NMN compound anti-aging preparation for activating sirt1 pathway and preparation thereof

Abstract

The application discloses an NMN compound anti-aging preparation for activating a SIRT1 channel and a preparation method thereof, relates to the technical field of pharmaceutical preparations, and discloses the preparation which is composed of active ingredients and pharmaceutically acceptable adjuvants, wherein the active ingredients account for 100 parts in total in terms of weight fraction and include the following components: 30-60 parts of beta-nicotinamide mononucleotide; 8-20 parts of trans-resveratrol; 2-10 parts of quercetin; 0.5-3 parts of piperine; and the rest is trimethylglycine. The application forms a synergistic regulation system targeting the SIRT1 channel by compounding five active ingredients, combines with a time sequence synergistic release dual-phase preparation design and adjuvant collocation, realizes continuous and efficient activation of the channel, solves the problems of asynchronous release of components in traditional preparations, poor absorption of poorly soluble components, improves the bioavailability of components and the storage stability of the preparation, reduces the metabolic burden of the body, and the preparation process is suitable for conventional pharmaceutical equipment, so that the quality consistency between batches can be ensured.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical formulation technology, specifically to an NMN complex anti-aging formulation that activates the SIRT1 pathway and its preparation. Background Technology

[0002] Aging is a progressive decline in the function of cells, tissues, and organs that occurs with age. The NAD+-dependent SIRT1 pathway is a core molecular pathway regulating this aging process. SIRT1, an NAD+-dependent deacetylase, participates in aging-related physiological processes such as DNA damage repair, mitochondrial energy metabolism, and oxidative stress response by deacetylifying downstream target proteins. Its activity is directly regulated by intravenous NAD+ levels. With increasing age, the body's NAD+ synthesis capacity declines and its degradation rate accelerates, leading to a continuous decrease in NAD+ levels and a corresponding decline in SIRT1 activity, exacerbating age-related phenotypes. β-Nicotinamide mononucleotide (NMN), as a direct precursor to NAD+, can be rapidly converted into NAD+, making it a core raw material for anti-aging formulation development. The industry commonly employs a technical route combining NMN with SIRT1 activators and NAD+ degradation inhibitors.

[0003] Existing anti-aging formulations have several shortcomings in practical applications. Nicotinamide, produced from NMN metabolism, inhibits SIRT1 activity, and long-term use can lead to nicotinamide accumulation, negating the activation effect. Traditional compound formulations lack targeted nicotinamide metabolism-regulating components. The physicochemical properties of different active ingredients vary greatly; NMN is absorbed rapidly, while poorly soluble components such as trans-resveratrol and quercetin dissolve slowly, resulting in asynchronous release and absorption of each component, failing to achieve synergistic effects. Furthermore, conventional excipient combinations lack specificity, poorly soluble components have low bioavailability, and some formulations lack stability, easily degrading during storage. This prevents sustained activation of the SIRT1 pathway and fails to meet the needs of long-term anti-aging use. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of existing technologies and provide an NMN complex anti-aging formulation that activates the SIRT1 pathway and its preparation. By precisely combining five active ingredients, including β-nicotinamide mononucleotide and trans-resveratrol, a synergistic regulatory system targeting the SIRT1 pathway is constructed. Simultaneously, by combining a time-sequential synergistic release biphasic formulation design with a scientific combination of proprietary pharmaceutical excipients, the invention effectively solves the problems of nicotinamide accumulation, asynchronous component release, and low bioavailability of poorly soluble components in traditional anti-aging formulations. This invention achieves continuous and efficient activation of the SIRT1 pathway, stably maintains intracellular NAD+ levels, and improves the storage stability and long-term safety of the formulation. Its preparation process is compatible with conventional pharmaceutical equipment, batch quality is controllable, and different component ratios can be adapted to various application scenarios.

[0005] To solve the above-mentioned technical problems, the present invention provides the following technical solution: In one aspect, an NMN complex anti-aging preparation that activates the SIRT1 pathway, the preparation comprising an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient comprises a total of 100 parts by weight, and includes the following components: 30-60 parts of β-nicotinamide mononucleotide; 8-20 parts of trans-resveratrol; Quercetin 2-10 parts; Piperine 0.5-3 parts; The balance is trimethylglycine.

[0006] Furthermore, the trimethylglycine is a natural betaine extract with an extraction purity of ≥99%, wherein the free betaine content is ≥98.5% and the moisture content is ≤0.5%.

[0007] Furthermore, the β-nicotinamide mononucleotide is β-configuration crystal form I, with a particle size range of 10-50 μm, a purity ≥99.5%, and α-configuration impurities ≤0.1%; the trans-resveratrol has a trans isomer content ≥98% and a melting point of 254-256℃; the quercetin is anhydrous quercetin aglycone, with a particle size ≤20 μm, a purity ≥98%, and glycoside impurities ≤1%; and the piperine has a purity ≥95% and a melting point of 128-130℃.

