Gluconic acid derivatives for use in the treatment and / or prevention of microbial infections

Abstract

The present invention relates to a pharmaceutical composition comprising a compound of formula i for use in the treatment and / or prevention of microbial infections. Furthermore, the present invention relates to a method for preventing and / or reducing biofilm formation.

Description

1 / 40 “GLUCONIC ACID DERIVATIVES FOR USE IN THE TREATMENT AND / OR PREVENTION OF MICROBIAL INFECTIONS” TECHNICAL FIELD

[0001] The present invention relates to a compound of Formula I for use in the treatment and / or prevention of microbial infections. Furthermore, the present invention relates to a method for preventing and / or reducing biofilm formation. BACKGROUND

[0002] Antimicrobial agents have been used in recent years to treat patients with infectious diseases. Since the 1940s, these medications have greatly reduced illness and death from infectious diseases. However, these medications have been used so widely and for so long that the infectious organisms that the antimicrobial agents are designed to kill have adapted to them. As a result, the medications become ineffective and infections persist in the body, increasing the risk of spreading to others.

[0003] Antimicrobial resistance threatens the effective prevention and treatment of an ever-increasing range of infections caused by bacteria, parasites, viruses, and fungi.

[0004] Thus, antimicrobial resistance is an increasingly serious threat to global public health that requires action across all government sectors and in society.

[0005] All classes of microbes develop resistance: fungi develop antifungal resistance, viruses develop antiviral resistance, protozoa develop antiprotozoal resistance, and bacteria develop antibiotic resistance.

[0006] The problem with antibiotic resistance is a common phenomenon. Treatment-resistant fungal infections are an emerging public health problem. Overall, antifungal resistance is still relatively uncommon, but the problem will likely continue. Petition 870200044173, dated 06 / 04 / 2020, page 132 / 201 2 / 40 evolving, unless more is done to prevent the development of further resistance and stop the spread of these infections. Although most antifungal resistance occurs in Candida species, resistance in other types of fungi, such as Aspergillus, is also an emerging issue.

[0007] Because fungi are eukaryotes, just like the human hosts they infect, there are only a few distinct targets that can be employed in the development of antifungal drugs. For this reason, antifungals are mainly restricted to drugs targeting certain metabolic pathways.

[0008] A target for antimicrobial agents is the biofilm, a product of microbial growth. Biofilms are formed by microbial cells stuck together and surrounded by a self-produced extracellular polymeric matrix. Biofilm formation is a survival strategy for bacteria and fungi to adapt to their living environment, especially in a hostile environment. When a cell switches to biofilm growth mode, it undergoes a phenotypic change in behavior, in which large sets of genes are differentially regulated. Biofilms can contain many different types of microorganisms, for example, bacteria, archaea, protozoa, fungi, and algae.

[0009] Biofilms are estimated to be associated with 80% of microbial infections, and the growth of microorganisms in biofilms can increase their resistance to antimicrobial agents. Furthermore, biofilm bacteria are up to 1000 times more tolerant and / or resistant to antibiotics than planktonic cells.

[00010] In most fungi, hyphae are the primary mode of vegetative growth and are collectively called mycelium. For example, the virulence of Candida albicans is mediated by a transformation of planktonic cells into hyphae. The hyphal form, that is, filamentous cells, has Petition 870200044173, dated 06 / 04 / 2020, page 133 / 201 3 / 40 the ability to invade tissues and induce inflammation, mediated by candidalisin, a cytotoxic peptide toxin that destroys epithelial cells (Moyes et al., Nature, 2016, 532, 64).

[00011] Antimicrobial resistance is not only problematic in primary infections, but also in secondary infections. Without effective antimicrobials for the prevention and treatment of infections, medical procedures such as organ transplantation, cancer chemotherapy, diabetes management, and major surgeries become a very high risk. Furthermore, fungal infections have become a major cause of morbidity and mortality in immunocompromised patients, such as those with HIV / AIDS, tuberculosis, or undergoing chemotherapy. Despite increased awareness and improved treatment strategies, the frequent development of resistance to antifungals used in clinical settings contributes to the increasing number of mycoses.

[00012] Thus, there is an emerging need for new antimicrobial agents. SUMMARY

[00013] The present inventors have developed a method for preventing and / or reducing biofilm formation. When biofilm formation is reduced or prevented, individual microbial cells can no longer attach to the surface. Therefore, further infections are avoided and microbial cells that no longer form a biofilm are discarded. The present inventors have shown that treatment with a compound of Formula I, or a lactone thereof, characterized by the fact that Petition 870200044173, dated 06 / 04 / 2020, p. 134 / 201 4 / 40 R is selected from the group consisting of -H, -alkyl, -C(O)alkyl and phenyl; R' is selected independently of the group consisting of -OR, -H, and halogen; en is an integer 1, 2, or 3; It reduces the presence of biofilm of fungal species and exerts a cytotoxic effect on various fungal species. Furthermore, the present inventors have shown that a compound of formula I is useful as an antibacterial agent.

[00014] Thus, in one aspect, the present invention relates to a compound of Formula I or a pharmaceutical composition comprising a compound of Formula I for use in the treatment and / or prevention of microbial infections, provided that, if the microbial infection is a fungal infection, then the compound of Formula I is not a compound of Formula (XIV) OH OH O or a lactone from it. XIV

[00015] In one embodiment, the present invention relates to glucono-Delta-lactone for use in the treatment of a bacterial infection or a mixed bacterial and fungal infection.

[00016] In another aspect, the present invention relates to a method for preventing and / or reducing biofilm formation, wherein the method comprises administering a compound of Formula I.

[00017] In a further aspect, the present invention relates to a compound of Formula I for use in the prevention of premature births. Petition 870200044173, dated 06 / 04 / 2020, p. 135 / 201 5 / 40 DESCRIPTION OF THE FIGURES

[00018] Figure 1. Standardized biofilm formation of Candida albicans treated with different acids. GlyA = glyceric acid (pH = 7), XA = xylonic acid (pH = 7), CA = citric acid (pH = 4.6), GA = gluconic acid (pH = 6.5), LA = lactic acid (pH = 4.9), the biofilm was measured after 24h.

[00019] Figure 2. Changes in optical rotation during the hydrolysis of glucono-Delta-lactone (GDL) in distilled water (unfilled circles), pH 4 buffer (filled squares), pH 5 buffer (unfilled squares), and pH 7 buffer (filled circles).

[00020] Figure 3. Standardized biofilm formation of Candida albicans in minimal media at pH 2.6 to 6.6 with phosphate buffer (unfilled circles, dotted line) or glucono-Delta-lactone (filled squares, solid line). Biofilm thickness was measured after 24 hours and staining was performed with crystal violet.

[00021] Figure 4. (A) Normalized biofilm formation of (A) Normalized biofilm formation of Candida glabrata treated with lactonized / oligomerized GA. One granule of lactonized / oligomerized gluconic acid was added to a pH 3.71 buffer solution (10 ml) at 37°C. Samples (4 ml) were collected hourly (at different time intervals) and new buffer solution (4 ml) was added. The samples were diluted 50 times with biofilm medium and the amount of biofilm formation was measured after 24h. (B) Normalized biofilm formation of Candida glabrata treated with lactonized / oligomerized GA. One granule of lactonized / oligomerized gluconic acid was added to a pH 3.71 buffer solution (10 ml) at 37°C. Samples (4 ml) were collected hourly (at different time intervals) and new buffer solution (4 ml) was added. The samples were diluted 50 times with biofilm medium, and the amount of biofilm formation was measured after 24 hours. Petition 870200044173, dated 06 / 04 / 2020, page 136 / 201 6 / 40

[00022] Figure 5. (A) Normalized biofilm formation of Candida albicans treated with glucono-Delta-lactone (GDL). One GDL bead was added to a pH 3.71 buffer solution (10 ml) at 37 °C. Samples (4 ml) were collected after 1, 2, 3, 4, 5, 6, and 24 h, and a new buffer solution (4 ml) was added. The samples were diluted 50 times with biofilm medium, and the amount of biofilm formation was measured after 24 h. (B) Normalized biofilm formation of Candida glabrata treated with GDL. One GDL bead was added to a pH 3.71 buffer solution (10 ml) at 37 °C. Samples (4 ml) were collected after 1, 2, 3, 4, 5, 6, and 24 h, and a new buffer solution (4 ml) was added. The samples were diluted 50 times with biofilm medium, and the amount of biofilm formation was measured after 24 hours.

