POLYMERIC SPONGES INCORPORATED WITH ANDIROBA OIL AND MALEATE AND CU(II) CRYSTALS

Description

Polymeric sponges incorporated with andiroba oil and Cu(II) maleate crystals Field of invention

[001] This product is a polymeric maleate sponge with Cu2+ ions incorporated with andiroba oil (Carapa guianensis), based on a porous polymeric mixture of alginate / cellulose, for application with antibacterial activity. Fundamentals of the invention

[002] Cellulose is the most abundant natural polymer in the biosphere, consisting of linked β-D-glucopyranose monosaccharide units. It is found mainly in plant cells, but can also be produced by algae, fungi, and bacteria. Alginate is a linear anionic copolymer consisting of β-D-mannuronate (M) and α-L-guluronate (G) units linked by glycosidic bonds. The physicochemical and biological properties of alginates depend on their source. This polymer is usually extracted from brown algae and used in biomedical applications because its degradation products are biocompatible. Alginate and cellulose-based materials can be processed into different structures such as gels, polymer blends, films, sponges, and porous membranes, which have potential applications in bone regeneration, as dressings, and in the release of bioactive compounds such as drugs, enzymes, cells, and growth factors. Petition 870250005159, dated 01 / 22 / 2025, page 8 / 23 2 / 10 (BERNARDO, Marcela Piassi et al. Processing and application of polymeric biomaterials: recent advances and perspectives. Química Nova, v. 44, p. 1311-1327, 2021).

[003] Due to bacterial infections and drug resistance, the healing of chronic wounds is impaired, and thus, severe skin wounds are a major cause of morbidity and mortality. The available treatments are, in some cases, limited in effectiveness and generally do not restore the structure and function of the damaged skin, inevitably leading to dehydration and wound infection due to the need for constant dressing changes and cleaning. Polymeric sponges have the advantage of absorbing exudates and fluids from the wound without the need for constant dressing changes and cleaning, thus reducing risks. Therefore, polymers have been widely effective in regenerative medicine and tissue engineering in the treatment of difficult-to-heal skin ulcers, with the ability to significantly accelerate the regeneration process (DA NÓBREGA, Maria Eduarda Alves et al.).Applications of polymer-based membranes in the treatment of cutaneous wounds: An integrative review. Research, Society and Development, v. 11, n. 15, p. e597111537583-e597111537583, 2022).

[004] The pharmaceutical industry is constantly advancing in the search for more effective treatments with a better cost / benefit ratio. Therefore, the incorporation of principles Petition 870250005159, dated 01 / 22 / 2025, page 9 / 23 3 / 10 active ingredients in polymeric membranes to amplify their pharmacological effects is one of these innovations. Active ingredients include: crystalline grains and andiroba oil (Carapa guianensis).

[005] Andiroba oil, derived from the seeds of the Carapa guianensis tree, is traditionally used in Amazonian medicine for its anti-inflammatory, analgesic, antimicrobial, and antimalarial properties. Recent research has explored its therapeutic potential in various medical applications, including the treatment of oral mucositis, cancer, and its antioxidant properties. The yield and quality of andiroba oil are influenced by the moisture content and drying temperature of the seeds, with optimal conditions for maximum oil extraction being identified (PORFÍRIO-DIAS, Clara L. et al. Andiroba oil (Carapa guianensis Aubl) shows cytotoxicity but no mutagenicity in the ACPP02 gastric cancer cell line. Journal of Applied Toxicology, vol. 40, no. 8, p. 1060-1066, 2020).

[006] Non-volatile oils, often derived from natural sources, play a significant role in various industries, including food, cosmetics, and pharmaceuticals. These oils are characterized by their high boiling points and low vapor pressures, making them stable and effective vehicles for bioactive compounds (SADGROVE, Nicholas John et al. Pharmacology of natural Petition 870250005159, dated 01 / 22 / 2025, p. 10 / 23 4 / 10 volatiles and essential oils in food, therapy, and disease prophylaxis. Frontiers in Pharmacology, vol. 12, p. 740302, 2021).

[007] Milhomem-Paixão and collaborators (2017) observed that the development of Andiroba oil nanoemulsions ensures the stability and efficacy of the product, as well as biocompatibility, analyzing its effects on different toxicity parameters, such as cytotoxicity, genotoxicity, and hematotoxicity, as a contribution to future applications (MILHOMEM-PAIXÃO, Susana Suely Rodrigues et al. Andiroba oil (carapa guianensis aublet) nanoemulsions: development and assessment of cytotoxicity, genotoxicity, and hematotoxicity. Journal of Nanomaterials, v. 2017, n. 1, p. 4362046, 2017).