[0008] Furthermore, the formulation is a time-dependent synergistic release biphasic formulation, consisting of an immediate-release phase and an enteric-coated sustained-release synchronous phase, wherein the weight ratio of the immediate-release phase to the enteric-coated sustained-release synchronous phase is 1:4-6.

[0009] Furthermore, the immediate-release phase comprises 10%-20% of the total weight of the β-nicotinamide mononucleotide, as well as a filler and a disintegrant. The filler is microcrystalline cellulose, and the disintegrant is crospovidone. The enteric-coated sustained-release phase comprises 80%-90% of the total weight of the β-nicotinamide mononucleotide, all of the trans-resveratrol, all of the quercetin, all of the piperine, all of the trimethylglycine, as well as sustained-release materials and enteric coating materials.

[0010] Furthermore, the pharmaceutically acceptable excipient comprises hydroxypropyl-β-cyclodextrin with a degree of substitution of 4-9, and its mass ratio to trans-resveratrol, quercetin, and piperine is 4-8:1.

[0011] Furthermore, the pharmaceutically acceptable excipients include sustained-release materials and enteric coating materials; the sustained-release material is hydroxypropyl methylcellulose K4M, used in an amount of 3%-8% of the total weight of the enteric sustained-release phase; the enteric coating material is a mixture of acrylic resins II and III, with a weight ratio of 1:1-3, and the coating weight gain is 3%-8% of the total weight of the enteric sustained-release pellets.

[0012] On the other hand, the preparation of an NMN complex anti-aging agent that activates the SIRT1 pathway includes: S1, Preparation of trans-resveratrol-quercetin-piperine complex inclusion complex: The prescribed amounts of trans-resveratrol, quercetin, piperine and hydroxypropyl-β-cyclodextrin were mixed, purified water was added, the amount of purified water was 30%-50% of the total mass of solid materials, the mixture was ground and included at 300-600 r / min for 20-60 min, vacuum dried at 40-60℃ for 8-12 h, and passed through an 80-100 mesh sieve to obtain the complex inclusion complex powder; S2, Preparation of enteric-coated sustained-release synchronous phase: The composite inclusion powder obtained in step S1, trimethylglycine, 80%-90% of β-nicotinamide mononucleotide, hydroxypropyl methylcellulose K4M, and microcrystalline cellulose are mixed, purified water is added to make a soft material, and sustained-release microspheres are prepared by extrusion and spheroidization. The extrusion speed is 50-80 rpm, the spheroidization speed is 600-900 rpm, and the microspheres are dried at 40-50℃ until the moisture content is ≤3%. The sustained-release microspheres are coated with a mixed coating solution of acrylic resins II and III, and cured at 40-45℃ for 2-4 hours after coating to obtain enteric-coated sustained-release microspheres. S3, Preparation of immediate-release phase: Mix the remaining 10%-20% of β-nicotinamide mononucleotide with microcrystalline cellulose and cross-linked polyvinylpyrrolidone evenly, dry granulate and pass through a 20-mesh sieve to obtain immediate-release granules. S4, Total Mixing and Molding: Enteric-coated sustained-release microspheres and immediate-release granules are mixed evenly at a weight ratio of 4-6:1, and then filled into capsules or compressed into tablets to obtain an anti-aging preparation.

[0013] Compared with existing technologies, this NMN complex anti-aging agent that activates the SIRT1 pathway and its preparation method have the following beneficial effects: I. This invention utilizes a compound of β-nicotinamide mononucleotide, trans-resveratrol, quercetin, piperine, and trimethylglycine to form a synergistic regulatory system targeting the SIRT1 pathway, achieving sustained activation of this pathway. β-nicotinamide mononucleotide replenishes NAD+ required for SIRT1 activation; quercetin reduces NAD+ degradation to maintain its homeostasis; trans-resveratrol directly enhances SIRT1 catalytic activity; trimethylglycine eliminates nicotinamide produced by NMN metabolism, relieving its inhibition of SIRT1 and preventing long-term efficacy attenuation; piperine improves the oral absorption efficiency of each component, allowing each component to fully exert its effect. This compounding method forms a functional closed loop, improving age-related cellular function decline. Simultaneously, the synergistic effect of multiple components reduces the dosage of any single component, lessening the metabolic burden on the body and improving the safety of long-term use of the formulation.

