Polycarboxylated metal tetrapyrrolic macrocycle, compositions, formulations, production processes and uses

Abstract

The present patent application discloses novel combinations of polycarboxylated metal tetrapyrrolic macrocyclic compounds with one or more derivatives of iminomethanediamine compounds, related compositions and formulations thereof, production processes therefor, and related uses and methods.

Description

Tetrapyrrolic metallic polycarboxylated macrocycle, compositions, formulations, production processes and uses. Technical Field of the Patent Application

[0001] This patent application presents novel associations of polycarboxylated metallic tetrapyrrolic macrocyclic compounds with one or more derivatives of imimomethanediamine compounds, their related compositions and formulations, as well as their production processes, uses and related methods. State of the Art

[0002] The term "supramolecular synthon" was first used in 1995 to describe structural units that comprise the formation of supermolecules through synthetic operations conceivable by means of intermolecular interactions. Most synthons described in the literature are formed by non-covalent interactions between chemical species, predominantly hydrogen bonds, but strong enough to direct the formation of well-defined compounds and structural units with reflections on the physicochemical characteristics and new properties resulting from these interactions. (DESIRAJU, Gautam R. Supramolecular synthons in crystal engineering — a new organic synthesis. Angewandte Chemie International Edition, v. 34, n. 21, p. 2311-2327, 1995. DOI: 10.1002 / anie.199523111).

[0003] Despite the consolidation of the supramolecular synthon concept and the recurring use of non-covalent interactions (e.g., hydrogen bonds and electrostatic interactions) to direct the formation of supermolecules with emergent properties, the rational application of these principles to polycarboxylated metallic tetrapyrrolic macrocyclic systems still faces significant technical limitations, especially when such species exhibit high catalytic / redox activity. Under these conditions, the presence of oxygen and neutral to moderately alkaline pH can induce the generation of reactive oxygen species that attack the macrocycle itself, resulting in self-degradation and loss of spectral / functional characteristics (e.g., with an appreciable reduction in absorption band in accelerated assays). In other words, it compromises the physicochemical stability, reproducibility, and shelf life of aqueous compositions and dispersions intended for practical applications.

[0004] Thus, a gap remains in the state of the art regarding the availability of sufficiently robust and reproducible strategies that minimize the intrinsic self-degradation of these macrocycles without requiring complex covalent modifications or the use of additives that may impact solubility, performance, and / or safety profile. Summary of the Patent Application

[0005] This patent application thus presents novel polycarboxylated metallic tetrapyrrolic macrocyclic compounds in association with one or more imimomethanediamine compound derivatives.

[0006] These new molecular associations surprisingly result in new entities that exhibit low toxicity and surprising stability to self-degradation, including in aqueous solutions and at neutral to slightly alkaline pH (7 < pH < 9).

[0007] In one embodiment of the present patent application, the said compound can be prepared in different types of compositions and formulations such as tinctures, suspensions, gels, pastes and creams, especially aqueous solutions and dispersions.

[0008] Furthermore, new uses and biocidal methods derived from these compounds, compositions, and formulations are presented. Description of the Figures

[0009] Figure 1: UV-Vis spectrum of the guanidine compound iron phthalocyanine octacarboxylate in 0.1 mol.L⁻¹ sodium hydroxide solution. 1 .

[0010] Figure 2: Infrared spectrum of the guanidine compound of iron phthalocyanine octacarboxylate obtained by the ATR technique.

[0011] Figure 3: Elemental analysis of the guanidine compound of iron phthalocyanine octacarboxylate.

[0012] Figure 4: Thermogravimetric curve of the guanidine compound of iron phthalocyanine octacarboxylate.

[0013] Figure 5: Comparison of mouthwash formulations with guanidine iron phthalocyanine octacarboxylate compound (left) and iron phthalocyanine octacarboxylic acid (i.e., in its protonated form) (right) after 120 days. Initially, both solutions showed the same color intensity (optical density).

[0014] Figure 6: Monitoring of the UV-vis spectral profile of the phthalocyanine-2,3,9,10,16,17,23,24-octacarboxylic acid iron (II) complex in carbonate buffer pH 9, temperature of 50 °C and saturation with air, for 120 minutes. The arrows indicate the direction of the curve's evolution as a function of time.

[0015] Figure 7: Self-degradation curve of phthalocyanine-2,3,9,10,16,17,23,24-octacarboxylic acid iron (II) complex in carbonate buffer pH 9, temperature 50 °C and saturation with air, for 120 minutes.

[0016] Figure 8: Monitoring of the UV-Vis spectral profile of the supramolecular complex of phthalocyanine-2,3,9,10,16,17,23,24-octacarboxylic acid iron (II) complex with guanidine in carbonate buffer pH 9, temperature of 50 °C and saturation with air, for 120 minutes. The arrows indicate the direction of the curve's evolution as a function of time.

[0017] Figure 9: Self-degradation curve of the supramolecular phthalocyanine-2,3,9,10,16,17,23,24-octacarboxylic acid iron (II) complex with guanidine in carbonate buffer pH 9, temperature of 50 °C and saturation with air, for 120 minutes. Detailed Description of the Patent Application

[0018] This detailed description sets forth some, non-limiting definitions of the main terminology and technical features employed throughout this patent application, as well as providing examples of some embodiments of the present invention so that it may be reproduced by a person skilled in the art.

