Hyper-ozonised fatty acids and production method thereof

Abstract

The present invention relates to a method for producing hyper-ozonised fatty acids from the high-grade ozonization of extra virgin sunflower oil comprising a high concentration of polyunsaturated fatty acids. Likewise, the present invention relates to the hyper-ozonised fatty acids produced by means of said method and to the cosmetic and medical use thereof, particularly due to their restorative effect on skin damage.

Description

[0001] HYPEROZONIZED FATTY ACIDS AND THEIR OBTAINING PROCESS

[0002] The present invention relates to a process for obtaining hyperozonized fatty acids from the high-degree ozonization of extra virgin sunflower oil containing a high concentration of polyunsaturated fatty acids. The present invention also relates to the hyperozonized fatty acids obtained by this process and their cosmetic and medical use, particularly for their skin-damaging repair effect. Therefore, the invention could be classified within the fields of chemistry, pharmaceuticals, and cosmetics.

[0003] STATE OF THE ART

[0004] Ozonized oils do not contain ozone as such, but rather possess by-products derived from the reaction of ozone with unsaturated compounds present in vegetable oils and fats, such as aldehydes and peroxy compounds (hydroperoxides, ozonides, diperoxides, polyperoxides, among others). The mechanism of this reaction is well defined, as well as the conditions under which the reaction must develop to favor the preferential formation of one of these peroxy compounds (Ozonation in organic chemistry, Vols. 1 and 2, Philip S. Bailey, Editors: Alfred T. Blomquist and Harry H. Wasserman, Academic Press, New York, 1978).

[0005] These peroxy compounds, mainly stable ozonides (1,2,4-thoxolanes) are responsible for a series of biological functions (antimicrobial effect, healing, cell regeneration, antioxidant effect of tissues, among others) widely studied (Martínez Sánchez, Gregorio, 2021. Scientific rationale for the medical applications of ozonized oils, update. Ozone Therapy Global Journal o. 1 1 , n 9 1 , pp. 201 -237).

[0006] In the state of the art it has been described that ozonized vegetable oils have been used in the treatment of burns, ulcers of the lower limb due to venous insufficiency, bedsores, acne, fistulas and surgical wounds (see, for example, patent applications US4451480, US45916021, US984722, US2356062, US1210949). On the other hand, the cosmetic use related to the stimulation of tissue renewal, oxygenation of cells and tissues can be consulted in patent GB Pat. No. 820,463).

[0007] In general, ozonized oils are safe and harmless products of commercial interest. Toxicological studies have been carried out that demonstrate this (see, for example, CENIC Magazine, Vol. 26, Special No., p. 105, 1995), histological and mutagenic studies (see, for example, CENIC Magazine, Biological Sciences 20 (1 -2-3), 1 -4 and 23, 1989), genotoxic studies (see, for example, CENIC Magazine Biological Sciences 29 (3): 200, 1998) and teratogenic studies (see, for example, Proceedings of the 1st Ibero-American Congress on Ozone Applications, Havana City, Ozone in Medicine 11, 1990; and WO 03 / 085072 A1).

[0008] Within these ranges, ozonized oils have been described for medical and cosmetic use (see, for example, WO 01 / 37829 A1 and WO 03 / 085072 A1) in rapid skin regeneration and healing.

[0009] Given the medical and cosmetic interest of said compounds, production methods have been described which, to date, allow them to be obtained by, for example, ozonation of a 1 - 50% by volume water - oil emulsion, by bubbling an ozone - oxygen mixture (ozone - air) whose ozone concentration is between 1 - 15% by volume of gas by means of a scheme of 100 Liters / hour of the ozone - oxygen mixture per 500 Liters of emulsion, obtaining high levels of peroxides without the need to ozonize the unsaturated compounds until complete saturation, favoring the formation of -hydroxy-hydroperoxides, delaying the formation of polymers with a stability and achieving a stability of the final product greater than one year for sunflower oil and 24 months for theobroma oil (see WO 03 / 085072 A1).

[0010] Therefore, it would be desirable to have this new matrix of hyperozonized fatty acids that, by having a high concentration of stable ozonides, allow to improve the effects of ozonized oils in addition to greater stability, together with a simple and harmless production procedure, which allows to obtain said stable compounds without increasing viscosity or altering safety parameters, such as a high acidity index, presence of formaldehyde.

