Nanotechnology product formulation in liposomal form based on extra virgin olive oil, olive stem and leaf alcoholic extract and trace minerals from the gulf of california

Abstract

The present invention relates to a nanotechnology product formulation in liposomal form based on extra virgin olive oil, olive stem and leaf alcoholic extract and trace minerals from the Gulf of California. The formulations are highly efficient carriers for any vitamin, nutrient, drug or food supplement, ensuring the quality of the manufactured products since the invention relates to a smart drug administration system based on nano-scale particles, wherein it comprises: An oral extra virgin olive oil nanoemulsion produced from seeds with an Oleuropein content between 35% and 40% as an antioxidant agent; emulsifying agents; Gulf of California water extract; and optionally a sweetener, a flavouring and a preservative.

Description

Title of the invention: FORMULATION OF NANOTECHNOLOGICAL PRODUCTS IN LIPOSOMAL FORM BASED ON EXTRA VIRGIN OLIVE OIL, ALCOHOLIC EXTRACT OF OLIVE STEM AND LEAVES AND TRACE MINERALS FROM THE SEA OF CORTES I TECHNICAL FIELD OF THE INVENTION

[0001] The present invention is related to the field of health and nutrition, since lately technological advances have been developed in food supplements such as liposomes, which give advantage in the absorption in the body of nutritional components and / or medications, through the use of nanotechnology, that is why the present invention combines these elements in addition to being based on components of natural origin, unexpectedly solving the absorption of components such as vitamin C, since to date there is no presentation of it that allows assimilating doses beyond 30% of current pharmaceutical compositions, and as this technological advance is detailed below for this composition, we can see that this technology is viable to be applied to other products, since the way of obtaining the liposomes of the present invention, makes them highly efficient carriers for any other vitamin,nutrient or medication, ensuring the quality of the manufactured products; since the invention corresponds to an intelligent drug delivery system based on nanoscale particles.

[0002] Nanotechnology has been defined as “interested in materials and systems whose structures and components exhibit significantly enhanced physical, chemical, and biological properties, as well as phenomena and processes, due to their nanoscale size” (National Nanotechnology Initiative, 2000).

[0003] Nanomedicine, the application of nanotechnology in the delivery of bioactive agents (drugs) [2], In practical terms, nanomedicine involves the development of nano-scale devices (<100 nm in diameter), referred to as “nano vectors”.

[0004] In 1995, the inventor of this application, in his company, made the decision to begin the development of nanotechnological formulations, mainly food supplements and cosmetics, during the first 10 years, tests were carried out with different types of vegetable oils such as soy lecithin, sunflower, extra virgin olive oil and jojoba, as well as other solvents and surfactants such as SPAM and Tween 80 were used.

[0005] Following these tests, it was decided to use what is mentioned further ahead in this same document, which constitutes the invention and which corresponds to nanoliposomes not previously made and which offer enormous technical advantages, with the aim of guaranteeing greater stability, flavour and pleasant appearance for the consumer and, above all, providing the nutrients, which, with this technology, we achieve greater absorption and rapid effect in the body. BACKGROUND OF THE INVENTION

[0006] Document US11066355B2 Branched tail lipid compounds and compositions for intracellular delivery of therapeutic agents with the patent family EP3736261A1 , EP3596041 B1 , US20200069599A1 , US10799463B2, describes liquid dosage forms for oral and parenteral administration including, but not limited to, pharmaceutically acceptable emulsions, micro emulsions, nano emulsions, solutions, suspensions, syrups and / or elixirs.In addition to the active ingredients, the liquid dosage forms may comprise inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing and emulsifying agents such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (particularly cottonseed, peanut, corn, germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols, and sorbitan fatty acid esters, and mixtures thereof. In addition to the inert diluents, the oral compositions may. include additional therapeutic and / or prophylactic, additional agents such as wetting agents, emulsifying and suspending agents, sweeteners, flavorings and / or perfuming agents. In certain embodiments for parenteral administration, the compositions are mixed with solubilizing agents such as Cremophor®, alcohols, oils, modified oils, glycols, polysorbates, cyclodextrins, polymers and / or combinations thereof. This document and its family of patents refer to the use of olive oil or other types of oils, in our case the invention is that the extra virgin olive oil used is acquired from the seeds of olives processed in the month of December, because at these dates the % Oleuropein content is at its maximum, which is between 35% and 40%. Oleuropein is the main antioxidant component.In addition, this prior art references the use of polysorbates, among others, as surfactants, but primarily for parenteral formulations. In our case, our nanoemulsions are oral.

[0007] In patents CN104127326A, WO2021072139A1, CN104146883A the use of surfactants or emulsifiers is mentioned, however it does not mention which ones; in our formulations Polysorbate (60 or 80) is used. In patent US20150231069A1, the use of the surfactant Tween 80 or polysorbate 80 is mentioned, but the proportion used is not indicated; in our formulations it is between 7% and 10% of the formulation. For a technician with average knowledge in the field, it is not obvious to reach the ideal formulation from what is already known and disclosed in these backgrounds, since, as detailed later in the description of the invention, there are several parameters that had to be considered to reach an ideal formulation.

[0008] Patents CN104127326A, WO2021072139A1 , CN104146883A mention the use of emulsifying agents, such as the liquid dosage form for oral administration includes, but is not limited to, pharmaceutically acceptable lotion, micro emulsion, solution, suspension, syrup and elixir. Except active component (i.e. particle, nanoparticle, liposome, micelle, polynucleotides / lipid complex) In the exterior, the liquid dosage form may also contain inert diluent common in the fields, such as water or other solvents, solubilizing and emulsifying agent, such as ethyl alcohol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-BDO, two Methylformamide, oil (especially cottonseed oil, peanut oil, corn oil, embryo oil, olive oil, castor oil and sesame oil), glycerol, Tetrahydrofurfuryl alcohol, polyethylene glycol and sorbitan fatty acid esters and their mixture. In addition to inert diluents, the orally administered composition may also include an adjuvant, such as wetting agent, emulsifying and suspending agent, sweetener, flavoring and aromatic agent. But none of these antecedents makes any reference to polysorbate (60 or 80).

[0009] In patents US11066355B2 and its patent families (EP3736261A1, EP3596041 B1, US20200069599A1 and US10799463B2), CN104910025B, CN101990425B, US20070059248A1, CN114728886A, ethyl alcohol is mentioned as an emulsifying agent but not for the preparation of extracts.

[0010] Patent US20090252796 mentions the use of olive leaf extract, however, in our process we use olive leaves and stems.

[0011] In patent IN264 / MUM / 2015, the use of seawater extracts with Calcium, Lead, Iron and Zinc is mentioned, unlike our process for trace minerals from the Sea of ​​Cortez, the elements found are: Magnesium, Potassium, Boron, Iodine, Molybdenum, Cobalt, Zinc and 72 more minerals, which are essential minerals for the cellular functions of the body.

[0012] Patents CN104127326A and CN104146883A refer to the use of inorganic salts in a proportion of 0.02 to 1%, but are only limited to Sodium Chloride, Potassium Chloride or Calcium Chloride, our process provides more essential minerals than those indicated in said patents.

[0013] Patent KR 10-2020-0032092 mentions the use of saline solution, but does not specify the formulation. Our process defines the use of trace minerals from the Sea of ​​Cortez, which is detailed later in the description of the invention.

[0014] Chinese patent application, CN113116824A describes the method of preparing a nano emulsion by means of stirring, heating, incubation and ultrasonic dispersion. In our invention, the method for obtaining the nanoemulsion is through micronization, which allows the nanoemulsions to remain stable for up to 18 months (particle size distribution and z-potential studies are attached in accelerated stability studies, as mentioned in the NOM-073-SSA1-2015 standard, Stability of Drugs and Medications, as well as Herbal Remedies).

[0015] Patent application US20230023038A1 establishes a method for producing liposomes using a pressurized mixture based on supercritical carbon dioxide. As mentioned in our invention, the method for producing the nanoemulsion is micronization.

[0016] Patent application CN102743325A and its patent families CN102743324, CN102743326 and CN102743327) use biological methods. They also differ from our invention, where the method for obtaining the nanoemulsion is through micronization.

[0017] The granted US patent US11066355B2 and its patent families (EP3736261A1 , EP3596041 B1 , US20200069599A1 and US10799463B2), CN104910025B, CN101990425B, US20070059248A1 , CN114728886A, IN264 / MUM / 2015, CN104127326A, WO2021072139A1 , KR10-2020-0032092, US20090252796A1 , CN104146883A, US20150231069A1 , AU2018304530A1 and US10632205B2, do not mention what technology they use to obtain the nano emulsions.

[0018] The Korean precedent, KR10-2020-0032092 mentions that the emulsion was stable for a week or more, patents US11066355B2 and its patent families (EP3736261A1 , EP3596041 B1 , US20200069599A1 and US10799463B2), CN104910025B, CN101990425B, US20070059248A1 , CN114728886A, CN113116824A, IN264 / MUM / 2015, CN104127326A, CN102743325A and their patent families CN102743324, CN102743326 and CN102743327), US20230023038A1, WO2021072139A1, US20090252796A1, CN104146883A, US20150231069A1, AU2018304530A1 and US10632205B2, do not mention the time during which the nano emulsion remained stable, our invention, In contrast, it has established a maximum period of 18 months based on the results obtained in the stability tests.