[0019] In the context of this patent application, the terms “comprises,” “comprising,” “includes,” “including,” and “contains” should be interpreted as open terms, that is, they indicate the presence of the elements / steps / ingredients mentioned, without excluding the presence of other additional elements / steps / ingredients, explicit or implicit. Thus, an embodiment that “comprises” a given component may include additional components not listed, provided they are compatible with the scope of the claims. Furthermore, unless expressly indicated otherwise, the term “or” is used in an inclusive sense (A or B, or both), and the expression “and / or” indicates any possible combinations of the listed items. Unless expressly indicated otherwise, the articles “a,” “an,” and “the” may mean “one or more.” The expression “at least one” means “one or more,” and “plurality” means “two or more.”The term “optionally” indicates that the characteristic may be present or absent without departing from the scope of the claims. Expressions such as “in one embodiment,” “in another embodiment,” “in an alternative embodiment,” and similar terms are non-limiting examples; characteristics described in different embodiments may be combined, when technically compatible, without departing from the claimed scope. “Effective quantity” refers to a quantity sufficient to produce the intended technical effect (e.g., biocidal / antiseptic / antiviral action), and may vary according to the route of administration, pharmaceutical form, application regimen, biological target, and environmental conditions. The term “excipient” encompasses vehicles, diluents, stabilizers, thickeners, preservatives, buffers, surfactants, and other components conventionally used to obtain compositions / formulations, provided they are compatible with the active ingredient(s) and the intended technical effect.

[0020] For the purposes of this application, the term "association" refers to the formation and / or existence of an entity resulting from intimate contact and intermolecular interaction between at least two components, which may occur in solution, dispersion and / or solid state, with defined or variable stoichiometry, and without the need for the formation of new covalent bonds between such components; the term "supramolecular association" designates an association in which the components are held together predominantly by non-covalent interactions (including, without limitation, electrostatic / ion-ion interactions — including proton transfer and salt formation —, hydrogen bonds, TT-TT and / or cation-rr interactions, van der Waals forces and hydrophobic interactions), preserving the structural identity of each component;The term "supramolecular complexation" refers to the process and / or result of the formation of a supramolecular complex (or adduct) through molecular recognition and non-covalent interactions between two or more components, generating an associated entity with physicochemical and / or functional properties distinct from those of the isolated components, regardless of isolation or complete characterization.

[0021] For the purposes of this application, the terms “for prophylaxis” and “for treatment” shall be interpreted broadly: “prophylaxis” includes use intended to prevent, reduce the likelihood of occurrence, delay onset, decrease susceptibility, prevent recurrence and / or reduce colonization / microbial load associated with infections or lesions; “treatment” includes use intended to cure, control, mitigate, alleviate signs and symptoms, reduce severity and / or duration, inhibit progression, promote resolution and / or reduce or eradicate the microbial load, including in persistent infections or those associated with biofilms;and the expression “infections or injuries caused by microorganisms” encompasses any pathological conditions in which microorganisms are involved as a cause or contributing factor, including, without limitation, infections and lesions of the skin, mucous membranes, wounds, burns, ulcers, surgical sites and colonized surfaces / tissues, as well as local or systemic infections, with “microorganisms” being understood as including, without limitation, bacteria (including Gram-positive and Gram-negative), mycobacteria, fungi / yeasts and protozoa (and, when applicable to the intended scope, viruses), in a subject (human or animal) that receives an effective amount of the composition by any appropriate route.

[0022] For the purposes of this application, the term "imimomethanediamine compound derivatives" covers compounds containing the imimomethanediamine / guanidine structural motif (i.e., a -N=C(NR) type group). 1 R 2 )-NR 3 R 4, including protonated forms) and resulting from substitutions, functionalizations, or structural variations of a reference imimomethanediamine compound, including, without limitation, N-substituted derivatives (mono-, di-, or tri-substituted with alkyl, cycloalkyl, aryl, heteroaryl, aralkyl, heteroaralkyl groups and / or functionalized groups), N-acylated, N-alkoxycarbonylated, N-sulfonylated derivatives, derivatives containing spacers and / or additional groups (e.g., charged, hydrophilic, lipophilic, or polymeric groups), as well as salts (including acid and / or base addition salts), ion pairs, solvates / hydrates, tautomers, stereoisomeric forms (where applicable), mixtures of any of the foregoing forms, and isotopically labeled analogs, provided they retain the ability to participate in the chemical / supramolecular interactions attributed to the imimomethanediamine / guanidine motif within the scope of the claims.Additionally, for the purposes of this application, it is understood that "imimomethanediamine compound derivatives" constitute a subclass of guanidine compound derivatives, since the described imimomethanediamine motif (group of the type -N=C(NR. 1 R 2 )-NR 3 R 4 (including protonated forms) corresponds functionally and structurally to a guanidine group and its variations; thus, the expression "derivatives of guanidine compounds" includes, without limitation, compounds that contain such a guanidine / iminomethanediamine motif and / or are derived from it by substitutions and functionalizations, and such derivatives may optionally be derived from (i.e., obtained from, contain as a structural unit, or be analogous / substituted to) arginine, biguanide, creatine, creatinine, guanidinoacetate, metformin and / or chlorhexidine, as well as their salts, solvates / hydrates, tautomers and other equivalent forms as applicable.