[0014] II. This invention addresses the problems of asynchronous component release and poor absorption of poorly soluble components in traditional formulations through a time-sequential synergistic release biphasic formulation design and a specific combination of excipients. The immediate-release phase rapidly releases a portion of β-nicotinamide mononucleotide, promptly replenishing basal NAD+; the enteric-coated sustained-release phase releases all remaining active ingredients simultaneously in the intestine, ensuring matching peak plasma concentrations of each component and guaranteeing effective synergistic effects. Hydroxypropyl-β-cyclodextrin improves the water solubility and dissolution rate of poorly soluble components, while the combination of hydroxypropyl methylcellulose K4M and acrylic resin ensures targeted sustained-release in the intestine, enhancing the bioavailability of poorly soluble components. Simultaneously, this design enhances the storage stability of the formulation, prevents degradation of active ingredients, ensures stable efficacy within the shelf life, and the preparation process is compatible with conventional pharmaceutical equipment, guaranteeing consistent quality between batches.

[0015] Other advantages, objectives and features of the invention will be set forth in part in the description which follows, and in part will be apparent to those skilled in the art from the following examination or study, or may be learned from the practice of the invention. Attached Figure Description

[0016] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the accompanying drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are merely some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without any creative effort.

[0017] Figure 1 This is a process flow diagram for preparing the NMN complex anti-aging agent that activates the SIRT1 pathway according to the present invention. Figure 2 This is a schematic diagram illustrating the mechanism of action of the active ingredients in this invention in synergistically activating the SIRT1 pathway; Figure 3 This is a schematic diagram of the structure of the time-sequential synergistic release biphasic formulation of the present invention. Detailed Implementation

[0018] To further illustrate the technical means and effects of the present invention in achieving its intended purpose, the following detailed description of the specific implementation methods, structures, features, and effects of the present invention, in conjunction with the accompanying drawings and preferred embodiments, is provided below.

[0019] In all embodiments and comparative examples of this invention, all raw materials used are pharmaceutical grade. β-nicotinamide mononucleotide is β configuration crystal form I with a purity ≥99.5%, trans-resveratrol trans isomer content ≥98%, quercetin is anhydrous quercetin aglycone with a purity ≥98%, piperine purity ≥95%, and trimethylglycine is natural betaine extract with a purity ≥99%. All excipients are pharmaceutical grade. All preparation processes are carried out in a Class D cleanroom with a temperature of 20-25℃ and a relative humidity of 30%-40%. All purified water used is pharmaceutical grade purified water.

[0020] Example 1 This embodiment describes a time-dependent synergistic release biphasic anti-aging formulation with optimal active ingredient ratios, and its structure is as follows: Figure 3 As shown, the specific components of the active ingredients are: 50 parts β-nicotinamide mononucleotide, 15 parts trans-resveratrol, 5 parts quercetin, 1 part piperine, and 29 parts trimethylglycine.

[0021] The specific components of the excipients are: 84 parts of hydroxypropyl-β-cyclodextrin, 60 parts of microcrystalline cellulose, 8 parts of crospovidone, 12 parts of hydroxypropyl methylcellulose K4M, 6 parts of acrylic resin II, and 12 parts of acrylic resin III; purified water is used as a process solvent and is not included in the total amount of the formula.

[0022] like Figure 1 As shown, the specific preparation method is as follows: Preparation of trans-resveratrol-quercetin-piperine complex inclusion complex: The prescribed amounts of trans-resveratrol, quercetin, piperine, and hydroxypropyl-β-cyclodextrin were added to a colloid mill and mixed evenly. Purified water was added, with the amount of purified water being 40% of the total mass of the above solid materials. The speed of the colloid mill was adjusted to 450 r / min, and the inclusion complex was continuously ground and mixed for 40 min, with the temperature controlled at ≤30℃ throughout the process. The ground inclusion slurry was transferred to a vacuum drying oven, and the temperature was set at 50℃ and the vacuum degree at -0.09 MPa for 10 h. After drying, the material was pulverized by a universal pulverizer and passed through a 90-mesh standard sieve to obtain the complex inclusion powder, which was collected, sealed, and stored for later use.

[0023] Preparation of enteric-coated sustained-release synchronous phase: Take the above-mentioned composite inclusion complex powder, the prescribed amount of trimethylglycine, 90% of the prescribed amount of β-nicotinamide mononucleotide, hydroxypropyl methylcellulose K4M, and 40 parts of microcrystalline cellulose, and put them into a three-dimensional motion mixer. Adjust the speed to 18 r / min and mix for 15 min until the material is uniform. Add an appropriate amount of purified water to make a soft mass. After passing the soft mass through a 16-mesh sieve, put it into an extrusion spheronizer. Set the extrusion speed to 65 rpm, the spheronizer speed to 750 rpm, and the spheronizer time to 5 min to prepare the desired product. Spherical microspheres were prepared by transferring the spherical microspheres into a hot air drying oven and drying them at 45°C until the moisture content of the material was ≤3%. The microspheres were then granulated through a 20-40 mesh standard sieve to obtain sustained-release microspheres. The sustained-release microspheres were placed in a fluidized bed coating machine and coated with a mixed coating solution prepared with acrylic resins II and III. The inlet air temperature was set at 45°C, the bed temperature at 38°C, and the atomization pressure at 0.2 MPa. After coating, the microspheres were placed in an electric constant temperature oven and cured at 42°C for 3 hours to obtain enteric-coated sustained-release microspheres, which were then collected, sealed, and stored for later use.

