarotene on the metabolic activity of C. glabrata biofilm is evident. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM). AmB was used as a positive control drug. The metabolic activity of the biofilm was assessed by XTT tetrazolium salt reduction assay and quantified by measuring the optical density (O.D.) using a spectrophotometer at 490 nm. The results are the mean of the O.D. values ± SD of three independent experiments conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001.

[0235] As shown in Figure 11, trifarotene has an inhibitory effect on the biofilm biomass of C. albicans. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM). AmB was used as a positive control drug. Biofilm growth was assessed as total biomass (A) by staining with crystal violet (CV) and quantified by measuring optical density (OD) using a spectrophotometer at 595 nm. The results are the mean of the O.D. values ± SD of three independent experiments conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001.

[0236] As shown in Figure 12, trifarotene has an inhibitory effect on the metabolic activity of C. albicans biofilm. The Candida cells were cultured for 24 hours in the absence or presence of trifarotene at various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM). AmB was used as a positive control drug. The metabolic activity of the biofilm was assessed by XTT tetrazolium salt reduction assay and quantified by measuring the optical density (O.D.) using a spectrophotometer at 490 nm. The results are the mean of the O.D. values ± SD of three independent experiments conducted in triplicate. * p <0.05, ** p <0.01 ; *** p <0.001 .Quantification of cell viability of C. albicans

[0237] In order to assess whether the inhibitory effect of trifarotene on Candida growth and biofilm formation was due to a fungistatic or fungicidal effect, the viability of planktonic cells of C. albicans was assessed by double immunofluorescence staining with CW and PI. Calcofluor White colours the cell wall of fungi blue, regardless of their metabolic state, while PI emits red fluorescence when binding to the DNA of dead cells. Microscopic images show that exposure to trifarotene 1.0 mM produced 55% non-viable Pl-positive cells, respectively. In contrast, at concentrations < 0.5 mM, the percentage of Pl-positive cells was less than 2%. This value was very similar to that of the Candida control cells. Overall, the results suggest that trifarotene is able to inhibit growth and biofilm formation in C. albicans by acting as a fungicide or fungistatic agent, depending on the concentration used.

[0238] As shown in Figure 13, the viability assessment of C. albicans is shown by double staining with Calcofluor White (CW) and propidium iodide (PI); planktonic cells of C. albicans were incubated in the absence or presence of various concentrations (1 mM, 0.5 mM, 0.25 mM, 0.12mM and 0.06 mM corresponding to the final concentrations of 300 pg / mL, 150 pg / mL, 75 pg / mL, 37.5 pg / mL and 18.75 pg / mL of trifarotene) of trifarotene. After 24 hours of incubation, Candida cells were double-stained with CW and PI.Fluorescent images of the labelled cells show: in the left column, the blue fluorescence represents the CW-stained cell wall; in the middle column, the red fluorescence represents the Pl-stained nucleic acids of the dead cells; in the right column, the merge of images shows the double-labelled cells, stained with CW and PI (blue and red). The column on the left shows Pl-labelled cells, and these are live cells. In the middle column, the fluorescent cells are dead cells; in the right column, the image merge shows the double-marked cells, stained with CW and PI. All images were obtained at 100x magnification, using a fluorescence microscope. The results, represented graphically with histograms, show the percentage of dead cells. At least 10 fields per slide were counted. Chi-square test; * p <0.05, ** p <0.01 ; ***p<0.001.Assessment of the biofilm biomass of Trichophyton spp.

[0239] The impact of trifarotene on biofilm production by Trichophyton mentagrophytes complex was evaluated. For this purpose, 2x105conidia of Trichophyton mentagrophytes complex were cultured in 96-well plates in 200 pL of RPM1 1640 medium with 10% FCS and incubated at 30°C for 24 h in the absence or presence of different concentrations of trifarotene. As shown in Figure 18, there is a potent inhibitory effect of trifarotene, at concentrations between 1.0 mM and 0.75 mM, on the biofilm production of Trichophyton mentagrophytes complex, evaluated in terms of biomass by light microscopy after staining with crystal violet.