[0023] For the purposes of this patent application, the term "polycarboxylated metallic tetrapyrrolic macrocycle" designates any macrocyclic compound comprising four pyrrole or pyrrole-derived units arranged in a ring, coordinating a central metal ion, and having a plurality of carboxyl (-COOH) and / or carboxylate (-COO) groups. -(linked to the macrocyclic skeleton, including its protonated, deprotonated forms, salts, and corresponding ion pairs. Without limitation, the expression "polycarboxylated metallic tetrapyrrolic macrocycle" encompasses compounds belonging to the classes of porphyrins, porphinazines (also called porphyrazines or tetraazaporphyrins), phthalocyanines, benzoporphyrins, tetrabenzoporphyrins, naphthalocyanines, and structurally analogous compounds that share the aforementioned polycarboxylated metallic tetrapyrrolic core, regardless of the nature of the bridges between the pyrrole units (methine, aza, or mixed), the presence or absence of carbocyclic or heterocyclic rings fused to the pyrrole units, and the oxidation state of the central metal ion. The term "polycarboxylated" indicates the presence of two or more carboxyl and / or carboxylate groups in the macrocycle.)

[0024] For the purposes of this patent application, the term "polycarboxylated metallophthalocyanine" designates a compound belonging to the class of phthalocyanines—tetrapyrrolic macrocycles consisting of four isoindolic units linked by aza (-N=) bridges—containing a coordinated metal ion at the center of the macrocyclic ring and exhibiting a plurality of carboxyl (-COOH) and / or carboxylate (-COO) groups. - ) linked to the peripheral positions of the phthalocyanine skeleton, including its protonated, deprotonated forms, salts, and corresponding ion pairs. Polycarboxylated metallophthalocyanines constitute a subclass of the polycarboxylated metallic tetrapyrrolic macrocycles as defined above.

[0025] For the purposes of this patent application, the term "octocarboxylated metallophthalocyanine" designates a polycarboxylated metallophthalocyanine as defined above that specifically has eight carboxyl (-COOH) and / or carboxylate (-COO) groups. -Octacarboxylated metallophthalocyanines are a subclass of methylcellulose (McC) and polycarboxylated metallophthalocyanines. These are groups of metallophthalocyanines linked to the phthalocyanine skeleton, corresponding to two groups per isoindolic unit, including their totally or partially protonated and / or deprotonated forms, salts and corresponding ion pairs, and, where applicable, species that may exhibit axial coordination by ligands present in the medium. Octacarboxylated metallophthalocyanines include their organoderivatives and may optionally be functionalized by one or more organic substituents selected from alkyl, aryl, and N-heterocyclic groups, without altering their octacarboxylated metallophthalocyanine nature. Octacarboxylated metallophthalocyanines constitute a subclass of polycarboxylated metallophthalocyanines and polycarboxylated metallic tetrapyrrolic macrocycles as defined above.

[0026] In one embodiment, the ratio between the amount of imimnomethanediamine compound derivatives (e.g., guanidine species) and the polycarboxylated metallic tetrapyrrolic macrocycle can be conceptually defined by the acid-base balance and the supramolecular / ion-ion interactions between (i) the peripheral carboxyl / carboxylate groups of the macrocycle and (ii) the cationizable / protonable centers of the imimnomethanediamine derivative, with the association having a defined or variable stoichiometry depending on pH, ionic strength, and preparation conditions.Thus, the ratio “imimomethanediamine derivatives per macrocycle” can be expressed as w, corresponding to the effective number of imimomethanediamine units associated per macrocycle; in embodiments where the macrocycle is octacarboxylated metallophthalocyanine, there are eight carboxyl / carboxylate groups available, so w can reach 8 when maximum neutralization / ion pairing is sought, and a partial association is equally possible (e.g., w = 4) when only part of these sites are paired.

[0027] In one embodiment, the polycarboxylated metallic tetrapyrrolic macrocycle is a polycarboxylated metallophthalocyanine. In another embodiment, it is an octacarboxylated metallophthalocyanine and may optionally be functionalized with organic substituents selected from alkyl, aryl, and N-heterocyclic groups.

[0028] In one embodiment, the association is a supramolecular association or supramolecular complexation.

[0029] In one embodiment, the derivatives of imimomethanediamine compounds are derivatives of guanidine compounds.

[0030] A new composition is also presented, comprising one or more polycarboxylated metallic tetrapyrrolic macrocycles and further comprising at least one excipient.

[0031] In one embodiment, the polycarboxylated metallic tetrapyrrolic macrocycle comprises the general formula I:

[0033] in which

[0034] M = transition metal ion; and;

[0035]

[0036] where R, R1, R2 and R3 can be individually H, alkyl substituents such as CH3, CH2CH2CH2C(NH2)COOH, CH2COOH, C(O)CH2-R, CH2COOH, C(NH)NH2, or C(NH)N(CH3)2; and

[0037] where men can vary between 1 and 2.

[0038] In one embodiment, X is guanidine (R = R1 = R2 = R3 = H), arginine (R = R1 = R2 = H; R3 = CH2CH2CH2C(NH2)COOH), creatine (R = CH3; R1 = R2 = H; R3 = CH2COOH), creatinine (R1 = C(O)CH2-R; R2 = H; R3 = CH3), guanidinoacetate (R = R1 = R2 = H; R3 = CH2COOH), biguanide (R = R1 = R2 = H; R3 = C(NH)NH2), metformin (R = R1 = R2 = H; R3 = C(NH)N(CH3)2), or a combination thereof.

[0039] In one realization, if m=1, n=2, or m=1, n=1, or if m=2, n=1.

[0040] In one embodiment, M = transition metal ion selected from either Fe or Co.

[0041] Also presented is a composition comprising one or more polycarboxylated metallic tetrapyrrolic macrocycles as defined in this patent application and further comprising at least one excipient.

