An NMN composite Anti-aging formulation for activating the SIRT1 pathway and a preparation thereof

Abstract

The present invention discloses an NMN composite anti-aging formulation for activating the SIRT1 pathway and a preparation thereof, relating to the technical field of pharmaceutical preparations. The formulation consists of active ingredients and pharmaceutically acceptable excipients. The active ingredients comprise β-nicotinamide mononucleotide, trans-resveratrol, quercetin, piperine, and trimethylglycine in specified weight ratios. The formulation forms a synergistic system targeting the SIRT1 pathway through a biphasic design enabling time-sequential release of components. The invention improves the solubility, stability, and bioavailability of active ingredients, and addresses asynchronous release and poor absorption of poorly soluble components in conventional formulations. The preparation process is compatible with conventional pharmaceutical equipment and ensures consistent batch quality.

Description

[0001] DESCRIPTION

[0002] An NMN composite anti-aging formulation for activating the SIRT1 pathway and a preparation thereof TECHNICAL FIELD

[0003] The present invention relates to the technical field of pharmaceutical preparations, specifically to an NMN composite anti-aging formulation for activating the SIRT1 pathway and a preparation thereof.

[0004] BACKGROUND

[0005] Aging is a process of progressive decline in cell, tissue, and organ function with increasing age. Among these, the NAD+-dependent SIRT1 pathway is a core molecular pathway regulating the aging process. SIRT1, as an NAD+-dependent deacetylase, participates in physiological processes closely related to aging, such as DNA damage repair, mitochondrial energy metabolism, and oxidative stress response, through deacetylation modification of downstream target proteins. Its activity is directly regulated by the level of NAD+ in the body. With increasing age, the body's NAD+ synthesis capacity decreases and its degradation rate accelerates, leading to a continuous decline in NAD+ levels and consequently a decrease in SIRT1 activity, exacerbating aging-related phenotypes. P-nicotinamide mononucleotide, as a direct precursor of NAD+, can be rapidly converted to NAD+, making it a core raw material for the research and development of anti-aging formulations. The industry commonly adopts a technical route of compounding NMN with SIRT1 activators and NAD+ degradation inhibitors.

[0006] Existing anti-aging formulations have many defects in practical application. Nicotinamide produced by NMN metabolism inhibits SIRT1 activity, and long-term administration can easily lead to nicotinamide accumulation, counteracting the activation effect. However, traditional compounding solutions do not include specific components for regulating nicotinamide metabolism. The physicochemical properties of different active ingredients vary greatly; NMN is absorbed quickly, while poorly soluble components such as trans-resveratrol and quercetin dissolve slowly, leading to asynchronous release and absorption of the components, preventing a synergistic effect. Furthermore, conventional excipient combinations lack specificity, resulting in low bioavailability of poorly soluble components, and some formulations have insufficient stability, with active ingredients prone to degradation during storage. This makes it impossible to achieve continuous activation of the SIRT1 pathway and difficult to meet the needs of long-term anti-aging use.

[0007] SUMMARY

[0008] An objective of the present invention is to address the shortcomings of the prior art by providing an NMN composite anti-aging formulation for activating the SIRT1 pathway and a preparation thereof. By precisely compounding five major active ingredients including P-nicotinamide mononucleotide and trans-resveratrol, a synergistic regulatory system targeting the SIRT1 pathway is constructed. Combined with a time-sequential synergistic release biphasic formulation design and the scientific combination of specific pharmaceutical excipients, the invention effectively solves the problems of nicotinamide accumulation, asynchronous release of components, and low bioavailability of poorly soluble components in traditional anti-aging formulations. The present invention achieves continuous and efficient activation of the SIRT1 pathway, stably maintains intracellular NAD+ levels, and also improves the storage stability and long-term administration safety of the formulation. The preparation process is compatible with conventional pharmaceutical equipment, batch quality is controllable, and different component ratio schemes can be adapted to various application scenarios.

[0009] To solve the above technical problems, the present invention provides the following technical solutions: In one aspect, an NMN composite anti-aging formulation for activating the SIRT1 pathway, the formulation consists of active ingredients and pharmaceutically acceptable excipients, the active ingredients totaling 100 parts by weight, comprising the following components:

[0010] 30-60 parts by weight of P-nicotinamide mononucleotide;

[0011] 8-20 parts by weight of trans-resveratrol;

[0012] 2-10 parts by weight of quercetin;

[0013] 0.5-3 parts by weight of piperine; and the balance being trimethylglycine.