[0019] In addition to all of the above, none of the patents analyzed mention stevia as a sweetener; furthermore, none of the patents analyzed mention the use of flavorings; nor does the use of potassium sorbate as a preservative. Existing Technology

[0020] In Mexico and around the world, different technologies have been proposed to generate liposomes, as we have mentioned in the background, but none of them have achieved what our invention achieves; since, although some of them use olive oil and nanoemulsion generation, they do so using different technologies and do not achieve what we achieved in terms of particle size (< 100 nm), z potential (positive "+"), and stability of the nanoemulsions (18 months). Technical Problem and Proposed Solution

[0021] Some of the problems with the state of the art are that drugs, vitamins and nutrients have a Low Bioavailability (Poor absorption) i side effects and adverse reactions, problems with food intake, high doses (large numbers of tablets), and consequently, treatment abandonment. In addition, for laboratories, they seek to achieve longer-term stability and bioavailability of the resulting product. Our invention achieves stability for 18 months, including particle size < 100 nm, as indicated herein, which had not been achieved to date. This is achieved thanks to the method and components used, which are detailed later in the description of the invention. Description of the invention

[0022] The invention is based on the use of liposomes of extra virgin olive oil (including stem and leaves) since the principle of the benefits of fats was considered, which among others are: Hepatoprotectors. Bile production. ■ Fluid exchange in the lung and intestine. ■ Detox of the gastric mucosa. ■ Lipoprotein synthesis. ■ Cognition, learning and memory. ■ Decreases adrenocorticotropic hormones (ACTH) and cortisol responses. ■ Improvement of psychiatric disorders. ■ Prevention of neurodegenerative and inflammatory events.

[0023] Where the assimilation of fats occurs as follows: The fats in food are emulsified by bile salts to form micelles. Fatty acids enter the epithelial cells and form triglycerides. Triglycerides combine with proteins within the Golgi apparatus to form chylomicrons. Chylomicrons enter the lacteal and are transported to the subclavian vein.

[0024] Why olive oil and olive leaf? • High content of polyphenolic compounds or Oleuropein (more abundant) or Hydroxytyrosol or Oleocanthal • Synergistic action that enhances the antioxidant effect. • Antihypertensive, antithrombotic, anti-inflammatory, hepatoprotective, anticancer and neuroprotective action.

[0025] Benefits of extra virgin olive oil

[0026] Olive oil is a functional food because, apart from its high oleic acid content, it contains many highly bioactive compounds. Only present in 1-2% of olive oil, it includes more than 230 compounds: J tocopherols, J squalene, J fatty alcohols, J triterpene alcohols, J phyto-sterols, J oleuropein, J hydroxytyrosol and J tyrosol J Oleocanthal

[0027] Polyphenols and Oleic Acid • Polyphenols, particularly hydroxytryrosol and tryrosol, have been shown to have potent anti-inflammatory effects and may also influence cell proliferation, cell cycle progression, apoptosis, and arachidonic acid metabolism in cancer cells. • Oleic acid has the ability to alter the expression of certain oncogenes (Her-2 / neu) associated with aggressive breast cancer tumors. (Source: Visioli F & Bernardini E. 2011. Polyphenols from extra virgin olive oil: biological activities. Curr Pharm Des 17, 786–804)

[0028] What are liposomes? • They are microspheres. • Oil phase + aqueous phase. • Retains the active ingredients inside (aqueous phase). • Retains the active ingredients on the outside (oily phase).

[0029] What are Nano-Emulsions? Nutrient / drug carriers. • Smart management. • Substrate that improves selectivity, efficacy and safety of the drug. • Control of nutrient release into the circulation. • Improves pharmacokinetics and / or bioavailability.

[0030] Drug carriers. The development of a technique or technology (smart delivery) that improves the effectiveness of a group of nutrients or drugs.

[0031] LIPOSOMES (Nano Emulsions) The trapped nutrients acquire the pharmacokinetic characteristics of the carrier.

[0032] Benefits of nano emulsions: 1. Increases bioavailability. 2. It is non-toxic. 3. Increases solubility. 4. Biocompatible. 5. Protects the active ingredients. 6. Biodegradable. 7. Non-immunogenic. 8. Minimizes side effects.

[0033] Bioavailability: Bioavailability is understood as the percentage of a nutrient that is detected in the circulation after a certain time of having been administered, this is given by the following factors: Chemical nature. • Pharmaceutical form (carrier). • Metabolism of the individual.

[0034] All these elements were considered to arrive at the formulation of the invention described and claimed in this application.

[0035] The technical advantages that our invention provides are: 1. Physicochemical stability, which represents a long shelf life. 2. It is functional, which implies a therapeutic and safe effect. 3. It is reproducible on a small and large scale. 4. It is administered orally, which is practical and allows for the addition of flavor. 5. It is accessible since its costs can be low. 6. And Innovative, since it is the result of an evolution in formulations and is adaptable.

[0036] As we will see in detail, the process to obtain our final product consists of several stages, which are the following, and later we will detail each of them. A. Process for producing alcoholic extract from olive leaves and stems. B. Crystallization process of trace minerals from the Sea of ​​Cortez. C. Emulsification process. Brief description of the figures

[0037] Figure 1 is a representative HPLC Chromatogram of Finished Product (PT) of COENZYME Q10. Where your QUANTITATIVE ANALYSIS OF COENZYME Q10 reports: Project Name IQK Instrument Name HPLC-Quality Sequence Date acquired 2021-11-22 14:50:47-08:00 Injection Acq Method Name COENZYME QW.amx Agilent OpenLab Acquisition Version 2.4 Acq Injection Software - Build 2.4.0-REL 2.4.0.628 Injection Volume 20,000 μL Final Time Signal 5.999 TOOLS Module Type Part. Serial No. Firmware Grad. Pump Pumpo G4294B DECAH00336 B.07.26

[0001] Sampler Samper G4294B DECAH00336 B.07.26

[0001] Column Oven Column Compartment G4294B DECAH00336 B.07.26

[0001] DAD Detector G4294B DECAH00336 B.07.26

[0001] CHROMATOGRAPHIC CONDITIONS Acquisition method: COENZIMA QW.amx Path: C:\CDSProjects\IQK\Results\CQENZIMAQ10\PT ANTIOXIDANT W.sirslt Method information Last saved as: C:\CDSProjects\IQK\Methods\COENZYME QW.amx Modified: 2021 -03-17 12:43:11 -07:00 Modifier: RICARDO CAZARES Created: 2020-01-20 08:53:12-08:00 Creator: RICARDO CAZARES Description: Version: 2021-0317-1943-11696 DAD Method Peak width: > 0.1 min (2 s response time) (2.5 Hz) Slot: 4 nm UV lamp required: Yes Requires Vis lamp: No Module name: DAD Module type G4294B Order 1 Column furnace method Module Display Name: Column Furnace Module type G4294B Order 1 Grad. Pumping Method Flow rate: 1,500 mL / min Low pressure limit: 0.00 bar High pressure limit: 600.00 bar Maximum flow gradient: 100,000 mL / min 2 . Primary Channel: Automatic Module Screen Name: Pump Grad. Module Type: G4294B Order 1 Solvent composition Sampler Method Module Display Name: Sampler Module type G4294B Order 1 Method properties Instrument technique: Liquid chromatography Scheme version Scheme version: 2.3

[0038] Figure 2 is a representation of the HPLC of Curcumin, showing 3 peaks, representing each of the curcuminoids (Bis-Demethoxy Curcumin (BDMC), Demethoxy Curcumin (DMC) and Curcumin), which provide the desired effect in the body.