[0042] In one embodiment, the polycarboxylated metallic tetrapyrrolic macrocycle is an octacarboxylated metallophthalocyanine comprising general formula II:

[0044] where Y and Z represent axial ligands of coordinating molecules present in the reaction medium and are selected from water, acetate, chloride, ammonia, hydroxyl molecules, or a combination thereof.

[0045] A new formulation is also presented, comprising at least one polycarboxylated metallic tetrapyrrolic macrocycle as defined in this patent application, or at least one composition as defined in this patent application and prepared in the form of a suspension, gel, ointment, cream, tincture, solution, or dispersion.

[0046] A novel use is also presented for at least one polycarboxylated metallic tetrapyrrolic macrocycle as defined in this patent application, or at least one composition as defined in this patent application, or the formulation as defined in this patent application, for the production of a biocidal composition or formulation.

[0047] In one embodiment, the use is for prophylaxis, treatment of infections or injuries caused by microorganisms, antiseptic, healing agent, or as a medication for the inactivation of microorganisms such as bacteria, fungi, and / or viruses, and for viruses that are respiratory viruses, such as enveloped viruses like influenza, herpes, monkeypox (MPX), RSV, and / or coronaviruses like SARS-CoV-2.

[0048] A novel biocidal method is also presented comprising the application of at least one compound as defined in this patent application, or at least one composition as defined in this patent application, or a formulation as defined in this patent application, to an individual or animal in an amount effective for the manifestation of the biocidal action.

[0049] In one embodiment, the method is for prophylaxis, treatment of infections or lesions caused by microorganisms, antiseptic, healing and / or inactivation of microorganisms, such as bacteria, fungi and / or viruses, and for viruses that are respiratory and / or mucocutaneous viruses, such as enveloped viruses, such as influenza, herpes, monkeypox (MPX), RSV and / or coronaviruses, such as SARS-CoV-2.

[0050] A process for producing a polycarboxylated metallic tetrapyrrolic macrocycle is presented, comprising association with one or more imimomethanediamine compound derivatives, as defined in this patent application, comprising the following steps:

[0051] (a) to disperse or dissolve a polycarboxylic metal tetrapyrrolic macrocycle acid in an alkaline aqueous medium;

[0052] (b) add at least one imimomethanediamine compound derivative, or a salt thereof, to the solution obtained in step (a);

[0053] (c) heat the mixture resulting from step (b) to a temperature between 40 °C and 120 °C for a period between 15 minutes and 180 minutes.

[0054] In one embodiment, there are additional steps (d) concentrating the solution resulting from step (c) by partially reducing the volume of solvent; and; (e) isolating the product by precipitation, filtration and drying.

[0055] In one embodiment, the metal tetrapyrrolic polycarboxylic macrocycle acid of step (a) is metallophthalocyanine octacarboxylic acid. In one embodiment, the concentration of step (d) is carried out by distillation, optionally under vacuum, until the volume is reduced to approximately half the initial volume. In one embodiment, the precipitation of step (e) comprises the addition of an organic antisolvent to the concentrated solution obtained in step (d), followed by filtration of the precipitate and drying. In one embodiment, the organic antisolvent is ethanol. In one embodiment, the aqueous alkaline medium of step (a) comprises ammonium hydroxide or sodium hydroxide, and the imimomethanediamine compound derivative of step (b) is guanidine hydrochloride or guanidine carbonate. In one embodiment, step (c) is carried out at a temperature of approximately 80 °C for approximately 60 minutes, and the drying of step (e) is carried out in an oven at a temperature of approximately 120 °C.In one embodiment, it further comprises step (f) incorporating the polycarboxylated metallic tetrapyrrolic macrocycle obtained in step (e) into at least one excipient to obtain a composition as defined in the present patent application.

[0056] Degradation profile

[0057] The iron(II) phthalocyanine-2,3,9,10,16,17,23,24-octacarboxylic acid complex, or simply iron phthalocyanine-octacarboxylic acid, in slightly to moderately alkaline aqueous solution (7 < pH < 9) and in the presence of oxygen in the reaction medium (such as air) exhibits a prominent self-degradation profile (which can be observed by the loss of its characteristic color), as monitored by UV-Vis spectroscopy (Figures 6 and 7). It is postulated that the intense catalytic activity of iron octacarboxyphthalocyanine in the presence of oxygen produces reactive oxygen species (ROS) that attack the catalyst's own molecular structure, self-degrading completely and forming much simpler molecular species, evidenced by the appearance of a single absorption band in a high-energy region, below 250 nm.The degradation profile approximates a simple exponential, with a reduction of approximately 25% in the absorption intensity of the band at 346 nm after 120 min, at pH 9 and 50 °C.

[0058] The degradation experiment under the same experimental conditions described above was performed with the new supramolecular compound, resulting from the interaction between the phthalocyanine-2,3,9,10,16,17,23,24-octacarboxylic acid iron (II) complex and guanidines. Figure 8 shows the degradation profile of the new supramolecular compound performed at pH 9 and 50 °C over a period of 120 minutes. In Figure 9, the decay of the two main bands of the complex at 336 and 681 nm can be observed. Surprisingly, the degradation profile changes and decreases drastically in the supramolecular complex compared to the original phthalocyanine, remaining practically stable at 681 nm and with a slight 5% drop in the band at 336 nm.