[0011] DESCRIPTION OF THE INVENTION In a first aspect, the invention relates to a process for obtaining hyperozonized fatty acids comprising the steps of: i. concentrating the polyunsaturated fatty acid fraction of sunflower oil to obtain a sunflower oil matrix with a concentration of at least 90% polyunsaturated fatty acids, keeping the minority fraction containing polyphenols, phytosterols and tocopherols unaltered, by means of dry crystallization fractionation; and

[0012] i. Ozonation of the sunflower oil matrix of step (i) in a bubble reactor with a gas flow of between 30% and 35% ozone from pure oxygen, at a temperature of between 25 °C and 30 °C, and using a gas flow ratio of between 150 L / h and 500 L / h to obtain the hyperozonized fatty acids.

[0013] The process for preparing the hyperozonized fatty acids of the present invention prioritizes the formation of stable ozonides (1,2,4-trioxolanes) that are attributed with greater biological activity than the "-hydroxy-hydroperoxides", also achieving high levels of peroxides without ozonizing the unsaturated compounds to complete saturation without the need to be an emulsion, avoiding a previous step (that of producing an emulsion) as in the case of patent application WO 03 / 085072 A1 , since it works directly with an extra virgin sunflower oil matrix; in addition, it exceeds the stability of the final product in both ozonized sunflower oil and theobroma, since the hyperozonized fatty acids achieve a stability of 36 months; finally, the hyperozonized fatty acids obtained by the ozonation process maintain the extra virgin nature of the sunflower oil and in order to maintain their stability they require to be stored at no more than 30.q C.

[0014] Thus, through the process of the invention, hyperozonized fatty acids are obtained after concentrating linoleic acid by means of a simple and harmless process known as dry crystallization fractionation, which allows filtering by melting points the presence of saturated fatty acids and a percentage of oleic acid present in the matrix until reaching a concentration greater than 90% of linoleic acid without affecting the presence of phytosterols, polyphenols, vitamins or the organoleptic qualities of extra virgin sunflower oil, which has a lower melting point.

[0015] In particular, the process of the invention is carried out by adding the extra virgin sunflower oil to a stainless steel container that is maintained at -1 eC for 3 hours, bringing the entire saturated fatty acid content to the melting point. The contents are then poured into another steel container after sieving, preferably 1.41 mm, which retains the oil in a solid state and extracts the liquid oil corresponding to the unsaturated fatty acids and the minor fraction (composed of polyphenols, tocopherols, and phytosterols). This second matrix is ​​the one that will be ozonized.

[0016] The percentages indicated in the process steps refer to both the weight and volume of the total sunflower oil matrix.

[0017] In another embodiment the invention relates to the process defined above, where the polyunsaturated fatty acids are linoleic acid.

[0018] In another embodiment, the invention relates to the process defined above, where the sunflower oil is extra virgin sunflower oil.

[0019] In the present invention, the term “hyperozonized fatty acids” refers to an extra virgin matrix of sunflower oil or other vegetable oil that, after a high-degree ozonization process, presents peroxide levels greater than 200 mEq, primarily of stable ozonides (1,2,4-trioxolanes) immersed in it.

[0020] Furthermore, the present invention relates to the fact that the extra virgin matrix, after the high-grade ozonization process, still has a high concentration of linoleic acid, as well as the intact and unaltered presence of the minority fraction composed of polyphenols, sterols, and tocopherols. The latter (linoleic acid and minority fraction) exhibit a synergistic effect with the hyperozonized fatty acids, improving their medical and cosmetic properties.

[0021] The process of the invention comprises a dry crystallization fractionation step that allows the presence of saturated fatty acids and a percentage of oleic acid present in the matrix to be filtered by melting points until reaching a concentration greater than 90% of linoleic acid without affecting the presence of phytosterols, polyphenols, vitamins or the organoleptic qualities of extra virgin sunflower oil.

[0022] The process of the invention, by ozonizing a matrix rich in unsaturations, in this case greater than 90% linoleic acid, with a gas flow of 30-35% ozone, allows reaching high levels of peroxides of 200 to 1200 mEq O2 / Kg, and mainly of secondary ozonides or Creigee ozonides, delaying the formation of polymers.

[0023] On the other hand, the 30-35% ozone flow applied by bubbling passes through the sunflower oil matrix highly concentrated in linoleic acid (greater than 90%) during the entire ozonation process which lasts 3 hours, using a gas flow (L / h) - Oil volume (L) ratio of between 150 and 500. This high ratio allows reducing the concentration of the most volatile components during the ozonation process, as they are dragged and removed from the reactor by the gas stream and prioritizes the formation of stable ozonides.