[0024] Preparation of the immediate-release phase: Take the remaining 10% of β-nicotinamide mononucleotide in the formula, add it to a high-speed mixing granulator along with 20 parts of microcrystalline cellulose and cross-linked polyvinylpyrrolidone, mix for 5 minutes until the material is uniform, granulate using a dry granulation process, and granulate the obtained particles through a 20-mesh standard sieve to obtain immediate-release particles, which are then collected, sealed, and stored for later use.

[0025] Total mixing and molding: The above enteric-coated sustained-release microspheres and immediate-release granules were added to a three-dimensional motion mixer at a weight ratio of 5:1. The mixing speed was adjusted to 20 r / min and mixed for 10 min until the materials were uniform. Using a fully automatic capsule filling machine, the mixture was filled into No. 0 hard capsule shells at a dosage of 0.5 g / capsule to obtain the NMN complex anti-aging preparation of the present invention that activates the SIRT1 pathway. Figure 2 As shown.

[0026] In this embodiment, there was no material loss exceeding the standard throughout the preparation process, and the test indicators of each intermediate product met the preset quality standards: the dissolution test of the composite inclusion powder showed that the dissolution rate of trans-resveratrol reached 92.5% after 15 minutes, and the dissolution rate of quercetin reached 90.8% after 15 minutes, which is more than 80% higher than that of the uninclusion raw material; the moisture content of the slow-release microspheres after drying was 2.5%, the particle size distribution was uniform, and the proportion of 20-40 mesh particles was 98.2%; the dissolution rate of the enteric-coated slow-release microspheres was ≤2.8% after standing in pH 1.2 hydrochloric acid medium for 2 hours, and the cumulative dissolution rate reached 88.6% after shaking in pH 6.8 phosphate buffer medium for 4 hours, which fully meets the requirements of the enteric-coated slow-release design; the dissolution rate of the immediate-release particles reached 95.3% after 15 minutes in pH 6.8 phosphate buffer medium, which can achieve rapid release and effect. In vitro cell experiments showed that the formulation in this embodiment increased the activation rate of the SIRT1 pathway by 5.2 times compared with the blank group, the nicotinamide clearance rate reached 98.3%, the intracellular NAD+ level was maintained for more than 48 hours, and all efficacy indicators reached the optimal level. In addition, the formulation had a smooth appearance, uniform particles, and no adhesion or clumping.

[0027] Example 2 This embodiment is a time-dependent synergistic release biphasic anti-aging formulation with a lower limit ratio of active ingredients. The specific components of the active ingredients are: 35 parts of β-nicotinamide mononucleotide, 10 parts of trans-resveratrol, 3 parts of quercetin, 0.8 parts of piperine, and 51.2 parts of trimethylglycine.

[0028] The specific components of the excipients are: 55 parts of hydroxypropyl-β-cyclodextrin, 50 parts of microcrystalline cellulose, 6 parts of crospovidone, 8 parts of hydroxypropyl methylcellulose K4M, 4 parts of acrylic resin II, and 8 parts of acrylic resin III; purified water is used as a process solvent and is not included in the total amount of the formula.

[0029] The specific preparation method is as follows: Preparation of trans-resveratrol-quercetin-piperine complex inclusion complex: The prescribed amounts of trans-resveratrol, quercetin, piperine, and hydroxypropyl-β-cyclodextrin were added to a colloid mill and mixed evenly. Purified water was added, with the amount of purified water being 35% of the total mass of the above solid materials. The speed of the colloid mill was adjusted to 350 r / min, and the inclusion complex was continuously ground and mixed for 30 min, with the temperature controlled at ≤30℃ throughout the process. The ground inclusion slurry was transferred to a vacuum drying oven, and the temperature was set at 45℃ and the vacuum degree at -0.08 MPa for 11 h. After drying, the material was pulverized by a universal pulverizer and passed through an 80-mesh standard sieve to obtain the complex inclusion powder, which was collected, sealed, and stored for later use.