[0014] Further, the trimethylglycine is a natural betaine extract with an extraction purity >99%, wherein the free betaine content is >98.5% and the water content is <0.5%.

[0015] Further, the P-nicotinamide mononucleotide is P-configuration crystal form I, with a particle size range of 10-50 pm, purity >99.5%, and a-configuration impurity <0.1%; the trans-resveratrol has a trans-isomer content >98% and a melting point of 254-256°C; the quercetin is anhydrous quercetin aglycone, with a particle size <20 pm, purity >98%, and glycoside impurities <1%; the piperine has a purity >95% and a melting point of 128-130°C.

[0016] Further, the formulation is a time-sequential 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.

[0017] Further, the immediate-release phase comprises 10%-20% of the total weight of the P-nicotinamide mononucleotide, along with a filler and a disintegrant, the filler being microcrystalline cellulose and the disintegrant being crospovidone; the enteric-coated sustained-release synchronous phase comprises 80%-90% of the total weight of the P-nicotinamide mononucleotide, all of the trans-resveratrol, all of the quercetin, all of the piperine, all of the trimethylglycine, along with a sustained-release material and an enteric coating material.

[0018] Further, the pharmaceutically acceptable excipients comprise hydroxypropyl-P-cyclodextrin, the degree of substitution of the hydroxypropyl-P-cyclodextrin is 4-9, and the ratio of its mass to the total mass of trans-resveratrol, quercetin, and piperine is 4-8:1.

[0019] Further, the pharmaceutically acceptable excipients comprise a sustained-release material and an enteric coating material; the sustained-release material is hypromellose K4M, used in an amount of 3%-8% of the total weight of the enteric-coated sustained-release synchronous phase; the enteric coating material is a mixture of acrylic resin II and acrylic resin III, the weight ratio of the two being 1:1-3, and the coating weight gain is 3%-8% of the total weight of the enteric-coated sustained-release pellets.

[0020] In another aspect, a preparation of an NMN composite anti-aging formulation for activating the SIRT1 pathway, the preparation method comprises:

[0021] 51. Preparation of trans-resveratrol-quercetin-piperine complex inclusion compound: mixing the prescribed amounts of trans-resveratrol, quercetin, piperine with hydroxypropyl- P-cyclodextrin, adding purified water in an amount of 30%-50% of the total mass of the solid materials, grinding and including at a speed of 300-600 r / min for 20-60 min, vacuum drying at 40-60°C for 8-12 h, passing through an 80-100 mesh sieve to obtain a complex inclusion compound powder;

[0022] 52. Preparation of enteric-coated sustained-release synchronous phase: mixing the complex inclusion compound powder obtained in SI, the prescribed amount of trimethylglycine, 80%-90% of the prescribed amount of P-nicotinamide mononucleotide with hypromellose K4M and microcrystalline cellulose, adding purified water to make a soft material, extruding and spheronizing to prepare sustained-release pellets, with an extrusion speed of 50-80 rpm and a spheronization speed of 600-900 rpm, drying at 40-50°C to a moisture content <3%; coating the sustained-release pellets with a mixed coating solution of acrylic resin II and acrylic resin III, and after coating, curing at 40-45 °C for 2-4 h to obtain enteric-coated sustained-release pellets;

[0023] 53. Preparation of immediate-release phase: uniformly mixing the remaining 10%-20% of the prescribed amount of P-nicotinamide mononucleotide with microcrystalline cellulose and crospovidone, dry granulating and passing through a 20-mesh sieve to obtain immediate-release granules;

[0024] 54. Total mixing and forming: uniformly mixing the enteric-coated sustained-release pellets and the immediate-release granules in a weight ratio of 4-6:1, and filling into capsules or compressing into tablets to obtain the anti-aging formulation.

