Active agents for use in the treatment of diseases caused by the yeast strains candida albicans ATCC 10231, candida krusei ATCC 6258, and candida tropicalis y-12968

Abstract

The present invention relates to new active agents for use in the treatment of diseases caused by the yeast strains C. albicans ATCC 10231, C. krusei ATCC 6258, and C. tropicalis Y-12968.

Description

[0001] DESCRIPTION

[0002] ACTIVE AGENTS FOR USE IN THE TREATMENT OF DISEASES CAUSED BY THE YEAST STRAINS Candida albicans ATCC 10231, Candida krusei ATCC 6258, AND Candida tropicalis Y-12968

[0003] Technical Field

[0004] The present invention relates to new active agents for use in the treatment of diseases caused by the yeast strains C. albicans ATCC 10231, C. krusei ATCC 6258, and C. tropicalis Y-12968.

[0005] Prior Art

[0006] Yeasts, particularly C. albicans, are considered a significant cause of healthcare- associated infections. Candida species are currently the fourth most common cause of bloodstream infections among hospitalized patients in the United States2and are particularly common pathogens in intensive care units (ICUs). Yeast bloodstream infections (candidemia) are associated with significant morbidity and mortality, with an attributable mortality risk approaching 49%.4An increase in the incidence of yeast bloodstream infections is driven by species such as C. krusei and C. albicans.6,10These species tend to be more resistant to commonly used triazole agents, such as fluconazole.7,12Therefore, they pose a particular challenge for clinical management.

[0007] The most commonly used antifungal drugs are clotrimazole, miconazole, or nystatin. For severe infections, the most commonly used antifungal drug administered orally or intravenously is fluconazole.

[0008] One of the significant health problems of our time is infectious diseases. In the treatment of infectious diseases caused by the uncontrolled proliferation of pathogens, various antibiotics are used. However, it has recently been determined that there is an increase in the antibiotic resistance of pathogens. Furthermore, changes in the structure of pathogens are causing antimicrobial resistance. This situation necessitates the development of new antimicrobial agents. The yeast strains against which the Octabutylamino(sbs)-cyclotriphosphazenes were determined to be active are Candida albicans, Candida knisei, and Candida tropicalis.

[0009] Candida species are the most common cause of fungal infections worldwide. They are a component of the normal microbiota in the gastrointestinal tract, respiratory tract, vaginal region, and mouth.8Candida is a fungal genus found in all women. Candida species differ in terms of their antifungal susceptibility and virulence factors. The genus consists of a heterogeneous group of organisms, and it is known that more than 17 different Candida species are the etiological agents of human infections. However, more than 90% of invasive infections are caused by C. albicans, C. glabrata, C. parapsilosis, C. tropicalis, C. krusei, C. dubliniensis, and C. lusitaniae The yeast begins to invade and colonize body tissues by releasing potent chemicals into the bloodstream, causing symptoms such as lethargy, chronic diarrhea, yeast vaginitis, bladder infections, muscle and joint pain, menstrual problems, constipation, and severe depression.13This condition, which is responsible for 90% of infectious vaginitis cases, is called candidiasis. C. albicans is the most common fungal pathogen in humans, the fourth most common cause of hospital-acquired infectious diseases, and the primary cause of systemic candidiasis, with mortality rates approaching 50%.11The long-term use of antifungals in the treatment of infections caused by C. albicans has led to the emergence of azole resistance. This acquired azole resistance in clinical isolates of C. albicans often results in cross-resistance to many unrelated drugs. This phenomenon is referred to as multidrug resistance. Multidrug resistance is a serious complication during the treatment of opportunistic fungal infections, causing significant concern given that the number of clinically useful antifungal drugs is limited. Fungal diseases of the gastroduodenum are reported less frequently. They most often occur as a secondary infection in individuals with tumors in this region, and they infiltrate benign or malignant ulcers that have a diminished healing capacity. Endoscopically, this resembles a white or grayish deposit located at the base of the ulcer, which is easily separated from the mucosa. The ulcer usually heals with anti -ulcer therapy.1The interaction of C. albicans as a pathogen with the intestinal mucosa occurs via adhesion, invasion, damage, and apoptosis. The main role in the infection, and therefore in pathogenicity, is performed by the fungal hyphae.5C. albicans can cause folliculitis, typically in the beard region, in an adult male with severe immunosuppression. In such cases, it can involve the deep layers of the skin and may present with the clinical finding of deeper nodules around the hairs and pustules.9

[0010] C. albicans is the most common fungus (or yeast) that lives on the skin, in the mouth, vagina, and gastrointestinal tract of humans. One of the most important reasons for the increase in Candida infections is the development of resistant strains due to the drugs used in the treatment of candidiasis.11They can be significant pathogens causing infections in patients with human immunodeficiency virus (HIV) and in vulnerable intensive care unit patients. Among hospital-acquired infections, Candida infections are the most common.

