BR-102024018098-A2 - Calcium/Alginate Hydrogel Containing 3-Hydroxytyrosol: Preparation Method, Pharmaceutical Composition, and Treatment Method for Diabetic Lesions

Abstract

The present invention relates to the method of preparation, the pharmaceutical composition, and the method of treatment of diabetic skin lesions with a hydrogel ointment containing 3-hydroxytyrosol. The pharmaceutical composition is a calcium-associated alginate hydrogel containing 3-hydroxytyrosol. The healing capacity of the alginate hydrogel containing 3-hydroxytyrosol was tested considering its local and systemic effects using the following parameters, such as: cell viability; cell migration; oxidative damage; gene and protein production of growth factors and pro- and anti-inflammatory cytokines; lesion closure; number of blood vessels; number of inflammatory cells; keratinocyte proliferation; epidermal thickness; collagen fiber deposition; number of myofibroblasts; among other parameters.

Inventors

• BRUNA ROMANA DE SOUZA

Assignees

• UNIVERSIDADE DO ESTADO DO RIO DE JANEIRO

Dates

Publication Date

20260317

Application Date

20240903

Claims (3)

1. 1. Topical pharmaceutical composition for the treatment of diabetic lesions, CHARACTERIZED by: (i) Sodium alginate at 5-10% in aqueous solution; (ii) Sodium sulfate at 1-3 mM in aqueous solution; (iii) 3-Hydroxytyrosol at 0.02-0.2 mg/mL or 10-1000 μM in aqueous solution.
2. 2. Method of preparing the topical pharmaceutical composition for the treatment of diabetic lesions, CHARACTERIZED by: (i) Preparation of a 5-20% sodium alginate solution in sterile ultrapure water or sterile distilled water; (ii) Preparation of a 10-40 mM sodium sulfate solution in sterile ultrapure water; (iii) Preparation of 1-10 mg/mL 3-hydroxytyrosol dissolved in sterile 100% ethyl alcohol; (iv) Preparation of the alginate hydrogel was carried out in a Class II A2 safety cabinet in the dark; (v) Storage of the sodium alginate, sodium sulfate, and 3-hydroxytyrosol solutions at 3-8°C; (vi) Addition of 4 volumes of sterile 5-20% sodium alginate to an insulin syringe or 1-10 mL syringe; (vii) Add 0.5 volume of sterile 10-40 mM sodium sulfate and 0.5 volume of 0.1-0.3 mg/mL 3-hydroxytyrosol diluted in sterile distilled water or sterile ultrapure water to another insulin syringe or 1-10 mL syringe; (viii) Connect the syringes with a connector and mix the solutions 10-30 times to obtain the hydrogel; (ix) Store the hydrogel in a sterile tube and protect from light with aluminum foil at 3-8°C.
3. 3. Topical treatment method for diabetic lesions, CHARACTERIZED by: (i) Topical application of 10-200 μL/cm2 of hydrogel ointment containing 3-hydroxytyrosol; (ii) Application of the hydrogel ointment for 3-7 days (from the day the lesion occurs until 2 or 7 days after).

Description

FIELD OF THE INVENTION [1] This refers to the method of preparation, the pharmaceutical composition and the method of treatment of diabetic lesions with a calcium/alginate hydrogel containing 3-hydroxytyrosol. [2] The present invention relates to the major fields of medicine, biomedicine and pharmacology. FUNDAMENTALS OF THE INVENTION [3] The process of cutaneous tissue repair or scarring occurs in four overlapping phases: homeostasis, inflammation, proliferation, and remodeling. During homeostasis, blood vessels constrict, and the activation and aggregation of platelets form a clot that prevents bleeding. Inflammation begins with the secretion of cytokines and chemokines that recruit inflammatory cells. Initially, neutrophils migrate to the lesion and remove cellular debris and microorganisms. Then, monocytes invade the lesion and differentiate into macrophages that remove apoptotic cells and tissue debris and stimulate the formation of new vessels from pre-existing vessels (angiogenesis). In addition, they promote the proliferation and migration of keratinocytes that form a new epidermis or neo-epidermis (re-epithelialization). Fibroblasts are also stimulated to proliferate, migrate to the injured area, and deposit a new matrix that is rich in collagen. Many fibroblasts, and other cell types, differentiate into myofibroblasts, which are responsible for lesion contraction. After complete lesion closure (by re-epithelialization and/or contraction), most cells (macrophages, endothelial cells, myofibroblasts, etc.) die by apoptosis, and the remaining fibroblasts reorganize the extracellular matrix. In some cases, the repair process does not occur correctly, forming chronic lesions, such as diabetic lesions. Diabetes mellitus is a chronic metabolic disease characterized by insufficient insulin production by the pancreas (type I diabetes) or by the body's unresponsiveness to insulin (type II diabetes), leading to hyperglycemia. It is one of the most important public health problems in the world. According to the International Diabetes Federation (IDF), diabetes already affects more than 400 million individuals worldwide, and this number could increase to 500 million in 2030 and 700 million in 2045. Brazil is already the fifth country in the IDF ranking in the number of adults with diabetes. In 2019, it was estimated that more than 16 million Brazilians had diabetes, comprising 9% of the population. If the current rate of diabetes growth is maintained, Brazil could have a prevalence similar to that of developed countries by 2045, with 26% of the population suffering from diabetes. [4] Diabetes is the leading risk factor for cardiovascular disease and is directly associated with reduced life expectancy. Diabetic patients have a 132% increased risk of death due to cardiovascular diseases such as heart attacks and hemorrhagic strokes. Chronic hyperglycemia also leads to various complications such as strokes, neuropathy, nephropathy, peripheral arterial disease, retinopathy and poor skin healing. [5] Chronic lower extremity lesions (or diabetic feet), which affect between 40 and 60 million diabetic patients worldwide, are one of the main factors contributing to increased diabetes-related morbidity and mortality. Between 4 and 27% of diabetic patients who develop ulcers undergo amputations of the lower limbs or parts thereof. [6] Even after decades of studies, the mechanism leading to impaired skin healing related to diabetes remains complex and multifactorial. The main factors related to the inability of diabetic skin to heal are: poor angiogenesis, exacerbated inflammation, and elevated production of reactive oxygen species (ROS) and/or free radicals. In diabetic lesions, prolonged inflammation characterized by a persistence of chronically activated pro-inflammatory macrophages and an elevated secretion of proteases and pro-inflammatory cytokines leads to a delay in angiogenesis and lesion closure. The excessive production of ROS stimulated by hyperglycemia also causes damage to lipids, proteins, and nucleic acids, compromising the activity of various cell types and impairing the skin repair process. [7] Prevention is the main treatment to avoid the formation of these ulcers, and among the various methods, the main ones are: blood glucose control, periodic examinations, and early detection of lesions. However, after the ulcer has formed, there are several methods aimed at stimulating the closure of these lesions, such as: debridement, systemic antibiotics, surgical revascularization procedures, negative pressure, specialized dressings, and the application of bioactive products. However, many of these methods are expensive, and none of them are completely effective. Thus, there is a need for the development of new therapies that promote the healing of diabetic lesions. [8] Several studies have shown that bioactive components derived from extra virgin olive oil, such as hydroxytyrosol, can be a therapy for the treatm