[0039] Figure 3 corresponds to HPLC Chromatogram of EXTRACT OF OLIVE LEAVES AND STEMS, representing the peak of the element Oleuropein. QUANTITATIVE ANALYSIS OF OLEUROPEIN. Project name IQK Instrument Name HPLC-Quality Sequence acquisition date 2023-05-04 12:44:12-07:00 Injection Acq Method Name OLEU.MP.amx Injection Acq Software Agilent OpenLab Acquisition Version 2.4 - Build 2.4.0- REL_2.4.0.628 Injection volume 20,000 μL EndTime signal 29.997. TOOLS Module Type Part. Serial No. Firmware No. Grad. Yam Pump G4294B DECAH00336 B.07.26

[0001] Sampler Name G4294B DECAH00336 B.07.26

[0001] Column Furnace Column Comparison G4294B DECAH00336 B.07.26

[0001] DAD Detector G4294B DECAH00336 B.07.26

[0001] CHROMATOGRAPHIC CONDITIONS Acquisition method: OLEU.MP.amx Path C:\CDSProjects\IQK\Results\OELUROPEINA MAYO 2023\EXTRACT.sirslt Information about the method Last saved as: C:\CDSProjects\IQK\Methods\OLEU.MP.amx Modified: 2023-05-03 16:34:37-07:00 Modifier: RICARDO CAZARES Created: 2023-05-03 16:34:37-07:00 Creator: RICARDO CAZARES Description: Version: 2023-0503-2334-37028 Properties of the Method Instrument Technique: Liquid Chromatography Scheme version Scheme version: 2.3 Grad. Pump method Flow: 1,000 mL / min Low limit pressure: 0.00 bar High pressure limit: 600.00 bar Maximum flow gradient: 100,000 mL / min 2 . Primary Channel: Automatic Module Screen Name: Grad. Pump Module Type: G4294B Order 1 Solvent composition Sampler Method Module Display Name: Sampler Module type G4294B Order 1 Column furnace method Module Display Name: Column Furnace Module type G4294B Order 1 DAD Method Peak width: > 0.1 min (2 s response time) (2.5 Hz) Slot: 4 nm UV lamp required: Yes Requires Vis lamp: Yes Module name: DAD Module type G4294B Order 1 TUECTION RESULTS

[0040] Figure 4 Representative HPLC Chromatogram of EXTRACT OF OLIVE LEAVES AND STEMS QUANTITATIVE ANALYSIS OF OLEUROPEIN Project name IQK Instrument Name HPLC-Quality Sequence acquisition date 2023-05-04 12:12:09-07:00 Injection Acq Method Name OLEU.MP.amx Injection Acq Software Agilent OpenLab Acquisition Version 2.4 - Build 2.4.0- REL_2.4.0.628 Injection volume 20,000 μL Signal end time 29.996 TOOLS Module Type Part. No. Serial No. Firmware Grad. Pump Pump G4294B DECAH00336 B.07.26

[0001] Sample Sample G4294B DECAH00336 B.07.26

[0001] Column Oven Column Compartment G4294B DECAH00336 B.07.26

[0001] DAD Detector G4294B DECAH00336 B.07.26

[0001] CHROMATOGRAPHIC CONDITIONS Acquisition method: OLEU.MP.amx Path: C:\CDSProjects\IQK\Results\OELUROPEINA MAYO 2023WITRA 1.sirslt Information about the method Last saved as: C:\CDSProjects\IQK\Methods\OLEU.MP.amx Modified: 2023-05-03 16:34:37-07:00 Modifier: RICARDO CAZARES Created: 2023-05-03 16:34:37-07:00 Creator: RICARDO CAZARES Description: Version: 2023-0503-2334-37028 Properties of the Method Instrumental Technique: Liquid Chromatography Scheme version Scheme version: 2.3 Grad. Pump method Flow: 1,000 mL / min Low limit pressure: 0.00 bar High pressure limit: 600.00 bar Maximum flow gradient: 100,000 mL / min 2 . Primary Channel: Automatic Module Screen Name: Grad. Pump Module type G4294B Order 1 Solvent composition Sampler Method Module Display Name: Sampler Module type G4294B Order 1 Column furnace method Module Display Name: Column Furnace Module type G4294B Order 1 DAD Method Peak width: > 0.1 min (2 s response time) (2.5 Hz) Slot: 4 nm UV lamp required: Yes Requires Vis lamp: Yes Module name: DAD Module type G4294B Order 1

[0041] Figure 5 Corresponds to the HPLC Chromatogram of D Standard - Mandelonithlo QUANTITATIVE ANALYSIS OF D-MANDELONITRILE Project name IQK Instrument Name HPLC-Quality Sequence acquisition date 2021 -06-08 14:47:30-07:00 Injection Acq Method Name B-17.amx Injection Acq Software Agilent OpenLab Acquisition Version 2.4 - Build 2.4.0-REL_2.4.0.628 Injection volume 20,000 μL Final signal time 5.995 TOOLS Module Type Part. Serial No. Firmware No. Pump Grad. Pump G4294B DECAH00336 B.07.26

[0001] Sampler Sampler G4294B DECAH00336 B.07.26

[0001] Column furnace Column compartment G4294B DECAH00336 B.07.26

[0001] DAD Detector G4294B DECAH00336 B.07.26

[0001] CHROMATOGRAPHIC CONDITIONS Acquisition method: B-17.amx Path: C:\CDSProjects\IQK\Results\B- 17\PT208.06.2021 .sirslt Method information Last saved as: C:\CDSProjects\IQK\Methods\B-17.amx Modified: 2021-04-16 12:39:05-07:00 Modifier: RICARDO CAZARES Created: 2021-03-09 10:17:09-08:00 Creator: RICARDO CAZARES Description: Version: 2021-0416-1939-05260 DAD Method Peak width: > 0.1 min (2 s response time) (2.5 Hz) Slot: 4 nm UV lamp required: Yes Requires Vis lamp: Yes Module name: DAD Module type G4294B Order 1 Column furnace Method module display name: Column Homo Module type G4294B Order Grad. Pumping Method Flow rate: 1,100 mL / min Low pressure limit: 0.00 bar High pressure limit: 600.00 bar Maximum flow gradient: 100,000 mL / min 2 . Primary Channel: Automatic Module Screen Name: Grad. Pump Module type G4294B Order 1 Solvent composition Sampler Method Module Display Name: Sampler Module type G4294B Order 1 Method properties Instrument technique: Liquid chromatography Scheme version Scheme version: 2.3 INJECTION RESULTS Signal: DAD1 B, Sig=215.4 Ref=off Peak Area Width Peak Height RT [min] plate Name Area Area ure s by % Max% Type meter (50%) USP D- 1695.474 1.115 11157.4267 91.4821 100.000 0.104 MANDELONITRILE MM m 284,708 1.455 1038.8743 8.5179 9.311 0.067 MB m Sum 12196.3010

[0042] Figure 6 Corresponds to: Representative HPLC Chromatogram of Curcumin (BAS-NA LIPOSOMAL:1004-19) RT (min) 2,604 Area 980. 9348 BDMC, RT (min) 2,950 Area 1319.6872 DMC, RT (min) 3,353 Area 6829.0605; Dilution factor 20:50 QUANTITATIVE ANALYSIS OF CURCUMIN Project name IQK Instrument Name HPLC-Quality Sequence Acquisition Date 2020-01-09 08:47:36-08:00 Injection Acq Method Name CURCUMINA LUIS.amx Injection Acq Software Agilent OpenLab Acquisition Version 2.4 - Build 2.4.0- REL_2.4.0.628 Injection volume 20,000 μL EndTime Signal 4.996 TOOLS Module Type Part. Serial No. Firmware No. Pump Grad. Pump G4294B DECAH00336 B.07.26

[0001] Sampler Sampler G4294B DECAH00336 B.07.26

[0001] Column oven Column compartment G4294B DECAH00336 B.07.26

[0001] DAD Detector G4294B DECAH00336 B.07.26

[0001] CHROMATOGRAPHIC CONDITIONS Acquisition method: CURCUMINA LUIS.amx Path: C:\CDSProjects\IQK\Results\HPLC-Quality-2020- 01-09 08-47-34-08-00. rsl Method Information Last Saved As: C:\CDSProjects\IQK\Methods\CURCUMINA LUIS.amx Modified: 2019-12-28 14:39:57-08:00 Modifier: RICARDO CAZARES Created: 2019-07-13 09:40:19-07:00 Creator: RICARDO CAZARES Description: CURCUMINA LUIS Version: 2019-1228-2239-57131 DAD Method Peak width: > 0.1 min (2 s response time) (2.5 Hz) Slot: 4 nm UV lamp required: Yes Requires Vis lamp: No Module name: DAD Module type G4294B Order 1 Column furnace method Module Display Name: Column Furnace Module Type: G4294B Order 1 Grad. Pumping Method Flow rate: 1,500 mL / min Low pressure limit: 0.00 bar High pressure limit: 600.00 bar Maximum flow gradient: 100,000 mL / min 2 . Primary Channel: Automatic Module Screen Name: Grad. Pump Module type G4294B Order 1 Solvent composition Sampler Method Display Module Name: Sampler Module Type: G4294B Order: 1 Method properties Instrument technique: Liquid chromatography Scheme version Scheme version: 2.3 INJECTION RESULTS Signal: DAD1 B, Sig=425.4 Ref=off RT [min] Name Area Area% Max Peak% Height Type Width (50%) 1.184 28.9307 0.3130 0.423 5.544 VV 1.438 19.5400 0.2114 0.286 2.278 VV 1.629 15.2067 0.1645 0.222 1.789 VV 1.772 15.0410 0.1627 0.220 2.726 VV 1.959 22.0063 0.2381 0.322 4.036 VB 2,595 981.9250 10.6243 14,361 188,393 BB 2,941 1322.1636 14.3057 19,337 236,015 BV 3,340 6837.4003 73.9801 100,000 1110,993 VB Total 9242.2136 Table 1. HPLC Curcumin Results RT (minutes), Area and Height.