[0025] Compared with the prior art, this NMN composite anti-aging formulation for activating the SIRT1 pathway and its preparation scheme have the following beneficial effects:

[0026] I. The present invention, by compounding P-nicotinamide mononucleotide, trans-resveratrol, quercetin, piperine, and trimethylglycine, forms a synergistic regulatory system targeting the SIRT1 pathway, achieving continuous activation of this pathway. P-nicotinamide mononucleotide supplements the NAD+ required for SIRT1 activation, quercetin reduces NAD+ degradation to maintain its homeostasis, trans-resveratrol directly enhances SIRT1 catalytic activity, trim ethyl glycine scavenges nicotinamide produced by NMN metabolism, relieving its inhibition on SIRT1 and avoiding attenuation of the effect during long-term administration, and piperine enhances the oral absorption efficiency of each component, allowing their effects to be fully exerted. This compounding method forms a functional IT] ST, improving cell function decline caused by aging, while also reducing the dosage of individual components through multi-component synergy, lessening the metabolic burden on the body, and enhancing the safety of long-term administration of the formulation.

[0027] II. The present invention, through the time-sequential synergistic release biphasic formulation design and the combination of specific excipients, solves the problems of asynchronous release of components and poor absorption of poorly soluble components in traditional formulations. The immediate-release phase rapidly releases a portion of P-nicotinamide mononucleotide, promptly supplementing the basal NAD+; the enteric-coated sustained-release synchronous phase releases all other active ingredients simultaneously in the intestine, matching the peak plasma concentrations of each component and ensuring the effective exertion of the synergistic effect. Hydroxypropyl-P-cyclodextrin improves the water solubility and dissolution rate of poorly soluble components; the combination of hypromellose K4M and acrylic resins ensures intestinal targeted sustained-release, enhancing the bioavailability of poorly soluble components. Simultaneously, this design enhances the storage stability of the formulation, preventing degradation of active ingredients, ensuring stable efficacy within the shelf life. The preparation process is compatible with conventional pharmaceutical equipment and ensures consistent quality between batches.

[0028] Other advantages, objectives, and features of the present invention will be set forth in part in the following description, and in part will become apparent to those skilled in the art upon examination of the following or may be learned from practice of the invention.

[0029] BRIEF DESCRIPTION OF DRAWINGS

[0030] To explain the technical solutions in the embodiments of the present invention or the prior art more clearly, the following briefly introduces the drawings required for describing the embodiments or the prior art. Obviously, the drawings in the following description are only some embodiments of the present invention. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without inventive effort.

[0031] Figure 1 is a process flow diagram of the preparation of the NMN composite anti-aging formulation for activating the SIRT1 pathway according to the present invention;

[0032] Figure 2 is a schematic diagram illustrating the mechanism of synergistic activation of the SIRT1 pathway by the active ingredients of the present invention;

[0033] Figure 3 is a schematic structural diagram of the time-sequential synergistic release biphasic formulation of the present invention.

[0034] DETAILED DESCRIPTION

[0035] To further elaborate on the technical means and effects adopted by the present invention to achieve the intended invention objectives, the following, in conjunction with the accompanying drawings and preferred embodiments, provides a detailed description of the specific implementation, structure, features, and effects of the present invention.

[0036] In all embodiments and comparative examples of the present invention, the raw materials used are all of pharmaceutical grade standard. The P-nicotinamide mononucleotide is P-configuration crystal form I with purity >99.5%. The trans-resveratrol has a trans-isomer content >98%. The quercetin is anhydrous quercetin aglycone with purity >98%. The piperine has purity >95%. The trimethylglycine is a natural betaine extract with purity >99%. All excipients are of pharmaceutical grade standard. The preparation procedures are all carried out in a D-level clean workshop with a temperature of 20-25 °C and relative humidity of 30%-40%. The purified water used is all pharmaceutical grade purified water.

[0037] Embodiment 1

[0038] This embodiment is a time-sequential synergistic release biphasic anti-aging formulation with an optimal ratio of active ingredients, its structure is shown in Figure 3. The specific composition of active ingredients is: P-nicotinamide mononucleotide 50 parts, trans-resveratrol 15 parts, quercetin 5 parts, piperine 1 part, trimethylglycine 29 parts.

[0039] The specific composition of excipients is: hydroxypropyl- P-cyclodextrin 84 parts, microcrystalline cellulose 60 parts, crospovidone 8 parts, hypromellose K4M 12 parts, acrylic resin II 6 parts, acrylic resin III 12 parts; purified water is a process solvent and its amount is not counted in the total formula.