[00023] Figure 6. (A) Viability of C. albicans biofilms and (A) Viability of C. albicans and C. glabrata biofilms after treatment with glucono-Delta-lactone (GDL) at different concentrations for 24 h. Biofilm staining was performed with XTT. Optical density measured at 485 nm. Diagonal stripes indicate data for C. albicans. Filled black columns indicate data for C. glabrata. (B) Viability of C. albicans and C. glabrata biofilms after treatment with GDL at different concentrations for 48 h. Biofilm staining was performed with XTT. Optical density measured at 485 nm. Diagonal stripes indicate data for C. albicans. Filled black columns indicate data for C. glabrata.

[00024] Figure 7. Effect of glucono-Delta-lactone (GDL) on mature biofilm of C. albicans and C. glabrata. Mature biofilm (cultured for 48 h) was incubated with GDL for 5 ha at 37 °C and then cells with serial dilution were plated on YPD plate to estimate cell survival.

[00025] Figure 8. (A) Microfluidic study of the development of untreated C. albicans biofilm in minimal medium pH 7.0. Untreated cells mainly form hyphae. (B) Microfluidic study of Petition 870200044173, dated 06 / 04 / 2020, page 137 / 201 7 / 40 Development of C. albicans biofilm treated in minimal medium with a glucono-Delta-lactone (GDL) hydrolysate at a final concentration of x50 pH 3.8. The addition of GDL caused C. albicans to grow predominantly as a yeast form, but not as hyphae.

[00026] Figure 9. Biofilm formation of Escherichia coli K12 treated with phosphate buffer (filled squares), citric acid (unfilled squares), lactic acid (filled triangles), gluconic acid (filled circles) or glucono-Delta-lactone (unfilled circles) to obtain media with different pH (2.6 to 6.6). The biofilm was stained with crystal violet. DETAILED DESCRIPTION COMPOUNDS

[00027] In one aspect, the present invention relates to a pharmaceutical composition comprising a compound of Formula I, R' o or a lactone thereof, characterized by the fact that R is selected from the group consisting of -H, -alkyl, -C(O)alkyl and phenyl; R' is selected independently of the group consisting of -OR, -H and halogen; en is an integer 1, 2 or 3, for use in the treatment and / or prevention of microbial infections, provided that, if the microbial infection is a fungal infection, the compound of Formula I will not be a compound of Formula (XIV) Petition 870200044173, dated 06 / 04 / 2020, p. 138 / 201 8 / 40 OH OH O or a lactone from it. XIV

[00028] In one embodiment, the alkyl group is a C1 to C20 aliphatic chain. The term aliphatic chain, as used herein, refers to non-aromatic hydrocarbons. Said aliphatic chains may be linear, branched, and / or cyclic. Said aliphatic chains may be saturated or unsaturated. In said aliphatic chains, one or more hydrogens may be substituted by an aromatic moiety, comprising one or more benzene moieties.

[00029] In one embodiment, an alkyl is an aliphatic chain from C1 to C20 in which one or more hydrogens are optionally substituted by -OH, =O or phenyl and in which one or more of the CH2 groups of the aliphatic chain are optionally substituted by O, S or NH. Non-limiting examples of alkyls in which one or more of the CH2 groups of the aliphatic chain are optionally substituted by O, S or NH are ethers, thioethers and tertiary amines. In one embodiment, alkyl is an aliphatic chain from C1 to C20, such as an aliphatic chain from C1 to C15, an aliphatic chain from C1 to C10, an aliphatic chain from C1 to C5, such as an aliphatic chain from C5 to C20, such as an aliphatic chain from C5 to C15, such as an aliphatic chain from C5 to C10, such as an aliphatic chain from C10 to C20, such as an aliphatic chain from C10 to C15. In a preferred embodiment, alkyl is selected from the group consisting of methyl, ethyl, and propyl.

[00030] In one embodiment, at least one of R' is -OR. In one embodiment, at least two, as well as at least three, as well as at least four, as well as at least five, as well as at least six of R' are OR. In a preferred embodiment, R' is -OR. Petition 870200044173, dated 06 / 04 / 2020, p. 139 / 201 9 / 40

[00031] In one embodiment, at least one of R' is -OH. In another embodiment, no more than one of R' is -H, just as no more than two of R' are -H. In a preferred embodiment, R is -H.

[00032] In one embodiment, -OR is acetate or lactate.

[00033] In aqueous solution, compounds according to Formula I may be in equilibrium with the corresponding lactones, for example, Delta-lactone or Gamma-lactone. In one embodiment, the compound of Formula I is a lactone of this type. Said lactone may preferably be a Delta-lactone or a Gamma-lactone.

[00034] In one configuration, the Formula 1 compound is selected from the group consisting of another modality, the IV The compound of Formula I is selected from the group consisting of Petition 870200044173, dated 06 / 04 / 2020, p. 140 / 201 10 / 40

[00036] In a preferred embodiment, n is 2.

[00037] In one modality, the Formula 1 compound is OH OH O OH OH XIV

[00038] In one embodiment, the Formula I compound is a Formula XV compound, OH OH XV

[00039] In one embodiment, the Formula I compound is selected from the group consisting of Petition 870200044173, dated 06 / 04 / 2020, p. 141 / 201 11 / 40

[00040] In one embodiment, the Formula I compound is selected from the group consisting of

[00041] In one embodiment, compounds XIX, XX, and XXI are in equilibrium in aqueous solution.

[00042] In a preferred embodiment, the compound of Formula I is glucono-Delta-lactone (GDL, Formula XIX),

[00043] In another embodiment, the compound is not glucono-Deltalactone (Formula XIX). Thus, in one aspect, the present invention relates to a compound of Formula I for use in the treatment and / or prevention of microbial infections, provided that the compound of Formula I is not glucono-Delta-lactone (Formula XIX).

[00044] In one embodiment, the present invention relates to a compound of Formula I for use in the treatment and / or prevention of microbial infections, provided that, if the compound of Formula I is Petition 870200044173, dated 06 / 04 / 2020, p. 142 / 201 12 / 40 the compound of Formula XIX, the microbial infection will not be a fungal infection.

[00045] In one embodiment, the present invention relates to a compound of Formula I for use in the treatment and / or prevention of microbial infections, provided that, if the compound of Formula I is the compound of Formula XIX, the microbial infection will not be a urogenital fungal infection.

[00046] In one embodiment, the present invention relates to a compound of Formula I for use in the treatment and / or prevention of microbial infections, provided that, if the compound of Formula I is the compound of Formula XIX, the microbial infection will not be vulvovaginal candidiasis.

[00047] In one embodiment, the present invention relates to a compound of Formula I for use in the treatment and / or prevention of microbial infections, provided that if the microbial infection is a urogenital fungal infection, the compound of Formula I will not be a compound of Formula XIV.

[00048] In one embodiment, the compound is an acetal of a compound of Formula I. In another embodiment, the oxo group of the compound is a corresponding acetal, that is, the compound of Formula I is selected from the group consisting of

[00049] In one embodiment, the Formula I compound is selected from the group consisting of Petition 870200044173, dated 06 / 04 / 2020, p. 143 / 201 13 / 40 XXV POLYMER / OLIGOMER

[00050] In one embodiment, the compound of Formula I is oligomerized to form an oligomer. In another embodiment, the compound of Formula I is polymerized to form a polymer.

[00051] In one embodiment, the oligomer or polymer comprises a compound of Formula I, that is, it is a homo-oligomer / polymer in which a compound of Formula I is the monomer. In another embodiment, the oligomer or polymer is a mixed oligomer / polymer, that is, a heterooligomer / polymer. In one embodiment, the oligomer comprises at least two different compounds of Formula I.

[00052] In one embodiment, the oligomer or polymer further comprises lactic acid, that is, a lactic acid oligomer / polymer.

[00053] In one embodiment, two compounds of Formula I are linked to form a dimer. In one embodiment, the dimer comprises two compounds of Formula XIV. In one embodiment, the dimer comprises two compounds other than Formula I. INFECTIONS

[00054] In one aspect, the present invention relates to a compound of Formula I or to a pharmaceutical composition comprising a compound of Formula I for use in the treatment and / or prevention of microbial infections. In one embodiment, the present invention relates to glucono-Delta-lactone for use in the treatment of a bacterial infection or a mixed bacterial and fungal infection. Petition 870200044173, dated 06 / 04 / 2020, p. 144 / 201 14 / 40

[00055] In one embodiment, the microbial infection is a urogenital infection. In another embodiment, the microbial infection is a vaginal infection.

[00056] In one embodiment, the infection is an infection in a mammal, that is, the patient who needs the said treatment is a mammal. Preferably, the mammal is a human. In one embodiment, the human is a woman. Said woman may be a pregnant woman.

