A complex, a method of producing the complex, a micropellet comprising the complex, and a method of producing the micropellet

Abstract

The subject of the invention is a complex comprising resveratrol and leucine, as well as a method of producing the complex according to the invention. The invention also relates to a micropellet comprising the complex according to the invention and a method of producing the micropellet according to the invention.

Description

[0001] A complex, a method of producing the complex, a micropellet comprising the complex, and a method of producing the micropellet

[0002] The present invention relates to the field of sirtuin activators. The invention relates to a complex comprising resveratrol and leucine as well as a method of producing such a complex. The invention further relates to a micropellet comprising the complex according to the invention and a method of producing such a micropellet. The invention is particularly applicable in dietetics, supplementation, and broadly understood medicine.

[0003] Among the intensively studied sirtuin activators, resveratrol has attracted particular attention. However, its poor solubility in gastrointestinal fluids and lipophilic nature result in low bioavailability from solid product formulations. Attempts are known in the state of the art to increase the solubility of resveratrol by complexing it with trimethylglycine (TMG). However, this ingredient is not regarded as compliant with the principles of safe food design and may negatively affect food quality and safety in product formulation and technological process optimization. This eliminates TMG as a complexing agent.

[0004] Considering the above state of the art and its limitations, the objective of the invention was to increase the solubility and bioavailability of resveratrol. Another objective of the invention was to provide sirtuin activators in a micropellet form with improved properties. Finally, the aim was to ensure an improved product form and process optimization.

[0005] These and other technical problems were solved by the complex according to the invention and the micropellet comprising said complex.

[0006] Furthermore, contrary to literature reports and general knowledge, it was found that the expected (low) moisture content of the product could adversely affect system stability (usually water and / or its excess is considered destabilizing). This observation was found to be true not only for the complexes but - unexpectedly - also for the micropellet. Results demonstrated that the water content is crucial, since its deficiency led to undesirable loosening of the complex structure and formation of leucine dimers.

[0007] Equally unexpected was the stabilizing influence on the micropellet of two active substances - quercetin and epigallocatechin gallate (EGCG) - which additionally limit water evaporation from the micropellet. Through optimization of the composition, the use of solvents and energy was minimized (e.g., minimal required water for the process and minimal energy input).

[0008] The subject of the invention is a resveratrol complex characterized in that it comprises resveratrol and leucine. Preferably, the complex according to the invention is characterized in that the molar ratio of resveratrol to leucine is from 1 :1 to 1 :3. More preferably, the molar ratio of resveratrol to leucine is 1 :1.625. Preferably, the complex according to the invention further comprises up to 2% by weight of PEG-3000 (polyethylene glycol), preferably up to 1% by weight of PEG-3000. Preferably, the complex according to the invention is characterized in that the bioavailability of resveratrol from the resveratrol complex is from 90 pg / ml. Preferably, the complex according to the invention is characterized in that the resveratrol is in the trans-resveratrol form.

[0009] The invention also relates to a method of producing the complex according to the invention, characterized in that:

[0010] K1. resveratrol is mixed with leucine until a homogeneous mixture is obtained,

[0011] K2. the mixture is homogenized,

[0012] K3. the mixture is gradually wetted to a water content of 27% ± 4%,

[0013] K4. the mixture is blended,

[0014] K5. the mixture is dried to a water content of 12% ± 3%, K6. the mixture is milled.

[0015] Preferably, the method according to the invention is characterized in that at least one of steps K1 - K3 is carried out in a high-shear granulator. More preferably, in step K2, the homogenization is carried out for 30 minutes with the lower impeller speed of approx. 80 rpm and the chopper speed of approx. 1500 rpm. Preferably, in step K3, the wetting is carried out with the lower impeller speed of approx. 120 rpm and the chopper speed of approx. 3000 rpm, slowly adding a water-ethanol solution of PEG-3000 at maximum spraying. Preferably, in step K4, the mixing is performed with the lower impeller speed of approx. 120 rpm and the chopper speed of approx. 3000 rpm. More preferably, step K4 is carried out for at least 2 hours, and most preferably for at least 4 hours. Preferably, in step K3 maximum spraying is continued for at least 2 hours, more preferably for at least 4 hours. Preferably, in step K5, the drying is carried out in a fluidized-bed dryer. Preferably, the method according to the invention is characterized in that the resveratrol is previously homogenized in a high-shear granulator for 15 minutes with the lower impeller speed of approx. 50 rpm and the chopper speed of approx. 500 rpm. Preferably, the method according to the invention further comprises the steps: K7. adding maltodextrin to the mixture, K8. homogenizing the mixture.

[0016] Preferably, in step K7 maltodextrin is added in an amount of up to 2% by weight. Preferably, in step K8 the homogenization is carried out for 15 minutes at the lower impeller speed of approx. 50 rpm and the chopper speed of about 500 rpm.

[0017] The subject of the invention is a micropellet characterized in that it comprises the complex according to the invention. Preferably, the micropellet according to the invention is characterized in that it further comprises an NAD precursor, curcumin, quercetin, and EGCG. Preferably, the micropellet according to the invention is characterized in that the NAD precursor is selected from nicotinamide adenine dinucleotide (NAD), reduced nicotinamide adenine dinucleotide (NADH), nicotinic acid (NA), nicotinamide riboside (NR), tryptophan (Trp), nicotinamide (N), and nicotinamide mononucleotide (NMN) or combinations thereof. Preferably, the micropellet according to the invention is characterized in that it further comprises at least one food- and / or pharmaceutically-acceptable excipient. More preferably, the micropellet according to the invention is characterized in that it comprises cellulose, maltodextrin, PEG-3000 and sucralose. Most preferably, the micropellet according to the invention is characterized in that the cellulose content is from 15% to 20% by weight. Preferably, the cellulose used in the micropellet according to the invention is 101 grade microcrystalline cellulose. Preferably, the micropellet according to the invention is characterized in that the PEG-3000 content does not exceed 1 %. Preferably, the micropellet according to the invention is characterized in that its size does not exceed 1000 pm, more preferably does not exceed 500 pm. Most preferably, the micropellet according to the invention is characterized in that its size is from 20 pm to 200 pm. Preferably, the micropellet according to the invention is characterized in that the water content in the final micropellet is 5% ± 2% by weight, more preferably at least 4.85%. Preferably, the micropellet according to the invention is characterized in that it is a part of a powder, granulate, micropellets, microtablet, capsule, tablet, food product, dietary supplement, and / or food additive.

[0018] The subject of the invention is also a method of producing a micropellet according to the invention, characterized in that:

[0019] M1. the resveratrol-leucine complex is mixed with at least one active substance and at least one food- and / or pharmaceutically-acceptable excipient until a homogeneous mixture is obtained,

[0020] M2, the mixture is gradually wetted to achieve a water content of 20% to 43%,

[0021] M3, the mixture is aggregated, M4. the mixture is milled,

[0022] M5. the mixture is dried.