[0059] Preparation and characterization procedures: example 1

[0060] To prepare a supramolecular embodiment of the compound, 4.6 g of iron octacarboxylic phthalocyanine acid (which can be prepared by various processes, for example, as described in patent BR102016023307) were added to 40 g of water containing 2.5 g of ammonium hydroxide. Then, 4.78 g of guanidine hydrochloride dissolved in 10 g of water were added to the solution containing the phthalocyanine and heated to 80 °C for 60 min. The volume was reduced by half by distillation, resulting in a dark green solution that was added to 50 g of ethanol. The precipitate was isolated by filtration, air-dried, and oven-dried at 120 °C. The compound was characterized by UV-Vis spectroscopy (Figure 1), infrared spectroscopy (Figure 2), elemental analysis (Figure 3), and thermogravimetry (Figure 4).

[0061] In an embodiment of the present patent application, the empirical molecular formula obtained for the compound C44H4iFeN2iOi7, where (n= 4) is:

[0062] Preparation and characterization procedures: Example 2.

[0063] Production scaling up: 10 kg of iron octacarboxylic phthalocyanine acid was dissolved in 70 kg of water and added to the reactor; 3.2 kg of sodium hydroxide dissolved in 10 kg of water was added, adjusting the pH to 10. Then, 7.0 kg of biguanidine carbonate previously dissolved in 20 kg of water was added to the phthalocyanine solution and heated to 80 °C for 1 hour. The initial total volume of the solution was reduced to about half by vacuum distillation and cooled to 50 °C. 75 kg of absolute ethanol was added, maintaining agitation for 30 minutes. The precipitate formed was removed by filtration, washed with absolute ethanol and dried in an oven at 120 °C. 12.5 kg of the compound with formula C48HeiFeN33Oi7 and molecular mass 1476 g.mol⁻¹ were obtained. 1corresponding to the supramolecular complex with 8 guanidine groups associated with iron phthalocyanine.

[0064] In this embodiment of the present patent application, the empirical molecular formula obtained for the compound C48HeiFeN330i7, where (n= 8) is:

[0065] Guanidinium iron octacarboxylate phthalocyanine (guanidinium iron octacarboxylate phthalocyanine) in aqueous solution exhibited high efficacy against the SARS-CoV-2 virus.

[0066] Guanidinium iron octacarboxylate phthalocyanine in aqueous solution showed high efficacy against SARS-CoV-2 viruses variant Omicron strain XBB.1.16 (Arcturus). The experiments were performed in the BSL3 Laboratory (Biosafety Level 3) of the Microbiology Department of the Institute of Biomedical Sciences of the University of São Paulo, which follows all WHO (World Health Organization) Biosafety standards and complies with Good Laboratory Practices (GLP). The Vero CCL-81 and MDCK cell culture seeded 1x10 5 cells / well in a 24-well plate and the SARS-CoV2 Variant Omicron strain XBB.1.16 (Arcturus) was tested at a concentration of 1x10 3 TCID50 / mL. Based on the observation of the cytopathic effect, cytotoxicity, and comparison of the results of Real-Time RT-PCR, we obtained a 99.93% reduction in viral load for SARS-CoV2 Variant Omicron strain XBB.1.16 (Arcturus), without showing a cytotoxic effect on the cell.

[0067] Guanidinium iron octacarboxylate phthalocyanine in aqueous solution showed high efficacy against the Monkeypox virus.

[0068] The efficacy of guanidinium iron octacarboxylate phthalocyanine in aqueous solution showed high efficacy against the Monkeypox MPXV virus (Genbank accession number ON751962.1). The experiments were performed in the BSL3 Laboratory (Biosafety Level 3) of the Microbiology Department of the Institute of Biomedical Sciences of the University of São Paulo, which follows all WHO (World Health Organization) Biosafety standards and complies with Good Laboratory Practices (GLP). Vero CCL-81 and MDCK cell cultures were seeded 1x10 5 cells / well in a 24-well plate, and Monkeypox MPXV virus (Genbank accession number ON751962.1) were tested at a concentration of 1x10 3TCID50 / mL. Based on the observation of the cytopathic effect, cytotoxicity, and comparison of the results of Real-Time RT-PCR, we obtained a 99.74% reduction in viral load for the Monkeypox MPXV virus strain (Genbank accession number ON751962.1) without showing a cytotoxic effect on the cell.

[0069] Guanidinium iron octacarboxylate phthalocyanine in aqueous solution exhibited high efficacy against RSV virus, respiratory syncytial virus, and human metapneumovirus.

[0070] Guanidinium iron phthalocyanine octacarboxylate in aqueous solution showed high efficacy against respiratory syncytial virus (RSV). This study evaluated the anti-RSV activity of guanidinium iron phthalocyanine octacarboxylate in aqueous solution in in vitro and in vivo models. RSV strains (A and B) were isolated from children with associated symptoms. Cytotoxicity was assessed by the MTT colorimetric assay, and antiviral activity was evaluated by titration of the supernatant after incubation in the presence of the compound and RSV. RESULTS: To evaluate the cytotoxic potential of guanidinium iron phthalocyanine octacarboxylate in aqueous solution, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium reduction technique was used, which measures the metabolic viability of the cell. The cytotoxic activity test of the compound showed that the dose required to inhibit cell monolayer viability by 50% was 5.43 ± 0.6 mg / mL (Figure 1A).It was also observed that, from a dose of 1.5 mg / mL, there was no statistical significance in relation to untreated cells. The determination of CC50 allowed the selection of concentrations to be tested for antiviral activity. To do this, monolayers were exposed to various concentrations of guanidinium iron phthalocyanine octacarboxylate diluted in DMEM medium and RSV subtype A (MOI 1), starting at 500 pg / mL, followed by incubation for 72 hours. The supernatants were then titrated. To investigate the compound's potential to act at the viral adsorption stage, the monolayer was incubated at 4 °C to make the cell membrane less fluid, which does not prevent adsorption but prevents penetration, followed by exposure to various concentrations of guanidinium iron phthalocyanine octacarboxylate in the presence of RSV. There was a drastic reduction in the viral titer of the supernatants in the presence of the compound throughout the entire incubation period (full time).At the highest concentration, inhibition was complete. A total absence of cytopathic effect on the monolayer was observed after 72 hours of infection at a concentration of 500 pg / mL, and a significant reduction was observed at concentrations of 125 and 31.2 pg / mL compared to the infected and untreated control. Surprisingly, in the entry inhibition assay, all tested concentrations were 100% effective in inhibiting viral infectivity. This result strongly supports the hypothesis that the action of guanidinium iron octacarboxylate phthalocyanine occurs in the first stage of the viral replicative cycle. The results show the high tolerance of HEp2 cells and the antiviral nature of the compound in the RSV A replicative cycle under in vitro conditions, with this molecule being able to significantly interfere with viral infectivity at concentrations well below the CC50 demonstrated in HEp-2 cells.Furthermore, the investigation of the mechanism carried out so far has found potential for adsorption inhibition, demonstrating that the compound may be effective in preventing infections. Further in vitro investigations of the mechanism, as well as evaluation in an in vivo model, will provide the necessary results to classify the antiviral activity of guanidinium iron octacarboxylate phthalocyanine in aqueous solution and possibly list the molecule as a candidate in the management of RSV.

