Superozonated fatty acids and methods for producing the same

Abstract

This invention relates to a method for producing superozonated fatty acids by highly ozonating extra virgin oil containing high concentrations of polyunsaturated fatty acids. Similarly, this invention relates to the use of superozonated fatty acids produced by the above method, and their restorative effects on skin damage, in cosmetics and pharmaceuticals.

Description

Technical Field

[0001] The present invention relates to a method for obtaining hyper-ozonized fatty acids by the advanced ozonization of extra-virgin sunflower oil containing high concentrations of polyunsaturated fatty acids. Similarly, the present invention relates to the hyper-ozonized fatty acids obtained by the above procedure, and particularly to their use in cosmetics and pharmaceuticals with regard to their skin damage repair effect. Therefore, the present invention can be classified in the fields of chemistry, pharmaceuticals and cosmetics.

Background Art

[0002] Ozonized oils do not contain ozone itself, but contain by-products resulting from the reaction of ozone with unsaturated compounds present in vegetable oils, such as aldehydes and peroxidized compounds (especially hydroperoxides, ozonides, diperoxides, polyperoxides). The mechanism of this reaction is clearly defined, as are the conditions for causing the reaction to favor the selective formation of some of these peroxidized compounds (Non-Patent Document 1).

[0003] These peroxidized compounds, mainly stable ozonides (1,2,4-trioxolane), are involved in a series of widely studied biological functions (especially antibacterial action, scar formation, cell regeneration, tissue antioxidant action) (Non-Patent Document 2).

[0004] In the current state of the art, it is described that ozonized vegetable oils are used in the treatment of burns, lower limb ulcers due to venous insufficiency, pressure ulcers, acne, fistulas and surgical wounds (see, for example, Patent Document 1, Patent Document 2, Patent Document 3, Patent Document 4, Patent Document 5).

[0005] On the other hand, for the use as cosmetics related to the stimulation of tissue regeneration and the supply of oxygen to cells and tissues, reference can be made to Patent Document 6.

[0006] In general, ozonated oils are harmless and safe products that have attracted commercial interest. This has been demonstrated by toxicological studies (see, e.g., Non-Patent Document 3), histological and mutagenic studies (see, e.g., Non-Patent Document 4), genotoxicity studies (see, e.g., Non-Patent Document 5), and teratogenicity studies (see, e.g., Non-Patent Documents 6 and 7).

[0007] Within these scopes, ozonated oils have been described for their use in pharmaceuticals and cosmetics in rapid skin regeneration and scar formation (see, for example, Patent Documents 8 and 7).

[0008] Given the pharmaceutical and cosmetic interest in the above compounds, a manufacturing method has been described that allows obtaining the above compounds by ozonation of a 1% to 50% by volume water-oil emulsion. This method involves bubbling an ozone-oxygen mixture (ozone-air) with an ozone concentration of 1% to 15% by volume of the gas, using a scheme of 100 liters / hour of ozone-oxygen mixture per 500 liters of emulsion. This method achieves a high peroxide value without the need to ozonate the unsaturated compound to complete saturation, which is advantageous for the formation of β-hydroxyhydroperoxides, delays the formation of stable polymers, and achieves final product stability of over one year with sunflower oil and 24 months with cocoa oil (see Patent Document 7).

[0009] Therefore, a simple and harmless acquisition procedure is desired that allows for the production of the above-mentioned stable compounds without increasing viscosity or altering safety parameters such as a high acidity index or the presence of formaldehyde, along with this novel super-ozonated fatty acid matrix, which not only improves the effectiveness of ozonated oil but also enhances its stability by containing high concentrations of stable ozonides. [Prior art documents] [Patent Documents]

[0010]

Patent Document 1

Patent document 2

Patent document 3

Patent document 4

Patent document 5

Patent document 6

Patent document 7

Patent document 8

Non-licensed literature

[0011] [Non-licensed document 1] Ozonation in organic chemistry, Vols. 1 and 2, Philip S. Bailey, Editors: Alfred T. Blomquist and Harry H. Wasserman, Academic Press, New York, 1978 [Non-licensed document 2] Martinez Sanchez, Gregorio, 2021. Scientific rationale for the medical applications of ozonized oils, update. Ozone Therapy Global Journal Vol. 11, n° 1, pp 201-237 [Non-licensed document 3] Revista CENIC, Vol. 26, Special No., p 105, 1995

Non-licensed Document 4

[0012] In a first embodiment, the present invention relates to a procedure for obtaining superozonated fatty acids, i. A step of concentrating the polyunsaturated fatty acid fraction of sunflower oil by dry crystallization to obtain a sunflower oil matrix with a polyunsaturated fatty acid concentration of at least 90%, while maintaining the small fraction containing polyphenols, phytosterols, and tocopherols without alteration. ii. The sunflower oil matrix from step (i) is ozonated in a bubbling reactor using a gas flow of 30% to 35% ozone derived from pure oxygen, at a temperature of 25°C to 30°C, with a gas flow rate ratio of 150 L / h to 500 L / h, to obtain superozonated fatty acids. Regarding procedures, including those mentioned above.