[00057] In one embodiment, the infection is dermatitis and / or eczema. Said dermatitis and / or eczema may be seborrheic dermatitis. The infection may also be a secondary infection of said dermatitis or eczema.

[00058] The term secondary infection, as used herein, refers to a sequela or complication of a root cause. The root cause may be a primary infection.

[00059] In one form, the infection is acne of all severities or acneiform conditions, such as rosacea, perioral or periorbital dermatitis.

[00060] In one form, the infection is furunculosis, carbunculosis, or folliculitis.

[00061] In one form, the infection is cheilitis. This cheilitis may be angular cheilitis.

[00062] In one embodiment, the infection is an infection of the face, scalp, trunk, and / or groin. The infection may be an infection of infected skin wounds in the aforementioned areas. The infection may also be located in the skin folds of the body.

[00063] In one form, the infection is impetigo or erysipelas.

[00064] In one form, the infection is a foot infection. The aforementioned foot infection may be associated with diabetic foot ulcers. In one form, the foot infection is secondary to ingrown toenails or foot blisters. Petition 870200044173, dated 06 / 04 / 2020, p. 145 / 201 15 / 40

[00065] In one embodiment, the infection is a secondary infection that arises after an animal bite. The animal in question may be an insect. In one embodiment, the infection is a secondary infection that arises after insect bites, mosquito bites, tick bites, erythema migrans, or benign cutaneous lymphadenosis.

[00066] In one embodiment, the infection is a secondary infection of dermatological, oral, or genital herpes simplex, or a secondary infection of herpes zoster or varicella zoster.

[00067] In one embodiment, the infection is a secondary infection of a skin injury, such as burns or cuts.

[00068] In one form, the infection is a fungal, bacterial, or mixed blepharitis.

[00069] In one form, the infection is conjunctivitis.

[00070] In one form, the infection is vaginitis or cervicitis.

[00071] In one form, the infection is caused by Trichomonas vaginalis. In one form, the infection is trichomoniasis.

[00072] In one embodiment, the microbial infection is selected from the group consisting of fungal infections, bacterial infections, and mixed fungal and bacterial infections. Bacterial infections

[00073] In some forms, the microbial infection is a bacterial infection. In one form, the microbial infection is a mixed fungal and bacterial infection.

[00074] In one form, the bacterial infection is periodontitis.

[00075] In one form, the bacterial infection is bacterial vaginosis.

[00076] In one embodiment, the bacterial infection is selected from the group consisting of Gardnerella vaginalis, Chlamydia Petition 870200044173, dated 06 / 04 / 2020, p. 146 / 201 16 / 40 trachomatis-, Neisseria gonorrhoeae-, Treponema pallidum (syphilis)-, Atopobium vaginae-, Prevotella spp-, Mobiluncus spp-, Peptostreptococcus spp-, Poryphyromonas spp-, Mycoplasma hominis-, Bacteroides spp-, Ureaplasma urealyticum-, Streptococcus spp-, Enterobacteriaceae-, Enterococci-, Staphylococcus spp. and G- and / or Porphyromonas gingivalis infection.

[00077] In one embodiment, the infection is secondary to oral, nasal, or anogenital colonization by group A or group B streptococci or multidrug-resistant bacteria.

[00078] In one form, the infection is perianal streptococcal dermatitis. Mixed bacterial and fungal infections

[00079] In one embodiment, the said microbial infection is a mixed fungal and bacterial infection. The bacterial component of said mixed fungal and bacterial infection may be a bacterial infection as defined herein. The fungal component of said mixed fungal and bacterial infection may be a fungal infection as defined herein. In one embodiment, said fungal component of the mixed fungal and bacterial infection is candidiasis.

[00080] The aforementioned mixed fungal and bacterial infection may be intertriginous dermatitis or paronychia. FUNGAL INFECTIONS

[00081] In some embodiments, the microbial infection is a fungal infection. In one embodiment, the microbial infection is a mixed bacterial and fungal infection. In one embodiment, the fungal infection is a mycosis. The mycosis may be selected from the group consisting of Dermatophytosis, Candidiasis, Coccidioidomycosis, Histoplasmosis, Petition 870200044173, dated 06 / 04 / 2020, p. 147 / 201 17 / 40 Blastomycosis, Paracoccidioidomycosis, Sporotrichosis, Chromomycosis and Phaeomycotic Abscess, Aspergillosis, Cryptococcosis and Zygomycosis.

[00082] In a preferred embodiment, the mycosis is candidiasis. Said candidiasis may be selected from the group consisting of candidiasis of the vulva and vagina; candidiasis of the skin and nails; candidiasis of urogenital and gastrointestinal sites; Candida stomatitis; Candida of the breast and nipple; pulmonary candidiasis; Candida meningitis; Candida endocarditis; and Candida sepsis.

[00083] In one embodiment, mycosis is dermatophytosis. Said dermatophytosis can be selected from the group consisting of inguinal dermatophytosis; onychomycosis of toenails; Tinea inguinalis, Tinea pedis; Tinea manuum; Tinae barbae, Tinea amiantacea Tinea capitis; Tinea corporis; Tinea imbricata; and Tinea cruris.

[00084] In one embodiment, the fungal infection is selected from the group consisting of Candida species infections, such as Candida albicans, Candida krusei, Candida glabrata, Candida tropicalis infections; Trichophyton species infections, such as Trichophyton verrucosum, Trichophyton rubrum, Trichophyton violaceum, Trichophyton tonsurans; and Microsporum species infections, such as Microsporum canis infection; Aspergillus and Malassezia infections.

[00085] In one form, the infection is caused by Saccharomyces cerevisiae.

[00086] In one form, the fungal infection is pityriasis versicolor.

[00087] In one embodiment, the compound of Formula I is not the compound of Formula XIX and the fungal infection is not a urogenital fungal infection.

[00088] In another embodiment, the Formula I compound is not the Formula XIX compound and the fungal infection is not vulvovaginal candidiasis.

[00089] In yet another embodiment, the fungal infection is not a urogenital fungal infection and the compound of Formula I is not Petition 870200044173, dated 06 / 04 / 2020, p. 148 / 201 18 / 40 OH OH O OH OH XIV VIRAL INFECTIONS

[00090] In one embodiment, the microbial infection is a viral infection. In another embodiment, the viral infection is HIV. FORMULATIONS

[00091] In some embodiments, the present invention relates to a pharmaceutical composition comprising a compound of formula I for use in the treatment of a microbial infection. In one embodiment, the pharmaceutical composition is formulated as a tablet, orally disintegrating tablet (or orally dissolving tablet (ODT)), lozenge, gum, chewing gum, cream, lotion, gel, emulsion, solution, foam, ointment, spray, suspension, mouthwash, mouth rinse, mouthwash, mouth bath, nail polish, dermal patch or shampoo. In another embodiment, said solution is adapted for use in a dressing, bandage and / or compress.

[00092] In one embodiment, the pharmaceutical composition comprises at least 5% by weight, as at least 10% by weight, as at least 15% by weight, as at least 20% by weight, as at least 25% by weight, as at least 30% by weight, as at least 40% by weight, as at least 50% by weight, as at least 60% by weight of the compound of Formula (I).

[00093] In one embodiment, the pharmaceutical composition for use according to any of the preceding claims, characterized in that the pharmaceutical composition comprises not more than 99% by weight, as well as not more than 95% by weight, as well as not more than 90% by weight. Petition 870200044173, dated 06 / 04 / 2020, page 149 / 201 19 / 40 weight, as not more than 85% by weight, as not more than 80% by weight, as not more than 75% by weight of the compound of Formula (I).

[00094] In one embodiment, the pharmaceutical composition comprises 5 to 99% by weight, as in 10 to 95% by weight, as in 15 to 95% by weight, as in 20 to 90% by weight, as in 40 to 95% by weight, as in 40 to 95% by weight, as in 50 to 95% by weight of the compound of Formula (I).

[00095] In one embodiment, the pharmaceutical composition comprises not more than 10% by weight of water, and not more than 5% by weight of water.

[00096] An antimicrobial agent, as used herein, refers to an agent that is capable of reducing, eliminating, or inhibiting the growth of microorganisms, as the term is known in the state of the art (exemplary microorganisms include microbes such as bacteria, fungi, viruses, and other pathogens). Similarly, the term antifungal agent refers to an agent capable of reducing, eliminating, or inhibiting the growth of fungi, and the term antibacterial agent refers to an agent capable of reducing, eliminating, or inhibiting the growth of bacteria.