[0071] Guanidinium iron octacarboxylate phthalocyanine in aqueous solution exhibited high efficacy against herpes viruses.

[0072] The efficacy of guanidinium iron octacarboxylate phthalocyanine in aqueous solution demonstrated high efficacy against herpes viruses. Herpes simplex virus 1 (HSV-1) is a neurotropic viral agent transmitted by intimate contact, affecting mucocutaneous tissue such as the oral mucosa. In this study, we evaluated the anti-herpetic potential of guanidinium iron octacarboxylate phthalocyanine in aqueous solution. The results demonstrate a robust ability of the compound to inhibit infection in the model employed. Notably, the concentration required to inhibit cytopathic effects at an MOI of 2 was 75.8 times lower than the CC50 (4.7 pg / mL). Furthermore, a significant reduction in HSV-1 US3 gene expression was observed at an MOI of 0.01 with concentrations as low as 3.9 pg / mL. The activity of guanidinium iron octacarboxylate phthalocyanine in aqueous solution was also demonstrated in the absence of light, indicating that the compound functions independently of light exposure.Furthermore, this discovery raises questions about the potential light-mediated enhancement and underlying mechanisms of the molecule's antiviral action. The release of reactive oxygen species suggests a potential virucidal mechanism, involving direct destruction of the viral particle or essential components necessary for infectivity. Singlet oxygen is known to oxidize unsaturated bonds in lipids, leading to molecular destabilization. Given that HSV-1 is an enveloped virus, the saturation and subsequent destabilization of its lipid envelope may be responsible for the observed inhibition of infectivity. However, this hypothesis requires empirical validation.

[0073] The cream formulation incorporating guanidinium iron octacarboxylate phthalocyanine exhibited high stability and efficacy as an antiviral, antiseptic, and wound-healing agent.

[0074] The experiment was conducted in the NB-3 (Biosafety Level 3) Laboratory of the Microbiology Department at the Institute of Biomedical Sciences of the University of São Paulo, which follows all WHO (World Health Organization) biosafety standards and complies with Good Laboratory Practices (GLP). The experiment used Reconstructed Human Epidermis (SkinEthic™ RHE) from the company EPISKIN (Figure 1); the Vero CCL-81 cell culture seeded 1x10 5 cells / well in a 24-hole plate; SARS-CoV2 virus / human / Bra / SP02cc / 2020 (GeneBank accession number MT350282) at a concentration of 1x10 3 TCID50 / mL.

[0075] SkinEthic™ RHE cultures were received in the laboratory 24 hours before the experiment. Each transwell was removed from the solid medium, transferred to a 6-hole plate in liquid maintenance medium, and incubated at 37°C with a 5% CO2 atmosphere. On the same day, 24-hole plates with Vero CCL-81 cells were seeded and maintained at the same temperature and atmospheric conditions. Three different concentrations of guanidinium iron octacarboxylate phthalocyanine mixed with gel cream were analyzed (0.02%, 0.05%, and 0.25%) with three different exposure times (5 minutes, 30 minutes, and 60 minutes). Transwells containing SkinEthic™ RHE were treated with a gel layer, and after the time specified above, 30 µl of the viral solution were added. After 30 minutes, the supernatant was removed from the transwell and inoculated into the cell culture. After inoculation, the plate was kept in an incubator at 37 °C with 5% CO2 for 30 minutes for viral adsorption.After this period, the plates were observed under an optical microscope to verify the integrity of the cell mat after adsorption, in order to determine the cytotoxicity of the tested product. The inoculum was 200 µl of virus (1x10). 3TCID50 / ml) was added to 270 µl of binding solution and stored until real-time RT-PCR was performed. The wells were then filled with 600 µl of culture medium (DMEM) supplemented with 2.5% fetal bovine serum. The plate was incubated at 37 °C with a 5% CO2 atmosphere for 72 hours in a humid chamber. After 72 hours of incubation, the plates were observed again under an optical microscope to verify the integrity of the cell carpet compared to the negative control, and to determine the inhibition of the cytopathic effect (morphological changes in the host cell caused by the inoculated virus) compared to the positive control. Immediately after microscopic analysis, 200 µl of supernatant were collected from each well and added to 270 µl of Binding Solution for Real-Time RT-PCR. After collection, the plate was fixed and stained with Naphtol Blue Black (Sigma-Aldrich).