[0013] The preparation procedure of the super-ozonated fatty acids of the present invention prioritizes the formation of stable ozonides (1,2,4-trioxolanes) having higher biological activity than β-hydroxy hydroperoxides, achieves a higher peroxide value without ozonating unsaturated compounds until they are completely saturated and without the need for an emulsion, and works directly using the matrix of the extra-virgin sunflower oil, thus avoiding the pre-step (the step of manufacturing an emulsion) as in the case of Patent Document 7. Furthermore, this super-ozonated fatty acid exceeds the stability of the final product in both ozonated sunflower oil and cocoa butter in order to achieve a stability of 36 months. Finally, since the super-ozonated fatty acid obtained by this ozonation procedure retains the extra-virginity of the sunflower oil, it needs to be stored at 30°C or lower in order to maintain its stability.

[0014] Therefore, according to the procedure of the present invention, the super-ozonated fatty acid is obtained after concentrating linoleic acid using a simple and harmless process known as fractional separation by dry crystallization, which process reaches a concentration of more than 90% linoleic acid without affecting the presence of phytosterols, polyphenols, vitamins, or the functional properties of the extra-virgin sunflower oil having a lower melting point, by filtering the presence of saturated fatty acids and a certain proportion of oleic acid present in the matrix by melting point.

[0015] In particular, the procedure of the present invention is carried out by putting extra-virgin sunflower oil into a stainless steel container, maintaining it at -1°C for 3 hours to reach the melting point of the total content of saturated fatty acids. Subsequently, after preferably passing through a sieve of 1.41 mm, the content is poured into another stainless steel container to retain the solid-state oil and extract the liquid-state oil corresponding to unsaturated fatty acids and a minor fraction (composed of polyphenols, tocopherols, and phytosterols). This second matrix is the matrix to be ozonated.

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

[0017] In another embodiment, the invention relates to the procedure defined above, wherein the polyunsaturated fatty acid is linoleic acid.

[0018] In another embodiment, the invention relates to the procedure defined above, wherein the sunflower oil is extra-virgin sunflower oil.

[0019] In the present invention, the term "hyper-ozonated fatty acid" refers to a sunflower oil or other plant-derived extra-virgin matrix that exhibits a peroxide value of more than 200 mEq, in which a stable ozonide (1,2,4-trioxolane) is preferentially incorporated after a high-level ozonation process.

[0020] On the other hand, the invention also mentions that the extra-virgin matrix after the high-level ozonation process still shows the presence of a small fraction composed of polyphenols, sterols and tocopherols, which are intact and unchanged, in addition to a high concentration of linoleic acid. These latter components (linoleic acid and the small fraction) exhibit a synergistic effect with the hyper-ozonated fatty acid and improve its properties for pharmaceuticals and cosmetics.

[0021] The procedure of the present invention includes a fractionation step by dry crystallization that enables filtering of the saturated fatty acids and a certain proportion of oleic acid present in the matrix by melting point, without affecting the presence of phytosterols, polyphenols, vitamins or the functional properties of the extra-virgin sunflower oil, until a concentration of more than 90% linoleic acid is reached.

[0022] The procedure of the present invention achieves a high peroxide value of 200 mEq O2 / kg to 1200 mEq O2 / kg by ozonating an unsaturated-rich matrix (in this case, with more than 90% linoleic acid) with a gas stream of 30% to 35% ozone, and can delay the formation of polymers, mostly as secondary ozonides or Criegee ozonides.

[0023] On the other hand, a 30%–35% ozone flow applied by bubbling, using a gas flow (L / h)-oil volume (L) ratio of 150–500, passes through a sunflower oil matrix highly concentrated with linoleic acid (over 90%) during the entire 3-hour ozonation process. This high ratio allows for a reduction in the concentration of more volatile components during the ozonation process. This is because the more volatile components are dragged out of the reactor by the gas flow, prioritizing the formation of stable ozonides.