[0024] The process of the invention preferentially ozonizes the highly concentrated linoleic acid in the matrix to a high degree, reaching a concentration of between 200 and 1,200 mEq, depending on the formulation of the final product. The reaction does not occur at full saturation of the sunflower oil matrix, so the minority fraction of sterols, polyphenols, and tocopherols in the sunflower oil matrix remains unchanged during ozonation, due to the affinity of ozone for linoleic acid.

[0025] In the previous paragraph, total saturation is understood as total oxidation, or the complete breakdown of unsaturations present in the matrix by ozone. If we ozonize the matrix of the invention, which preferably contains 90% linoleic acid, to total saturation, after the process, we would have 0% of that 90% unsaturation (linoleic acid).

[0026] Thus, thanks to the process of the invention, the production of stable ozonides in high concentration is favored without altering the extra virgin nature of the sunflower oil matrix, and without producing polyperoxides.

[0027] In the ozonation stage of the invention, an ozone concentration of between 30 and 35%, obtained from pure (medicinal) oxygen, is used in order to prevent the formation of toxic by-products derived from the nitrogen present in the air, avoiding subsequent filtration processes to eliminate these toxic by-products, processes that degrade the secondary ozonides formed or that alter the extra virgin nature of the sunflower oil matrix. Therefore, the process of the invention has the additional advantage that it can be carried out without the need to include a filtration stage since it does not generate cytotoxic by-products.The reason the process of the invention does not generate these cytotoxic byproducts is because the oxygen used is of medicinal origin, not from air. Furthermore, under the saturation conditions at which it operates, other components are not oxidized; only part of the majority fraction, i.e., linoleic acid, is oxidized. Thus, by operating with a matrix with a high concentration of unsaturations, the oxidation of all matrix components and the formation of toxic products is prevented.

[0028] Another aspect of the invention relates to hyperozonized fatty acids obtained by the process described above, characterized in that they comprise peroxy compounds, preferably in a concentration of between 200 meq and 1200 meq, more preferably where the peroxy compounds are 1,2,4 trioxolanes (creigee ozonides), and even more preferably where the 1,2,4 trioxolanes (creigee ozonides) are in a concentration of between 200 meq and 1200 meq.

[0029] Another aspect of the present invention relates to a pharmaceutical composition comprising the hyperozonized fatty acids described above and one or more pharmaceutically acceptable excipients.

[0030] Another aspect of the invention relates to the hyperozonized fatty acids described above for use as a medicament.

[0031] Another aspect of the invention relates to the use of the hyperozonized fatty acids described above for the manufacture of a medicament.

[0032] Another aspect of the present invention relates to a method of treating a disease, in a subject in need thereof, especially in humans, which comprises administering to said subject an effective amount of the hyperozonized fatty acids described above.

[0033] Another aspect of the invention relates to the hyperozonized fatty acids described above for use in the treatment and / or prevention of a disease, for example: prevention and treatment of pressure injuries in their various stages, prevention and treatment of radiodermatitis, among other conditions requiring the repair of skin damage. Another aspect of the invention relates to the use of the hyperozonized fatty acids described above as a cosmetic.

[0034] In another embodiment, the invention relates to the use of the hyperozonized fatty acids described above, where the cosmetic use relates to skin hydration, recovery of its elasticity and repair of photodamage.

[0035] Throughout the description and claims, the word "comprises" and its variants are not intended to exclude other technical features, additives, components, or steps. For those skilled in the art, other objects, advantages, and features of the invention will be apparent partly from the description and partly from the practice of the invention. The following examples are provided by way of illustration and are not intended to be limiting of the present invention.

[0036] EXAMPLES

[0037] Example 1. Obtaining an extra virgin matrix with a concentration greater than 90% of linoleic acid and the subsequent ozonation process to produce hyperozonized fatty acids.

[0038] Step 1. Obtaining an extra virgin matrix with a high linoleic acid content.

[0039] 100 liters of extra virgin sunflower oil are placed inside a continuous stirring reactor implemented with cooling jackets that allow the temperature of the lipid matrix to decrease to -1 degrees in 3 hours; After 3 hours, the contents are passed through a sieve number 14 (1.41 mm) into a storage container, extracting in the process the solid material (mostly saturated fatty acids that have reached their melting point) and in the matrix storage container, an extra virgin sunflower oil with a content greater than 90% of linoleic acid, according to gas chromatography.

[0040] Step 2. Ozonation process to produce hyperozonized fatty acids.