[0023] Preferably, in step M2 the mixture is wetted to a water content of 20% to 28%. Preferably, in step M5 the mixture is dried to a water content of 5% ± 2%. Preferably, in step M3 the mixture is aggregated in an extruder until strands of a desired diameter are obtained. Preferably, in step M4 the mixture is milled in a spheronizer to form micropellets. Preferably, in step M5 the mixture is dried in a fluidized-bed dryer with an air inlet temperature of 45 °C ± 2 °C. Preferably, the method according to the invention is characterized in that the mixture is further:

[0024] M6. encapsulated and / or tabletted and / or microtabletted and / or mixed and / or packaged.

[0025] Molecular complexes are often significantly more soluble than the individual molecules forming their core - this property is utilized in producing the complex using the method of grinding assisted by a liquid additive. Furthermore, molecular components are highly mobile within the complex and on the surface of the micropellet, since they are held in place by a network of intermolecular interactions, primarily hydrogen bonds. This means that drastic changes in conditions, especially humidity and temperature, may lead to the breakdown of the complex. The strength and equilibrium state of these interactions depend on the number of trans-resveratrol (TR) molecules relative to the number of ligand molecules and stabilizing molecules in the system - water and PEG-3000. The hydrogen bonds formed are easily broken as the molecules “move,” which is possible due to the flexibility of the connections and the considerable degrees of freedom within such a system. Consequently, the complex is formed successively, while the substrates TR and leucine (LEU) are transformed. The process involves endothermic cleavage of bonds in TR and / or LEU dimers and the formation of a new, thermodynamically preferred system comprising an increasing percentage of the complex. This phenomenon accounts for most molecular processes occurring essentially in the solid state “via the catalyst solution.”

[0026] Ultimately, to produce the TR-LEU complex on a quarter-industrial scale, the solvent drop grinding (SDG) technique, involving the use of micro-amounts of solvent, was selected. This method is classified as a mechanochemical method, enabling the production of complexes essentially in the solid state, without the need to transfer substrates into the solution phase and subsequently dry them again, which in industrial conditions would represent a major challenge due to water consumption and significant energy requirements for heating and drying. For precise process design, data obtained via the slurry method at the laboratory scale was necessary. In the SDG method, the solvent acts only as a catalyst. The remaining components were then added to the formed stable complex according to the formulation presented in Table 1 , after which they were blended, and from the obtained powder - after wetting - micropellets were formed using extrusion and subsequent spheronization. Micro pel lets were produced using a plate spheronizer, and the final product was obtained in two grades - 500 pm and 1000 pm. After drying, the product was examined by infrared spectroscopy, and its moisture content was measured using a moisture analyzer. The final prototype comprised 1.183% water, and no signals above the wavenumber of 3400 cm-1were present in the IR spectrum. This indicates that all -OH groups (both from TR and water) were bound and participated in intermolecular hydrogen bonding. This occurs as a result of stabilization and rigidification of the TR I LEU I water molecular system, with the participation of PEG-3000 molecules, mainly due to crosslinking of the emerging complex structure with hydrogen bonds. The percentage content of TR, both in the complex and in the final product (micropellets), was determined using high- performance liquid chromatography (HPLC). The content per one dose (2000 mg) was no less than 500 mg of TR, in accordance with the specification presented in Table 1.

[0027] Table 1. Final formula according to the invention comprising the TR / LEU complex.

[0028] Using molecular modeling methods, the molar ratio of TR to LEU was determined as 1.000 to 1.625. HyperChem® software and the semi-empirical PM3 method were used to determine the positions of individual atoms in the TR and LEU molecules, taking into account the location of water molecules, with the assumption that the entire system is further stabilized by PEG-3000 mers. A fragment of the constructed complex was subjected to detailed analysis - it consisted of 8 TR molecules, 13 LEU molecules, 11 water molecules, and two PEG-3000 mers. The analysis involved additional semi-empirical simulations using molecular dynamics methods to identify the global energy minimum. The theoretically calculated infrared vibration frequencies, determined using HyperChem® for this refined molecular arrangement, facilitated the identification and assignment of experimentally measured wavenumbers in the spectra of the pure TR and LEU raw materials. T ransmission IR spectra confirmed the presence of fundamental molecular arrangements responsible for hydrogen bond formation. Overlaying the spectra of the complex with those of the pure TR and LEU raw materials allowed identification of regions in the spectra where the largest changes in wavenumbers and / or signal intensity occurred. The high correspondence between the calculated and experimentally measured vibrational frequencies for the final complex confirms that the atomic arrangement proposed in the molecular modeling process is correct, and the signals of newly formed hydrogen bonds are clearly visible and consistent in both the spectra and their simulations.

[0029] Based on these assumptions, potential thermal effects (AH) for the designed processes on a quarter-industrial scale were calculated, and the details of their execution were planned. The calculated thermal effects for individual stages of the entire technological sequence were highly useful in confirming the spontaneity of the processes through Gibbs free energy changes (AG). This parameter enabled the determination of equilibrium constants for successive technological stages, allowing estimation of complex formation efficiency. Estimation of activation energies at individual stages allowed assessment of the rate of subsequent processes and planning of potential process kinetics. Consequently, it was possible to estimate the durations of individual process stages, which is crucial for calculating total production costs. Determination of chemical potentials (partial derivatives of Gibbs free energy with respect to the molar amounts of system components) for individual stages allowed the selection of appropriate molar amounts of auxiliary substances for each stage of the technological process.

[0030] The starting point for detailed calculations was the prior quantitative estimation of the exothermic effect accompanying complex formation (Slurry method at laboratory scale) and the determination of the number of moles of substrates in the reaction vessel when heat evolution begins. This exothermic effect is the sum of two stages - the breaking of hydrogen bonds between substrate molecules (TR-TR and LEU-LEU) and the formation of new bonds between the components of the TR-LEU complex with the participation of water. At the end of the entire technological sequence, after drying and optional milling of the obtained complex, the remaining amounts of components were added to ensure their total quantity matched the values provided in the formulation presented in Table 1. After thorough mixing and wetting of the components to a state in which they begin to form a dry-plastic mass, the entire mixture was subjected to shaping (extrusion) on a perforated extruder (NICA® / GEA - Sweden) with a suitably selected mesh, allowing the formation of strands with a cross-section of 0.5-1.0 mm. Subsequently, the material was transferred in portions onto the rotating plate of a spheronizer (NICA® / GEA - Sweden), and the process continued for approximately 2-4 minutes until the material formed round and / or slightly oval micropellets. The subsequent steps of the described process were: weighing of components, mixing, extrusion, spheronization, and incorporation of the final micropellets into capsules.