[0240] Furthermore, as shown in Figure 19, trifarotene at concentrations at which it had shown no or reduced efficacy (0.25-0.5 mM) showed efficacy when administered in conjunction with terbinafine (0.5 mg / L) even when combined with lower doses of terbinafine than those usually used in the treatment of Trichophyton spp. This figure shows that the composition containing both trifarotene and terbinafine even at low concentrations is effective in inhibiting biofilm formation by Trichophyton mentagrophytes complex. This shows that trifarotene and terbinafine have a synergistic effect in the treatment of Trichophyton mentagrophytes complex and that together they can be used in smaller quantities than alone.EXAMPLESExample 1 : Composition according to the invention

[0241] A preparation for topical use was prepared containing water, PPG-3 benzyl ether myristate, glyceryl stearate, panthenol, diisopropyl adipate, PPG-15 stearyl ether, PEG-100 stearate, dibutyl adipate, salicylic acid, dimethicone, cetearyl alcohol, sodium polyacrylate, glycerine, hydrogenated polydecene, phenoxyethanol, sorbitan stearate,bisabolol, citric acid, hydroxyethyl cellulose, sodium gluconate, sodium sulphite, trideceth-6, ethylhexylglycerine, trifarotene, BHT, sodium hyaluronate, sodium hydroxide and between 0.001 wt% to 0.01 wt% trifarotene.Example 2: Composition according to the invention

[0242] A preparation for topical use was prepared containing Allantoin, acrylamide copolymer, sodium-Vp-crosspolymer acryloyldimethyltaurate, isohexadecane, cyclomethicone, ethanol (5%), medium-chain triglycerides, trifarotene between 0.001wt% and 0.01 wt%.Example 3: Composition according to the invention

[0243] A preparation for topical use was prepared containing sodium docusate, disodium edetate, glycerol, polyoxamer, propylene glycol (E1520), Simulgel 600 PHA (copolymer of acrylamide and sodium acryloyldimethyltaurate, isohexadecane, polysorbate 80, sorbitan oleate), trifarotene, purified water and between 0.001 wt% to 0.01 wt% trifarotene.Example 4: Composition according to the invention

[0244] A preparation for topical use was prepared containing cetyl alcohol, stearic acid, isopropyl myristate, polysorbate 60, sorbitan stearate, non-crystallisable sorbitol 70%, sorbic acid, sodium edetate, butylhydroxytoluene, purified water, and between 0.001 wt% to 0.01 wt% of Trifarotene.Example 5: Composition according to the invention

[0245] A preparation for topical use was prepared containing water, diisopropyl adipate, ethylhexyl cocoate, caprylic / capric triglyceride, niacinamide, alcohol denat, glyceryl stearate, cetearyl alcohol, ethoxydiglycol, potassium azeloyl diglycinate, ammonium acryloyldimethyltaurate / vp copolymer, cellulose glycerine, xylitol, dimethicone, peg-100 stearate, phenoxyethanol, glycine soybean oil, trifarotene, phospholipids, bisethylhexyl hydroxy benzylmalonate, alpha-glucan oligosaccharide, zinc pea, xanthan gum, allantoin, trisodium ethylenediamine disuccinate, glycyrrhetinic acid-5 -ol, calcium gluconate, lactic acid, pentaerythrityl tetra -di-t-butyl hydroxyhydrocinnamate, hippophae rhamnoides fruit oil, menthol, menthyl lactate, ascorbyl palmitate, and between 0.001 wt% to 0.01 wt% of Trifarotene.Example 6: Composition according to the invention

[0246] A preparation for topical use was prepared containing between 0.001 wt% to 0.01 wt% Trifarotene, Acido Glycol, Propylene Glycol, PEG-7 Glyceryl Cocoate,Aminomethyl Propanol, Hydroxyethylcellulose, Polyvinyl Alcohol, Imidazolidinyl Urea, Disodium EDTA, BHT, Aqua.Example 7: Composition according to the invention

[0247] A preparation for topical use was prepared containing between 0.001 wt% to 0.01wt% of Trifarotene, carbomer 940, edetate disodium, methylparaben, poloxamer 182 propylene glycol, purified water and sodium hydroxide.Example 8: Composition according to the invention

[0248] A preparation for topical use was prepared containing between 0.001 wt% to 0.01 wt% of Trifarotene, carbomer 934P, Peg-20 methyl glucose sesquistearate, glycerol, natural squalane, methyl parahydroxybenzoate, propyl parahydroxybenzoate, disodium edetate, methyl glucose sesquistearate, phenoxyethanol, cyclomethicone, sodium hydroxide, purified water.Example 9: Composition according to the invention