[0040] As shown in Figure 1, the specific preparation method is as follows:

[0041] Preparation of trans-resveratrol-quercetin-piperine complex inclusion compound: Put the prescribed amounts of trans-resveratrol, quercetin, piperine, and hydroxypropyl- P-cyclodextrin into a colloid mill and mix uniformly. Add purified water, the amount of purified water being 40% of the total mass of the solid materials. Adjust the colloid mill speed to 450 r / min, continuously grind and include for 40 min, controlling the temperature throughout to <30 °C. Transfer the ground inclusion slurry to a vacuum drying oven, set the temperature to 50°C and vacuum degree to -0.09 MPa, vacuum dry for 10 h. After drying, grind the material using a universal grinder, pass through a 90-mesh standard sieve to obtain the complex inclusion compound powder, collect and seal for later use. Preparation of enteric-coated sustained-release synchronous phase: Take the above complex inclusion compound powder, the prescribed amount of trimethylglycine, 90% of the prescribed amount of P-nicotinamide mononucleotide, along with hypromellose K4M and 40 parts of microcrystalline cellulose, put them into a three-dimensional motion mixer, adjust the speed to 18 r / min and mix for 15 min until the materials are uniform. Add an appropriate amount of purified water to make a soft material. Pass the soft material through a 16-mesh sieve and then put it into an extruder- spheronizer, set the extrusion speed to 65 rpm, spheronization speed to 750 rpm, and spheronization time to 5 min to prepare spherical pellets. Transfer the spherical pellets to a hot air drying oven, dry with air blast at 45°C until the material moisture content is <3%, and sieve using 20-40 mesh standard sieves to obtain sustained-release pellets. Place the sustained-release pellets in a fluid bed coater, coat with the mixed coating solution prepared from acrylic resin II and acrylic resin III, set the inlet air temperature to 45 °C, bed temperature to 38°C, and atomization pressure to 0.2 MPa. After coating, place the pellets in an electric thermostatic oven and cure at 42°C for 3 h to obtain enteric-coated sustained-release pellets, collect and seal for later use.

[0042] Preparation of immediate-release phase: Take the remaining 10% of the prescribed amount of P-nicotinamide mononucleotide, along with 20 parts of microcrystalline cellulose and crospovidone, put them into a high-speed mixing granulator, mix for 5 min until uniform. Use a dry granulation process for granulation. Sieve the obtained granules through a 20-mesh standard sieve to obtain immediate-release granules, collect and seal for later use.

[0043] Total mixing and forming: Put the above enteric-coated sustained-release pellets and immediate-release granules into a three-dimensional motion mixer in a weight ratio of 5:1, adjust the speed to 20 r / min and mix for 10 min until uniform. Use an automatic capsule filling machine to fill into size 0 hard gelatin capsules at a filling weight of 0.5 g / capsule, obtaining the NMN composite anti-aging formulation for activating the SIRT1 pathway of the present invention, as shown in Figure 2.

[0044] During the entire preparation process of this embodiment, there was no situation of excessive material loss, and the test indicators of all intermediate products met the preset quality standards: Dissolution testing of the complex inclusion compound powder showed a 15 min dissolution rate of 92.5% for trans-resveratrol and 90.8% for quercetin, an increase of over 80% compared to the un-included raw materials. The moisture content of the sustained-release pellets after drying was 2.5%, with uniform particle size distribution, and the proportion of pellets in the 20-40 mesh range was 98.2%. The dissolution rate of the enteric-coated sustained-release pellets in pH 1.2 hydrochloric acid medium after standing for 2 h was <2.8%, and the cumulative dissolution rate in pH 6.8 phosphate buffer medium after shaking for 4 h reached 88.6%, fully meeting the design requirements for enteric sustained-release. The 15 min dissolution rate of the immediate-release granules in pH 6.8 phosphate buffer medium reached 95.3%, enabling rapid release for onset of action. In vitro cell experiment tests showed that the formulation of this embodiment increased the activation rate of the SIRT1 pathway by 5.2 times compared to the blank group, the nicotinamide clearance rate reached 98.3%, and the intracellular NAD+ level was maintained for over 48 h. All efficacy indicators reached optimal levels. The formulation had a smooth appearance, uniform granules, and no adhesion or caking.