[0031] Attempts were made to estimate the number of LEU molecules required to saturate the hydrogen bonds in a TR molecule. Three variants of TR / LEU systems were prepared with different molar ratios of the added LEU ligand, considering molar ratios of 1 : 1 , 1 :2, and 1 :3. Preliminary evaluation of the complexation process efficiency was carried out based on the appearance of the reagents, supplemented by TLC chromatography to visualize and compare the reaction products.

[0032] Initially, resveratrol-leucine complexes were prepared using the slurry technique (Preparation via Slurrying), which allowed obtaining the first pilot samples of the complex for further studies. Based on these samples, analytical methods were selected, developed, and verified. Moreover, the properties of the complex, such as solubility, were evaluated, and, most importantly, the maximum water content at which complex decomposition begins was determined. This value was preliminarily estimated at 37% ± 6%. This parameter is critical and must be meticulously controlled during the wetting of the powdered mass in the extrusion and spheronization processes, to prevent exceeding the value at which secondary complex decomposition occurs. Pilot experiments using a rotary evaporator also revealed a significant influence of ethanol molecules on the formation of the TR-LEU complex according to the invention, providing essential guidance for designing the composition of the catalyst solution necessary for producing the complex using the Solvent Drop Grinding (SDG) method.

[0033] The catalyst solution was prepared based on molecular modeling methods. It comprised the entire allowable amount of PEG-3000 (1 % by weight of the final micropellet) and ethanol in an amount providing a final concentration in the catalyst solution of maximum 3.25% by weight. For production purposes, a recommended value of 3.00% is used, with minor adjustments possible by the operator within ±0.25%. Subsequently, the complexes were produced using the Solvent Drop Grinding (SDG) technique, which involves dispersing the components with micro-amounts of solvent. Optimal starting ratios of TR to LEU were confirmed by molecular modeling methods. The values providing the highest thermodynamic probability of complex formation were calculated as 1.000 to 1.625 in molar ratios. The optimal moisture content for the process in the high-shear granulator was determined to be 27% ± 4%.

[0034] The primary objective of this stage was to establish optimal conditions for complex formation in the high-shear granulator while maintaining the powder temperature during mixing below 46 °C. During the process, a volume reduction of the mixed powder by several percent was observed due to so-called volume contraction. This phenomenon began when the added water content reached approximately 12% ± 3% by weight. At this point, a sudden increase in temperature of several degrees was also recorded. Based on these experiments, the final formulation shown in Table 1 was established, yielding the final form of the complex.

[0035] Analytical methods, particularly HPLC, confirmed that a single dose (2000 mg) of the TR- LEU complex according to the invention comprises no less than 500 mg of pure transresveratrol. The solubility of trans-resveratrol in water was approximately 35 pg / ml. The bioavailability of trans-resveratrol from the formed complexes according to the invention was from around 90 pg / ml.

[0036] This value was estimated based on the second inflection point shown on the curve presented in Fig. 5. Without departing from the scope of the invention, release can be further modulated by selecting appropriate excipients, in particular the properties of cellulose and / or starch.

[0037] The stages of complex formation were also monitored using IR spectroscopy. Experimental wavenumbers were consistent with theoretically calculated vibrational frequencies in the infrared range. Transmission FTIR measurements confirmed the correctness of the designed complex geometry in the regions of hydrogen bond formation. Theoretical methods determined that the complex is most stable at a water content of 11.56% by weight, which was subsequently confirmed experimentally.

[0038] It was estimated that drying in a fluidized-bed dryer should proceed to a water content of 12% ± 3% to avoid removing bound water necessary for maintaining maximum complex stability. Meanwhile, during complex formation in the high-shear granulator, the optimal moisture content should be within 27% ± 4% by weight.

[0039] During the preparation of the final micropellet, the remaining ingredients listed in the formulation in Table 1 were added to the formed complex. For the final micropellet, the entire powder was wetted to 20-28%, a safe range for complex stability while allowing stable extrusion and spheronization processes.

[0040] The obtained mass was then directed to the extruder, and the extruded strands were transferred to the spheronizer. The fresh micropellets obtained were left to dry on trays, which were optionally placed in a convection and / or fluidized-bed dryer.

[0041] The final product was analyzed by ATR-IR spectroscopy to confirm the presence of peaks indicating complex formation, as shown in Fig. 4. The results demonstrated that water content is critical for the proper structure and stability of the complex. Using molecular modeling, confirmed by experimental results, it was established that the minimum water content in the final micropellet required to guarantee stability of the formed system is 5% ± 2%. Accordingly, the critical control point in the manufacturing process was set at a water content of 5%, which is also a safe value from a microbiological perspective.

[0042] The invention will be further illustrated in the following preferable and non-limiting examples, with reference to the attached drawings, in which:

[0043] Fig. 1 shows the chemical structure of the complex according to the invention;

[0044] Fig. 2 shows a comparison of I R wavenumbers measured for the TR and LEU raw materials versus the formed complex;

[0045] Fig. 3 shows SEM images (1 mm corresponds to 1 pm in reality), A - TR and LEU after mixing (many small irregular shapes), B - TR-LEU complexes according to the invention (more regular shapes), C - final micropellet according to the invention;

[0046] Fig. 4 shows the ATR spectrum of the micropellet surface structure according to the invention;

[0047] Fig. 5 shows the TR release curve for a pilot batch of micropellets according to the invention.

[0048] Example 1 - Grinding of Resveratrol and Leucine

[0049] TR (M = 228.25 g / mol) in an amount of 0.200 mol (45.65 g) was ground in a mortar with the addition of corn oil (M » 888 g / mol) in an amount of 0.020 mol (17.76 g). Then, 50 g of glycerol was added to the mortar, and the mixture was intensively ground. The resulting mixture was transferred to a 5 L vessel. Glycerol was added to bring the volume up to 500 mL, and a homogenizer operating at 4000 rpm was used. Three portions of this suspension were prepared.

[0050] LEU (M = 131.17 g / mol) was added in the following amounts: 0.200 mol (26.3 g) for variant 1 , 0.400 mol (52.47 g) for variant 2, and 0.600 mol (78.70 g) for variant 3. The prepared variants were dissolved in 1.5 L, 2.5 L, and 4 L of purified water, respectively, in separate 5 L vessels. The glycerol TR solution was then very slowly added to each aqueous LEU solution variant while stirring at 4000 rpm. Subsequently, the entire mixture was further homogenized at 6000 rpm for 50 min using the homogenizer.