[0030] Preparation of enteric-coated sustained-release synchronous phase: Take the above-mentioned composite inclusion complex powder, the prescribed amount of trimethylglycine, 85% of the prescribed amount of β-nicotinamide mononucleotide, hydroxypropyl methylcellulose K4M, and 35 parts of microcrystalline cellulose, and put them into a three-dimensional motion mixer. Adjust the speed to 18 r / min and mix for 15 min until the material is uniform. Add an appropriate amount of purified water to make a soft mass. After passing the soft mass through a 16-mesh sieve, put it into an extruder and rounding machine. Set the extrusion speed to 55 rpm, the rounding speed to 650 rpm, and the rounding time to 5 min to prepare the desired product. Spherical microspheres were prepared by transferring the spherical microspheres into a hot air drying oven and drying them at 42°C until the moisture content of the material was ≤3%. The microspheres were then granulated through a 20-40 mesh standard sieve to obtain sustained-release microspheres. The sustained-release microspheres were placed in a fluidized bed coating machine and coated with a mixed coating solution prepared with acrylic resins II and III. The inlet air temperature was set at 42°C, the bed temperature at 35°C, and the atomization pressure at 0.2 MPa. After coating, the microspheres were placed in an electric constant temperature oven and cured at 40°C for 4 hours to obtain enteric-coated sustained-release microspheres, which were then collected, sealed, and stored for later use.

[0031] Preparation of the immediate-release phase: Take the remaining 15% of β-nicotinamide mononucleotide in the formula, add it to a high-speed mixing granulator along with 15 parts of microcrystalline cellulose and cross-linked polyvinylpyrrolidone, mix for 5 minutes until the material is uniform, granulate using a dry granulation process, and granulate the obtained particles through a 20-mesh standard sieve to obtain immediate-release particles, which are then collected, sealed, and stored for later use.

[0032] Total mixing and molding: The above enteric-coated sustained-release microspheres and immediate-release granules were put into a three-dimensional motion mixer at a weight ratio of 4:1. The speed was adjusted to 20 r / min and mixed for 10 min until the materials were uniform. The mixture was then filled into No. 1 hard capsule shells at a dosage of 0.4 g / capsule using a fully automatic capsule filling machine to obtain the NMN complex anti-aging preparation of the present invention that activates the SIRT1 pathway.

[0033] This embodiment uses a slightly lower limit ratio of active ingredients for preparation. The process parameters are finely adjusted according to the amount of raw materials used. The material loss throughout the preparation process complies with pharmaceutical specifications, and the test indicators of each intermediate product meet the preset quality standards: The dissolution test of the composite inclusion powder showed that the dissolution rate of trans-resveratrol reached 89.2% after 15 minutes, and the dissolution rate of quercetin reached 87.5% after 15 minutes, which is more than 75% higher than that of the uninclusion raw materials; The moisture content of the sustained-release microcapsules after drying was 2.8%, and the particle size distribution was uniform, with 97.5% of the particles being 20-40 mesh; The dissolution rate of the enteric-coated sustained-release microcapsules was ≤3.2% after standing in a pH 1.2 hydrochloric acid medium for 2 hours, and the cumulative dissolution rate reached 85.3% after shaking in a pH 6.8 phosphate buffer medium for 4 hours, meeting the requirements of the enteric-coated sustained-release design; The dissolution rate of the immediate-release granules reached 93.8% after 15 minutes in a pH 6.8 phosphate buffer medium, enabling rapid release and efficacy. In vitro cell experiments showed that the formulation in this embodiment increased the activation rate of the SIRT1 pathway by 4.1 times compared with the control group, achieved a nicotinamide clearance rate of 95.6%, maintained intracellular NAD+ levels for over 36 hours, and demonstrated stable and controllable efficacy indicators. This embodiment effectively activates the SIRT1 pathway while reducing the amount of raw materials used, balancing formulation efficacy and raw material costs, and possesses good cost-effectiveness for industrial production.

[0034] Example 3 This embodiment is a time-dependent synergistic release biphasic anti-aging formulation with an active ingredient ratio close to the upper limit. The specific components of the active ingredients are: 55 parts of β-nicotinamide mononucleotide, 18 parts of trans-resveratrol, 8 parts of quercetin, 2 parts of piperine, and 17 parts of trimethylglycine.

[0035] The specific components of the excipients are: 108 parts of hydroxypropyl-β-cyclodextrin, 70 parts of microcrystalline cellulose, 10 parts of crospovidone, 15 parts of hydroxypropyl methylcellulose K4M, 7 parts of acrylic resin II, and 14 parts of acrylic resin III; purified water is used as a process solvent and is not included in the total amount of the formula.

[0036] The specific preparation method is as follows: Preparation of trans-resveratrol-quercetin-piperine complex inclusion complex: The prescribed amounts of trans-resveratrol, quercetin, piperine, and hydroxypropyl-β-cyclodextrin were added to a colloid mill and mixed evenly. Purified water was added, with the amount of purified water being 45% of the total mass of the above solid materials. The speed of the colloid mill was adjusted to 550 r / min, and the inclusion complex was continuously ground and mixed for 50 min, with the temperature controlled at ≤30℃ throughout the process. The ground inclusion slurry was transferred to a vacuum drying oven, and the temperature was set at 55℃ and the vacuum degree at -0.09 MPa for 9 h. After drying, the material was pulverized by a universal pulverizer and passed through a 100-mesh standard sieve to obtain the complex inclusion powder, which was collected, sealed, and stored for later use.