[0043] Figure 7, this refers to the analysis of olive oil reporting And corresponding to the following table: Sample from 6 / 21 / 2023 11:17:05 AM

[0044] Figure 8 refers to the analysis of vitamin C and states the following: Sample ID STD SODIUM ASCORBATQ Method Mame: C: \Usars\P ubkADocu menfetAgifendMicroLa b\Methods®STANQAR£SCAUDA£>¿2m Sample Ssw:8 Useradmin Background Soans:32 Qai®'Tífa®:0&19!202t W:28:33 AM Resolution : 4 Range 4000 - &50 System Status: Good Apodizsátón:Happ-Qen«el File Lccáfión:: C;\Users\PublVÁÜótu¡riér!teV^gi(erti\MícrcjLBb\Resijíí:s\£stBsid <sre5\ STÜ ASCOS BATO DE SODIUM„2Q21'O849TÍ0>23GS.a2r Where said sample was analyzed on June 21, 2023.

[0045] Figures 9, 10 and 11 refer to the analysis of particle size B of the complex with nanoliposomal iron, carried out by the National Technological Institute of Mexico in Tijuana, where it is reported:

[0046] Figures 12 and 13 refer to the particle size analysis of liposomal colostrum, prepared by the Technological Institute of Mexico, with good quality results, reporting the following: SAMPLE: LIPOSOMAL COLOSTRUM (KEMED / FOR ONLY). Analysis: Hydrodynamic diameter (Dh) Sample preparation: The sample was diluted in distilled water and measured after 24 h and 48 h. Measurement temperature: 25 °C. The analysis was performed in triplicate. For the LIPOSOMAL COLOSTRUM sample, the Dh (number %) is 75.05 nm (99.2%) with a unimodal distribution. Quality of the results: The polydispersity value of this sample is large. (Figure 12) SAMPLE: LIPOSOMAL COLOSTRUM (KEMED / SOLO PARA). Analysis: Hydrodynamic diameter (Dh) Sample preparation: The sample was diluted in distilled water and measured after 24 h and 48 h. Measurement temperature: 25 °C. The analysis was performed in triplicate. For the LIPOSOMAL COLOSTRUM sample, the Dh (% number) is 75.05 nm (99.2%) having unimodal distribution. Quality of the results: The value of polydispersity of this sample is large. Analysis: Hydrodynamic diameter (Da) Sample preparation: The sample was diluted in distilled water and measured after 24 h and 48 h. Measurement temperature:25 °C. The analysis was earned out in triplicate. For the sample CALOSTRO LIPOSOMAI .. she Dh (% number) is 75.05 nto having uaimodal distributhm. Result ípaaliíyt The pulydispersity value of this sample is large. Sssnlt Wtedfty ; hsiSs- : te steW repast In this graph we can see that it has a good result, reporting the following: Analysis: Zeta Potential (mV) (Figure 13) SAMPLE: LIPOSOMAL COLOSTRUM (KEMED / FOR ONLY). Sample preparation: The sample was diluted in distilled water and measured after 24 h. Measurement temperature: 25 °C. The analysis was performed in triplicate. The analysis result shows a zeta potential of -39.9 mV (100%). The figure shows the unimodal distribution peak of the analyzed sample. Result quality: good. Analysis: Zeta Potential (mV| SAMPLE: LIPOSOMAL COLOSTRUM (KEMEIW^LY H)«). Sample preparation: The sample wax diluted in distilled wter and measured after 24 h.. Méásisreméni temperature: 25 ®C- Th® analysis; was carried out in triplicate. The result of the analysis shows up to potential of -39.9 mV (100 %). Th® figure shows unimodal dist'-ibuíioti peak of the aaaiyxed sample, Result quality: good.

[0047] Figures 14 and 15 refer to the particle size of Proanselin, produced by the Tijuana Institute of Technology, reporting that it was produced under the following conditions, and considering the product to be of good quality: The dynamic light scattering (DLS) studies for the following nano liposomal sample are reported below: PROANSELIN (PILOT BATCH) ITFOSOMA1... The. analysis carried out was hydrodynamic diameter (Ds) and zeta potential. The equipment used ts a Maiveni Instruments Nano-ZS Nanosizer (ZEN 3OÜ). The mstrumeat is equipped with a helium neon laser (633 nm) with a size detection range of 0.6 nm -5 pm. DLS experiments were performed at the scattering angle of W ; and equilibrated for 2 mia before data collection. Áiiíüysis: íiyclmiiynsimk diameter (IM) Sample preparation:; The sampr? o a-, PJ4b\’ ¡a .h'\ k<? v.jk‘j .mt >v?jmred after 24 h. Measurement tsjmperaiure: 2? \ t he d\ih vs v. as ; n : K d eat m u .phcnte. For the sample PR0ANSEI JN S the D& (% number) is 89 nm (£§. I %) having nnitnodal distributiva. Résiiit quality : gaud. Analysis: Zeta Potencial (mV) Sample preparation:: The sample was diluted m distilled dressing and measured after 24 fa. Measurement temperature: 25 a C. Tire imalysis was carried mt ia tri plicate. The result of tire analysis stows a zeta potential Tire ñgiirs stews the great tolkridsl stability of the analyzed saEtiple.

[0048] Figures 16 to 22 refer to an analysis carried out by the National Technological Institute of Mexico in Tijuana, which reports the following: The dynamic light scattering (DLS) studies of the following nanoliposome samples are presented below: ANTIOXIDANT 10, CURCUFLEX, VITA C PLUS, POWER PEA PROTEIN, VITA+MINS, B COMPLEX WITH IRON, HEALTHY FISH MEAL LIPOSOMAL. The analyses performed were the hydrodynamic diameter (Dh) and zeta potential. The equipment used is a Malvern Instruments Nano-ZS Nanosizer (ZEN 3690). The instrument is equipped with a helium neon laser (633 nm) with a size detection range of 0.6 nm–5 Dm. DLS experiments were performed at a scattering angle of 90° and equilibrated for 2 minutes prior to data collection. An individual report for each sample is attached. Figure 16 refers to SAMPLE: VITA+MINS LIPOSOMAL Aíialysis: Hydrtídyssniii' diameter (1K) Sample preparation: The sample was diluted in distilled water and measured after 24 h. Measutmetn temperature. 25 °C The a»alysis was mmed out in triplieafe. M) Wviag La figura 17, corresponde a: MUESTRA: HARINA DE PESCADO SALUDABLE LIPOSOMAL. Analysis: llydr^ymunic diameter (IM Sa.tspk pi'.'p.Jt The sample was diluted in distilled water and measured after 24 h. MeasüfViiK'ht '.empsnítarS: 25 ®C, The -analy sis w» carried oúi isi triplicate. For the sample HEALTHY FISH MEAL LIPOSOMAL, the D Si (% uante) is 130 nm (100 %) having unimodai distribution. Result quality; gmxL SU8(á.aaik W tíesríW.' Sí Bes ÍOBpt Peats, t: 130 W.& SFS3 Peak A 0.0 ®.S 0.0 ¡W 3: & o Bh 8» La 18 corresponde a: MUESTRA: ANTIOXIDANTE 10 LIPOSOMAL. Analysis: Hydr&dystamíc diameter (EM Sample proparariten: The sample was dilute! m disriHed: waler and measured: after 24 k MessürsHíéíir temperature: 25 ®Q The analysis was carried »in in triplícate. Fm-thc sample ANTfOXIOANT 10 iAPOSOMAI^ the '& (% number) is 122.4 mn (W %) ttevmg uiúutódai di&tributkm. Result quality: gm>d« Bte (OW te itemtem Si W (On$ esu t t|íía y : La Figura 19 corresponde a: MUESTRA: CURCUFLEX LIPOSOMAL Analysis: Hydrodynamic diameter (Ba) Sample preparaiter: The sample was -diluted in distilled water and measured after 24 h. Measurement temperature: 25 ®C. The analysis was carried out its triplicate. For the siuiipls CIMCVFLEX UPOSOMAL^ the Ihs (% nambcr) is 45.71 M (99.11 %) having unisa&dal distribute». Result quality; gwt La figura 20 corresponde a: MUESTRA: COMPLEJO B CON HIERRO LIPOSOMAL Analysis: Hydrodynamic diameter (IM Sample preparatíen.- The sample was diluted in distilled water and mwamd after 24 h. Mestetrenwi temperature; 25 i Y'. The analysis was carried out in triplicate; Far the sample B COMPLEX WITH IRON LIPOSOMAL, site a (% uamber) is 59.13 am (W %) having unimodal distribution. Result quality: goat!» sus g&mh =% teteter.- & Bte temte La figura 21 corresponde a: SAMPLE: VITA C PLUS LIPOSOMAL Analysis: Itydr&dynsmle ditunefer (ESfc) Sampk pteparat-ont 1 to.- mmpk' toas dt ktivd in dt'dilk'd u a ter and n . »'i\ d ahss 24 h. Mcgsaremersi KmpcraHjn.: 25 T' Fisa íspaksís uss earned t»ui in tnpheaie. For tito sample VITA C FLUS LIPOSOMAL, the IA, (% number I is 45 SS UKS (9S5 having tmtmodnl distab«ito.to Result quality; gwd> La figura 22 corresponde a: MUESTRA: PROTEÍNA DE GUISANTE POWER LIPOSOMAL

[0049] Figures 23 to 28 refer to another analysis performed by the Technological Institute of Mexico Tijuana Campus, in reference to the DLS analysis of particle size, where this report presents the results performed on our liposomal OME-NA and liposomal SEL-NA samples. It is noted that hydrodynamic diameter (Dh) and zeta potential analyses were carried out on the liposomal OME-NA and liposomal SEL-NA samples. The dynamic light scattering (DLS) equipment was used to perform the measurements and has the following characteristics: Malvern Nano-ZS Nanosizer instruments (ZEN 3690). The instrument is equipped with a helium-neon laser (633 nm) with a detection range of 0.6 nm - 5 pm. DLS experiments were carried out with a scattering angle of 90° and equilibrated for 10 minutes before each data collection.