[0051] If precipitation and / or turbidity appeared, it was removed by the addition of small amounts of ethanol. A reference solution of TR was prepared by dissolving 15 g of TR in 100 g of anhydrous ethanol. After each addition of ethanol, the mixture was homogenized again for 2 min at 6000 rpm. Upon completion of homogenization, the three vessels (variants 1 , 2, and 3) were placed in a dark room until sediment appeared at the bottom. The supernatant was then decanted. Samples of 50 g of dense suspension from each variant were exposed to halogen lamp light (50 W) for 72 h. Next, the vessels were stored at room temperature in the dark until a solid residue was obtained. The material was rinsed with ethanol, and residual solvent was evaporated. Subsequently, 1 g of the dry residue was weighed and extracted with 250 mL of an ethanol-water solution (5.2:8.9 v / v) for TR analysis.

[0052] Example 2 - Laboratory-Scale Complex Formation

[0053] Laboratory-scale complexes were produced using the slurry method (Preparation via Slurrying). The main objective of this stage was to determine the molar water content range in the reaction mixture in which TR / LEU complex formation according to the invention occurs and to estimate the thermal effect of this phase transition. A 135.21 g of natural Japanese knotweed extract (Polygonum cuspidatum) was weighed into the first 1000 mL beaker, considering a 6% technological excess (equivalent to 132.51 g of pure TR, 0.58 mol), and dissolved in 250.00 g of ethanol, stirred until a homogeneous suspension was obtained. The beaker was then placed under a homogenizer and homogenized for 40 min at 4000 rpm. After 30 min, the stirring rate was increased to 6000 rpm and maintained for another 10 min. Into the second 1000 mL beaker, 4.98 g of PEG-3000 (1.66 mmol) was weighed, followed by 150.00 g of purified water, heated to 35 °C, and stirred with a magnetic stirrer for 5 min or until PEG-3000 was completely dissolved. Then, 123.74 g of LEU (including a 6% technological excess, 0.94 mol) was added, and the beaker was placed under a homogenizer. The mixture was homogenized for 70 min at 6000 rpm. After 40 min, the speed was increased to 8000 rpm and homogenization continued for another 30 min.

[0054] The contents of beaker no. 1 were transferred to a 5 L flask and heated to 38 °C. The contents of beaker no. 2 were also heated to 38 °C and very slowly added to the flask while stirring intensively. Subsequently, 250.00 g of ethanol was added to the flask under continuous stirring.

[0055] After preparing the reaction solution, the flask was mounted on a rotary evaporator at 90 rpm. The contents were stirred intensively for 30 min before starting pressure reduction. The flask was monitored to prevent liquid transfer to the receiver. Simultaneously, the temperature of the mixture was increased on the thermostat to initiate boiling. Depending on atmospheric conditions and vacuum pump pressure, boiling occurred at 54-68 °C (approximate range, to be determined empirically, the aim of the experiment was to confirm the nature of effects). Initially, the mixture foamed strongly, and the condensate was minimal. After ~30 min of slow distillation of the water-ethanol azeotrope, a turning point was reached, where the ethanol molar content was sufficiently low for complex formation to begin. Complex formation according to the invention was also confirmed by I R spectroscopy (example spectrum shown in Fig. 2). This point was characterized by a sudden increase in condensate formation and release of ~10 kJ of heat per mole of TR, confirmed by computational and simple calorimetric measurements. These data served as the basis for calculating thermal effects and designing quarter-industrial-scale processes on a GSM high- shear granulator. From this information, the approximate molar fraction of the formed TR complex relative to unreacted substrate could be estimated (AG = -R T In K).

[0056] It is then possible to estimate the thermodynamic potential of the TR / LEU complex formation according to the invention in relation to the initial TR / TR and LEU / LEU dimers, as well as the equilibrium constant of such a process. After transforming the above formula, we obtain an equation showing that the equilibrium constant of the complexation process at a given AG value is approximately 40 (in our case estimated at approximately -10 kJ and an average process temperature of 50°C, i.e. 323 K).

[0057] It was demonstrated that, at equilibrium, the amount of TR / LEll complex according to the invention should be approximately 0.5 mol, and unreacted TR about 0.08 mol. After the rapid heat release, foaming ceased. Pressure was gradually reduced to 150 mmHg, and the flask temperature decreased to 46 °C. The flask rotation speed was reduced to 30 rpm, and the remaining solvent was gently evaporated. When no further condensate collected in the receiver, the flask temperature was lowered to 36 °C and pressure equalized to ambient. The flask was cooled to room temperature and left for 5 min. The thick slurry was then transferred to a stainless-steel tray, a sample was taken for moisture analysis, and the material was cooled at 8 °C. Meanwhile, the material was further dried at room temperature and milled.

[0058] After 2 h of cooling, the tray was transferred to a dryer, where residual solvent was removed at 25 °C. After 8 h, the complex was sieved through a 1 mm mesh, and maltodextrin was added to 2% by weight. The mixture was thoroughly blended, sieved again, and prepared for the addition of remaining ingredients according to the formulation in Table 1. After incorporating all final ingredients and forming the process mass, the mixture was directed to the formation of the final micropellets. Based on experimental studies and preliminary computer simulations, it was determined that the maximum moisture content during micropellet formation should not exceed 37% ± 6%. This value, representing the water content in the process mass, corresponds to the onset of TR / LEU complex decomposition. During micropellet production, this threshold must not be exceeded when wetting the raw materials. These data, along with the formulation in Table 1 , also served as the basis for preparing the catalyst solution sprayed during complex formation in the final SDG method. All of these parameters formed the foundation for developing the final technology for producing micropellets according to the invention.

[0059] Complexes according to the invention were produced using the SDG (Solvent Drop Grinding) method. Laboratory-scale trials were performed to determine the optimal chopper rotational speed necessary for the complex formation process, while preventing the powder temperature from exceeding 55 °C. Different chopper speeds were tested, and it was established that the optimal range should be 1000-2000 rpm. During the initial mixing phase of the substrates, the rotation speed should range between 1000-1200 rpm, and upon reaching a moisture content of 12% ± 3%, the speed should be increased to 1800-2000 rpm. At this point, a volume contraction of several percent and a sudden increase in temperature of a few degrees Celsius are expected. The exact moment to increase the rotation speed should be determined by the operator, who must closely observe the mixture and the powder volume. The moment of volume contraction corresponds to an increase in the density of the mixed system, causing higher resistance to the main mixer and a sharp increase in current draw. This parameter, along with the chopper and main impeller speeds, should be continuously monitored throughout the process. After this pseudo-phase transition, the rotational speed should be maintained at 1300-1500 rpm. At the laboratory scale, this provided optimal conditions for complex formation according to the invention, as confirmed by IR spectra collected during the process.