[0024] In the procedure of the present invention, it is preferable to highly ozonate the highly concentrated linoleic acid in the matrix to a concentration of 200 mEq to 1200 mEq, depending on the formulation of the final product. Since the reaction does not occur when the sunflower oil matrix is ​​completely saturated, the small fractions of sterols, polyphenols, and tocopherols in the sunflower oil matrix remain unchanged during ozonation due to the affinity between ozone and linoleic acid.

[0025] In the preceding paragraph, complete saturation is understood to mean that the unsaturated material present in the matrix is ​​completely oxidized, i.e., completely decomposed, by ozone. Preferably, when the matrix of the present invention containing 90% linoleic acid is ozonated to complete saturation, after this process, the unsaturated material (linoleic acid) of that 90% becomes 0%.

[0026] Therefore, the procedure of the present invention supports the production of highly concentrated and stable ozonides without altering the extra virginity of the sunflower oil matrix or generating polyperoxides.

[0027] In the ozonation process of the present invention, 30% to 35% ozone obtained from pure oxygen (medical grade) is used to avoid the formation of toxic byproducts derived from nitrogen present in the air, and to avoid subsequent filtration processes to remove these toxic byproducts, processes to decompose the formed secondary ozonides, or processes that alter the extra virginity of the sunflower oil matrix. Therefore, the procedure of the present invention has the additional advantage of being able to be carried out without including a filtration process because it does not generate cytotoxic byproducts.

[0028] The reason why the above-mentioned cytotoxic byproducts are not generated by the method of the present invention is that the oxygen used is derived from medical-grade oxygen rather than from air, and furthermore, under the saturated conditions under which the method is carried out, other components are not oxidized, and only a portion of the main fraction, namely linoleic acid, is oxidized. For this reason, by carrying out the method using a matrix with a high concentration of unsaturation, oxidation of all components of the matrix and the formation of toxic products can be avoided.

[0029] Another aspect of the present invention relates to a hyperozonated fatty acid obtained by the above procedure, characterized in that the peroxide compound is preferably contained in a concentration of 200 meq to 1200 meq, more preferably the peroxide compound is 1,2,4-trioxolane (creaggy ozonide), and even more preferably the concentration of 1,2,4-trioxolane (creaggy ozonide) is 200 meq to 1200 meq.

[0030] Another aspect of the present invention relates to a pharmaceutical composition comprising the above-mentioned hyperozonated fatty acid and one or more pharmaceutically acceptable additives.

[0031] Another aspect of the present invention relates to the above-mentioned hyperozonated fatty acids used as pharmaceuticals.

[0032] Another aspect of the present invention relates to the use of the above-mentioned hyperozonated fatty acids for the manufacture of pharmaceuticals.

[0033] Another aspect of the present invention relates to a method for treating a disease in a subject in need of treatment, particularly in humans, the method comprising administering an effective amount of the superozonated fatty acid to the subject.

[0034] Another aspect of the present invention relates to the above-mentioned superozonated fatty acids used for the treatment and / or prevention of diseases, such as the prevention and treatment of pressure wounds at various stages, and the prevention and treatment of radiation dermatitis, among other conditions requiring the repair of skin damage.

[0035] Another aspect of the present invention relates to the use of the above-mentioned superozonated fatty acids as cosmetics.

[0036] In another embodiment, the present invention relates to the use of the above-mentioned superozonated fatty acids, whose use as cosmetics refers to moisturizing the skin, restoring its elasticity, and repairing photodamage.

[0037] Throughout this specification and the claims, the word “comprises” and its variations are not intended to exclude other technical features, additives, components, or processes. Other objects, advantages, and features of the present invention will be apparent to those skilled in the art, partly from the description and partly from the practice of the invention. The following examples are provided for illustrative purposes only and are not intended to limit the invention. [Examples]

[0038] Example 1. An extra virgin matrix with a linoleic acid concentration exceeding 90% is obtained, and a subsequent ozonation process is used to produce superozonated fatty acids. Step 1. Obtain an extra virgin matrix with a high linoleic acid content. 100 liters of extra virgin sunflower oil are placed in a continuous stirring reactor equipped with a cooling jacket that allows the temperature of the lipid matrix to be reduced to -1 degree Celsius in 3 hours. After 3 hours, the contents are passed through a 1.41 mm sieve and transferred to a storage container. In this process, solid matter (mainly saturated fatty acids that have reached their melting point) is extracted, and extra virgin sunflower oil with a linoleic acid content exceeding 90% by gas chromatography is obtained in the matrix storage container.