[0037] Preparation of enteric-coated sustained-release synchronous phase: Take the above-mentioned composite inclusion complex powder, the prescribed amount of trimethylglycine, 88% of the prescribed amount of β-nicotinamide mononucleotide, hydroxypropyl methylcellulose K4M, and 45 parts of microcrystalline cellulose, and put them into a three-dimensional motion mixer. Adjust the speed to 18 r / min and mix for 15 min until the material is uniform. Add an appropriate amount of purified water to make a soft mass. After passing the soft mass through a 16-mesh sieve, put it into an extrusion spheronizer. Set the extrusion speed to 75 rpm, the spheronizer speed to 850 rpm, and the spheronizer time to 5 min to prepare the desired product. Spherical microspheres were prepared by transferring the spherical microspheres into a hot air drying oven and drying them at 48°C until the moisture content of the material was ≤3%. The microspheres were then granulated through a 20-40 mesh standard sieve to obtain sustained-release microspheres. The sustained-release microspheres were placed in a fluidized bed coating machine and coated with a mixed coating solution prepared with acrylic resins II and III. The inlet air temperature was set at 48°C, the bed temperature at 40°C, and the atomization pressure at 0.2 MPa. After coating, the microspheres were placed in an electric constant temperature oven and cured at 45°C for 2 hours to obtain enteric-coated sustained-release microspheres, which were then collected, sealed, and stored for later use.

[0038] Preparation of the immediate-release phase: Take the remaining 12% of β-nicotinamide mononucleotide in the formula, add it to a high-speed mixing granulator along with 25 parts of microcrystalline cellulose and cross-linked polyvinylpyrrolidone, mix for 5 minutes until the material is uniform, granulate using a dry granulation process, and granulate the obtained particles through a 20-mesh standard sieve to obtain immediate-release particles, which are collected, sealed and stored for later use.

[0039] Total mixing and molding: The above enteric-coated sustained-release microspheres and immediate-release granules were put into a three-dimensional motion mixer at a weight ratio of 6:1. The speed was adjusted to 20 r / min and mixed for 10 min until the materials were uniform. The mixture was then filled into No. 00 hard capsule shells at a dosage of 0.6 g / capsule using a fully automatic capsule filling machine to obtain the NMN complex anti-aging preparation of the present invention that activates the SIRT1 pathway.

[0040] This embodiment uses a ratio of active ingredients slightly above the upper limit for preparation. Process parameters are adjusted accordingly as the amount of raw materials increases. Material loss throughout the preparation process meets pharmaceutical standards, and the test indicators of each intermediate product are better than the preset quality standards: The dissolution test of the composite inclusion powder showed that trans-resveratrol achieved a dissolution rate of 93.1% after 15 minutes, and quercetin achieved a dissolution rate of 91.5% after 15 minutes, which is more than 82% higher than the dissolution rate of the uninclusion raw materials; The moisture content of the sustained-release microcapsules after drying was 2.3%, and the particle size distribution was uniform, with 98.6% of the particles being 20-40 mesh; The enteric-coated sustained-release microcapsules had a dissolution rate of ≤2.5% after standing in a pH 1.2 hydrochloric acid medium for 2 hours, and a cumulative dissolution rate of 90.2% after shaking in a pH 6.8 phosphate buffer medium for 4 hours, which is better than the design requirements for enteric-coated sustained-release; The immediate-release granules achieved a dissolution rate of 96.1% after 15 minutes in a pH 6.8 phosphate buffer medium, demonstrating a significant rapid release effect. In vitro cell experiments showed that the formulation in this embodiment increased the activation rate of the SIRT1 pathway by 5.5 times compared with the blank group, and the nicotinamide clearance rate reached 98.8%. The intracellular NAD+ level increased rapidly in a short period of time and was maintained for more than 48 hours, showing outstanding short-term activation effect. It is suitable for rapid intervention on the decline of age-related physiological functions. The formulation has a smooth appearance, with only a small amount of microspheres adhering slightly, which does not affect the dissolution effect and performance of the formulation.

[0041] Comparative Example This comparative example is a traditional conventional compound anti-aging preparation, which does not contain trimethylglycine and does not adopt a time-sequential synergistic release biphasic design, in contrast to the present invention. The total amount of active ingredients is 100 parts by weight, specifically: 60 parts of β-nicotinamide mononucleotide, 20 parts of trans-resveratrol, 10 parts of quercetin, and 10 parts of piperine.

[0042] The total excipients amount to 176 parts by weight, specifically: 200 parts of hydroxypropyl-β-cyclodextrin, 50 parts of microcrystalline cellulose, and 3 parts of magnesium stearate; there are no sustained-release materials, enteric coating materials, or disintegrants, and purified water is used as a process solvent and is not quantitatively included in the total formulation.

