Spheres with antitumor properties

Description

1 / 8 SPHERES WITH ANTITUMOR PROPERTIES Field of invention

[01] The present invention relates to spheres synthesized from sodium alginate solution supplemented with concentrations of 1%, 3% and 5% (w / w) of copper(II) crystal, L-proline and 1,10-phenanthroline with experimentally proven antitumor properties against PC-3 cells (prostate cancer) and SNB-19 cells (glioblastoma). Encapsulation in the sodium alginate polymer matrix forms carriers, providing protection and controlled, prolonged release of the drug in the body. Fundamentals of the invention

[02] Copper(II) complexes with 1,10-phenanthroline and amino acids have been reported to have great biological potential, due to their ability to interact with DNA, causing a slowdown in the replication rate of cancerous cells (Singh, NK, AA Kumbhar, YR Pokharel, and PN Yadav (2020), Anticancer potency of copper(II) complexes of thiosemicarbazones, Journal of Inorganic Biochemistry, 210, 111-134, doi: https: / / doi.org / 10.1016 / j.jinorgbio.2020.111134.)

[03] The copper(II), L-proline and 1,10-phenanthroline crystal was synthesized by Venkatraman and collaborators and its ability to bind to DNA and undergo oxidative cleavage confirmed (Venkatraman, R., JD Zubkowski, and EJ Valente (1999), Aqua(1,10-phenantrlina)(l-prolinato)cobre(II) nitrate monohidrato, Acta Crystallographica Section C, 55(8), 1241Petição 870250013306, dated 18 / 02 / 2025, pág. 2 / 8 1243, doi: https: / / doi.org / 10.1107 / S0108270199005570); (Rao, R., AK Patra, and PR Chetana (2007), DNA binding and oxidative cleavage activity of ternary (l-proline)copper(II) complexes of heterocyclic bases, Polyhedron, 26(18), 5331-5338, doi: https: / / doi.org / 10.1016 / j.poly.2007.07.040.)

[04] In this context, the anticancer properties of the copper(II), L-proline and 1,10-phenanthroline complex can be potentiated by encapsulation in sodium alginate, since this system facilitates transport, increases stability, bioavailability, regulates the concentration and release of the drug, reducing side effects.

[05] Sodium alginate is a polysaccharide extracted from seaweed or brown algae widely used in medicine due to its non-toxicity, encapsulation capacity, biocompatibility and ease of use in constructing matrices for the release of bactericidal or antineoplastic drugs (Guo, H., Q. Qin, J.-S. Chang, and D.-J. Lee (2023), Modified alginate materials for wastewater treatment: Application prospects, Bioresource Technology, 387, 129639, doi: https: / / doi.org / 10.1016 / j.biortech.2023.129639).

[06] The present invention relates to the production of spheres of the copper(II), L-proline and 1,10-phenanthroline complex encapsulated in sodium alginate facilitating transport in the body.

[07] The patent under registration number: BR 102021016255-4 A2, entitled: CRYSTALLINE FILM WITH BIOLOGICAL PROPERTIES, Petition 870250013306, dated 02 / 18 / 2025, page 8 / 19 Figure 3 / 8 presents chitosan films in association with a copper(II) complex with 1,10-phenanthroline and L-proline with the aim of optimizing the bioactivity of the crystal for biological application. However, in the present invention, the objective is to produce spheres, instead of films, encapsulated in sodium alginate and not chitosan, since this polysaccharide has special properties suitable for this purpose and favors the carrying of complexes in the organism, enhancing its application.

[08] The patent under registration number: BR 102022008725-3 A2, entitled: CHITOSAN MICROSPHERES WITH TERNARY COPPER (II) CRYSTAL COMPLEXED WITH 1,10-PHENANTHROLINE AND L-TYROSINE, presents chitosan microspheres, in association with a copper(II) complex with 1,10-phenanthroline and L-tyrosine for biological application. However, in the present invention, the spheres produced are made of sodium alginate, with properties and a different origin than chitosan, and the amino acid used is L-proline, which notably has different metabolic functions than L-tyrosine.