[008] Yanyi Wen and colleagues (2020) produced sodium alginate sponges and their antibacterial activity showed that the sponges exhibited a three-dimensional network structure with high porosity, as well as good antibacterial activity against Gram-positive and Gram-negative bacteria. Therefore, polymeric sponges incorporated with oils have attracted significant attention for their potential antibacterial properties. These materials are being explored for various applications, including oil-water separation, wound healing, and water remediation, due to their unique structural and functional properties (WEN, Yanyi et al.). Petition 870250005159, dated 01 / 22 / 2025, page 11 / 23 5 / 10 al. Synthesis of antibacterial gelatin / sodium alginate sponges and their antibacterial activity. Polymers, vol. 12, no. 9, p. 1926, 2020).

[009] Lyophilization, also known as freeze-drying, is a process used to remove water from a substance while preserving its structure and chemical properties. It is widely used in the pharmaceutical, food, and biotechnology industries. It is a gentle method that preserves molecular structure, making it ideal for sensitive products such as food, medicines, enzymes, and vaccines (Nowak, D., & Jakubczyk, E. (2020). The Freeze-Drying of Foods—The Characteristic of the Process Course and the Effect of Its Parameters on the Physical Properties of Food Materials. Foods, 9.). The sponges of this invention were obtained using the freeze-drying method.

[0010] In studies addressing materials previously mentioned in bibliographic references, alginate and cellulose stand out as natural biopolymers of great importance, widely used in various areas due to their favorable physicochemical properties, ease of handling, biocompatibility, biodegradability, and non-toxicity. Sponges incorporating andiroba oil (Carapa guianensis) and crystalline grains exhibit biological activities and potential use in treating skin wounds. Brief description of the drawings Petition 870250005159, dated 01 / 22 / 2025, page 12 / 23 6 / 10

[0011] The results obtained present the structural analysis of bioactive sponges, vibrational spectroscopy, heat flow and bactericidal activity.

[0012] Figure 1 shows a photograph of sponges with different concentrations (0.5%, 1%, and 2%) of crystalline grains dispersed in a polymeric matrix of alginate / cellulose and andiroba oil. Five sponges can be observed, the standard being only the polymer, polymer and andiroba oil, and the others varying the concentration of crystals in the polymeric matrix. As the concentration of crystalline grains increases, a stronger coloration is observed in the sponges incorporated with andiroba oil.

[0013] Figure 2 shows the X-ray diffraction pattern of polymeric sponges incorporated with andiroba oil and maleate crystals with Cu2+ ions in the angular range of 7 to 50°, 2θ. The diffractograms show that there were significant structural modifications. The main one was that in the range of 15° to 25° amorphous halos with low intensity are observed. Indicating an amorphous profile of these sponges.

[0014] Figure 3 shows the vibrational spectra in the infrared, observing CH, CO, C=O and C=C stretching possibly related to the complexes and triglycerides present in andiroba oil, which is rich in fatty acids, as well as the polymeric chains and coordination compounds related to Cu(II), maleate and phenanthroline. Petition 870250005159, dated 01 / 22 / 2025, p. 13 / 23 7 / 10

[0015] Figure 4 presents the heat flow graph of sponges incorporating andiroba oil and Cu(II) ions, showing endothermic and exothermic events. The first endothermic events are related to the enthalpy of vaporization of water absorbed by the sponges during storage time: without oil 84 kJ / mol, with oil 136 kJ / mol, 0.5%+oil+polymer 108 kJ / mol, 1%+oil+polymer 131 kJ / mol, and 2%+oil+polymer 180 kJ / mol. It is observed that the sponges without andiroba oil show a much lower enthalpy value compared to the sponges containing andiroba oil; the enthalpy of vaporization is higher. The exothermic events are related to the breaking of carbon chains as the temperature increases. Description of the invention

[0016] The invention can be better understood through the following detailed description, in accordance with the steps of the synthesis process from the preparation of sponges incorporated with oil and crystalline grains.

[0017] One aspect of the present invention relates to the process of obtaining the sponges, which comprises the following steps:

[0018] CRYSTAL SYNTHESIS: Initially, the masses of 1,10-phenanthroline (10 mmol), copper(II) nitrate trihydrate (10 mmol), and maleic acid (10 mmol) are determined and weighed to produce a saturated solution. The Petition 870250005159, dated 01 / 22 / 2025, page 14 / 23 8 / 10 Copper(II) nitrate trihydrate is dissolved in 40 mL of methanol under constant stirring at 400 rpm for 10 min, then 1,10-phenanthroline is slowly added to the methanolic solution of copper(II) nitrate trihydrate, in which the mixture has been kept under constant stirring for about 1 hour at 75 °C K. Maleic acid is dissolved with 10 mL of H2O treated with NaOH (20 mmol) and slowly added to the above reaction mixture with continuous stirring for 30 min at 75 °C. The final solution was filtered and subjected to slow solvent evaporation. The crystals were obtained after 30 days, macerated, and subsequently added to the alginate / cellulose polymer solution and andiroba oil (Carapa guianensis).