[00097] In one embodiment, the pharmaceutical composition further comprises one or more antifungal agents. In one embodiment, the present invention relates to a method of treating one or more microbial infections comprising the co-administration of a compound of Formula I and an antimicrobial agent to a patient in need thereof. In one embodiment, said antimicrobial agent is an antifungal agent or an antibacterial agent. Said antifungal agent may be selected from the group consisting of miconazole, terconazole, isoconazole, fenticonazole, fluconazole, nystatin, ketoconazole, clotrimazole, butoconazole, econazole, tioconazole, itraconazole, 5-fluorouracil and metronidazole.

[00098] In one embodiment, the pharmaceutical composition also includes one or more antibacterial agents. Antibiotics are one Petition 870200044173, dated 06 / 04 / 2020, pages 150 / 201 20 / 40 type of antimicrobial agents used in the treatment and prevention of bacterial infections. The aforementioned antibacterial agent may be selected from the group consisting of clindamycin, tetracycline, amoxicillin, ampicillin, erythromycin, doxycycline, lumefloxacin, norfloxacin, afloxam, ciprofloxacin, azithromycin, and cefltoxin.

[00099] In one embodiment, the pharmaceutical composition also includes a steroid. The said steroid may be cortisone. [000100] In one embodiment, the pharmaceutical composition is formulated as a tampon, vaginal spray, vaginal cup, vaginal gel, vaginal insert, vaginal patch, vaginal ring, vaginal sponge, vaginal suppository, vaginal cream, vaginal emulsion, vaginal foam, vaginal lotion, vaginal ointment, vaginal powder, vaginal shampoo, vaginal solution, vaginal spray, vaginal suspension, vaginal tablet, vaginal rod, vaginal disc, vaginal device and any combination thereof, or wherein the pharmaceutical composition is present in a sanitary article, such as a tampon, a sanitary napkin, an incontinence pad or diaper or an underwear liner. [000101] In one embodiment, the pharmaceutical composition is adapted for administration at least once a day, as well as at least twice a day, as well as at least three times a day. [000102] In one embodiment, the pharmaceutical composition is adapted for administration no more than every two days, or no more than every three days, or no more than once a week. [000103] In one embodiment, the pharmaceutical composition is adapted for administration for no more than six days. [000104] In one embodiment, the pharmaceutical composition is adapted for administration for at least one week, as for at least two weeks, as for at least three weeks, as for at least four weeks. Petition 870200044173, dated 06 / 04 / 2020, p. 151 / 201 21 / 40 [000105] In one embodiment, the pharmaceutical composition is adapted for administration at least once daily for at least one week. [000106] In one embodiment, the pharmaceutical composition is formulated to release the compound according to Formula I over an extended period of time, such as for at least 4 hours, such as for at least 6 hours, such as for at least 24 hours after administration. METHOD FOR PREVENTING AND / OR REDUCING BIOFILM FORMATION [000107] In one aspect, the present invention relates to a method for preventing and / or reducing biofilm formation, characterized in that the method comprises administering a compound of Formula I. The term biofilm, as used herein, refers to an aggregate of microorganisms in which microbial cells adhere to each other and / or to a surface. These adherent cells are frequently covered with a matrix of extracellular polymeric substance, for example, comprising extracellular DNA, proteins and polysaccharides, which is produced by the cells. The microbial cells growing in a biofilm are often physiologically distinct from the planktonic cells of the same organism. [000108] Such biofilms can form on any living or non-living surface. In one embodiment, the biofilm is a biofilm within or on a mammal. [000109] In one embodiment, the biofilm is a biofilm of an implant or prosthesis. The term implant or prosthesis, as used herein, refers to artificial substitutes for body parts and materials inserted into tissue for functional, cosmetic, or therapeutic purposes. Implants or prostheses may be functional, as in the case of artificial arms and legs, or cosmetic, as in the case of an artificial eye. Implants, all surgically inserted or grafted into the body, tend to be used therapeutically. Petition 870200044173, dated 06 / 04 / 2020, page 152 / 201 22 / 40 [000110] The aforementioned implant or prosthesis may be selected from the group consisting of catheters, peripheral venous catheters, central venous catheters, heart valves, ventricular assist devices, coronary stents, neurosurgical ventricular shunts, implantable neurological stimulators, arthroprostheses, fracture fixation devices, inflatable penile implants, breast implants, cochlear implants, intraocular lenses, dental implants, laryngectomy implants, tracheostomy implants, voice prostheses, jaw mobility implants, tympanostomy implants and dental implants. METHOD FOR USE IN THE PREVENTION OF PREMATURE BIRTH [000111] In one instance, the microbial infection is bacterial vulvovaginitis. With cervical ripening or cervical insufficiency, this infection can migrate to the uterus and cause chorioamnionitis and subsequently premature labor. There is evidence to support that excessive inflammation, such as vaginitis or cervicitis, through the production of prostaglandins, can cause premature contractions and premature labor, even in the absence of overt chorioamnionitis. The premature newborn may subsequently face invasive bacterial infection; pneumonia, meningitis, or sepsis, due to neonatal immunodeficiency in the premature infant. Group A and B streptococci and multidrug-resistant bacteria can cause severe perinatal infections in the infant and postpartum endometritis in the newborn, and be the cause of severe neonatal and maternal morbidity and mortality. [000112] In another form, the microbial infection is vulvovaginal candidiasis. Premature newborns may experience invasive Candida infection, one of the most serious hospital-acquired infections, causing greater morbidity and mortality than bacterial infection, particularly in neonatal intensive care units. [000113] Thus, in one aspect, the present invention relates to a compound of Formula I for use in the prevention of premature births. In a preferred embodiment, said compound is administered via Petition 870200044173, dated 06 / 04 / 2020, p. 153 / 201 23 / 40 vaginal. The compound may be formulated as a tampon, vagitorium, vaginal aerosol, vaginal cup, vaginal gel, vaginal insert, vaginal patch, vaginal ring, vaginal sponge, vaginal suppository, vaginal cream, vaginal emulsion, vaginal foam, vaginal lotion, vaginal ointment, vaginal powder, vaginal shampoo, vaginal solution, vaginal spray, vaginal suspension, vaginal tablet, vaginal rod, vaginal disc, vaginal device and any combination thereof, or where the compound is present in a sanitary article, such as a tampon, a sanitary napkin, pad or diaper for incontinence or underwear liner. EXAMPLES EXAMPLE 1: EFFECTS OF DIFFERENT HYDROXYLATED CARBOXYLIC ACIDS BIOFILM FORMATION ASSAY [000114] Yeast strains (Table 1) were cultured at °C in complete YPD medium (0.5% yeast extract, 1% peptone, 2% glucose) or minimal medium consisting of YNB (yeast nitrogen base without amino acids and ammonium sulfate, FORMEDIUM™, CYN0505) supplemented with 0.5% ammonium sulfate, 0.2% glucose, and 100 mM L-proline. If necessary, 2% agar was used to solidify the medium. The liquid minimal medium (YNB (yeast nitrogen base without amino acids and ammonium sulfate, FORMEDIUM™, CYN0505) supplemented with 0.5% ammonium sulfate, 0.2% glucose, and 100 mM L-proline) was used for the biofilm assay (biofilm medium). [000115] In experiments on the impact of pH on biofilm, pH values ​​(from 2.6 to 6.6) were obtained using different potassium phosphate buffers at a final concentration of 0.25 M or by adding citric acid, lactic acid, and gluconic acid to the biofilm medium. TABLE 1. YEAST STRAINS USED IN THIS STUDY. Original name Designation Description Strain reference of Petition 870200044173, dated 06 / 04 / 2020, p. 154 / 201 24 / 40 lab Candida SC5314 albicans Y775 Wild type, virulent in a mouse model with systemic infection, sequenced strain [AM Gillum, et al. Mol. O Gen. Genet. 