[0050] Figure 23 refers to SAMPLE: LIPOSOMAL OME-NA, reporting: Anafek: Hydrodynamic diameter (Dfe Figure 25 corresponds to SAMPLE: OME-NA and reports: Analysis: Pderód Team; fJLS Preparation: The rwfcstra was diiulda using distilled water. Measuring temperature: 25 ®C. The ;.ÍSU&Í5 was carried out by tópUcsde. The result of the attallsis shows a zeta of '9.59 jtnV (area i 00%). The figure shows the great colloidal stability of our analyzed sample. Figure 26 corresponds to the SEL-NA SAMPLE generating the following information: Array: Hydrodynamic diameter ID«) Preparation: The nwestfa fits diluted using distilled water. Measuring temperature. S -'C. The analysis, of redúd by tnpae.*do As can be seen, the Dti (H in number) is 79.B8 nm>(W0%) and the distribution wmtíKÍaL Ltisge is observed, the .LE (H in Irs tosidad) d cusí is ds 22115 nía (M.é %X Siss gUrnb b bwber; $iw lOsny s'.-AWí vyv glsss and DSS Pwk'E D : ss i».e 3S.43. Figure 27 also corresponds to the SEL-NA SAMPLE reporting: Figure 28 corresponds to: a SAMPLE: SEL-NA LIPOSOMAL giving the following data: Analysis; Potential zsia Es^iEpíi: PLS Preparation; U sample ¡Diluted using distilled water. Temperature Je i'sov ucr 25 §C The saáUsis of realizo pnrtnplíeadto> From the resuksde ee m pvteuc'ál zeta of -34.4 a¥ (100% area). From the figure are pwds : obsm ; ar la gmn tsiaNitdau c^Undai Ju la m «is saallized anger,

[0051] Figures 29 and 30 refer to SAMPLE: VITAMIN C WITH ZINC NANO LIPOSOMAL LOT No. 509-20, produced by the National Technological Institute of Mexico, Tijuana campus, which presents: Analysis: Hydrodynamic diameter (Dh) Sample preparation: The sample was diluted in distilled water and measured after 24 h. Measuring temperature: 25 °C. The analysis was performed in triplicate. For the sample VITAMIN C WITH NANO LIPOSOMAL ZINC (LOT #509-20), the Dh (% number) is 18.29 nm (100%) having a unimodal distribution. The Dh (% intensity) is 236.3 nm (81.7%) and 27.65 nm (18.3%) showing a bimodal distribution. Quality of the results: See the quality report. Figure 29 Figure 30

[0052] Figure 31 refers to: PARTICLE SIZE of Cardio Minerals Liposomal Support, where the distribution we can observe that: 80% measure between 10 nm - 66 nm 20% measures between 122nm - 293nm

[0053] Figure 32 represents the PARTICLE SIZE of Cardio Liposomal Support that reports: 90% measure between 10 nm - 62 nm 10% measure between 126 nm -139 nm

[0054] Figure 33 refers to the PARTICLE SIZE of Cardio Liposomal Support that reports: 90% measure between 10 nm - 62 nm 10% measure between 126 nm -139 nm

[0055] Figure 34 represents the PARTICLE SIZE Cardio Liposomal Support Minerals reporting: 80% measure between 10 nm - 66 nm 20% measures between 122nm - 293nm From all the figures previously reported we can see that these correspond to graphs that confirm the good quality of the nanoliposomes of the present invention, as well as their effective use as carriers for different active substances such as vitamins, supplements or drugs, for which reason we will proceed to claim the invention once we have described its details.

[0056] Figures 35 to 38 are very representative because the long-term Proanseline result (stability) shows what was mentioned about the particle size and z potential in stability studies. Dynamic light scattering (DLS) studies for the following nanoliposomal samples are reported below: PROANSELIN (PROANSELIN) (Long Term Stability and Accelerated Stability) LIPOSOMAL. The analysis performed was hydrodynamic diameter (Dh) and zeta potential. The equipment used is a Nano-ZS Nanosizer from Malvern Instruments (ZEN 3690). The instrument is equipped with a helium neon laser (633 nm) with a size detection range of 0.6 nm–5 pm. DLS experiments were performed at scattering angle of 90° and equilibrated for 2 min before data collection. An individual report is attached for each sample. These figures represent the following: Figure 35 SAMPLE: LIPOSOMAL PROANSELIN (long-term stability) Analysis: Hydrodynamic diameter (Dh) Sample preparation: The sample was diluted in distilled water and measured after 24 h. Measurement temperature: 25 °C. The analysis was performed in triplicate. For the PROANSELIN sample (long-term stability), the Dh (number in %) is 36.64 nm (100%) with a unimodal distribution. Result quality: good. that's qsrn ty í > Figure 36 Analysis: Zeta Potential (mV) Sample preparation: The sample was diluted in distilled water and measured after 24 h. Measurement temperature: 25 °C. The analysis was performed in triplicate. The analysis result shows a zeta potential of -41.7 mV (100%). The figure shows the high colloidal stability of the analyzed sample. Figure 37 SAMPLE: PROANSELIN LIPOSOMAL PILOT BATCH (accelerated stability) Analysis: Hydrodynamic diameter (Dh) Sample preparation: The sample was diluted in distilled water and measured after 24 h. Measurement temperature: 25 °C. The analysis was performed in triplicate. For the PROANSELIN sample (accelerated stability), the Dh (number %) is 46 nm (100%) with a unimodal distribution. Result quality: good. Figure 38 Analysis: Zeta Potential (mV) Sample preparation: The sample was diluted in distilled water and measured after 24 h. Measurement temperature: 25 °C. The analysis was performed in triplicate. The analysis result shows a zeta potential of -46.6 mV (100%). The figure demonstrates the high colloidal stability of the analyzed sample.

[0057] Figure 39 shows the intestinal permeability test obtained after ingesting the formulation of the present invention containing iron, which shows an increase in the absorption of said trace metal. That is, according to the results obtained, the amount of iron permeated compared to other pharmaceutical formulations is highly acceptable (85.39%). Detailed Description of the Invention

[0058] The following definitions are provided for the purpose of enabling a better understanding of the invention:

[0059] “Approximately.”—The use of the term “approximately” provides a certain additional range with respect to the numerical value to which it is being applied. This additional range is approximately ± 10%. For example, but not limited to, if “approximately 10 liters” is used, the exact range described and / or claimed is between 9 and 11 liters.

[0060] Trace minerals and their composition: Generally, between 72 and 76 trace minerals are present in the waters or salt of the Sea of ​​Cortez. The amount of trace minerals varies in composition and concentration depending on the time of collection, since evaporation from the sea is greater in the summer and the mineral contribution is greater during the rainy season, mainly due to the rainfall from the Colorado River, which passes through four states of the American Union and empties into the port of San Felipe in the Sea of ​​Cortez, where the Baja California Peninsula begins.

[0061] Minerals comprise 5% of the normal human diet, but are essential for health and optimal body function. Minerals can be classified into different groups. Macrominerals: Minerals that adults require in amounts greater than 100 mg / day or that constitute <1% of total body weight. Trace minerals: Minerals that adults require in amounts of 1 to 100 mg / day or that constitute less than 0.01% of total body weight. Ultratrace minerals: Minerals required in amounts less than 1 g / day.

[0062] Nanotechnology is the understanding and control of matter at the nanoscale, with dimensions between 1 and 100 µm. It is used to create and use structures, devices, and systems that have new properties and functions due to their size.

[0063] Nanos = Dwarf and Manometer = billionths of a meter

[0064] A liposome is a nanobubble made from natural oils that has the capacity to transport bioactive substances (vitamins, minerals, amino acids, etc.) inside or in its membrane. They have a Excellent tolerance thanks to its composition, identical to that of our cell membranes, and the low concentration of active ingredients. They allow for a controlled release of the active ingredients contained, enhancing the product's bioavailability (extended release). Their particle size, less than 100 nanometers, allows them to be rapidly absorbed by the intestinal wall, significantly increasing the bioavailability of nutrients. Their structure, almost identical to the cell wall, allows for more efficient fusion and delivery and absorption up to 9 times greater than tablets or capsules of similar formulas, which increases the beneficial effect of the supplement to a greater percentage.