[0060] During complex formation using the SDG method, the chopper supplies the additional energy necessary to initiate aggregation of molecules. This one-time, localized kinetic energy input allows TR and LEU molecules to overcome the activation energy barrier, significantly accelerating and / or even enabling complex formation. The complexation process organizes the molecules in the system, thereby reducing entropy (AS < 0). As a result, heat must be absorbed from the immediate environment of the complexing molecules, usually provided by the kinetic energy of the rotating chopper. During chopper operation, the total volume of substrates decreased by at least several percent, indicating additional interactions between TR, LEU, and water molecules, stabilized by PEG-3000 chains. The complex components also hinder water evaporation, and PEG-3000 polymer networks facilitate molecular ordering into regular systems. LEU present in the complex binds water effectively, slowing its evaporation and further stabilizing the TR / LEU complex. In the final micropellet, water loss is additionally limited by quercetin molecules and EGCG components, which also enhance the cross-linking of cellulose-101 and starch chains. The complex is further stabilized by the addition of 2% w / w maltodextrin immediately after complex formation.

[0061] In one embodiment, the complex comprised resveratrol and leucine. Preferably, the molar ratio of resveratrol to leucine was from 1 :1 to 1 :3. In a particularly preferred embodiment, the ratio was 1 :1.625.

[0062] In another embodiment, in addition to resveratrol and leucine, the complex comprised up to 2% w / w of PEG-3000. Preferably, the complex comprised up to 1% w / w of PEG-3000.

[0063] In a preferred embodiment, resveratrol bioavailability from the complex was from 90 pg / mL. In another preferred embodiment, resveratrol was in the trans-resveratrol form.

[0064] In one embodiment, the complex was produced according to the following steps:

[0065] K1. Mix resveratrol with leucine until a homogeneous mixture is obtained

[0066] K2. Homogenize the mixture

[0067] K3. Gradually wet the mixture to a water content of 27% ± 4%

[0068] K4. Stir the wetted mixture

[0069] K5. Dry the mixture to a water content of 12% ± 3%

[0070] K6. Mill the mixture

[0071] In this non-limiting example, 51.0211 kg of Polygonum cuspidatum raw material (98% transresveratrol), 45.1200 kg of pure leucine, and 12.4800 kg of total water were mixed. These quantities were determined by analyzing IR spectra of TR and leucine; the TR spectrum showed a strong signal at 965 cm-1. Moisture measurements enabled the calculation of water needed to achieve 12% ± 3% moisture in the final complex. Theoretical molar ratios of the complex at this stage were TR:LEU:WATER = 1.000 : 1.625 : 3.250, corresponding to 50.0000 kg TR, 45.1200 kg leucine, and 12.4800 kg water.

[0072] In another preferred embodiment, Polygonum cuspidatum and leucine were pre-ground. In a particularly preferred embodiment, resveratrol was homogenized in a high-shear granulator for 15 min at ~50 rpm of the lower impeller and -500 rpm of the chopper. At least one of steps K1-K3 was conducted in a high-shear granulator. In a particularly preferred embodiment, step K2 the homogenization lasted 30 min at -80 rpm lower impeller speed and -1500 rpm chopper speed. In another embodiment, during step K3, wetting was performed at -120 rpm lower impeller speed and -3000 rpm chopper speed, with water-ethanol PEG-3000 solution slowly added at maximum spray. In other embodiments, maximum spraying in step K3 lasted at least 2 hours, or at least 4 hours. Preferably, the water-ethanol solution was prepared by heating water to 50 °C, dissolving PEG-3000, cooling to 25 °C, and slowly adding ethanol azeotrope (96%) to achieve 3.01 % w / w ethanol. For this resveratrol and leucine amount, 1.9912 kg PEG-3000 and 0.9589 kg ethanol azeotrope were added. Water amount was determined using IR measurements of the starting materials. The optimal total water was 30.3661 kg, corresponding to process moisture of 27% ± 4%. This ensures optimal SDG processing and sufficient energy to overcome activation barriers for TR and LEU to form the complex. The theoretical molar ratios at this stage were TR:LEU:WATER = 1.000 : 1.625 : 7.000. In another embodiment, the water-ethanol solution was heated in a jacketed tank with a propeller, PEG-3000 added, and stirred for 15 min until completely dissolved.

[0073] In another embodiment, step K4 was carried out at -120 rpm of the lower impeller and -3000 rpm of the chopper. Preferably, step K4 lasted at least 2 hours, and more preferably, at least 4 hours. In one embodiment, step K5 the drying was performed in a fluidized-bed dryer; in another embodiment, drying was continued until 12% ± 3% moisture was achieved.

[0074] In yet another embodiment, the process further comprised:

[0075] K7. adding maltodextrin to the mixture

[0076] K8. homogenizing the mixture

[0077] In a preferred embodiment, maltodextrin was added up to 2% w / w during step K6. In another preferred embodiment, the homogenization in step K7 lasted 15 min at -50 rpm of the lower impeller and -500 rpm of the chopper speed.

[0078] Example 4 - Production of Micro pel lets

[0079] A total of 200 kg of micropellets according to the invention were produced. Each micropellet comprised the complex described above. In one embodiment, the micropellet comprised NAD precursor, curcumin, quercetin and EGCG. Various NAD precursors with essentially comparable efficacy were tested. In one embodiment, the NAD precursor used in the micropellet was nicotinamide adenine dinucleotide (NAD), in another reduced nicotinamide adenine dinucleotide (NADH), in another nicotinic acid (NA), in another nicotinamide riboside (NR), in another tryptophan (Trp), in another nicotinamide (N) and in yet another nicotinamide mononucleotide (NMN). A skilled person understands that combinations of the above substances may also be used. Particularly preferably precursors are nicotinic acid (NA) and nicotinamide mononucleotide (NMN) due to their relative bioavailability, costeffectiveness, thorough study, and approval for use in food. In another embodiment, the micropellet additionally comprised at least one food- and / or pharmaceutically acceptable excipient. In a preferred embodiment, the micropellet comprised cellulose, maltodextrin, PEG-3000 and sucralose. In one preferred embodiment, the cellulose content was 15-20% w / w. In another, grade 101 microcrystalline cellulose (MCC) was used, responsible for structural properties such as mass plasticity, typically achieved by using 15-20% MCC (usually Avicel PH-101) depending on observed flow-plastic properties. In another preferred embodiment, PEG-3000 content did not exceed 1%. In one embodiment, the micropellets were encapsulated. In another, they were incorporated into a food product (e.g., yogurt). In another, micropellets were tableted into microtablets. A skilled person understands that, without departing from the invention, micropellets can be incorporated into other forms such as powders, granules, tablets, dietary supplements, and / or food additives. In a particularly preferred embodiment, the micropellet size did not exceed 1000 pm. In a more preferred embodiment, it did not exceed 500 pm. In the most preferred embodiment, the size was from 20 pm to 200 pm. In one embodiment, water content in the final micropellet was 3%, and in another, 7%. Results indicated that water content is critical, as its deficiency leads to unwanted loosening of the complex structure and formation of leucine dimers. Molecular modeling and experiments established that the minimum water content to guarantee stability of the final micropellet is 4.87%. Accordingly, the critical control point in production was set at a minimum water content of 5%.