1984, 198, 179-182] Candida CBS138 glabrata Y1092 Wild type, isolated from human feces, sequenced strain [B. Dujon, et al. Nature, 2004, 430, 35-44] [000116] The biofilm was measured in liquid culture as described. [K. Scherz et al., G3 (Bethesda), 2014, 4, 1671-1680. I. Serrano-Fujarte et al. Biomed Res Int. 2015; 2015:783639] with some modifications. Prior to the biofilm assay, yeast cultures were grown in liquid YPD medium for 24 hours to the stationary phase (OD600 11-17)2, the cells were granulated by centrifugation (1699 g), washed with sterile water, and the cells were subsequently inoculated into the biofilm test medium (YNB (yeast nitrogen base without amino acids and ammonium sulfate) supplemented with 0.5% ammonium sulfate, 0.2% glucose, and 100 mM L-proline pH7.0) at a final concentration of 0.2 OD600 / ml and incubated in 96-well flat-bottom polystyrene microtiter plates (Sigma Aldrich, Corning®Costar® culture plates, CLS3596-50EA) for 72 hours at a thermostat of 37 °C. At defined time points, crystal violet (HT901-8FOZ; Sigma Aldrich) was added to the medium at a final concentration of 0.05%, and the total biomass was also measured.After 24 hours of cell staining, the plate wells were washed four times with 200 ml of water to remove planktonic cells. The biofilms were then dried and dissolved in 200 μL of 96% ethanol. Total biomass and crystal violet biofilm staining measurements were performed on the OD560 using the FLUOstar OPTIMA plate reader, BMG LABTECH. Crystal violet biofilm measurements were normalized to total biomass (OD560 Biofilm / OD560 total biomass). Petition 870200044173, dated 06 / 04 / 2020, page 155 / 201 25 / 40 EFFECTS OF DIFFERENT CARBOXYLIC ACIDS HYDROXYLATED [000117] To compare the effects of different hydroxylated carboxylic acids at low concentrations, biofilm formation was measured 24 h after the addition of 0.06 wt% of glyceric acid, xylonic acid, citric acid, gluconic acid, and lactic acid, under unbuffered conditions. The data are shown in Table 2 and Figure 1. TABLE 2. EFFECT OF LOW CONCENTRATIONS (0.06%) OF HYDROXYLATED CARBOXYLIC ACIDS ON CANDIDA ALBICANS BIOFILM FORMATION. BIOFILM WAS MEASURED AFTER 24 HOURS. Resulting pH of normalized biofilm (% of control): glyceric acid (GlyA) 7.0 40, xylonic acid (XA) 7.0 31, citric acid (CA) 4.6 14, gluconic acid (GA) 6.5 6, lactic acid (LA) 4.9 13 [000118] As can be seen in Table 2, gluconic acid is the most efficient compound, followed by lactic acid and citric acid. Replacing lactic acid with gluconic acid resulted in >50% less biofilm formation (6% vs. 13% in the control). Given that biofilm formation is very sensitive to low pH and that gluconic acid only induces a moderate decrease in pH compared to lactic acid and citric acid, the result is impressive. EXAMPLE 2: FORMATION OF CANDIDA ALBICANS BIOFILM AT DIFFERENT pH [000119] In order to better evaluate the effect of gluconic acid in preventing biofilm formation, biofilm formation at different pH levels was determined (as described in Example 1) for gluconic acid, lactic acid, and citric acid. Petition 870200044173, dated 06 / 04 / 2020, page 156 / 201 26 / 40 [000120] As can be seen in Table 3, gluconic acid shows strong effects on Candida albicans biofilm formation, while the effects of lactic acid and citric acid are much less pronounced. Furthermore, gluconic acid shows a strong effect even at pH values ​​of at least 6, while the effect observed with lactic acid and citric acid begins to diminish at pH 5. Additionally, gluconic acid results in a complete loss of biofilm formation at pH 2.6. The data are summarized in Table 3. TABLE 3. CANDIDA ALBICANS BIOFILM FORMATION BY ADDITION OF GLUCONIC ACID, LACTIC ACID OR CITRIC ACID (24H TREATMENT). gluconic acid lactic acid citric acid normalized biofilm buffer (OD560) at pH 6.1 0.17 3.1 4.6 3.2 biofilm 0 1.6 0.34 0.25 normalized (OD560) at pH 2.6 EXAMPLE 3: CANDIDA GLABRATA BIOFILM FORMATION AT DIFFERENT pH LEVELS [000121] Candida glabrata is much more complicated to treat compared to Candida albicans. However, a clear effect is obtained with longer treatment, i.e., 72 h with gluconic acid (Table 4). TABLE 4. BIOFILM FORMATION OF CANDIDA GLABRATA BY ADDITION OF GLUCONIC ACID, LACTIC ACID OR CITRIC ACID (72 H TREATMENT). gluconic acid lactic acid citric acid normalized biofilm buffer (OD560) at pH 6.1 1.6 2.8 2.9 4.7 Petition 870200044173, dated 06 / 04 / 2020, page 157 / 201 27 / 40 biofil 0.07 0.9 1.0 4, m and normalized 8 (OD560) at pH 2.6 [000122] Based on these results, it was concluded that gluconic acid has a superior effect in directing Candida biofilm formation compared to lactic acid and citric acid. In contrast to the effect observed for lactic acid and citric acid, the effect observed for gluconic acid is not merely a pH-related effect. The effect is present even at pH 6. [000123] It is thus concluded that gluconic acid is useful as an antifungal compound, as indicated by the reduction in biofilm formation. The compound is physiologically and pharmaceutically acceptable. Gluconic acid is therefore useful for providing pharmaceutical formulations for use in the treatment of vulvovaginal candidiasis. EXAMPLE 4: PREPARATION OF GLUCONIC ACID DERIVATIVES Gluconic acid lactonization / oligomerization: [000124] Gluconic acid (GA) (50% by weight of H2O, 4 g) was poured into an open flask and heated to 120 °C. After 24 h, the mixture was cooled to room temperature while it solidified. [000125] To analyze the composition of the aqueous gluconic acid (GA) solution, GA (50% by weight of H2O) was dissolved in DMSO-d6 and analyzed by 1H- and 13C-NMR. Lactonized / oligomerized gluconic acid (cf. above) was dissolved in DMSO-d6 and analyzed by 1H- and 13C-NMR, see Table 5. TABLE 5. COMPOSITION OF GA, LACTONIZED AND OLIGOMERIZED GA, ANALYZED BY 1H- AND 13C-NMR. DMSO-d6 compound (1H and 13C) GA [526-95-4] 70% gluconic acid 15% glucono-δ-lactone Petition 870200044173, dated 06 / 04 / 2020, pp. 158 / 201 28 / 40 15% γ-Gluconolactone Lactonized and oligomerized GA with 10% gluconic acid 25% δ-glucone lactone 50% γ Glucone lactone 15% oligomerized material [000126] It was concluded that GA forms a complex mixture of different lactones, as well as oligomerized material after prolonged dehydration. EXAMPLE 5: GLUCONE HYDROLYSIS --δ-LACTONE (GDL) [000127] In aqueous solution, glucono δ-lactone (GDL) is in equilibrium with gluconic acid (GA, CAS 526-95-4). GDL (200 mg) was added to distilled H2O (20 ml), pH 4 buffer, pH 5 buffer, or pH 7 buffer at 37 °C. Optical rotation and pH were measured over time. Optical rotation, measured at 37°C, sodium D-line, C = 10 mg / ml, path length = 10 cm. The optical rotation of GDL is approximately 66°. The optical rotation of gluconic acid is approximately 5° [DT Sawyer, JB Bagger, J. Am. Chem. Soc., 1959, 81, 5302-5306]. [000128] This experiment shows that GDL is slowly hydrolyzed to a mixture of GDL and GA (Figure 2). The equilibrium is pH dependent and relevant concentrations of GDL are present under all buffered conditions. EXAMPLE 6: BIOFILM FORMATION IN AN IN VIVO MODEL USING GLUCONIC ACID (GA) [000129] Granules of lactonized / oligomerized gluconic acid (1.3 g, duplicate samples) were added to pH 3.71 buffer solution (0.5 M KH2PO4 / orthophosphoric acid, 10 ml) at 37°C. Samples (4 ml) were collected every hour (1, 2, 3, 4, 5, 6, and 24 h) and new buffer solution (4 ml) was added. The samples were diluted 50 times with biofilm medium (see above) and the amount of biofilm formation was measured after 24 h, as Petition 870200044173, dated 06 / 04 / 2020, page 159 / 201 29 / 40 described above. As seen in Figure 4A, the released GA significantly reduces the amount of biofilm formation in Candida albicans. Furthermore, the hydrolysis of the granule is apparently slow enough to provide a preventive effect for at least up to 6 hours, probably much longer. The effect is less pronounced with Candida glabrata (Figure 4B). The data are summarized in Table 6. TABLE 6. BIOFILM FORMATION OF CANDIDA ALBICANS AND CANDIDA GLABRATA TREATED WITH LACTONIZED / OLIGOMERIZED GA IN AN IN VIVO MODEL. SAMPLES WERE COLLECTED AFTER 1 HOUR, DILUTED 50 TIMES WITH BIOFILM MEDIUM, AND THE AMOUNT OF BIOFILM FORMATION WAS MEASURED AFTER 24 HOURS. Normalized biofilm (% control), 1 h Candida albicans 6.5 Candida glabrata 42 EXAMPLE 7: BIOFILM FORMATION IN AN IN VIVO MODEL USING GLUCONA-DELTA-LACTONE [000130] Glucon-Delta-lactone (GDL) granules (2.5 g, duplicate samples) were added to pH 3.71 buffer solution (0.5 M KH2PO4 / orthophosphoric acid, 10 mL) at 37°C. Samples (4 mL) were collected at fixed time points (1, 2, 3, 4, 5, 6, and 24 h) and fresh buffer solution (4 mL) was added. Samples were diluted 50 times with biofilm medium (see above) and the amount of biofilm formation was measured after 24 h, as described above. As seen in Figure 5A, the released GDL significantly reduces the amount of biofilm formation in C. albicans. Furthermore, the hydrolysis of the granule is apparently slow enough to provide a preventive effect for at least 24 hours, probably much longer. The effect is less pronounced with C. glabrata (Figure 5B). Petition 870200044173, dated 06 / 04 / 2020, pages 160 / 201 30 / 40 TABLE 7. BIOFILM FORMATION OF C. ALBICANS AND C. GLABRATA TREATED WITH GDL IN AN IN VIVO MODEL. SAMPLES WERE COLLECTED AFTER 1 H, DILUTED 50 TIMES