[0065] An intestinal permeability study is a test performed in a New York laboratory called Creative Bioarray:

[0066] By assessing compound permeability in the early stages of drug discovery, compounds most likely to pose challenges during preclinical and clinical development can be identified. The permeability of compounds through cell membranes, such as the intestinal epithelium, is a critical characteristic that determines the rate and extent of human absorption and ultimately impacts the bioavailability of a drug candidate. Compounds with poor permeability are more likely to have poor absorption, distribution, metabolism, excretion (ADME), and pharmacokinetic (PK) properties.

[0067] Nanocarriers: These are capable of directing active substances to the site where they are required, with the goal of maintaining and controlling drug delivery in time and space. First-generation nanocarriers They comprise a passive delivery system that is localized within the target site where the contents are released. An example of this is nanoparticles or liposomes, which, when administered with the intention of reaching a tumor as the target tissue, are forced to accumulate at the site of action due to a "promoted permeation and retention" (EPR) effect. This effect occurs because tumors present fenestrations or interruptions in the cell lineage of the epithelium adjacent to the vascularization.

[0068] PROCESS FOR PRODUCTION OF ALCOHOLIC EXTRACT FROM OLIVE LEAVES AND STEMS 1. Add the crushed or powdered olive leaves and pure ethyl cane alcohol to a glass container in a ratio ranging from approximately 75:25 to approximately 25:75 respectively. 2. Shake vigorously for approximately 10 minutes. 3. Let stand for 14 days, shaking vigorously for approximately 10 minutes once a day during the stand time. 4. At the end of the resting time, filter through rapid filter paper to remove larger particles. 5. Pass the solution through a fiberglass filter. 6. Concentrate the volume of the solution obtained in step 5 in a rotary evaporator until it reaches approximately 20 to 30% of the initial volume. 7. Take a sample portion and analyze according to the Oleuropein analysis method, to verify that the solution obtained in step 6 contains between 30 to 35% v / v of said component. 8. If it is below the specification mentioned in step 7, evaporation is continued in relation to the % of the component until the required 30 to 35% v / v is reached. 9. Once the required % of Oleuropein has been reached, it is filtered through a sterile 0.45 micron filter to achieve sterility of the Olive Leaf and Stem Extract. 10. Store in innocuous containers that are selected from previously sanitized glass containers, identify with the name of the extract, the date of preparation and the % v / v of Oleuropein contained, 11. Use for a period of no more than 12 months, after this time, re-evaluate in HPLC.

[0069] CRYSTALLIZATION PROCESS OF TRACE MINERALS FROM THE SEA OF CORTEZ 1. Add trace minerals from the Sea of ​​Cortez to a container and pour hot purified water in a 25:75 ratio. 2. Mix with constant stirring and heating until a homogeneous solution is achieved. 3. Cool to room temperature. 4. Add pure ethyl cane alcohol and mix with constant stirring without heating. The ratio of alcohol to be added is approximately 50:50, with respect to the mixture of minerals and water. 5. Let the mixture rest for 18 to 24 hours. 6. Decant the mixture, discarding the precipitate (sodium salts). 7. Heat the mixture resulting from step 6 to a temperature ranging from approximately 55°C to approximately 60°C (boiling and distillation temperature) in order to evaporate the alcohol present in the mixture. 8. Filter the aqueous solution from step 7 containing the trace minerals through a glass fiber filter. 9. Finally, filter with the help of a vacuum, using a sterile 0.45 micron filter, to obtain a sterile solution of trace minerals from the Sea of ​​Cortez. 10. Take a portion of the sample and carry out the microbiological analysis. 11. Place the filtrate in a previously sanitized container. 12. Identify with the name of the solution and the date of preparation. 13. Use for a period of no more than 24 months; after this time, re-evaluate through microbiological analysis and assess the concentrations of each of the trace minerals using ICP or atomic absorption.

[0070] It is understood by a technician with average knowledge in the field that steps 10 and subsequent ones correspond to a standard quality control and are not part of the invention, but are listed herein. request, since they are elements that demonstrate the quality that we seek to preserve and obtain in our products.

[0071] EMULSIFICATION PROCESS

[0072] The emulsification process of water-soluble and fat-soluble ingredients to obtain a nanoemulsion is carried out as follows:

[0073] In the present invention, the method for obtaining the nanoemulsion is through micronization, which allows the nanoemulsions to remain stable for up to 18 months (particle size distribution and z potential studies are attached in accelerated stability studies (see figures and their description, especially figures 35 to 38), as mentioned in the NOM-073-SSA1 -2015 standard, Stability of drugs and medications, as well as herbal remedies).

[0074] Our invention has established a maximum shelf life of 18 months based on the results obtained in stability tests.

[0075] In the present invention, the method for obtaining the nanoemulsion is by means of immersion micronization with a head, formed by a rotor and a toothed stator made of 316 L stainless steel (pharmaceutical grade), which is driven by a 3.0 horsepower (HP) explosion-proof motor, our equipment reaches up to 20,000 revolutions per minute (RPM), regulated by an Ultra Turrax type central gearbox.

[0076] This process allows the nanoemulsions to remain stable for up to 18 months, obtaining a particle size between 5 nm and 100 nm, with a positive Z potential (+), a) as we mentioned, studies are attached Particle size distribution and z-potential in accelerated stability studies, as mentioned in the NOM-073-SSA1-2015 standard, Stability of drugs and medications, as well as herbal remedies, the evaluations were carried out by the National Technological Institute of Mexico, Tijuana campus.

[0077] It is understood by a technician with average knowledge in the area that this equipment is an illustrative example of how to put the invention into practice, but that any other existing technology that achieves the same effect of obtaining nanoemulsions with these characteristics is covered within the scope of the present invention.

[0078] The liposome preparation process consists of the following steps: 1. Dissolution Phase: i. Water-soluble active ingredients: Dissolve the water-soluble ingredients in 50% of the total solvent in the formula, dividing the 50% by the number of ingredients to achieve the dissolution independently. Finally, mix each of the solutions obtained, except for the flavoring solution. i. Fat-soluble active ingredients: Dissolve the fat-soluble ingredients in 7% of the extra virgin olive oil, dissolving the ingredients one by one. Where: WATER-SOLUBLE ACTIVE INGREDIENTS. - Vitamin C (Ascorbic Acid) - Vitamin C (Sodium Ascorbate) - Vitamin C (Calcium Ascorbate) - Vitamin B1 (Thiamine HCl) - Vitamin B3 (Pyrophosphate) - Vitamin B3 (Niacin / Niacinamide) - Vitamin B6 (Phydoxin HCl) - Vitamin B12 (Cyanocobalamin / Hydro / Methyl cobalamin) - Vitamin B9 (Folic Acid sodium salt) - Vitamin B17 (Amygdalin) - Glutathione HCL / Sodium Salt - L-Lysine HCL / Sodium Salt - L-Arginine HCL / Sodium Salt - Calcium (Citrate / Gluconate) - Magnesium (Citrate / Chloride) - Potassium (Chloride) - Zinc (Sulfate) - Iron (Sulfate) - EDTA (Disodium Salt) - Amygdalin - Herbal extracts (Hydroalcoholics) examples: olive, curcumin, silymarin etc. FAT-SOLUBLE ACTIVE INGREDIENTS - Coenzyme Q10 - Vitamin D3 - Vitamin E - Vitamin A - Lipoic acid - Omega 3 - Omega 6 - Vitamin K2

[0079] The quantities will depend on the active ingredients to be dissolved, that is, if 70% of the active ingredients in the formulation are water-soluble, it means that the formulation will have 70% water and only 7% extra virgin olive oil; otherwise, the formulation will contain 8% extra virgin olive oil and 60% purified water.

[0080] 2. Micronization process:

[0081] Pour the aqueous solution into a container with a maximum capacity of 90% of the batch size.

[0082] Place under the micronizer and turn on the equipment for approximately 15 minutes at a speed between 15,000 and 18,000 RPM (revolutions per minute).

[0083] In a separate container with constant stirring, add the surfactant to the oil mixture and stir for a period of 5 minutes.

[0084] With the micronizer on, add the oil-surfactant mixture slowly over a period of no more than 10 minutes.

[0085] 3. Microemulsion:

[0086] Micronize the aqueous-oil-surfactant mixture for a period of 15 minutes between 15,000 and 18,000 RPM.

[0087] 4. Stabilization:

[0088] With the micronizer on, slowly add the olive stem and leaf extract over a period of no more than 10 minutes. Continue with the equipment running at 15,000 to 18,000 RPM for approximately 5 minutes. Turn off the micronizer.

[0089] 5. Flavoring:

[0090] Turn on the micronizer and add the natural and artificial flavoring solution slowly over a period of no more than 10 minutes.

[0091] Obtaining the nanoemulsion:

[0092] Continue micronizing the total mixture for 5 more minutes, diluting with solvent to the batch size indicated in the master production order.