[0249] A medicated nail polish was prepared in the form of a filmogenic oil-in-water emulsion with acrylic polymer trifarotene at 0.001 -0.01 % wt%. Indicative composition: trifarotene (0.001 -0.01 wt%), film-forming acrylic copolymer in 30-40% aqueous dispersion (15-25%), carbomer 934P (0.3-0.8%), glycerol (3-8%), natural squalane (2- 5%), cyclomethicone (1 -3%), PEG-20 methyl glucose sesquistearate (1 -3%), methyl glucose sesqualistearate (0.5-2%), disodium edetate (0.05-0.1 %), methyl p- hydroxybenzoate (0.1 -0.2%), propyl p-hydroxybenzoate (0.02-0.1 %), phenoxyethanol (0.5-1 %), sodium hydroxide sufficient to adjust pH to 6-7, purified water q.s. to 100%.Example 10: Composition according to the invention

[0250] Nail varnish in the form of a film-forming hydroalcoholic solution containing trifarotene at 0.001 -0.01 wt%. Indicative composition: trifarotene (0.001 -0.01 wt%), filmforming polymer, e.g. acrylate copolymer or PVPA / A (10-20%), ethanol (30-50%), isopropanol (10-25%), purified water (5-15%), glycerol (2-5%), natural squalane (1 -3%), cyclomethicone (1 -3%), disodium edetate (0.05-0.1 %), methyl p-hydroxybenzoate (0.1 - 0.2%), propyl p-hydroxybenzoate (0.02-0.1 %), phenoxyethanol (0.5-1 %), plasticiser, e.g. triacetin or a citrate (2-8%), antioxidant, e.g. vitamin E or BHT (0.01 -0.1 %).Example 11: Composition according to the invention

[0251] Medicated nail varnish containing per 100 g: Terbinafine hydrochloride 8.0%, trifarotene 0.005%, Ammonium methacrylate copolymer type B (Eudragit RL 100) 8.0%, Hydroxypropyl methylcellulose 4.0%, Propylene glycol 6.0%, Thioglycolic acid 2.0%,Triacetin 3.0%, Butylhydroxytoluene (BHT) 0.05%, Ethyl acetate 25.0%, Anhydrous ethanol q.s. to 100%.Example 12: Composition according to the invention

[0252] Medicated nail varnish containing per 100 g: Terbinafine base 6.0%, trifarotene 0.005%, Nitrocellulose 12.0%, Toluenesulphonamide / formaldehyde resin 7.0%, Triethyl citrate 8.0%, Titanium dioxide 3.0%, Iron oxide pigments 2.0%, Ethyl acetate 35.0%, Butyl acetate 15.0%, Isopropyl alcohol 5.0%, Butylhydroxytoluene (BHT) 0.02%, Ethanol to 100%.Example 13: Composition according to the invention

[0253] Filmogenic / keratoprotective nail varnish containing: Hydroxypropyl chitosan 4.0%, Urea 10.0%, Lactic acid 5.0%, Methylsulphonylmethane 3.0%, Equisetum Arvense (Horsetail) extract 1.0%, Propylene glycol 7.0%, Panthenol 1.0%, Denatured alcohol 30.0%, Water q.s. to 100%, Sodium benzoate 0.30%, Potassium sorbate 0.20%, Citric acid q.s. pH 3.5-4.0.Example 14: Composition according to the invention

[0254] Nail varnish I barrier film containing: Cyclomethicone 25.0%, Dimethicone 30.0%, Isopropyl myristate 10.0%, Ethyl lactate 10.0%, Denatured alcohol 15.0%, Ammonium methacrylate copolymer type B 4.0%, Hydroxypropylated chitosan 3.0%, Tocopheryl acetate (Vitamin E) 1.0%, Perfume 0.3%, BHT (Butylated Hydroxytoluene) 0.05%, Water q.s. to 100%.Example 15: Micellar hydroalcoholic mouthwash:

[0255] Mouthwash containing: Trifarotene 0.001 %, Ethanol 96% 10.0%, Propylene Glycol 10.0%, Hydrogenated Castor Oil PEG-40 1.0%, Polysorbate 80 0.5%, Glycerin 10.0%, Sorbitol solution 70% 15.0%, Sodium Saccharin 0.10%, Mint Flavour 0.20%, Sodium Benzoate 0.15%, Potassium Sorbate 0.10%, Citric Acid 0.05%, Sodium Citrate 0.10%, Purified Water q.s. to 100%.Example 15: Mouthwash with cyclodextrin

[0256] Mouthwash containing: Trifarotene 0.001 %, Hydroxypropyl-[3-Cyclodextrin 5.0%, Glycerine 10.0%, Sorbitol solution 70% 20.0%, Propylene Glycol 5.0%, Polysorbate 80 0.3%, Sodium saccharin 0.10%, Mint flavour 0.20%, Sodium benzoate 0.15%, Potassium sorbate 0.10%, Citric acid 0.05%, Sodium citrate 0.10%, Purified water q.s. to 100%.

Claims

Claims1. Pharmaceutical composition comprising trifarotene for use in the treatment of fungal infections, wherein said fungal infections are fungal infections caused C. albicans or fungal infections caused by species of C. non-albicans in particular by one or more of C. krusei, C. tropicalis, C. auris, C. glabrata or fungal infections caused by Trychophyton spp, in particular by Trichophyton mentagrophytes complex, Trichophyton rubrum.

2. Pharmaceutical composition for use according to claim 1 wherein said composition comprises trifarotene in a percentage between about 0.001 wt% and about 10 wt%, preferably between about 0.005wt% and about 5wt%, more preferably between about 0.01 wt% and about 1wt%.

3. Pharmaceutical composition for use according to any one of claims 1 or 2 wherein said composition comprises trifarotene in a percentage between 0.1 and 2 mM, preferably between 0.25 mM and 1 .5 mM, more preferably between 0.5 mM and 1 mM.

4. Pharmaceutical composition for use according to one of the preceding claims and further comprising an additional antifungal agent.

5. Pharmaceutical composition for use according to claim 4, wherein the at least one additional antifungal agent is selected from the group consisting of azoles such as for example fluconazole, voriconazole, posaconazole, itraconazole, ketoconazole, isavuconazole, thioconazole, opelconazole, miconazole.

6. Pharmaceutical composition for use according to claim 4, or 5, wherein the at least one additional antifungal agent is selected from the group consisting of polyenes such as amphotericin B, nystatin or liposomal and lipid forms.

7. Pharmaceutical composition for use according to one of claims 4 to 6, wherein the at least one additional antifungal agent is selected from the group consisting of purine or pyrimidine nucleotide inhibitors such as flucytosine.

8. Pharmaceutical composition for use according to one of claims 4 to 7, wherein the at least one additional antifungal agent is selected from the group consisting of polyoxins, such as nikkomycins, preferably nikkomycin Z or other chitin inhibitors.

9. Pharmaceutical composition for use according to one of claims 4 to 8, wherein the at least one additional antifungal agent is selected from the group consisting of echinocandins such as micafungin and anidulafungin.

10. Pharmaceutical composition for use according to any one of the preceding claims and comprising one or more additional fungal agents, said one or more additional fungal agents being selected from azoles, polyenes, purine or pyrimidine nucleotide inhibitors, polyoxins, or other chitin inhibitors, echinocandins such as micafungin and anidulafungin or combinations thereof, from alilamines, preferably terbinafine.11 . Pharmaceutical composition for use according to one of claims 4 to 10 wherein said at least one additional antifungal agent is present in said composition in a percentage between about 0.01 wt% and about 10 wt%, preferably between about 0.05 wt% and about 5 wt% more preferably between about 0.02 wt% and about 2wt%.

12. Pharmaceutical composition for use according to one of the preceding claims and further comprising at least one compound selected from at least one excipient, at least one carrier, at least one adjuvant such as glycerin, gelatin, propylene glycol, butylene glycol, panthenol, sorbitol, urea, hyaluronic acid glycolic acid, lactic acid, sodium pyrrolidine carboxylic acid, cholesterol, squalene, linoleic acid, stearic acid, oleic acid, fatty alcohols, white / petrolatum soft paraffin, beeswax, mineral oil, dimethicone, lanolin, carnauba wax, cetyl alcohol, caprylic / capric triglyceride13. Pharmaceutical composition for use according to any one of the preceding claims, and comprising between about 1 pg / g and about 100 pg / g of trifarotene.