WITH BIOFILM ASSAY MEDIUM, AND THE AMOUNT OF BIOFILM FORMATION WAS MEASURED AFTER 24 H. Normalized biofilm (% control), 1 h Candida albicans Candida glabrata 8.3 71 [000131] The results show that biofilm formation of C. albicans and C. glabrata biofilm formation was reduced in the presence of GDL. In addition to decreasing biofilm formation, GDL may affect the viability of mature C. albicans and C. glabrata biofilms. EXAMPLE 8: VIABILITY OF MATURE BIOFILMS OF C. ALBICANS AND C. GLABRATA TREATED WITH GLUCONA-DELTA-LACTONE [000132] The viability of C. albicans and C. glabrata biofilms The effectiveness of glabrata after treatment with glucono-Delta-lactone (GDL) at different concentrations and for different periods of time was evaluated by staining the cells with XTT. XTT is a colorimetric assay for quantifying cell viability and cytotoxicity. The assay is based on the cleavage of the XTT tetrazolium salt, a conversion that occurs only in viable cells. Mature biofilm was exposed to GDL for 24 h. Then, the cells were washed twice with PBS, after which the XTT reaction mixture was added. After 30 min, the optical density was measured at 485 nm. [000133] The XTT assay showed a strong decrease in the viability of C. glabrata after 24h of incubation (Figure 6A). The effect was less pronounced for C. albicans, but clearly observed after 48 h (Figure 6B). [000134] In addition, the mature biofilm (cultivated for 48 h in A solution of YNB, 0.2% glucose, and 100 mM proline of C. albicans and C. glabrata was incubated with GDL of different concentrations (0.05–0.5 g / ml) at 37 °C. Petition 870200044173, dated 06 / 04 / 2020, p. 161 / 201 31 / 40 At this end, the biofilm medium (YNB, 0.2% glucose, 100 mM proline) was removed and GDL was added, which was dissolved in water at concentrations of 0.05, 0.1, 0.2, and 0.5 g / ml. After incubation with GDL for 5 h or 73 h, 5 μl of cells were plated in serial dilution (1:10 to 1:1000) on YPD agar medium to estimate cell survival. The plated cells were incubated for 24 hours at 37 °C and visually analyzed. Mature biofilm cells treated with water were used as a control. It was found that GDL decreases the cell viability of C. albicans and C. glabrata, particularly at high concentrations. At concentrations of 0.2 and 0.5 g / ml after 5 h of incubation, cell viability decreased approximately 100-fold for C. albicans and C. glabrata. After 73 h of incubation with 0.5 g / ml of GDL, the cell viability of C. albicans decreased approximately 1000-fold (data not shown). C. glabrata proved to be more sensitive to GDL (Figure 7). EXAMPLE 9: MICROFLUIDIC STUDY OF BIOFILM DEVELOPMENT [000135] To monitor the cell morphology of C. albicans, we also studied biofilm development using microscopy and microfluidics. After inoculation of yeast cells, hyphae began to form within the first hour of incubation in biofilm medium (YNB supplemented with 100 mM proline and 0.2% glucose, pH 7.0). Figure 8A shows untreated cells after 5 h. A glucono-Delta-lactone bead (2.5 g) was added to pH 3.71 buffer solution (0.5 M KH2PO4 / orthophosphoric acid, 10 mL) at 37 °C. A sample was collected after 1 h, diluted 50 times with biofilm medium, and added to C. albicans. After 5 h, most of the treated cells were planktonic (Figure 8B). EXAMPLE 10: VIABILITY OF DIFFERENT CANDIDA SPECIES IN THE PRESENCE OF GLUCONA-DELTA-LACTONE [000136] Other Candida sp. studied were also sensitive (e.g., cell viability measurement using the XTT assay, cf. Petition 870200044173, dated 06 / 04 / 2020, page 162 / 201 32 / 40 Example 8) Glucon-Delta-lactone (GDL). However, they exhibited different levels of sensitivity. Candida albicans SC5314 showed the lowest susceptibility and Candida krusei silicone isolate A4-1 showed the highest susceptibility. GDL toxicity is mediated through cell wall damage, as cells exposed to GDL showed lower viability in the calcofluor white medium compared to the medium supplemented with osmotic stabilizer (0.5 M sucrose) and compared to untreated cells in these media. Table 3 summarizes the qualitative effects shown by GDL. TABLE 8. SENSITIVITY OF DIFFERENT CANDIDATE SPECIES GDL Strain Sensitivity to GDL, 24h exposure, C. albicans SC5314 + C. glabrata CBS138 +++ + ++ silicone isolate U3-3 of C. tropicalis ++++++++ silicone isolate of C. krusei ++ silicone isolate A6-1 of C. tropicalis ++ + + silicone isolate U2-12 C. krusei ++++++++ Silicone isolate of C. krusei ++ silicone isolate A4-1 of C. krusei +++++++++ [000137] In conclusion, (i) GDL can break down the mature biofilm formed by C. albicans and C. glabrata, (ii) upon exposure to GDL, C. albicans transforms into yeast form, while the viability of C. glabrata decreases, (iii) the effect is clear even in other strains, for example, C. tropicalis and C. krusei. EXAMPLE 11: COOLIGOMERIZATION OF GLUCONIC ACID (GA) WITH LACTIC ACID (LA) LA: GA (MOLAR RATIO OF 4:1) [000138] DL-lactic acid (563 mg, 6.26 mmol) and D-gluconic acid (50% in water, 0.50 ml, 1.57 mmol) were mixed in a test tube and heated to 130 °C. After 4 h, the temperature was increased to 140 °C. Petition 870200044173, dated 06 / 04 / 2020, page 163 / 201 33 / 40 °C. After a total of 27 hours, the reaction mixture was allowed to reach room temperature. The reaction mixture solidified after cooling. LA: GA (MOLAR RATIO 8:1) [000139] DL-Lactic acid (569 mg, 6.32 mmol) and D-gluconic acid (50% in water, 0.25 ml, 0.78 mmol) were mixed in a test tube and heated to 130 °C. After 4 h, the temperature was increased to 140 °C. After a total of 27 h, the reaction mixture was allowed to reach room temperature. The reaction mixture solidified upon cooling. LA: GA (MOLAR RATIO OF 16:1) [000140] DL-Lactic acid (565 mg, 6.28 mmol) and acid Dgluconic acid (50% in water, 0.35 ml, 0.39 mmol) was mixed in a test tube and heated to 130 °C. After 4 h, the temperature was increased to 140 °C. After a total of 27 h, the reaction mixture was allowed to reach room temperature. The reaction mixture solidified upon cooling. LA:GA (MOLAR RATIO OF 10:1, DIRECT MIXTURE) [000141] DL-Lactic acid (1 g, 11 mmol) and D-gluconic acid (50% in water, 0.35 ml, 0.78 mmol) were mixed in a test tube and heated to 130 °C under vacuum. After a total of 22 h, the reaction mixture was allowed to reach room temperature. The reaction mixture became almost completely solid upon cooling. LA: GA (MOLAR RATIO OF 10:1, PREHEATING FOR 5 H) [000142] DL lactic acid (1 g, 11 mmol) was preheated to 130 °C under vacuum. After 5 h, D-gluconic acid (50% in water, 0.35 ml, 1.1 mmol) was added. After another 16 h under vacuum at 130 °C, the reaction mixture was allowed to reach room temperature. The reaction mixture solidified after cooling. BY WEIGHT % OF CA IN GA [000143] D gluconic acid (50% in water, 4 g) and citric acid monohydrate (20 mg, 1% by weight) were mixed in a test tube. Petition 870200044173, dated 06 / 04 / 2020, page 164 / 201 34 / 40 and heated to 120 °C. After a total of 15 h, the reaction mixture was allowed to reach room temperature. The reaction mixture did not solidify after cooling. By weight % of CA in ga [000144] D-gluconic acid (50% in water, 4 g) and citric acid monohydrate (105 mg, 5 wt%) were mixed in a test tube and heated to 120 °C. After a total of 15 h, the reaction mixture was allowed to reach room temperature. The reaction mixture did not solidify after cooling. BY WEIGHT % CA IN GA [000145] D-gluconic acid (50% in water, 4 g) and citric acid monohydrate (222 mg, 10% by weight) were mixed in a test tube and heated to 120 °C. After a total of 15 h, the reaction mixture was allowed to reach room temperature. The reaction mixture did not solidify after cooling. [000146] From these results, we show that gluconic acid can be oligomerized with lactic acid to form a solid, unlike pure gluconic acid or lactic acid, both of which are liquids. EXAMPLE 12: EFFECT OF GLUCONONE-DELTA-LACTONE AND SUGAR ACIDS ON E. COLI GROWTH CONDITIONS [000147] The bacterial strain Escherichia coli K12 was used for the biofilm study. This strain was maintained in LB medium at 37 °C. The biofilm was studied in synthetic medium M9 (minimal salts x1 M9 (Sigma M6030), 2 mM MgSO4, 0.1 mM CaCl2, and 0.2% glucose), which contained phosphate buffer, citric acid, lactic acid, gluconic acid, or glucono-Delta lactone to obtain media with different pH (2.6 to 6.6). BIOFILM [000148] The overnight culture of E. coli K12 (OD600 ~ 5.0) was washed with sterile water and inoculated to a final concentration of 0.2 OD / ml. Petition 870200044173, dated 06 / 04 / 2020, pp. 165 / 201 35 / 40 of M9 medium with different pH of different compounds. Biofilm development was studied in 96-well flat-bottom polystyrene microtiter plates (Sigma Aldrich, Corning® Costar® culture plates). The biofilm was stained with crystal violet. [000149] During the biofilm experiment (24 to 48 hours), the bacterial strain increased its biomass (2 to 3 times) only in the M9 medium supplemented with phosphate buffer at pH 6.1 and pH 6.6 (Figure 9). Biomass was lower in other media, and we observed a decrease in bacterial biomass with decreasing pH of the medium. Acids (citric acid, lactic acid, gluconic acid) and glucono-Delta-lactone had an inhibitory effect on growth. In addition to growth inhibition, reducing the pH of the medium resulted in decreased biofilm formation. The lowest amount of biofilm was observed in the medium supplemented with gluconic acid. The second most effective in inhibiting biofilm was lactic acid, followed by glucono-Delta-lactone. Compared to the phosphate medium, the biofilm in the gluconic acid medium was 38 times smaller at pH 2.6 and 15 times smaller at pH 3.0. At pH 6.6, gluconic and lactic acids reduced the biofilm by approximately 2 times.Glucono-Delta-lactone at pH 2.6 decreased biofilm formation 14.2 times at 24h and 30 times at 48h, respectively. [000150] Reducing the pH of the medium had a clear effect on the development of the E. coli biofilm, which is probably associated with the bactericidal effect (the lower biofilm thickness was accompanied by lower biomass). EXAMPLE 13: GLUCONE-DELTA-LACTONE MIC TEST IN GARDNERELLA VAGINALIS, LACTOBACILLUS CRSIPATUS, AND LACTOBACILLUS INERS STRAIN [000151] Gardnerella vaginalis CCUG 3717 [000152] Lactobacillus crispatus CCUG 44128 [000153] Lactobacillus iners CCUG 44025 Inoculum preparation Petition 870200044173, dated 06 / 04 / 2020, p. 166 / 201 36 / 40 [000154] Gardnerella vaginalis, Lactobacillus iners, and Lactobacillus crispatus were recovered by the CCUG (Culture Collection of the University of Gothenburg). Subculture plates were made for G. vaginalis on Chocolate-GL plates and Lactobacillus spp. on MRS agar plates at 5% CO2, 36 °C. Inocula were prepared from the subculture plates. Colonies were inoculated into culture tubes with 5 ml of test medium and vortexed for 2 minutes with approximately 10 glass beads of 3 mm diameter. Colonies were maintained until the turbidity of the solution was OD475 from 0.4 to 0.5. Bacterial solutions were checked by optical microscopy at x40 with phase contrast to ensure that the cells were dispersed. Each bacterial suspension was diluted in its test medium, 1:100, to equal 1-3 x 10⁶ CFU / ml. The microbial solutions were stored at 20 °C during the preparation of the inocula. Preparation for Microdilution and CIM Test [000155] The test substance solution was prepared aseptically at 1 g / ml in sterile H2O. The first row of wells was filled with 100 pL of substance, and then 2-fold dilutions were made vertically in 8 steps. Controls included: i) growth controls for each strain (+ctrl) = respective microorganism in the test medium without antimicrobial agent (AM); ii) no growth control (ctrl) = test medium and substance at the highest concentration tested; ensure that the substance alone does not cause a color change in the test medium; iii) gentamicin control for each strain [000156] Next, 100 µl of microorganisms were added and the plates were gently shaken at 500 rpm for 30 seconds before incubation at 35 °C and 90% RH in a CO2 incubator set at 5% CO2. After 48 and 72 hours of incubation, OD was measured and eye assessments were performed. pH was also measured for all dilutions, n = 1, pH = 6.8–7.1 Petition 870200044173, dated 06 / 04 / 2020, page 167 / 201 37 / 40 [000157] Microdilution MIC assays were performed in triplicate to evaluate MIC values ​​for GDL against G. vaginalis (CAMHB) and L. crispatus (IsoS-MRS) after 48 hours of incubation at 5% CO2 and L. iners cultured under anaerobic conditions after 120 hours (Table 7). TABLE 9. MINIMUM INHIBITORY CONCENTRATIONS (CIMS) FOR GDL AGAINST G. VAGINALIS (CAMHB), L. CRISPATUS (ISOS-MRS) AND L. INERS (CAMHB). Strain Identification CCUG MIC (g / ml) G. vaginalis CCUG 3717 0.001 L. crispatus CCUG 44128 0.0078 L. iners CCUG 44025 0.0078 [000158] The value of the minimum inhibitory concentration (MIC) of The GDL value against Gardnerella vaginalis is much lower than the MIC value of GDL against Lactobacillus iners and Lactobacillus crispatus. Therefore, GDL is more effective against Gardnerella vaginalis than against benign Lactobacillus iners and Lactobacillus crispatus. EXAMPLE 14: GDL MIC TEST IN ESCHERICHIA COLI, STAPHYLOCOCCUS EPIDERMIDIS, STAPHYLOCOCCUS AUREUS, PSEUDOMONAS AERUGINOSA, ACETINOBACTER BAUMANII, STREPTOCOCCUS PYOGENES, STREPTOCOCCUS AGALACTIAE, BETAHEMOLYTIC STREPTOCOCCI GROUPS CEGE PORPHYROMONES GINGIVALIS. STRAIN [000159] Aerobics: [000160] Escherichia coli CCUG 3274 / ATCC 10536 [000161] Staphylococcus epidermidis CCUG 23118 [000162] Staphylococcus aureus CCUG 15915 / ATCC 29213 [000163] Pseudomonas aeruginosa (PAO1) CCUG 56489 / ATCC 15692 [000164] Acetinobacter baumanii CCUG 57035 Petition 870200044173, dated 06 / 04 / 2020, p. 168 / 201 38 / 40 [000165] Demanding aerobics: [000166] Streptococcus pyogenes CCUG 47803 / ATCC 700294 [000167] Streptococcus agalactiae CCUG 29376 [000168] Beta-Hemolytic Streptococcal Groups C Streptococcus dysgalactiae ss equisimilis CCUG 4211 [000169] Group G Beta-Hemolytic Streptococci Streptococcus dysgalactiae ss equisimilis CCUG 7975 [000170] Anaerobic: [000171] Porphyromonas gingivalis CCUG 25893 / ATCC 33277 PREPARATION OF INOCULI [000172] All strains were retrieved from microbanks and made into strip plates and subcultures. Aerobes were placed on TSA plates, fastidious aerobes on horse blood plates, and anaerobes on FAA plates. Throughout the study, aerobes were cultured under aerobic conditions, the fastidious strain at 5% CO2, and the anaerobe under strict anaerobic conditions at 37 °C. At the time of testing, suspensions of selected colonies from 18-24 hour agar plates for aerobes and fastidious aerobes were suspended in 5 ml of saline solution in 10 ml tubes containing 10 glass beads with a diameter of 3 mm. [000173] Anaerobes were suspended in Concept 400 and the diluent was reduced with Brucella broth supplemented with hemin (5 μg / ml), vitamin K1 (1 μg / ml), and lysed horse blood (5%). The cell suspensions were then vigorously vortexed for 1 minute to obtain a turbid suspension. Each suspension of aerobes and fastidious aerobes was adjusted to OD equal to 0.28 at 475 nm using a spectrophotometer, which correlates approximately with 1-3 x 10⁸ CFU / ml with most species. For the anaerobe, OD measurement proved difficult due to the use of blood in the culture medium. Therefore, it was suspended to match the 0.5 McFarland standard, and the inoculum cell density was verified by plate counting. Aerobes and fastidious aerobes Petition 870200044173, dated 06 / 04 / 2020, pp. 169 / 201 39 / 40 were subsequently diluted in saline solution 10 times so that the inoculum concentration was equal to 1.5–3.0 x 10⁷ CFU / ml. The anaerobe was not diluted, but used directly as is. The different tepid aerobic bacterial suspensions were transferred to the respective wells of a 96-well plate. A. baumannii, E. coli, and P. aeruginosa were placed in one row, Staphylococcus spp. were placed in a second row, and the fastidious aerobes were placed in a third row of the plate. The anaerobe was transferred from a 25 ml multichannel pipette reservoir. Preparation for Microdilution and CIM Test [000174] Microdilution preparations and MIC tests were performed as in Example 13. [000175] MIC microdilution assays were performed in triplicate to evaluate MIC values ​​for glucono-Delta-lactone (GDL) (Table 10). TABLE 10. MINIMUM INHIBITORY CONCENTRATIONS (MICs) FOR GDL AGAINST ESCHERICHIA COLI, STAPHYLOCOCCUS EPIDERMIDIS, STAPHYLOCOCCUS AUREUS, PSEUDOMONAS AERUGINOSA, ACETINOBACTER BAUMANII, STREPTOCOCCUS PYOGENES, STREPTOCOCCUS AGALACTIAE, STREPTOCOCCUS AGALACTIAE, BETA HEMOLYTIC STREPTOCOCCI OF THE CEGE GROUPS PORPHYROMONES GINGIVALIS. Strain Identification CCUG MIC (g / ml) A. baumannii CCUG 57035 0.0031 E. coli CCUG 3274 0.0063 P. aeruginosa CCUG 56489 0.0031 S. aureus CCUG 15915 0.0063 S. epidermis CCUG 23118 0.0031 S. agalactiae CCUG 29376 0.0031 Group C Beta-Hemolytic Streptococci CCUG 4211 0.0016 Petition 870200044173, dated 06 / 04 / 2020, p. 170 / 201 40 / 40 Group G BetaHemolytic Streptococci CCUG7975 0.0031 S. piogenes CCUG 47803 0.0031 P. gingivalis CCUG 25893 0.0031 [000176] The minimum inhibitory concentration (MIC) value of The MIC value of GDL against Escherichia coli, Staphylococcus epidermidis, Staphylococcus aureus, Pseudomonas aeruginosa, Acetinobacter baumannii, Streptococcus pyogenes, Streptococcus agalactiae, Groups C and G of beta-hemolytic Streptococci, and Porphyromonas gingivalis is generally lower than the MIC value of GDL against Lactobacillus iners and Lactobacillus crispatus (Example 13). Thus, GDL is more efficient against these pathogens than against the benign Lactobacillus iners and Lactobacillus crispatus. Petition 870200044173, dated 06 / 04 / 2020, p. 171 / 201