[0041] 500 liters of extra virgin sunflower oil with a high linoleic acid content (greater than 90%) is placed inside a bubble reactor. A gas flow (ozone-oxygen) with a concentration of up to 35% ozone v / v at a rate of 150 liters per hour is passed through the oil, the reactor temperature is maintained at no more than 30 ° C. From this ozonation scheme, a peroxide index between 200-400 mEq is obtained in the first hour, mainly of stable ozonides (1,2,4-trioxolanes), in the second hour a peroxide index between 400-800 mEq of stable ozonides and in the third hour a peroxide index between 800-1200 mEq of stable ozonides. The product is stored at a temperature that does not exceed 30 ° C.

[0042] In relation to the quality of the hyperozonized fatty acids proposed in the present invention, the following can be used as indicators: the acid number (AI), the concentration of aldehydes, the determination of the viscosity and / or the peroxide number (PI) using potassium iodide according to USP XXIV and BP 2000. However, since the present proposed ozonation process has as its end point reaching parameters between 200 to 1200 mEq 02 / kg, since it is a simple procedure, the peroxide number (IP) and the acid number (AI) are used as indicators of progress and safety of the process, evaluated every hour until completing the 3-hour cycle.

[0043] Stability tests confirm that the hyperozonized fatty acids obtained by means of the present invention maintain their clinical and / or cosmetic properties, as the case may be, with no evidence of production of toxic derivatives for 36 months.

[0044] Example 2. Application of hyperozonized fatty acids (AGHOZ) in grade I and II pressure injuries.

[0045] The CIEC International Center for Clinical Studies conducted an exploratory study to verify the efficacy and safety of hyperozonized fatty acids (HAFAs) in the treatment of grade 1 and 2 LPP in institutionalized elderly patients in Santiago, Chile. To this end, a clinical protocol was approved by the Ethics Committee, nursing staff were trained, and informed consent or assent was signed as appropriate. This center performed all standard LPP prevention measures (positioning, orthoses, etc.), but lesions persisted. A total of 6 patients were included: 3 men and 3 women, aged 72 + / - 2.5 years, all bedridden, with comorbidities (83%).

[0046] Hypertension, 67% DM2, 33% kidney failure, 33% organic brain damage among others. The lesions were in bony prominences, sacrum 80% heels 30% elbows 10% (2 patients had more than 1 LPP) 70% lesions corresponded to grade I LPP and 30% LPP gil, none with signs of active infection. All patients were elderly, with multiple pathologies that delay healing, and had failed standard therapies for prevention and treatment of LPP. Hyperozonized fatty acids (AGHOZ) were used twice a day with gentle extension on healing plus standard measures against pressure injuries. The use of hyperozonized fatty acids daily led to a total improvement of grade I LPP in 10 ± 3 days and grade II LPP in 15 ± 3 days, with no related adverse events and with very good tolerance and satisfaction of the nurses in charge and the patients.The first signs of clear improvement were evident between the 3rd and 7th day in all patients, which is considered excellent given the population and conditions of the patients included in the study.

Claims

CLAIMS 1. Process for obtaining hyperozonized fatty acids comprising the steps of: i. concentrating the polyunsaturated fatty acid fraction of sunflower oil to obtain a sunflower oil matrix with a concentration of at least 90% polyunsaturated fatty acids, keeping the minority fraction containing polyphenols, phytosterols and tocopherols unchanged, by means of dry crystallization fractionation; and i. Ozonation of the sunflower oil matrix of step (i) in a bubble reactor with a gas flow of between 30% and 35% ozone from pure oxygen, at a temperature of between 25 °C and 30 °C, and using a gas flow ratio of between 150 L / h and 500 L / h to obtain the hyperozonized fatty acids. 2.- The method according to claim 1, wherein the polyunsaturated fatty acids are linoleic acid. 3.- The method according to any of claims 1 or 2, wherein the sunflower oil is extra virgin sunflower oil. 4.- Hyperozonized fatty acids obtained by the process described according to any of claims 1 to 3 and characterized in that they comprise peroxy compounds in a concentration of between 200 meq and 1200 meq. 5.- A pharmaceutical composition comprising the hyperozonized fatty acids according to claim 4 and one or more pharmaceutically acceptable excipients. 6.- Hyperozonized fatty acids according to claim 4, for use as a medicine. 7.- Hyperozonized fatty acids according to claim 4, for use in the treatment and / or prevention of a disease selected from pressure injuries in their different degrees and radiodermatitis in their different degrees. 8.- Use of hyperozonized fatty acids according to claim 4, as a cosmetic. 9.- The use according to claim 8, where the cosmetic use refers to the hydration of the skin, recovery of its elasticity and repair of photo damage.

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