[09] The patent under registration number: BR 102020000797-1 A2, entitled: NEW TERNARY CRYSTAL OF 1,10-PHENANTHROLINE AND L-METHIONINE COMPLEXED WITH COPPER (II) WITH INHIBITORY POTENTIAL FOR CANCER CELLS, refers to a new, unprecedented crystal with crystallographic data and structure different from the present crystal; furthermore, the above document does not mention alginate. Petition 870250013306, dated 02 / 18 / 2025, page 9 / 19 4 / 8 sodium, the production of spheres or the carrying process in none of its processes, as occurs in the present invention. Brief description of the drawings

[010] The results obtained demonstrate that spheres of the copper(II), L-proline and 1,10-phenanthroline complex encapsulated in sodium alginate were obtained at concentrations of 1; 3 and 5%.

[011] Figure 1 shows spheres of the copper(II) complex, L-proline and 1,10-phenanthroline encapsulated in sodium alginate at concentrations a) 1% [Cu(II)-(Pro)(1,10phen)]1%; b) 3% [Cu(II)-(Pro)(1,10-phen)]3%) and c) 5% [Cu(II)(Pro)(1,10-phen)]5% and standard average size of 3.0 mm in diameter.

[012] Figure 2 shows the X-ray diffraction of spheres of the copper(II) complex, L-proline and 1,10-phenanthroline encapsulated in sodium alginate at concentrations a) 1%[Cu(II)-(Pro)(1,10-phen)]1%; b) 3%[Cu(II)-(Pro)(1,10-phen)]3% and c) 5%[Cu(II)-(Pro)(1,10-phen)]5%, making it possible to confirm the presence of crystalline peaks incorporated in the amorphous polymeric matrix.

[013] Figure 3 shows the results obtained in the thermal analyses performed on the spheres: a) [Cu(II)(Pro)(1,10-phen)]1%, b) 3%[Cu(II)-(Pro)(1,10-phen)]3% and c) 5%[Cu(II)-(Pro)(1,10-phen)]5%. From the analyses, it is possible to see that up to 200°C, endothermic decomposition events occur in the DTA curve and a decline in the TG curve, referring to Petition 870250013306, dated 02 / 18 / 2025, page 10 / 19 5 / 8 loss of free water and coordinated water present in the system. At higher temperatures, there are exothermic degradation events of the organic compounds that constitute the spheres of the copper(II), L-proline, and 1,10-phenanthroline complex. The presence of the metal Cu(II) in the complex is observed by the presence of exothermic events at high temperatures related to the oxidation process of the metallic ion.

[014] Figure 4 presents the dose-response curve results obtained in the cytotoxicity analysis of the [Cu(II)-(Pro)-(1,10-phen)] complex in prostate cancer (PC3) and glioblastoma (SNB-19) cells. From the analysis, it is possible to state that the studied compound inhibits cancer cells proportionally to the amount of dose applied. For prostate cancer cells, the test indicated IC50 = 1.5 μM (95% CI = 0.8-2.8 μM) and for glioblastoma (SNB-19) it was IC50 = 2.7 (95% CI = 1.9-3.8 μM). Compared to normal cells, the cytotoxic effect of the complex showed an IC50 = 8.7 μM (95% CI = 8.1-9.3 μM), less harmful than to cancerous cells, a positive factor for potential pharmacological application. Therefore, sodium alginate spheres with concentrations of the complex will favor the transport of the material to these regions of the body, which may enhance its pharmacological application. Description of the invention

[015] The present invention relates to spheres made from a polymeric solution of sodium alginate in Petition 870250013306, dated 02 / 18 / 2025, page 11 / 19 6 / 8 combined with the Cu(II), L-proline and 1,10-phenanthroline complex.