[0043] The specific preparation method is as follows: Preparation of simple inclusion complex: The prescribed amounts of trans-resveratrol, quercetin, piperine and hydroxypropyl-β-cyclodextrin were put into a glass mortar and ground and mixed for 10 minutes until initially uniform. A small amount of purified water was added and the mixture was ground until it became a paste. The paste was placed in a 40°C hot air drying oven and dried. After drying, the material was pulverized by a universal pulverizer and passed through an 80-mesh standard sieve to obtain simple inclusion complex powder, which was collected for later use.

[0044] Total mixing and molding: The above simple inclusion complex powder, along with all β-nicotinamide mononucleotide and microcrystalline cellulose, are put into a trough mixer and mixed for 10 minutes until the material is roughly uniform. Magnesium stearate is added and mixed again for 5 minutes. Using a semi-automatic capsule filling machine, the mixture is filled into No. 00 hard capsule shells at a dosage of 0.6g / capsule to obtain a traditional conventional compound anti-aging preparation.

[0045] This comparative example is a conventional compound formulation without added trimethylglycine. The total active ingredient content is 100 parts by weight, and each component is at its upper limit. The preparation process is simple, with few types of excipients and no targeted process parameter control. Material loss during preparation is relatively high, and the test indicators of each intermediate product are far below the standards of the formulation of this invention: the dissolution rate of the simple inclusion complex powder was only 45.2% after 15 minutes, the dissolution rate of quercetin was only 41.8% after 15 minutes, and the dissolution rate of piperine was only 43.5%, indicating poor dissolution effect; the formulation lacks an enteric coating design, and after standing in hydrochloric acid medium at pH 1.2 for 2 hours, the dissolution rate of β-nicotinamide mononucleotide reached 98.5%, which is easily released and degraded rapidly in gastric acid; there is no dual-phase design of immediate and sustained release, and the dissolution of each component is irregular. The dissolution rate of trans-resveratrol in phosphate buffer medium at pH 6.8 was only 62.3% after 4 hours, which is more than 3 hours different from the dissolution time of β-nicotinamide mononucleotide. In vitro cell experiments showed that the comparative formulation only increased the activation rate of the SIRT1 pathway by 1.2 times compared with the blank group, had no nicotinamide clearance ability, and the in vivo accumulation rate of nicotinamide reached 89.5%. The intracellular NAD+ level only increased briefly in the early stage and quickly dropped within 12 hours, failing to achieve sustained activation of the SIRT1 pathway. All efficacy indicators did not meet the basic requirements for the use of anti-aging formulations. Furthermore, the formulation was prone to component degradation and particle agglomeration during storage, and the material loss was more than 15% higher than that of the formulation of this invention.

[0046]

[0047] As shown in the table above, Examples 1, 2, and 3 are all NMN compound anti-aging preparations that activate the SIRT1 pathway, meeting the design requirements of this invention. The process parameters were adjusted according to the differences in the active ingredient ratios, achieving a dual-phase release design of enteric-coated sustained release and immediate release. All test indicators met relevant pharmaceutical industry quality standards, with low material loss and no process defects or efficacy shortcomings. Example 1, with its optimal ratio, achieved the best balance between core efficacy indicators such as SIRT1 pathway activation rate, nicotinamide clearance rate, and NAD+ maintenance time, and the formulation process indicators. It exhibited excellent storage stability and component dissolution, making it the best implementation method of this invention. Example 2, with its lower-limit ratio, reduced raw material usage while maintaining core efficacy, offering significant cost advantages and suitable for general consumption scenarios. Example 3, with its upper-limit ratio, demonstrated outstanding short-term SIRT1 pathway activation effects, rapidly improving age-related physiological function decline, making it suitable for targeted and rapid intervention scenarios. Both examples can adapt to different consumer needs and application scenarios. Comparative Example 1, as a traditional conventional compound formulation, lacks the addition of trimethylglycine for nicotinamide metabolism regulation, does not employ a time-sequential synergistic release biphasic design, and has a simple preparation process without precise parameter control. As a result, its core efficacy and process indicators are still far lower than those of the formulation of this invention. It not only fails to achieve continuous and efficient activation of the SIRT1 pathway, but also suffers from poor component dissolution, low storage stability, and high material loss. This fully demonstrates the innovation of this invention in active ingredient compounding, formulation structure design, and preparation process control.

[0048] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the above-disclosed technical content to create equivalent embodiments without departing from the scope of the present invention. Any simple modifications, equivalent changes and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. An NMN complex anti-aging agent that activates the SIRT1 pathway, characterized in that, This formulation comprises an active ingredient and pharmaceutically acceptable excipients, wherein the active ingredient comprises 100 parts by weight and includes the following components: 30-60 parts of β-nicotinamide mononucleotide; 8-20 parts of trans-resveratrol; Quercetin 2-10 parts; Piperine 0.5-3 parts; The balance is trimethylglycine.