[0093] Take samples of the nanoemulsion by quality control to perform the following analyses: 2. Organoleptic tests: taste, color, texture, appearance. Physicochemical analysis: pH, density, microscopy, ingredient titration by HPLC, particle size and Z potential. 4. Microbiological analysis: aerobic mesophiles, fungi and yeasts, pathogens (E. Coli, Staphylococcus spp and Salmonella spp). 1. When it comes to a modification to the ingredients, change of supplier of inputs, new formulation or new product, stability tests are used based on the NOM-073-SSA1-2015 standard, Stability of drugs and medicines, as well as herbal remedies, upon compliance with the time established by the standard (6 months for accelerated stability), the product is marketed.

[0094] Examples

[0095] The following are some technical examples of how to carry out the invention, but it will be understood that one skilled in the art understands that obvious variations thereon are covered within the protection of the present invention, since these are illustrative but not limiting.

[0096] Liposomal B complex • Only 15 mg of Iron (low doses) • Ideal for anemia • No side effects With olive leaf extract

[0097] Using the World Health Organization (WHO) hemoglobin cutoff points, it is estimated that anemia affects more than 2 billion people worldwide.

[0098] Iron deficiency anemia (IDA) remains the most common type of anemia worldwide.

[0099] In Figure 39 we can observe the permeability obtained when ingesting the formulation of the present invention and which allows achieving high assimilation of Fe with low doses.

[0100] According to the Cenetec clinical guide, elemental iron is required in adults at a dose of 180 mg / day divided into three doses. This is related to the problems with current iron formulations, which are: • Low bioavailability • High doses are required • Long-term treatments (up to 6 months) • Low patient compliance and • Various side effects such as gastrointestinal pain, constipation, diarrhea, nausea, vomiting, heartburn, fatigue, redness, etc.

[0101] Our formulation reduces side effects, shortens treatment time, requires a lower dose of approximately 30 mg, increases bioavailability, is stable for 18 to 24 months, is versatile, low-cost, and increases compliance. This formulation is achieved with iron-containing nanoparticles, with particle sizes ranging from 5 to 75 nm, and is suitable for oral administration.

[0102] EDTA

[0103] It is a molecule used to detoxify heavy metal contamination in contact with humans. Its target is minerals in the bloodstream and binds to them through four carboxylate groups and two metallic amine groups that can act as electron pair donors or Lewis bases to form covalent bonds combined with cations, making it a hexadentate ligand and the most important of the chelate ligands.

[0104] FORMING LINK: □ Mn (II) □ Cu (II) □ Fe (III) □ Co (III).

[0105] This shows the Lewis structure, where the free electron pairs of oxygen and nitrogen are seen.

[0106] EDTA AS DISODIUM SALT

[0107] The reagent commonly used is its disodium salt, since in practice EDTA is usually partially ionized, forming fewer than six coordinated covalent bonds with metal cations.

[0108]

[0109] It has 6 bonds, as it forms a hexaprotic system, giving rise to a complex with an octahedral structure.

[0110] DISODIUM EDTA Vs EDTA

[0112] Disodium EDTA SOLUBILITY: 100 g / l DENSITY: 700 g / MI MELTING POINT: 110 °C pH: 4-5 MOLECULAR WEIGHT: 372.24 g / mol EDTA SOLUBILITY: 0.5 g / l DENSITY: 700 g / MI MELTING POINT: 220 °C (DECOMPOSITION) pH: 2.5 MOLECULAR WEIGHT: 292.24 g / mol

[0113] From the previous examples we can see the advantages of using each of our components and their practical application in each formulation. Another example of application is in formulations with vitamin C.

[0114] Vitamin C has multiple health benefits, such as antioxidant, anti-tumor, anti-aging and helping collagen; Vitamin C can prevent, help and even cure several viral and bacterial diseases, since the action of vitamin C is dual since on the one hand it strengthens the immune system and on the other it limits inflammatory damage, which begins when the defenses detect an infectious agent, so that the symptoms are milder, and recovery faster, unfortunately, vitamin C has low intestinal absorption and rapid elimination in urine, reasons why it is a challenge to reach the doses required to obtain all its benefits.

[0115] Its arrival to the cells is an obstacle course, since the vitamin C that we ingest in foods, supplements, capsules, gummies, solutions, effervescent tablets, lyophilized powders, etc., are absorbed in the body through specific transporters, located in the intestine, these transporters have the ability to introduce only a certain amount of vitamin C per minute, so only the amount of vitamin C that is capable of being transported will be absorbed, it is known that of a gram of vitamin C, only half is absorbed (500mg), the rest is discarded; once in the body, it has a first transformation in the liver, generating another loss when it reaches blood circulation and is distributed to the organs of the body, and begins its elimination process through urine, the entry of vitamin C into the cells will depend again on the quantity and functionality of the transporters located on the cell surface;It is unknown how much vitamin C reaches the cell, although it is estimated to be less than a third of what is ingested, which is why conventional formulations do not reach the concentrations that the body requires to experience all its benefits.

[0116] With nanotechnology, the effectiveness of orally ingested vitamin C can be achieved, since with this technology nanocarriers can be developed for various substances such as vitamin C, and more specifically thanks to the nanoliposomes of the present invention, which are obtained from vegetable fats such as olive oil, and more specifically from olive oil leaves and stems.

[0117] Due to their size and composition, the nanoliposomes of the present invention penetrate directly into the intestine, without requiring any carrier. specific, and its absorption is close to 100%; when the nanoliposome measures less than 100 nm, it is not detected by the liver, passing directly into the bloodstream and organs, distributing throughout the body.

[0118] The surface of nanoliposomes is very similar to the cell membrane, and upon contact, it recognizes it and introduces it directly through a process called endocytosis. This process occurs anywhere on the cell surface, without relying on any specific transporter. Once inside the cell, the nanoliposome opens, releasing the administered substance. As we mentioned for the present invention, this substance may not only be vitamin C, but any other substance such as vitamins, drugs, or nutrients in general. In the case of vitamin C, we can say that practically 100% of the vitamin C administered via this route is absorbed, so one gram of liposomal vitamin C will have a much greater effect compared to any other conventional form of administration.The formulation of our example with vitamin C shows that thanks to this form of administration with the formulation of the liposomes of the present invention, it is possible to reach the concentrations required to strengthen the immune system and maintain the protection of the individual against bacterial and viral diseases, such as the flu, influenza and coronavirus, among others.

[0119] Nanoliposomes are not eliminated through urine because they are not water-soluble, so they remain in the circulation for as long as necessary to be utilized by cells.

[0120] In addition, the nanoliposomes of this invention, made from olive oil, leaves and stems, and minerals from the Sea of ​​Cortez, provide the anti-inflammatory and antioxidant benefits of these substances.

[0121] In general, the percentages of the essential components of the formulations sought to be protected in this patent application are: Main components per dose (15 mL): • Extra virgin olive oil: between 7% and 8%. • Purified water: between 60% and 70%. • Trace minerals: between 0.50% and 1%. • Sweetener: <0.1%. • Flavoring: <1% • Conservative: <0.1% • Olive leaf extract: between 0.50% and 1%. • Active ingredients: between 25% and 30%

[0122] Example 1: Liposomal Vitamin C

[0123] Manufacturing process for 30 kg of new nanoemulsion 1. Dissolve approximately 6 kg of sodium ascorbate (Vitamin C) in approximately 10 liters of purified water at room temperature and stir until completely dissolved and empty everything into the stainless steel production drum. 2. Add the sodium polysorbate with gentle stirring, stir for approximately 5 minutes at 5000 rpm and stop for approximately 5 minutes. 3. Slowly add, with light stirring, the mixture of flavors and preservatives and stir for approximately 5 minutes at approximately 10,000 rpm and stop for approximately 5 minutes. 4. Add, while stirring, approximately 1.5 kg of extra virgin olive oil, stir for approximately 5 minutes at approximately 10,000 rpm and let stand for approximately 5 minutes. 5. Add approximately 180 g of citric acid dissolved in purified water and stir for approximately 5 minutes at approximately 14,000 rpm. Adjust the pH to approximately 4.5 - 4.7. 6. Weigh the total volume and top up the weight to approximately 30 kg with purified water, stir for approximately 5 minutes at approximately 14,000 rpm, and let stand for approximately 24 hours. 7. A retention sample is taken for quality control for physical, chemical and microbiological analysis; once approved by quality control, it is conditioned.

[0124] Example 2: Preparation of liposomal proanselin:

[0125] The active ingredient called D-Mandelonithlo or Amygdalin, is a water-soluble ingredient, as defined above, which is strictly soluble in water.

[0126] The amygdalin content is approximately 1 g per 15 mL (dose). This means that for a batch of approximately 30 L, approximately 2 kg of raw material was weighed.

[0127] To do this, the active ingredient was solubilized in approximately 50% of the total purified water to be used was then micronized for approximately 15 min at approximately 16,000 RPM (revolutions per minute) to achieve the desired particle size. (less than 100 nanometers). The microemulsion, stabilization, and nanoemulsion production processes continued.

[0128] This description has been presented for purposes of illustration and disclosure. The foregoing is not intended to limit the disclosure to the form or forms disclosed herein. In the detailed description above, for example, various features of the disclosure are grouped into one or more aspects, embodiments, and / or configurations for the purpose of simplifying the description. Features of the aspect, embodiments, and / or configurations of the description may be combined into alternative embodiments and configurations other than those discussed above. This description should not be construed as reflecting an intention that the claimed subject matter requires more features than are expressly recited in each claim.