[0076] Supernatants collected after 72 h were subjected to Real-Time RT-PCR for SARS-CoV-2 virus detection. Genetic material extraction was performed using the NucliSENS® easyMag® ​​automated system (BioMerieux, Lyon, France), following the manufacturer's instructions. Viral RNA detection was performed using the AgPath-ID One-Step RT-PCR Kit (Applied Biosystems Inc., USA) on a 7500 Real-Time PCR machine (Applied Biosystems, Weiterstadt, Germany), according to protocol, primers, and probe for identification of the viral E gene (Corman et al., 2020).

[0077] Based on observation of the cytopathic effect, cytotoxicity, and comparison of real-time RT-PCR results, we obtained the following percentages of SARS-CoV-2 virus inhibition: Cream 0.25% and 0.05% - 1 / 2 dilution - 100% viral reduction and: Cream 0.25% and 0.05% - 1 / 8 dilution - 100% viral reduction.

[0078] In a second experiment, the efficacy of an antiseptic cream containing guanidinium iron octacarboxylate phthalocyanine as an antiseptic was tested against coronavirus b, leading to a reduction of more than 99% in viral load at concentrations below 0.05 pg / mL. Guanidinium iron octacarboxylate phthalocyanine was described as having the ability to reduce COVID-19 symptoms 5, 6, 7, and 72 hours after infection using a mouthwash containing this compound. The efficacy of a hand sanitizing cream with two different concentrations of the supramolecular compound (0.05% and 0.25%) and the duration of this efficacy at five different times (5 minutes, 1, 2, 3, and 4 hours) were also evaluated on Reconstructed Human Epidermis (SkinEthic™ RHE) from the company EPISKIN LTDA.In this study, we evaluated the antiviral capacity of a hand sanitizing cream with two different concentrations of guanidinium iron octacarboxylate phthalocyanine (0.05% and 0.25%) and the duration of this capacity at five different times (5 minutes, 1, 2, 3, and 4 hours) in Reconstructed Human Epidermis (SkinEthic™ RHE) from the company EPISKIN LTDA.

[0079] Guanidinium iron octacarboxylate phthalocyanine has shown high efficacy against influenza viruses.

[0080] Guanidinium iron octacarboxylate phthalocyanine in aqueous solution showed high efficacy against the pandemic INFLUENZA A virus H1N1, strain ICB-2022 / 0042. The experiments were performed in the BSL3 Laboratory (Biosafety Level 3) of the Microbiology Department of the Institute of Biomedical Sciences of the University of São Paulo, which follows all WHO (World Health Organization) Biosafety standards and complies with Good Laboratory Practices (GLP). Vero CCL-81 and MDCK cell cultures seeded 1x10 5 cells / well in a 24-well plate and pandemic Influenza A H1N1 virus, strain ICB-2022 / 0042, were tested at a concentration of 1x10 3TCID50 / mL. Based on the observation of the cytopathic effect, cytotoxicity, and comparison of real-time RT-PCR results, we obtained a 99.93% reduction in viral load for the H1N1 pandemic influenza A virus strain, ICB-2022 / 0042, without showing a cytotoxic effect on the cell.

[0081] Mouthwash formulations incorporating guanidinium iron phthalocyanine octacarboxylate exhibited high stability and efficacy as an antiviral antiseptic and wound healing agent.

[0082] In this study, the objective was to perform laboratory tests to evaluate the antiviral and cytotoxic properties of a mouthwash containing the compound. Our results, obtained by real-time RT-PCR, confirmed viral reduction with the use of the antiseptic mouthwash compared to the positive control used in the study in Vero CCL-81 cells. Based on the results obtained in this study, it was possible to assess that the antiseptic mouthwash with the compound was able to reduce the viral load when compared to the positive control at titers of 1:2 (99.96%), 1:4 (99.88%), 1:8 (99.84%), and 1:16 (92.65%), while partial viral neutralization was observed at titers of 1:32 (77.42%) and 1:64 (11.06%). No viral neutralization was observed below the titer of 1:128.

[0083] Formulations of quanidinium iron octacarboxylate phthalocyanine have shown high efficacy against microorganisms.

[0084] The efficacy of guanidinium iron phthalocyanine octacarboxylate as an antiseptic was tested against the following microorganisms: Staphylococcus aureus ATCC6538; Pseudomonas aeruginosa ATCC9027; Escherichia coli ATCC8739; Salmonella choleraesuis ATCC10708 and Leptospira biflexa (Patoc strain 1), leading to a reduction of more than 99.98% at concentrations below 0.05 pg / mL. This study demonstrated the efficacy of guanidinium iron phthalocyanine octacarboxylate in reducing microbial activity, particularly against the microorganisms Acinetobacter baumannii ATCC 19606, Candida auris CDBD 11903 and Clostridioides difficile ATCC 9689, with a reduction of up to 99.99% of the microbial load at application.In a Time Kill test conducted by ProLab Biotechnology to determine microbial effectiveness, a 99.98% reduction in viable microorganisms (Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, Salmonella choleraesuis, and Candida albicans) was observed in the first 10 minutes (except for Candida albicans), demonstrating efficacy in microbial reduction for 240 minutes, thus showing satisfactory efficacy results for guanidinium iron octacarboxylate phthalocyanine. Based on the tests performed, we can observe a 99.98% reduction in viable bacteria (Staphylococcus aureus; Pseudomonas aeruginosa; Escherichia coli; Salmonella choleraesuis; and Leptospira biflexa).