Claims

1 / 5 CLAIMS 1. Use of a compound of Formula XX, OH OH O HO. .Ã. 4L ÕH ÕH XX J or a lactone thereof, wherein the lactone is a compound of Formula XIX or Formula XXI, characterized by the fact that it is used in the manufacture of a medicament for the treatment and / or prevention of a bacterial infection caused by a bacterium selected from the group consisting of Gardnerella vaginalis-, Mobiluncus spp-, Ureaplasma urealyticum-, Mycoplasma hominis-, Prevotella spp-, Enterococci-, Bacteroides spp-, Peptostreptococcus spp-, Porphyromonas gingivalis-, Escherichia coli-, Pseudomonas aeruginosa-, Acetinobacter baumanii-, Streptococcus pyogenes-, Group C beta-hemolytic streptococci-, Group G beta-hemolytic streptococci- and / or Streptococcus agalactiae.

2. Use according to claim 1, characterized in that the compound is a compound of Formula XIX.

3. Use, according to any of the preceding claims, characterized by the fact that the bacterial infection is a urogenital infection. Petition 870260067831, dated 09 / 07 / 2026, p. 8 / 17 2 / 5 4. Use, according to any of the preceding claims, characterized by the fact that the bacterial infection is a vaginal infection.

5. Use, according to any of the preceding claims, characterized by the fact that the bacterial infection is bacterial vaginosis.

6. Use, according to any of the preceding claims, characterized in that the infection is selected from the group consisting of: a. dermatitis and / or eczema; b. a secondary infection of dermatitis or eczema; c. acne or acneiform states; d. furunculosis; e. carbunculosis; f. folliculitis; g. impetigo; h. erysipelas; i. periodontitis; j. an infection secondary to colonization by multidrug-resistant group A or B streptococcal bacteria; k. perianal streptococcal dermatitis; l. intertriginous dermatitis; m. paronychia; n. an infection of infected skin wounds; o. an infection secondary to ingrown toenails or blisters on the feet; an infection associated with diabetic foot ulcers; p. a secondary infection arising after an animal bite; q. a secondary infection that arises after insect bites, mosquito bites, tick bites, erythema migrans, or benign lymphadenosis cutis; r.a secondary infection of herpes simplex; a secondary infection of herpes zoster or varicella zoster; Petition 870260067831, dated 09 / 07 / 2026, p. 9 / 17 3 / 5 s. blepharitis; t. conjunctivitis; u. vaginitis; and v. cervicitis.

7. Use, according to any of the preceding claims, characterized by the fact that the infection is an infection in a woman.

8. Use, according to any of the preceding claims, characterized by the fact that the infection occurs in a pregnant woman.

9. Use of a compound of Formula XX, xx or a lactone thereof, wherein the lactone is a compound of Formula XX characterized by being used in the manufacture of a medicament for the treatment of bacterial vaginosis.

10. Use according to claim 9, characterized in that the compound is a compound of Formula XIX. Petition 870260067831, dated 09 / 07 / 2026, p. 10 / 17 4 / 5 11. Use, according to any of the preceding claims, characterized in that the compound is oligomerized to form an oligomer or polymerized to form a polymer.

12. Use according to claim 11, characterized in that the oligomer or polymer further comprises lactic acid.

13. Use, according to any of the preceding claims, characterized in that the compound is in a pharmaceutical composition and in that said pharmaceutical composition comprises 5 to 99% by weight of the compound and / or in that the pharmaceutical composition comprises not more than 10% by weight of water.

14. Use according to claim 13, characterized in that the pharmaceutical composition is formulated as a tampon, vaginal spray, vaginal cup, vaginal gel, vaginal insert, vaginal patch, vaginal ring, vaginal sponge, vaginal suppository, vaginal cream, vaginal emulsion, vaginal foam, vaginal lotion, vaginal ointment, vaginal powder, vaginal shampoo, vaginal solution, vaginal spray, vaginal suspension, vaginal tablet, vaginal rod, vaginal disc, vaginal device and any combination thereof, or in that the composition is present in a sanitary article, a tampon, a sanitary pad, an incontinence pad or diaper or an underwear liner.

15. Use of a compound of Formula XX, xx or a lactone thereof, wherein the lactone is a compound of Formula XIX or Formula XXI, Petition 870260067831, dated 09 / 07 / 2026, page 11 / 17 5 / 5, characterized by the fact that it is used in the manufacture of a medicament for the prevention of premature births caused by bacterial vulvovaginitis.

16. Use according to claim 15, characterized in that the compound is administered vaginally. Petition 870260067831, dated 09 / 07 / 2026, p. 12 / 17

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