[0080] Micropellets were produced using the following method:

[0081] M1. Mix the resveratrol-leucine complex with at least one active substance and at least one food- and / or pharmaceutically acceptable excipient until a homogeneous mixture is obtained

[0082] M2. Gradually wet the mixture to a water content of 20% to 43%

[0083] M3. Aggregate the mixture M4. Mill the mixture M5. Dry the mixture

[0084] In one embodiment, step M2 was conducted to achieve a water content of 20% to 28%. In another embodiment, in step M5 the drying was performed to achieve 5% ± 2% water content. In another, in step M3 the aggregation was performed in an extruder until strands of desired diameter were obtained. In another embodiment, in step M4 the milling was performed in a plate spheronizer to produce micropellets. In another, in step M5 the drying was carried out in a fluidized-bed dryer at 45 °C ± 2 °C. As before, micropellets were either encapsulated, added to food products (e.g., yogurt), and / or tableted into microtablets. A skilled person understands that micropellets may be processed into other acceptable forms such as powder, granules, tablets, dietary supplements, and / or food additives. Example 5 - Principal Analytical Methods and Quality Control

[0085] The formation, morphology, and size of the complexes and micropellets were visually confirmed using SEM (NOVA NANOSEM 450). Micropellets were produced in two sizes: 500 pm and 1000 pm. SEM images (Fig. 3A) showed that loosely mixed TR and LEU raw materials had less organized morphology compared to complexes crosslinked with water and PEG-3000. Electron micrographs of the final complex demonstrated greater structural order compared to loosely mixed raw materials (Fig. 3A). Complex particles exhibited cleaner, glossier surfaces than the raw mixture (Fig. 3B). Regular structures confirmed successful complexation and correctness of design assumptions. Fig. 3C shows the final micropellet, which has a regular shape. Regular shape improves flow properties and visual appeal. Polyphenols and amino acid complexes significantly influenced the functional properties of crosslinked materials, such as 101 grade cellulose, facilitating micropellet formation. Water content and material structure must be controlled at each stage using SEM and IR methods.

[0086] At various stages, moisture was measured using a moisture balance (Radwag MA 50.X7.IC.A). In one example, micropellets comprised 1.183% of water.

[0087] Due to the speed and simplicity of the measurements, at various stages of the production of the complex according to the invention and the micropellet according to the invention, the product was examined using IR as a primary method for characterizing intermolecular interactions in the designed systems, particularly confirming the formation of hydrogen bonds, which is crucial when studying the interactions of OH groups, which are abundant in polyphenols, and NH3 groups from amino acids, for example, after drying the complexes and the micropellet according to the invention. The IR spectrum of the final micropellet showed no signals above the wavenumber of 3400 cm-1 . This indicates that all -OH groups (from both TR and water) were bound and participating in intermolecular bonds. It also confirms that the carbonyl group was deprotonated. Signals around 3289 cm-1 indicated the involvement of OH and NH3 groups in the formation of hydrogen bonds. Intense bands at 2957 cm-1 and 2876 cm-1 indicated the lack of involvement of methyl groups in intermolecular interactions. Signal shifts for the N-H groups of NH3 indicated their participation in hydrogen bonds, as confirmed by the values measured between 3019 and 2995 cm-1 . Therefore, IR spectroscopy confirmed the formation of hydrogen bonds and the formation of the complexes proposed by the invention through intermolecular interactions, stabilized by water molecules and PEG-3000 polymer chains. The IR data obtained for the Polygonum cuspidatum extract and the prepared micropellet helped verify the accuracy of the calculations in line with theoretical assumptions and reduce random errors, which was a significant step in ensuring quality and repeatability. Meanwhile, a special IR spectroscopy method, Attenuated Total Reflectance (ATR), allows for the analysis of molecular absorption with the quenched water signal. The FTIR-ATR technique (Shimadzu FTIR 8400S with MIRacle Single Reflection ATR) is dedicated to providing information on the structure and intermolecular interactions, and in particular provides a wealth of structural information if the systems contain strong intermolecular interactions, such as hydrogen bonds, especially those involving COO- groups. Therefore, the spectrum recorded using the ATR technique enables rapid analysis of the structure located on the surface of the produced micropellet - the relevant spectrum is shown in Fig. 4. The signals of free methyl groups at 2964 cm-1 and 2870 cm-1 are clearly visible. The signal at 3279 cm-1 indicates strong interactions of NH3 groups and their involvement in interactions with oxygen atoms in the complex. The spectrum was recorded for a dried sample with a moisture content of 1.187%. It is clearly visible that the signal of the CH group linked to COO- and NH3+, as well as the NH in NH3, returns to 3062 cm-1 , confirming the significant contribution of water molecules to the formation of the complex structure. Using molecular modeling methods, it was determined that the final micropellets according to the invention should not be dried beyond a moisture content of 5% + / - 2%. This moisture content ensures the stability of the complex according to the invention during storage. In the IR spectrum of the final micropellets according to the invention (ATR technique), a rounded signal was measured at a wavenumber of 3293 cm-1 , indicating the involvement of the -OH and -NH3+ groups (proton donors) in the formation of hydrogen bonds with the -OH and COO- groups (proton acceptors). The intense ATR band at 2957 cm-1 and 2871 cm-1 indicates the lack of involvement of methyl groups in intermolecular interactions - these frequencies are identical in both the LEU spectrum and the complex according to the invention. The calculated wavenumbers for the methyl groups of an isolated single LEU molecule, obtained in vacuum, and for the TR / LEU complex according to the invention, taking into account hydrogen bonds, also lead to the same results. In turn, the involvement of COO- groups in the formation of the complex according to the invention is most evident in the shift in the vibrational frequency of the C-C group located immediately adjacent to the carbonyl group. Also in the spectrum recorded for the micropellets according to the invention using the ATR technique (Fig. 4), the C-C vibrational wavenumber shifts from 855 cm-1 in LEU to 827 cm-1 in the complex according to the invention. This indicates that water is involved in hydrogen bonds primarily through the NH3+ group.

[0088] The final product (a micropellet according to the invention) should have a water content within the range of 5% + / - 2% by mass. This value was obtained based on a more detailed analysis using molecular dynamics methods for the starting geometry. The extensive model system consisted of 16 TR molecules and 26 LEU molecules. The amount of water was selected using the periodic box tool available in HyperChem® 8.1 , and the generated values were then verified using semi-empirical methods available in this program.

[0089] The molar ratio of water relative to the remaining components of the complex according to the invention were determined using molecular modeling methods, and the theoretically estimated values were verified by moisture measurements at each stage of the process. The theoretically estimated moisture values demonstrated very good agreement with previous theoretical calculations. The molecular modeling results enabled the precise location of some water molecules relative to the TR and LEU molecules. These locations and the contribution of water molecules to the hydrogen bonding network were verified and confirmed by infrared measurements. The obtained intermediate values were used for further calculations and the final design of quarter-scale processes.