2. The NMN complex anti-aging agent that activates the SIRT1 pathway according to claim 1, characterized in that, The trimethylglycine is a natural betaine extract with an extraction purity of ≥99%, of which the free betaine content is ≥98.5% and the moisture content is ≤0.5%.

3. The NMN complex anti-aging agent for activating the SIRT1 pathway according to claim 1, characterized in that, The β-nicotinamide mononucleotide is β-configuration crystal form I, with a particle size range of 10-50 μm, purity ≥99.5%, and α-configuration impurities ≤0.1%; the trans-resveratrol has a trans isomer content ≥98% and a melting point of 254-256℃; the quercetin is anhydrous quercetin aglycone, with a particle size ≤20 μm, purity ≥98%, and glycoside impurities ≤1%; the piperine has a purity ≥95% and a melting point of 128-130℃.

4. The NMN complex anti-aging agent that activates the SIRT1 pathway according to claim 1, characterized in that, The formulation is a time-dependent synergistic release biphasic formulation, consisting of an immediate-release phase and an enteric-coated sustained-release synchronous phase, wherein the weight ratio of the immediate-release phase to the enteric-coated sustained-release synchronous phase is 1:4-6.

5. The NMN complex anti-aging agent that activates the SIRT1 pathway according to claim 4, characterized in that, The immediate-release phase comprises 10%-20% of the total weight of the β-nicotinamide mononucleotide, as well as a filler and a disintegrant. The filler is microcrystalline cellulose, and the disintegrant is crospovidone. The enteric-coated sustained-release phase comprises 80%-90% of the total weight of the β-nicotinamide mononucleotide, all trans-resveratrol, all quercetin, all piperine, all trimethylglycine, as well as sustained-release materials and enteric coating materials.

6. The NMN complex anti-aging agent for activating the SIRT1 pathway according to claim 1, characterized in that, The pharmaceutically acceptable excipient comprises hydroxypropyl-β-cyclodextrin, wherein the degree of substitution of the hydroxypropyl-β-cyclodextrin is 4-9, and the mass ratio of the hydroxypropyl-β-cyclodextrin to the total mass of trans-resveratrol, quercetin, and piperine is 4-8:

1.

7. The NMN complex anti-aging agent for activating the SIRT1 pathway according to claim 1, characterized in that, The pharmaceutically acceptable excipients include sustained-release materials and enteric coating materials; the sustained-release material is hydroxypropyl methylcellulose K4M, and the amount used is 3%-8% of the total weight of the enteric sustained-release phase; the enteric coating material is a mixture of acrylic resins II and III, with a weight ratio of 1:1-3, and the coating weight gain is 3%-8% of the total weight of the enteric sustained-release pellets.

8. Preparation of an NMN complex anti-aging agent that activates the SIRT1 pathway, the method being used to prepare the NMN complex anti-aging agent that activates the SIRT1 pathway as described in any one of claims 1-7, characterized in that, The preparation method includes: S1, Preparation of trans-resveratrol-quercetin-piperine complex inclusion complex: The prescribed amounts of trans-resveratrol, quercetin, piperine and hydroxypropyl-β-cyclodextrin were mixed, purified water was added, the amount of purified water was 30%-50% of the total mass of solid materials, the mixture was ground and included at 300-600 r / min for 20-60 min, vacuum dried at 40-60℃ for 8-12 h, and passed through an 80-100 mesh sieve to obtain the complex inclusion complex powder; S2, Preparation of enteric-coated sustained-release synchronous phase: The composite inclusion powder obtained in step S1, trimethylglycine, 80%-90% of β-nicotinamide mononucleotide, hydroxypropyl methylcellulose K4M, and microcrystalline cellulose are mixed, purified water is added to make a soft material, and sustained-release microspheres are prepared by extrusion and spheroidization. The extrusion speed is 50-80 rpm, the spheroidization speed is 600-900 rpm, and the microspheres are dried at 40-50℃ until the moisture content is ≤3%. The sustained-release microspheres are coated with a mixed coating solution of acrylic resins II and III, and cured at 40-45℃ for 2-4 hours after coating to obtain enteric-coated sustained-release microspheres. S3, Preparation of immediate-release phase: Mix the remaining 10%-20% of β-nicotinamide mononucleotide with microcrystalline cellulose and cross-linked polyvinylpyrrolidone evenly, dry granulate and pass through a 20-mesh sieve to obtain immediate-release granules. S4, Total Mixing and Molding: Enteric-coated sustained-release microspheres and immediate-release granules are mixed evenly at a weight ratio of 4-6:1, and then filled into capsules or compressed into tablets to obtain an anti-aging preparation.

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