[0011] C. krusei is able to grow in a vitamin-free environment. This pathogen, which emerges in individuals with immunodeficiency and in HIV patients, has developed resistance to the triazole antifungal drug fluconazole due to its frequent use.3

[0012] C. tropicalis is seen more frequently in adult individuals. C. tropicalis has developed resistance to antifungal drugs such as azole derivatives, amphotericin B, and echinocandins.

[0013] Trimeric phosphazene derivatives with various properties can be obtained as a result of nucleophilic substitution reactions of Cl atoms in the hexachlorocyclotriphosphazene (trimer, N3P3CI6) with different groups. Thanks to the important properties gained from these different groups, phosphazenes can be used in chemical, technological, and biological applications. Some of these are used as anticancer and antimicrobial agents, flame retardant additives, liquid crystals, support materials in dyes and catalysts, and in the production of materials such as organic light-emitting diodes (OLEDs) and fluorescence sensors. Furthermore, due to the development of resistance by bacterial and yeast strains to existing drugs, the production of new active pharmaceutical ingredients is highly important. It is known that cyclotriphosphazene compounds, their metal complexes, and some phosphazene polymers possess antibacterial and antifungal activities. In particular, the pyrrolidine-, piperidine-, and l,4-dioxa-8-azaspiro[4.5]decane (DASD)- substituted derivatives of cyclotriphosphazenes have been found to possess antimicrobial activities.

[0014] JPH01175999A discloses cyclotriphosphazene derivatives.

[0015] Upon reviewing the existing studies in the prior art, it has been determined that there is a need for new active agents that can be used in the treatment of diseases caused by C. albicans ATCC 10231, C. krusei ATCC 6258 and C. tropicalis Y- 12968 yeast strains.

[0016] The Object of the Invention

[0017] The object of the present invention is to develop new active agents for use in the treatment of diseases caused by the yeast strains C. albicans ATCC 10231, C. krusei ATCC 6258, and C. tropicalis Y-12968.

[0018] Detailed Description of the Invention

[0019] The present invention relates to new active agents for use in the treatment of diseases caused by the yeast strains C. albicans ATCC 10231, C. krusei ATCC 6258, and C. tropicalis Y-12968. The new active agents subject to the invention are compounds of Formula A,

[0020] (Formula A) wherein, n is 0 or 1; R is CFF or C2H5; and R' is butylamine. The IUPAC names and structures of the compounds subject to the invention are specified below.

[0021] Compound 1 : Bis(7,7,9,9-tetra(butylamino)-(l-methyl-4-[3-(4-methyl-l,4,6,8,10- pentaaza-5,7,9-triphosphaspiro[4.5]deca-5,7,9-trien-l-yl-)benzyl]

[0022] Compound 2: Bis(7, 7, 9, 9-tetra(buthylamino)-(l-ethyl-4-[3-(4-ethyl-l, 4,6,8,10- pentaaza-5,7,9-triphosphaspiro[4.5]deca-5,7,9-triene-l-yl-)benzyl] Compound 3 : Bis(8,8, 10, 10-tetra(buthylamino)-(l-methyl-5-[4-(5-methyl- l,5,7,9,l l-pentaaza-6,8,10-triphosphaspiro[5.6]deca-6,8,10-triene-l-yl-)benzyl]

[0023] The synthesis steps for the compounds of the invention (1, 2, 3) are specified below. Firstly, the condensation reaction of isophthalaldehyde with N-methyl-1,2- diaminoethane, N-ethyl-l,2-diaminoethane, and N-methyl-l,3-diaminopropane was carried out in an ethyl alcohol medium to form the corresponding Schiff bases. The resulting Schiff bases were then reduced with NaBFU in the same solvent medium to obtain a tetradentate benzyltetraamine with N2N2 donor atoms. The spiro-bino-spiro (sbs)-methyl / ethylcyclotriphosphazene derivative was synthesized by reacting the benzyltetraamine with the trimer in a tetrahydrofuran (THF) medium in the presence of triethylamine.

[0024] Subsequently, the target compounds (1), the octabutylamino-sbs- methyl / ethylcyclotriphosphazene derivatives, were obtained from the reactions of the sbs-methylcyclotriphosphazene with butylamine in a 1 :8 stoichiometric ratio, in a THF medium, and in the presence of tri ethyl amine.