[0129] Although the description of the invention has included one or more features, embodiments or configurations and certain variations, other embodiments, combinations and / or modifications are within the scope of the disclosure, for example, as they may be within the knowledge of the person skilled in the art, after understanding the present invention.

[0130] Having sufficiently described the nature of the invention, as well as the manner of putting it into practice, it should be noted that the provisions indicated above are susceptible to modifications in detail, as long as they do not alter their fundamental principle.

Claims

Claims

1. i A process for obtaining a nanotechnological formulation in liposomal form that improves the absorption in the body of nutritional components and medications characterized in that it comprises the following steps: a. Preparation of the alcoholic extract of olive leaves and stems b. Extraction of trace minerals from the Sea of ​​Cortez c. Emulsification of the mixture d. Preparation of liposomes

2. The process according to claim 1, characterized in that section a) comprises the following steps: a. Add crushed or pulverized olive leaves and stems and pure ethyl cane alcohol to a container, in a ratio ranging from approximately 75:25 to approximately 25:75 respectively; b. Shake vigorously for a time interval of approximately 10 minutes; c. Let it stand for 14 days, shaking vigorously for a time interval of approximately 10 minutes, once a day during the resting time; d. At the end of the resting time, filter through filter paper to eliminate larger particles and obtain a solution; e. Pass the solution through a fiberglass filter; f. Concentrate the volume of the solution obtained in step e) in a rotary evaporator until reaching approximately 20 to 30% of the initial volume; g.Take a sample portion and analyze according to the Oleuropein analysis method, to verify that the solution obtained in step f) contains between 30 to 35% v / v of said component. h. If the content is below the specification mentioned in step g), evaporation is continued in relation to the component percentage until the required 30 to 35% v / v is reached; i. Once the required percentage of Oleuropein is reached, the extract is filtered through a sterile filter of approximately 0.45 microns to achieve sterility. j. Store in innocuous containers selected from previously sanitized glass containers, and label with the name of the extract, the date of preparation, and the v / v percentage of Oleuropein contained. k. Use for a period of no more than 12 months. After this time, reassess by HPLC.

3. The process according to claim 1 characterized in that section b) comprises the following steps: a. Add trace minerals from the Sea of ​​Cortez to a container and pour hot purified water, in a ratio ranging from approximately 25:75 % m / v; b. Mix with constant stirring with heating until a homogeneous solution is achieved; c. Cool to room temperature; d. Add pure ethyl cane alcohol and mix with constant stirring without heating, the ratio of alcohol to be added is approximately 50:50 % v / v, with respect to the mixture of minerals and water; e. Let the mixture stand for a range between 18 and 24 hours; f. Decant the mixture, discarding the precipitate (sodium salts); g. Heat the mixture resulting from step f to a temperature ranging from approximately 55 ° C to approximately 60 ° C in order to evaporate the alcohol present in the mixture; h. Filter the aqueous solution from step g containing the trace minerals through a fiberglass filter; i. Finally, filter using a 0.45-micron sterile vacuum filter to obtain a sterile solution of trace minerals from the Sea of ​​Cortez.

4. The process according to claim 3, characterized in that optionally, item b) includes the steps of: a) Taking a portion of the sample and carrying out the microbiological analysis; b) Placing the filtrate in a previously sanitized container; c) Identifying the solution with the name and the preparation date; d) Using it for a period of no more than 24 months, after which time, re-evaluating it through microbiological analysis and assessing the concentrations of each of the trace minerals by ICP or atomic absorption.

5. The process according to claim 1, characterized in that section c) for obtaining the nanoemulsion comprises immersion micronization with a head, formed by a rotor and a toothed stator of 316 L stainless steel (pharmaceutical grade), which is driven by a 3.0 horsepower (HP) explosion-proof motor, the equipment reaches up to 20,000 revolutions per minute (RPM), regulated by an Ultra Turrax type central gearbox.

6. The process according to claim 5, characterized in that the process allows the nanoemulsions to remain stable for up to 18 months, obtaining a particle size between 5 nm and 100 nm, with a positive Z potential (+).

7. The process according to claim 1, characterized in that item d) consists of the following steps: a. Dissolution phase; b. Micronization process; c. Microemulsion; d. Stabilization; e. Obtaining the nanoemulsion

8. The process according to claim 7, characterized in that the dissolution phase consists of: a) Water-soluble active ingredients: Dissolve the water-soluble ingredients in 50% of the total solvent volume of the formula, dividing 50% by the number of ingredients to perform the dissolution independently. At the end, mix each of the solutions obtained, except for the solution of the flavorings; b) Fat-soluble active ingredients: Dissolve the fat-soluble ingredients in 7% of the volume of the extra virgin olive oil, dissolving the ingredients one by one.

9. The process according to claim 8, wherein the water-soluble active ingredients are selected from: ascorbic acid, sodium ascorbate, calcium ascorbate, thiamine HCl (vitamin B3), niacin (vitamin B3), niacinamide (vitamin B3), Pyhdoxine HCl (vitamin B6), cyanocobalamin (vitamin B12), hydrocobalamin (vitamin B12), methyl cobalamin (vitamin B12), folic acid sodium salt (vitamin B9), Amygdalin (vitamin B17), glutathione HCL / sodium salt, L-lysine HCL / sodium salt, L-arginine / sodium salt, calcium citrate, calcium gluconate, magnesium citrate, magnesium chloride, potassium chloride, zinc sulfate, ferrous sulfate, EDTA disodium salt and herbal extracts in olive alcohol, curcumin, silimahna.

10. The process according to claim 9, wherein the fat-soluble active ingredients are selected from: coenzyme Q10, Vitamin D3, vitamin E, vitamin A, lipoic acid, Omega 3, Omega 6 or vitamin K2.

11. The process according to claim 7, characterized in that the micronization process consists of: a. Pour the aqueous solution into a container with a maximum capacity of 90% of the batch size; b. Place it under the micronizer and turn on the equipment for approximately 15 minutes at a speed between 15,000 and 18,000 RPM (revolutions per minute); c. In a separate container with constant stirring, add the surfactant to the oily mixture and stir for approximately 5 minutes; d. With the micronizer on, add the oil-surfactant mixture slowly over a period of no more than 10 minutes.

12. The process according to claim 7, characterized in that it consists of: Micronizing the aqueous-oil-surfactant mixture for a period of 15 minutes between 15,000 and 18,000 RPM.

13. The process according to claim 7, characterized in that the stabilization stage is carried out with the micronizer on, where the extract of olive stems and leaves is added slowly in a time not exceeding 10 minutes, continuing with the equipment on between 15,000 and 18,000 RPM for a time of 5 minutes, and finally the micronizer is turned off.

14. The process according to claim 7, characterized in that the emulsion is obtained by continuing the micronization of the total mixture for approximately 5 more minutes, filling with the solvent to the batch size indicated in the master production order.

15. The process according to claim 7, characterized in that it optionally has the following steps: a. Add Flavoring: Turn on the micronizer and add the solution of the natural and artificial flavorings slowly in a time not exceeding 10 minutes. b. Take samples of the nanoemulsion by quality control to perform the following analyses: a) Organoleptic tests b) Physicochemical analysis c) Microbiological analysis

16. A nanotechnological formulation in liposomal form based on extra virgin olive oil, alcoholic extract of the stem and leaves of olive tree and trace minerals from the Sea of ​​Cortez obtained with the process of claim 1 characterized in that it is constituted by: a) An oral nanoemulsion of extra virgin olive oil produced from the seeds with an Oleuropein content between 35% and 40%, as antioxidant b) Emulsifying agents c) Extract of trace minerals from the Sea of ​​Cortez d) Optionally a sweetener, a flavoring and a preservative

17. A nanotechnological formulation in liposomal form according to claim 16, characterized in that the method for obtaining the nanoemulsion is by means of micronization, which allows the nanoemulsions to remain stable for up to 18 months.

18. A nanotechnological formulation in liposomal form according to claim 16, characterized in that the emulsifying agent is Polysorbate (60 or 80).

19. A nanotechnological formulation in liposomal form according to claim 18, characterized in that the Polysorbate (60 or 80) is between 7% and 10%.

20. A nanotechnological formulation in liposomal form according to claim 16, characterized in that the salts of the Sea of ​​Cortez water are selected from: Magnesium, Potassium, Boron, Iodine, Molybdenum, Cobalt, Zinc and 72 other minerals.

21. A nanotechnological formulation in liposomal form according to claim 16, characterized in that stevia is used as a sweetener.

22. A nanotechnological formulation in liposomal form according to claim 16, characterized in that potassium sorbate is used as a preservative.

23. A nanotechnological formulation in liposomal form according to claims 16 to 22, characterized in that it is viable for oral administration.

24. A nanotechnological formulation in liposomal form according to claims 16 to 23, characterized in that they are highly efficient carriers for any vitamin, nutrient or medicine or food supplement, i.e.

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