[0045] Embodiment 2

[0046] This embodiment is a time-sequential synergistic release biphasic anti-aging formulation with a lower limit ratio of active ingredients. The specific composition of active ingredients is: P-nicotinamide mononucleotide 35 parts, trans-resveratrol 10 parts, quercetin 3 parts, piperine 0.8 parts, trimethylglycine 51.2 parts. The specific composition of excipients is: hydroxypropyl- P-cyclodextrin 55 parts, microcrystalline cellulose 50 parts, crospovidone 6 parts, hypromellose K4M 8 parts, acrylic resin II 4 parts, acrylic resin III 8 parts; purified water is a process solvent and its amount is not counted in the total formula.

[0047] The specific preparation method is as follows:

[0048] Preparation of trans-resveratrol-quercetin-piperine complex inclusion compound: Put the prescribed amounts of trans-resveratrol, quercetin, piperine, and hydroxypropyl- P-cyclodextrin into a colloid mill and mix uniformly. Add purified water, the amount of purified water being 35% of the total mass of the solid materials. Adjust the colloid mill speed to 350 r / min, continuously grind and include for 30 min, controlling the temperature throughout to <30 °C. Transfer the ground inclusion slurry to a vacuum drying oven, set the temperature to 45°C and vacuum degree to -0.08 MPa, vacuum dry for 11 h. After drying, grind the material using a universal grinder, pass through an 80-mesh standard sieve to obtain the complex inclusion compound powder, collect and seal for later use.

[0049] Preparation of enteric-coated sustained-release synchronous phase: Take the above complex inclusion compound powder, the prescribed amount of trimethylglycine, 85% of the prescribed amount of P-nicotinamide mononucleotide, along with hypromellose K4M and 35 parts of microcrystalline cellulose, put them into a three-dimensional motion mixer, adjust the speed to 18 r / min and mix for 15 min until the materials are uniform. Add an appropriate amount of purified water to make a soft material. Pass the soft material through a 16-mesh sieve and then put it into an extruder- spheronizer, set the extrusion speed to 55 rpm, spheronization speed to 650 rpm, and spheronization time to 5 min to prepare spherical pellets. Transfer the spherical pellets to a hot air drying oven, dry with air blast at 42 °C until the material moisture content is <3%, and sieve using 20-40 mesh ii standard sieves to obtain sustained-release pellets. Place the sustained-release pellets in a fluid bed coater, coat with the mixed coating solution prepared from acrylic resin II and acrylic resin III, set the inlet air temperature to 42°C, bed temperature to 35°C, and atomization pressure to 0.2 MPa. After coating, place the pellets in an electric thermostatic oven and cure at 40°C for 4 h to obtain enteric-coated sustained-release pellets, collect and seal for later use.

[0050] Preparation of immediate-release phase: Take the remaining 15% of the prescribed amount of P-nicotinamide mononucleotide, along with 15 parts of microcrystalline cellulose and crospovidone, put them into a high-speed mixing granulator, mix for 5 min until uniform. Use a dry granulation process for granulation. Sieve the obtained granules through a 20-mesh standard sieve to obtain immediate-release granules, collect and seal for later use.

[0051] Total mixing and forming: Put the above enteric-coated sustained-release pellets and immediate-release granules into a three-dimensional motion mixer in a weight ratio of 4:1, adjust the speed to 20 r / min and mix for 10 min until uniform. Use an automatic capsule filling machine to fill into size 1 hard gelatin capsules at a filling weight of 0.4 g / capsule, obtaining the NMN composite anti-aging formulation for activating the SIRT1 pathway of the present invention.