[0019] PREPARATION OF POLYMERIC SOLUTIONS: Three solutions are prepared, consisting of 0.3% (W / W) sodium carboxymethylcellulose and 0.3% (W / W) sodium alginate. The mixture of the two reagents results in a 1.2% (W / V) solution, which is kept under constant stirring at 300 RPM for 24 h. The mixture is prepared in triplicate.

[0020] FORMATION OF THE SPONGES: With the three polymer solutions prepared previously, 0.5, 1 and 2% (W / W) of the crystalline powder prepared in section

[0018] is added to each 1.2% (W / V) polymer solution, resulting in three solutions of 1.4% (W / V), 1.8% (W / V) and 2.4% (W / V). Then, 1 mL of oil is added to each concentration, which remain Petition 870250005159, dated 01 / 22 / 2025, page 15 / 23 9 / 10 under constant stirring at 300 RPM for 24 h. Afterwards, the solutions are cooled to low temperatures, and subsequently lyophilized at -40 °C.

[0021] Another aspect of the present invention features polymeric sponges incorporated with andiroba oil and crystals of maleate, 1,10-phenanthroline and Cu(II), characterized by the following formulation: 1% to 2% (W / V) of sodium carboxymethylcellulose (CH2OCH2COONa) and sodium alginate (C6H7O6Na)x, preferably a mixture of 1.2% (W / V) of (CH2OCH2COONa) and (C6H7O6Na)x, 0.5%, 1% and 2% (W / V) of crystalline powder and andiroba oil (Carapa guianensis) between 0.5% and 2% (V / V), preferably 1%.

[0022] The sponge formed in this invention is a porous, lightweight, and flexible material that has the ability to absorb liquids. It is composed of a network of interconnected cavities, which make the material permeable and soft. Unlike film, which consists of a thin, continuous layer of material, generally uniform and smooth. Thus, although some parts of the production procedure of these materials have similarities, the final product is totally different with distinct characteristics and applications. Examples of how the invention can be implemented

[0023] The polymeric sponges were obtained in 6 days, with a white coloration corresponding to the polymers and blue Petition 870250005159, dated 01 / 22 / 2025, page 16 / 23 10 / 10 strength increases with increasing concentration of crystalline grains (Figure 1).

[0024] Structural characterization (XRD at room temperature) was performed to analyze the amorphous phase belonging to the polymer / oil mixture and the crystalline phase belonging to the incorporated crystalline grains (Figure 2). The diffractograms at concentrations of 0.5%, 1% and 2% of crystalline Cu(II) grains are observed, as well as the polymer / oil diffractogram.

[0025] Vibrational characterization of polymeric sponges incorporated with andiroba oil and crystalline grains was performed for each formulation, and IR spectra were observed.

[0026] Differential scanning calorimetry (DSC) analysis was performed to observe the melting points, dehydration, decomposition ranges, and structural phase transition of the sponges.

[0027] Table 1 presents the statistical data related to bacterial inhibition. Table 1 - Diameters (mm) of the bacterial inhibition halos of sponges against Gram-positive and Gram-negative bacteria. Gram-positiva 0,5% 1% 2% Staphylococcus aureus ATCC 6538 15 mm 18.3 mm 23.3 mm Enterococcus faecalis ATCC 29212 20.7 mm 26.7 mm 29.6 mm Gram-negativa 0,5% 1% 2% Escherichia coli ATCC 25922 19.1 mm 21.9 mm 22.8 mm Pseudomonas aeruginosa ATCC 27853 11.1 mm 18.7 mm 19.3 mm

Claims

CLAIMS 1. POLYMERIC SPONGES, characterized by being incorporated with andiroba oil and crystals of maleate, Cu(II) and 1,10-phenanthroline, wherein the formulation comprises: 1% to 2% (W / V) of sodium carboxymethylcellulose (CH2OCH2COONa) and sodium alginate (C6H7O6Na)x, preferably a mixture of 1.2% (W / V) of (CH2OCH2COONa) and (C6H7O6Na)x; 0.5%, 1% and 2% (W / V) of crystalline powder and andiroba oil (Carapa guianensis) between 0.5% and 2% (V / V), preferably 1%.

2. PROCESS FOR OBTAINING THE POLYMERIC SPONGES defined in claim 1, characterized by the addition of 0.5, 1 and 2% (W / W) of the crystalline powder to each 1.2% (W / V) polymeric solution, forming at the end three solutions of 1.4% (W / V), 1.8% (W / V) and 2.4% (W / V), subsequently adding between 0.5 and 2% (V / V), preferably 1% of andiroba oil to each concentration, which remain under constant stirring at 300 RPM for 24 h; after which, the solutions are subjected to cooling at low temperatures, to then be lyophilized at -40 °C.

Images