[0025] The final compounds were purified by column chromatography using silica gel as the stationary phase. Antimicrobial Activities of the Compounds:

[0026] The compounds were prepared by being dissolved in dimethyl sulfoxide (DMSO) to a concentration of 2500 pM. Ketoconazole was used as the control for the yeasts. Minimum Inhibitory Concentration (MIC) is defined as the lowest concentration of an antimicrobial agent that inhibits the visible growth of a microorganism. Minimum Fungicidal Concentration (MFC) is defined as the lowest concentration of an antimicrobial agent that reduces the viability of the initial microbial inoculum by 99.9%. The MIC determination was performed by the broth microdilution method in 96-well microplates. While the pathogenic microorganisms used in the study (the yeast strains C. albicans ATCC 10231, C. krusei ATCC 6258, and C. tropicalis Y- 12968) were found to be susceptible to the compounds at certain concentrations, the compounds were more effective than the standard antifungal when compared with the positive control (Ketoconazole for yeasts). The experiments were performed in triplicate.

[0027] The antimicrobial activities of the inventive compounds were investigated against the yeast strains C. albicans ATCC 10231, C. tropicalis (ATCC 13803), and C. krusei (ATCC 14243). The minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values of the compounds against these pathogens were determined. Considering the MIC values against the C. albicans strain, it was observed that compounds 1 (9.77 pM) and 3 (9.77 pM) were approximately 32 times more effective than the reference antifungal Ketoconazole (312.5 pM), while compound 2 (78.15 pM) was 4 times more effective. Furthermore, compounds 1, 2, and 3 were found to be 8 times more effective against the C. tropicalis yeast strain compared to the same reference. When considering the Minimum Fungicidal Concentration (MFC) values, it was determined that compounds 1, 2, and 3 were much more effective than Ketoconazole. The MFC values against the yeast strains were as follows; C. albicans'. 9.77 pM (for compounds 1 and 3) and 78.15 pM (for compound 2), compared to 1250 pM for Ketoconazole. C. tropicalis'. 156.8 pM (for compound 1), 39.08 pM (for compound 2), and 312.5 pM (for compound 3), compared to 1250 pM for Ketoconazole. C. krusei'. 78.15 pM (for compound 1), 39.08 pM (for compound 2), and 153.3 pM (for compound 3), compared to 156.3 pM for Ketoconazole.

[0028] References:

[0029] 1. Castelo Branco S., Ferreira A.T., Saraiva S., Silva M.J., Garcia T. Fungal Gastroduodenitis. Eur. J. Case Rep. Intern. Med. 2017;2.

[0030] 2. Edmond, M. B., S. E. Wallace, D. K. McClish, M. A. Pfaller, R. N. Jones, and R. P Wenzel. 1999. Nosocomial bloodstream infections in United States hospitals: a three-year analysis. Clin. Infect. Dis. 29:239-244.

[0031] 3. Gomez-Gaviria M, Mora-Montes HM. Current Aspects in the Biology, Pathogeny, and Treatment of Candida krusei, a Neglected Fungal Pathogen. Infect Drug Resist. 2020; 13: 1673- 1689.

[0032] 4. Gudlaugsson, O., S. Gillespie, K. Lee, J. Vande Berg, J. Hu, S. Messer, L. Herwaldt, M. Pfaller, and D. Diekema. 2003. Attributable mortality of nosocomial candidemia, revisited. Clin. Infect Dis. 37: 1172-1177.

[0033] 5. Moyes D.L., Richardson J.P., Naglik J.R. Candida albicans-epithelial interactions and pathogenicity mechanisms: Scratching the surface. Virulence. 2015;6:338-346.

[0034] 6. Pfaller, M. A., D. J. Diekema, R. N. Jones, S. A. Messer, and R J Hollis 2002 Trends in antifungal susceptibility of Candida spp. isolated from pediatric and adult patients with bloodstream infections: SENTRY Antimicrobial Surveillance Program, 1997 to 2000. J. Clin. Microbiol. 40:852-856.

[0035] 7. Pfaller, M. A., S. A. Messer, R. J. Hollis, R. N. Jones, and D. J. Diekema. 2002. in vitro activities of ravuconazole and voriconazole compared with those of four approved systemic antifungal agents against 6,970 clinical isolates of Candida spp. Antimicrob. Agents Chemother. 46: 1723-1727.

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Claims

CLAIMS1. A compound of formula A,(Formula A) wherein; n; 0 or 1,R; CH3or C2H5andR'; butylamine.

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