[016] Crystal synthesis: The crystal is synthesized by the slow solvent evaporation technique through the following steps: a) Prepare a 20 mL solution of 1,10-phenanthroline (0.5 mmol) in a volume containing 70% methanol and 30% water, using magnetic stirring; b) Add to the solution from step a) a 10 mL aqueous solution of copper(II) nitrate (0.5 mmol); c) The new solution formed in step b) is subjected to magnetic stirring for 30 minutes; d) After the process in step c), add proline (0.5 mmol) and sodium bicarbonate (0.5 mmol) and stir magnetically at a temperature of 70°C for 3 hours; e) The solution formed in step d) is kept in a beaker sealed with plastic film with small holes and placed in an oven at a constant temperature of 35 °C, being obtained after 10 days.

[017] Sphere production: the spheres are obtained by incorporating the crystal in powder form into a sodium alginate polymer solution through the following steps: a) The crystal was pulverized and concentrations of 1%, 3% and 5% (w / w) were used; Petition 870250013306, dated 02 / 18 / 2025, page 12 / 19 7 / 8 b) Sodium alginate concentrations (99%, 97% and 95%) are prepared and the crystal powder is added to them in 1%, 3% and 5% (w / w) concentrations, respectively, and then kept under magnetic stirring for 24 h; c) After the period described in step b), calcium chloride solutions (1%) are prepared; d) The solutions from step b) are added drop by drop to the solutions from step c, forming the spheres.

[018] Another aspect of the present invention is that they are characterized by the following formulations: - Formulation 1: 95% sodium alginate; 5% crystalline powder of Cu(II), L-proline and 1,10-phenanthroline and 1% calcium chloride (CaCl2); - Formulation 2: 97% sodium alginate; 3% crystalline powder of Cu(II), L-proline and 1,10-phenanthroline and 1% calcium chloride (CaCl2); - Formulation 3: 99% sodium alginate; 1% crystalline powder of Cu(II), L-proline and 1,10-phenanthroline and 1% calcium chloride (CaCl2). Examples of how the invention can be implemented

[019] From Figure 1 it is possible to see that the ternary complex of Cu(II), L-proline and 1,10-phenanthroline was incorporated into the polymeric matrix, resulting in homogeneous spheres with a blue color. The standard average size of 3.0 mm in diameter obtained by encapsulation favors the controlled release of the drugs and absorption by the body. Figure 2 Petition 870250013306, dated 02 / 18 / 2025, page 13 / 19 Figure 8 / 8 demonstrates through X-ray diffraction that the polymer matrix has crystalline peaks, confirming the presence of the complex.

[020] The TG-DTA curves shown in Figure 3 present the decomposition process of the spheres in a temperature range that varies from 30° to 900°, and it is possible to observe endothermic and exothermic events related to the release of water and organic compounds present in the matrix.

[021] Figure 4 demonstrates the cytotoxic activity against cancer cells of the ternary complex of Cu(II), L-proline and 1,10-phenanthroline, which combined with encapsulation in sodium alginate can potentiate this biological activity, increased by the bioactivity and biocompatibility of the polymer. Petition 870250013306, dated 02 / 18 / 2025, page 14 / 19

Claims

1 / 1 CLAIMS 1. SPHERES WITH ANTITUMOR PROPERTIES, characterized by comprising a solution of sodium alginate and copper(II) crystal, 1,10-phenanthroline and L-proline in powder form, specifically defined by the following formulations: - Formulation 1: 95% sodium alginate; 5% crystalline powder of Cu(II), L-proline and 1,10-phenanthroline and 1% calcium chloride (CaCl2); - Formulation 2: 97% sodium alginate; 3% crystalline powder of Cu(II), L-proline and 1,10-phenanthroline and 1% calcium chloride (CaCl2); - Formulation 3: 99% sodium alginate; 1% crystalline powder of Cu(II), L-proline and 1,10-phenanthroline and 1% calcium chloride (CaCl2).

2. SPHERES WITH ANTITUMOR PROPERTIES, according to claim 1, characterized by the average size of the spheres being 3.0 mm in diameter. Petition 870250013306, dated 02 / 18 / 2025, page 15 / 19

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