[0090] Molecular modeling methods were used to determine the optimal geometry of the studied fragment of the system, and then theoretical infrared resonance values were calculated. These values were compared with measured wavenumber values for samples collected at various stages of the complex formation process according to the invention to determine potential hydrogen bond formation sites. The final geometry of the fragment of the model complex system (TR / LEU) according to the invention was determined using the semi- empirical PM3 method.

[0091] Theoretical wavenumbers were calculated for individual functional groups of the fragment of the complex system according to the invention, including O-H, N-H, N-H2, C-H, C-H2, C- H3, and C-C-COO, and were used for further model calculations. Functional group vibration frequencies: for the O-H groups of TR and water, and for the N-H groups of LEU, wavenumber shifts were observed, indicating the formation of hydrogen bonds between them. The absence of a typical band for the C=O group indicates charge delocalization between the oxygen atoms.

[0092] Calculated vibration frequencies indicate the existence of symmetric and asymmetric vibrations for the C-0 groups, at frequencies vasym = 1548 cm-1 and vsym = 1445 cm-1. Vibrations of the trans H-C=C-H system at a wavenumber of 965 cm-1 confirm the presence of the (E) TR stereoisomer. Calculations of IR vibration frequencies at the theoretical stage enabled the identification of key signals in the IR spectra, confirming the formation of hydrogen bonds in the TR / LEU complexes. The TR / LEU complex is stabilized by water molecules and PEG-3000 polymer chains, as confirmed by, among other things, wavenumber shifts for the vibrations of the C-COO- groups from 855 cm-1 to 830 cm-1 . At each stage of the calculations, the assumptions were verified by actual IR measurements. An example spectrum confirming the above calculations is shown in Fig. 2. The geometry optimized using the semi-empirical PM3 method for the selected fragment of the TR and LEU complex according to the invention provided key data on binding energies and heats of formation. The calculations showed that a ligand such as LEU has a high affinity for TR, forming stable complexes within a certain range of coordination numbers. During complex formation, a significant amount of heat is released, which outweighs the entropy reduction and enables the spontaneity of the process. Molecular dynamics at the PM3 level was used as a supporting method to search for the global minimum for the designed geometries of TR-ligand complexes. Molecular modeling results also show that the addition of PEG-3000 polymer increases the stability of the complexes proposed in the invention, and the heat release during hydrogen bond formation confirms the spontaneous nature of the process. The use of these theoretical models enabled the effective design of a high-shear granulator process. Furthermore, the interaction of water molecules with TR and LEU molecules significantly stabilizes the designed complex, albeit within a narrow range of molar proportions. Detailed results are summarized in Table 2.

[0093] Table 2. Amounts of heat released during the interactions of 8 TR molecules with 13 LEU ligands and 11 water molecules in the presence of 2 mers of PEG-3000.

[0094] In the next stage of the molecular modeling process, a system was constructed consisting of 16 TR molecules, 26 LEU molecules, and 22 water molecules. It was also observed that, at these proportions in the complex according to the invention, the system is already stabilized, and further increasing the percentage of LEU in the complex does not provide a significant exothermic contribution to the spontaneity of the process. Similarly, increasing the amount of water no longer produced any additional stabilizing effect. Therefore, for further analysis, a system consisting of 8 TR molecules, 13 LEU molecules, and 11 water molecules was selected. Its size accurately represents the interactions occurring in nature, while also allowing rapid calculation of theoretical infrared frequencies, which will be important for future monitoring of technological processes. This number of molecules was also sufficient for reliable simulation of the amount of heat released and / or absorbed by the system. Based on these assumptions, it was determined that eight complexed TR molecules release a total of 399 kcal of heat due to the formation of hydrogen bonds with 13 LEU molecules and 11 water molecules. Detailed data are summarized in Table 3.

[0095] Table 3. Amounts of heat released as a result of intermolecular interactions between 8 TR molecules, 13 LEU ligands, and 11 water molecules.

[0096] This translates to approximately 49 kcal / mol per 1 mole of TR. Calculations indicate that the highest stability of the complex according to the invention occurs at molar ratios of TR:LEU:H2O of 1.000:1.625:1.375. In the next stage, this system was studied for increased stability through the addition of PEG-3000. Theoretical calculations of the final thermal effects also confirm the spontaneity of the TR / LEU complex formation according to the invention within the estimated molar ratio ranges.

[0097] Example 6 - Analysis of Resveratrol Complexation and Its Presence in the Micropellet

[0098] Glass TLC plates coated with silica gel modified with an RP-18 phase (reversed-phase system) were used. Three repetitions were performed for each tested variant. Chromatograms were developed in vertical chambers. The atmosphere was saturated for 20 minutes with vapors of an ethanol and water mixture (5.2:3.7 v / v). Chromatograms were developed to a height of 12 cm and then dried. Visualization was achieved using anise aldehyde. The developed chromatograms were sprayed with this reagent, then the plates were air-dried for 30 minutes and subsequently heated at 90°C for 1 minute. Brown bands appeared on the chromatograms in the locations corresponding to TR. Measurements were made using a reflective densitometer (X-Rite 404). The reflected light intensity was compared to a series of three standard solutions developed on the plates. The resulting TR content after extraction is summarized in Table 4.

[0099] Table 4. Concentrations of TR extracted from LEU-complexed samples for variants 1 , 2 & 3

[0100] The results indicate that the saturation of the TR molecule in the complex formed with LEU according to the invention occurs already at a stoichiometric ratio of 1 :2. In view of this result, further design of TR complexes with various ligands was carried out assuming a maximum stoichiometric ratio of TR / ligand of 1 :2. Based on the outcome of this experiment, initial assumptions for designing starting geometries were adopted taking this into account.

[0101] Example 7 - Assessment of Resveratrol Content and Release from the Micropellet According to the Invention

[0102] The TR content in intermediates was determined using high-performance liquid chromatography (HPLC). Samples for measurement were taken directly from the packaging and then filtered through a 0.45 pm filter. If necessary, the samples were diluted. All analyses were repeated at least three times. Results are presented as concentrations in the format: measurement ± SD (standard deviation).

[0103] Samples were analyzed for their TR content (pure reference substance - CAS number 501- 36-0). Determinations were carried out chromatographically using a Hitachi Chromaster HPLC system with a DAD detector and autosampler. The system was equipped with a reversed-phase column from Merck, Purospher STAR RP-18e (5 pm, 250 mm x 4.6 mm), operating at 30°C. Detection was performed at a wavelength of 305 nm. The eluent consisted of 0.1% formic acid (A) and acetonitrile (B). Substance content was determined based on a calibration curve in pg / mL - the limit of detection (LCD) was 0.81 pg / mL, and the limit of quantification (LOQ) was 2.40 pg / mL. HPLC-grade standards of the pure substance were purchased from PhytoLab GmbH & Co. KG with purity above 99.0%. Concentrations in the samples were as follows: sample 1 : 10.15 ± 0.04 mg / mL (10,147 ± 38 pg / mL), sample 2: 35.25 ± 0.49 pg / mL and sample 3: 35.54 ± 0.29 pg / mL.