[0039] Step 2. Ozonation process for producing hyperozonated fatty acids. 500 liters of extra virgin sunflower oil with a high linoleic acid content (over 90%) is placed in a bubbling reactor. A gas flow (ozone-oxygen) with a maximum ozone concentration of 35% (v / v) is passed through the oil at a rate of 150 liters per hour, maintaining the reactor temperature below 30°C. From this ozonation scheme, after 1 hour, a peroxide value of 200 mEq to 400 mEq of stable ozonide (1,2,4-trioxolane) is obtained, after 2 hours, a peroxide value of 400 mEq to 800 mEq of stable ozonide is obtained, and after 3 hours, a peroxide value of 800 mEq to 1200 mEq of stable ozonide is obtained. The product is stored at a temperature not exceeding 30°C.

[0040] Regarding the quality of the superozonated fatty acids proposed in this invention, the acid value (AI), aldehyde concentration, viscosity, and / or the peroxide value (PI) using potassium iodide in accordance with USP XXIV and BP 2000 can be used as indicators. However, since the proposed ozonation process needs to achieve parameters of 200 mEq O2 / kg to 1200 mEq O2 / kg as the endpoint and is a simple procedure, the peroxide value (PI) and acid value (AI) are used as indicators of process progress and safety, and are evaluated every hour until the 3-hour cycle is completed.

[0041] Stability tests have confirmed that the superozonated fatty acids obtained by this invention maintain their properties for clinical and / or cosmetic applications for 36 months without any evidence of the formation of toxic derivatives.

[0042] Example 2. Application of hyperozonated fatty acids (AGHOZ) to Grade I and Grade II pressure wounds. The CIEC International Clinical Research Center (Centro Internacional de Estudios Clinicos) conducted an exploratory study in Santiago, Chile, to investigate the efficacy and safety of hyperozonated fatty acids (AGHOZ) in the healing of grade I and grade II pressure wounds in elderly adult patients residing at its facility. For this purpose, a clinical protocol was approved by the ethics committee, nursing staff were trained, and informed consent or signed consent forms were obtained as needed. Although all standard pressure wound prevention measures (positioning, braces, etc.) were taken at the center, lesions still occurred. A total of six patients were included, three males and three females, aged 72 ± 2.5 years, all were bedridden, and had comorbidities, particularly 83% with high neurological disorders (HTN), 67% with diabetes mellitus 2, 33% with renal failure, and 33% with organic brain damage. Lesions were found on bony prominences, namely the sacrum (80%), heel (30%), and elbow (10%) (two patients had two or more pressure ulcers). 70% of the lesions were grade I pressure ulcers and 30% were grade II, none showing signs of active infection. All patients were frail, had multiple conditions that delayed healing, and had failed to respond to usual prophylaxis and treatment for pressure ulcers. Standard treatment for pressure ulcers involved lightly spreading hyperozonated fatty acids (AGHOZ) on a bandage and applying it twice daily.

[0043] Daily use of superozonated fatty acids resulted in overall improvement of Grade I pressure wounds at 10±3 days and Grade II pressure wounds at 15±3 days, with no associated adverse events. Tolerability and satisfaction from both the nurses and patients were very good. The first clear signs of improvement were observed in all patients between days 3 and 7. This is considered excellent given the patient population and conditions studied.

Claims

1. A method for producing superozonated fatty acids, i. A step of concentrating the polyunsaturated fatty acid fraction of sunflower oil by dry crystallization to obtain a sunflower oil matrix with a polyunsaturated fatty acid concentration of at least 90%, while maintaining the small fraction containing polyphenols, phytosterols, and tocopherols without alteration. ii. A step to obtain superozonated fatty acids by ozonating the sunflower oil matrix of step (i) in a bubbling reactor using a gas flow of 30% to 35% ozone derived from pure oxygen, at a temperature of 25°C to 30°C, and using a gas flow rate ratio of 150 L / h to 500 L / h. Methods that include...

2. The method according to claim 1, wherein the polyunsaturated fatty acid is linoleic acid.

3. The method according to claim 1 or 2, wherein the sunflower oil is extra virgin sunflower oil.

4. A superozonated fatty acid obtained by the method described in any one of claims 1 to 3, characterized in that it contains a peroxide compound at a concentration of 200 meq to 1200 meq.

5. A pharmaceutical composition comprising the superozonated fatty acid described in claim 4 and one or more pharmaceutically acceptable additives.

6. The superozonated fatty acid according to claim 4, used as a pharmaceutical product.

7. The superozonated fatty acid according to claim 4, used for the treatment and / or prevention of a disease selected from various grades of pressure wounds and various grades of radiation dermatitis.

8. Use of the superozonated fatty acid as a cosmetic according to claim 4.

9. The use according to claim 8, wherein the cosmetic is a cosmetic for moisturizing the skin, restoring its elasticity, and repairing photodamage.

Images