14. Pharmaceutical composition for use according to any one of the preceding claims for use in the veterinary treatment of fungal infections caused by C. albicans or fungal infections caused by species of non-albicans Candida in particular by one or more of C. krusei, C. tropicalis, C. auris, C. glabrata, or by Trychophyton spp. in particular Trichophyton mentagrophytes complex, Trichophyton rubrum.

15. Pharmaceutical composition comprising trifarotene in a percentage comprised between about 0.001 wt% and about 10 wt%, preferably between about 0.005wt% and about 5wt%, more preferably between about 0.01 wt% and about 1wt% for use in the treatment and / or prevention of recurrences of fungal infections caused by C. albicans or fungal infections caused by species of non-albicans Candida in particular by one or more of C. krusei, C. tropicalis, C. auris, C. glabrata, Trychophyton spp, in particular Trichophyton mentagrophytes complex and Trichophyton rubrum.

16. Pharmaceutical composition for use according to any one of the preceding claims for topical use.

17. Pharmaceutical composition for use according to any one of the preceding claims, wherein said composition is in the form of an ointment, topical cream, topical gel, liniment, paste, film, solution, hydrogel, liposome, transferable vesicle, cream, lotion, dermal patch, transdermal patch, transdermal spray.

18. Pharmaceutical composition for use according to any one of the preceding claims, wherein said fungal infections are fungal infections of the skin or skin adnexa, in particular of the nails.

19. Pharmaceutical composition for use according to any one of the preceding claims, wherein said fungal infections are fungal infections of the mucous membranes.

20. Pharmaceutical composition for use according to any of the preceding claims, wherein said fungal infections are chronic mucocutaneous candidiasis (ORPHA:1334).21 . Pharmaceutical composition for use according to any one of the preceding claims wherein said fungal infections are fungal infections of the vaginal, nasal, oral, anal mucosa.

22. Pharmaceutical composition for use according to any one of the preceding claims, wherein said fungal infections are fungal nail infections.

23. Pharmaceutical composition for use in the treatment of fungal infections of Trichophyton spp., in particular Trichophyton mentagrophytes complex, Trichophyton rubrum.

24. comprising trifarotene and terbinafine, wherein said composition preferably comprises between about 1 mg / L and about 0.06 mg / L of terbinafine and between about 1 mM and about 0.25 mM of trifarotene.

25. Pharmaceutical composition for use according to the preceding claim, wherein said composition is in the form of a cream, ointment, glaze, tablet, lacquer, solution, powder.

26. Pharmaceutical composition for use according to one of the preceding claims wherein said composition is in the form of an ointment, pessary, extra-amniotic infusion, intravesical infusion, nasal spray, ear drops, eye drops, ointment, insufflation, mucoadhesive microdisk.

27. Pharmaceutical composition for use according to any one of claims 1 to 18, characterised in that it is contained in a medical device for local and / or controlledrelease of the composition in a body district, wherein said medical device is an insertable or implantable device selected from devices for gynaecological, urological, rectal, gastrointestinal, cutaneous, intranasal, otological, ophthalmic administration and combinations thereof.

28. Pharmaceutical composition for use according to any one of claims 1 to 18, characterised in that it is inserted into a medical device, wherein said medical device is selected from vaginal ring, intrauterine device (IUD).

29. Pharmaceutical composition for use for the treatment of fungal infections comprising at least one compound suitable for binding the HSP90 protein, wherein said fungal infections are fungal infections sustained by one or more of the following species: C. albicans, C. auris, C. tropicalis, C. glabrata and C. krusei.

30. Pharmaceutical composition for use according to the preceding claim wherein said compound suitable for binding the HSP90 protein of a fungus is selected from the group of fourth generation retinoids, which include trifarotene and seletinoid G.31 . Pharmaceutical composition for use in the treatment of fungal infections of the skin sustained by species of Candida and / or Trichophyton mentagrophytes complex, Tricophyton rubrum, characterised in that it is contained in a medical device, said medical device comprising: a support or substrate configured to be applied to the skin; and a layer or coating on the support comprising a composition comprising trifarotene according to any one of the preceding claims; wherein said medical device is selected from medicated patch, occlusive bandage, adherent film or sheet hydrogel.

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