[0085] Cream / dental floss formulations incorporating quanidinium iron phthalocyanine octacarboxylate have exhibited high stability and efficacy as an antiviral antiseptic and wound healing agent.

[0086] Stable toothpaste gels and creams were prepared by dispersing guanidinium iron octacarboxylate phthalocyanine in standard formulations, and tests were performed indicating effectiveness in virus inactivation. Furthermore, a faster healing rate of wounds and canker sores was observed, in addition to improved bad breath.

[0087] Saline solution formulations incorporating guanidinium iron phthalocyanine octacarboxylate exhibited high stability and efficacy as an antiviral antiseptic and wound healing agent.

[0088] Also, stable formulations were developed by dissolving guanidinium iron octacarboxylate phthalocyanine in water and saline, which, when applied as a nasal or oral spray, showed efficacy as an antiseptic and antiviral, with excellent acceptance, and which also increased the speed of wound healing, suggesting antiseptic activity combined with action in the re-epithelialization process, accelerating the wound healing process.

[0089] The examples disclosed here demonstrate only some of the embodiments and uses of the present invention. Alternative embodiments and variations of the invention may be devised by a person skilled in the art based on the technical knowledge described in this patent application and are defined from the claims described herein.

Claims

Claims 1. Polycarboxylated metallic tetrapyrrolic macrocycle, characterized by comprising an association with one or more derivatives of imimomethanediamine compounds.

2. Polycarboxylated metallic tetrapyrrolic macrocycle, according to claim 1, characterized in that the polycarboxylated metallic tetrapyrrolic macrocycle is a polycarboxylated metallophthalocyanine.

3. Polycarboxylated metallic tetrapyrrolic macrocycle, according to claim 1, characterized in that the polycarboxylated metallic tetrapyrrolic macrocycle is an octacarboxylated metallophthalocyanine.

4. Polycarboxylated metallic tetrapyrrolic macrocycle, according to claim 1, characterized by the association being supramolecular.

5. Polycarboxylated metallic tetrapyrrolic macrocycle, according to claim 3, characterized by comprising the general formula I: in what M = transition metal ion; and; where R, R1, R2 and R3 can be individually H, alkyl substituents such as CH3, CH2CH2CH2C(NH2)COOH, CH2COOH, C(O)CH2-R, CH2COOH, C(NH)NH2, or C(NH)N(CH3)2; and where men can vary between 1 and 2.

6. A polycarboxylated metallic tetrapyrrolic macrocycle, according to claim 5, characterized in that X is guanidine (R = R1 = R2 = R3 = H), arginine (R = R1 = R2 = H; R3 = CH2CH2CH2C(NH2)COOH), creatine (R = CH3; R1 = R2 = H; R3 = CH2COOH), creatinine (R1 = C(O)CH2-R; R2 = H; R3 = CH3), guanidinoacetate (R = R1 = R2 = H; R3 = CH2COOH), biguanide (R = R1 = R2 = H; R3 = C(NH)NH2), metformin (R = R1 = R2 = H; R3 = C(NH)N(CH3)2), or a combination thereof.

7. Polycarboxylated metallic tetrapyrrolic macrocycle, according to claim 5, characterized in that m=1, n=2; or; m=1, n=1; or; if m=2, n=1.

8. Polycarboxylated metallic tetrapyrrolic macrocycle, according to claim 5, characterized in that M = transition metal ion selected from Fe or Co.

9. Composition, characterized by comprising one or more polycarboxylated metallic tetrapyrrolic macrocycles as defined in any one of claims 1 to 8 and further comprising at least one excipient.

10. Composition, according to claim 9, characterized in that the polycarboxylated metallic tetrapyrrolic macrocycle is an octacarboxylated metallophthalocyanine comprising General Formula II: "" where Y and Z represent axial ligands of coordinating molecules present in the reaction medium and are selected from water, acetate, chloride, ammonia, hydroxyl molecules or a combination thereof.

11. Formulation, characterized by comprising at least one polycarboxylated metallic tetrapyrrolic macrocycle as defined in any of claims 1 to 8 or at least one composition as defined in claim 9 or 10 prepared in the form of a suspension, gel, ointment, cream, tincture, solution or dispersion.

12. Use of at least one polycarboxylated metallic tetrapyrrolic macrocycle as defined in any one of claims 1 to 8, or at least one composition as defined in claim 9 or 10, or the formulation as defined in claim 11, characterized in that it is for the production of a biocidal composition or formulation.

13. Biocidal method characterized by comprising the application of at least one compound as defined in any of claims 1 to 8 or at least one composition as defined in claim 9 or 10 or of a formulation as defined in claim 11 to an individual or animal in an amount effective for the manifestation of the biocidal action.

14. Process for producing a polycarboxylated metallic tetrapyrrolic macrocycle comprising association with one or more derivatives of imimomethanediamine compounds, as defined in any of claims 1 to 8, characterized by comprising the steps of: (a) to disperse or dissolve a polycarboxylic metal tetrapyrrolic macrocycle acid in an alkaline aqueous medium; (b) add at least one imimomethanediamine compound derivative, or a salt thereof, to the solution obtained in step (a); (c) heat the mixture resulting from step (b) to a temperature between 40 °C and 120 °C for a period between 15 minutes and 180 minutes.

15. Process according to claim 14, characterized in that the metal tetrapyrrolic polycarboxylic macrocycloalic acid of step (a) is metallophthalocyanine octacarboxylic acid.

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