[0104] Sample 1 represented a complex prepared using the Slurry method. The observed significant increase in solubility is due to the formation of the complex according to the invention and the presence of ethanol. Micropellets according to the invention were also tested for TR release. The concentrations in these samples were analyzed using high- performance liquid chromatography, following the same procedure as described above. Figure 5 shows the TR concentration in the analyzed sample as a function of time after sampling from the release apparatus.

[0105] According to the measurements, the predicted bioavailability of TR from the final product is increased almost threefold. A saturated solution of pure TR in water has a concentration of 35 pg / mL, while the achieved bioavailability of this active substance from the developed micropellet comprising the complex according to the invention is estimated from 90 pg / mL.

Claims

Claims:

1. A resveratrol complex characterized in that it comprises resveratrol and leucine.

2. The complex according to claim 1, characterized in that the molar ratio of resveratrol to leucine is from 1 :1 to 1:3.

3. The complex according to claim 2, characterized in that the ratio of resveratrol to leucine is 1 :1.625.

4. The complex according to any of the preceding claims, characterized in that it further comprises up to 2% by weight of PEG-3000, preferably up to 1% PEG-3000.

5. The complex according to any of the preceding claims, characterized in that resveratrol bioavailability from the resveratrol complex is from 90 pg / ml.

6. The complex according to any of the preceding claims, characterized in that resveratrol is in the trans-resveratrol (TR) form.

7. A method of producing the complex defined in any of the preceding claims, characterized in that:K1. resveratrol is mixed with leucine until a homogeneous mixture is obtained,K2. the mixture is homogenized,K3. the mixture is gradually wetted to a water content of 27% ± 4%,K4. the mixture is stirred,K5. the mixture is dried to a water content of 12% ± 3%,K6. the mixture is milled.

8. The method according to claim 7, characterized in that at least one of steps K1 - K3 is carried out in a high-shear granulator.

9. The method according to claim 8, characterized in that in step K2, the homogenization is carried out for 30 min, with the lower impeller speed of approx. 80 rpm and the chopper speed of approx. 1500 rpm.

10. The method according to any of claims 8-9, characterized in that in step K3, the wetting is carried out with the lower impeller speed of approx. 120 rpm and the chopper speed of approx. 3000 rpm, slowly adding a water-ethanol solution of PEG-3000 at maximum spraying.

11. The method according to any of claims 8-10, characterized in that in step K4, the stirring is carried out with the lower impeller speed of approx. 120 rpm and the chopper speed of approx. 3000 rpm.

12. The method according to claim 11 , characterized in that step K4 is carried out for at least 2 hours, preferably for at least 4 hours.

13. The method according to any of claims 7-12, characterized in that step K3 is carried out at maximum spraying for at least 2 hours, preferably at least 4 hours.

14. The method according to any one of claims 7-13, characterized in that in step K5, the drying is carried out in a fluidized bed dryer.

15. The method according to of claims 7-14, characterized in that resveratrol is previously homogenized in a high-shear granulator for 15 min with the lower impeller speed of approx. 50 rpm and the chopper speed of approx. 500 rpm.

16. The method according to any of claims 7-15, characterized in that it further comprises the steps:K7. adding maltodextrin to the mixture,K8. homogenizing the mixture.

17. The method according to claim 16, characterized in that in step K7, maltodextrin is added an amount up to 2% by weight.

18. The method according to any of claims 16-17, characterized in that in step K8, the homogenization is carried out for 15 min with the lower impeller speed of approx. 50 rpm and the chopper speed of approx. 500 rpm.

19. A micropellet characterized in that it comprises the complex defined in any of claims 1- 6.

20. The micropellet according to claim 19, characterized in that it further comprises a NAD precursor, curcumin, quercetin, and EGCG.

21. The micropellet according to any of claims 19-20, characterized in that the NAD precursor is selected from nicotinamide adenine dinucleotide (NAD), reduced nicotinamide adenine dinucleotide (NADH), nicotinic acid (NA), nicotinamide riboside (NR), tryptophan (Trp), nicotinamide (N), and nicotinamide mononucleotide (NMN), or their combinations.

22. The micropellet according to any of claims 19-21 , characterized in that it further comprises at least one food- and / or pharmaceutically acceptable excipient.

23. The micropellet according to claim 22, characterized in that it comprises cellulose, maltodextrin, PEG-3000, and sucralose.

24. The micropellet according to claim 23, characterized in that the cellulose content is from 15% to 20% by weight.

25. The micropellet according to any one of claims 23-24, characterized in that the cellulose is 101 grade microcrystalline cellulose.

26. The micropellet according to any one of claims 23-25, characterized in that the PEG- 3000 content does not exceed 1 %.

27. The micropellet according to any of claims 19-26, characterized in that the micropellet size does not exceed 1000 pm, preferably does not exceed 500 pm.

28. The micropellet according to claim 27, characterized in that the micropellet size is from 20 pm to 200 pm.

29. The micropellet according to any of claims 19-28, characterized in that the water content in the final micropellet is 5% ± 2% by weight, preferably at least 4.85%.

30. The micropellet according to any of claims 19-29, characterized in that it is part of a powder, granulate, micropellets, microtablet, capsule, tablet, food product, dietary supplement, and / or food additive.

31. A method of producing the micropellet defined in any of claims 19-30, characterized in that:M 1. the resveratrol-leucine complex is mixed with at least one active substance and at least one food- and / or pharmaceutically acceptable excipient until a homogeneous mixture is obtained,M2, the mixture is gradually wetted to a water content of 20% to 43%,M3, the mixture is aggregated,M4. the mixture is milled,M5. the mixture is dried.

32. The method according to claim 31, characterized in that in step M2, the mixture is wetted to a water content of 20% to 28%.

33. The method according to any of claims 31-32, characterized in that in step M5, the mixture is dried to a water content of 5% ± 2%.

34. The method according to any of claims 31-33, characterized in that in step M3, the mixture is aggregated in an extruder until strands of a desired diameter are obtained.

35. The method according to any of claims 31-34, characterized in that in step M4, the mixture is milled in a plate spheronizer until micropellets are obtained.

36. The method according to any of claims 31-35, characterized in that in step M5, the mixture is dried in a fluidized-bed dryer with an air inlet temperature of 45°C ± 2°C.

37. The method according to any one of claims 31-36, characterized in that the mixture is further:M6. encapsulated and / or tabletted and / or microtabletted and / or mixed and / or packaged.

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