[0052] This embodiment was prepared using a lower limit ratio of active ingredients, with process parameters fine-tuned according to the raw material dosage. Material loss during the entire preparation process complied with pharmaceutical manufacturing specifications, and the test indicators of all intermediate products met the preset quality standards: Dissolution testing of the complex inclusion compound d powder showed a 15 min dissolution rate of 89.2% for trans-resveratrol and 87.5% for quercetin, an increase of over 75% compared to the un-included raw materials. The moisture content of the sustained-release pellets after drying was 2.8%, with uniform particle size distribution, and the proportion of pellets in the 20-40 mesh range was 97.5%. The dissolution rate of the enteric-coated sustained-release pellets in pH 1.2 hydrochloric acid medium after standing for 2 h was <3.2%, and the cumulative dissolution rate in pH 6.8 phosphate buffer medium after shaking for 4 h reached 85.3%, meeting the design requirements for enteric sustained-release. The 15 min dissolution rate of the immediate-release granules in pH 6.8 phosphate buffer medium reached 93.8%, enabling rapid release for onset of action. In vitro cell experiment tests showed that the formulation of this embodiment increased the activation rate of the SIRT1 pathway by 4.1 times compared to the blank group, the nicotinamide clearance rate reached 95.6%, and the intracellular NAD+ level was maintained for over 36 h. All efficacy indicators were stable and controllable. While reducing the amount of raw materials, this embodiment still achieves effective activation of the SIRT1 pathway, balancing formulation efficacy and raw material cost, and possesses good industrialization cost-effectiveness.

[0053] Embodiment 3

[0054] This embodiment is a time-sequential synergistic release biphasic anti-aging formulation with an upper limit ratio of active ingredients. The specific composition of active ingredients is: P-nicotinamide mononucleotide 55 parts, trans-resveratrol 18 parts, quercetin 8 parts, piperine 2 parts, trimethylglycine 17 parts.

[0055] The specific composition of excipients is: hydroxypropyl- P-cyclodextrin 108 parts, microcrystalline cellulose 70 parts, crospovidone 10 parts, hypromellose K4M 15 parts, acrylic resin II 7 parts, acrylic resin III 14 parts; purified water is a process solvent and its amount is not counted in the total formula.

[0056] The specific preparation method is as follows:

[0057] Preparation of trans-resveratrol-quercetin-piperine complex inclusion compound: Put the prescribed amounts of trans-resveratrol, quercetin, piperine, and hydroxypropyl- P-cyclodextrin into a colloid mill and mix uniformly. Add purified water, the amount of purified water being 45% of the total mass of the solid materials. Adjust the colloid mill speed to 550 r / min, continuously grind and include for 50 min, controlling the temperature throughout to <30 °C. Transfer the ground inclusion slurry to a vacuum drying oven, set the temperature to 55 °C and vacuum degree to -0.09 MPa, vacuum dry for 9 h. After drying, grind the material using a universal grinder, pass through a 100-mesh standard sieve to obtain the complex inclusion compound powder, collect and seal for later use.

[0058] Preparation of enteric-coated sustained-release synchronous phase: Take the above complex inclusion compound powder, the prescribed amount of trimethylglycine, 88% of the prescribed amount of P-nicotinamide mononucleotide, along with hypromellose K4M and 45 parts of microcrystalline cellulose, put them into a three-dimensional motion mixer, adjust the speed to 18 r / min and mix for 15 min until the materials are uniform. Add an appropriate amount of purified water to make a soft material. Pass the soft material through a 16-mesh sieve and then put it into an extruder- spheronizer, set the extrusion speed to 75 rpm, spheronization speed to 850 rpm, and spheronization time to 5 min to prepare spherical pellets. Transfer the spherical pellets to a hot air drying oven, dry with air blast at 48°C until the material moisture content is <3%, and sieve using 20-40 mesh standard sieves to obtain sustained-release pellets. Place the sustained-release pellets in a fluid bed coater, coat with the mixed coating solution prepared from acrylic resin II and acrylic resin III, set the inlet air temperature to 48°C, bed temperature to 40°C, and atomization pressure to 0.2 MPa. After coating, place the pellets in an electric thermostatic oven and cure at 45 °C for 2 h to obtain enteric-coated sustained-release pellets, collect and seal for later use.

[0059] Preparation of immediate-release phase: Take the remaining 12% of the prescribed amount of P-nicotinamide mononucleotide, along with 25 parts of microcrystalline cellulose and crospovidone, put them into a high-speed mixing granulator, mix for 5 min until uniform. Use a dry granulation process for granulation. Sieve the obtained granules through a 20-mesh standard sieve to obtain immediate-release granules, collect and seal for later use.

[0060] Total mixing and forming: Put the above enteric-coated sustained-release pellets and immediate-release granules into a three-dimensional motion mixer in a weight ratio of 6:1, adjust the speed to 20 r / min and mix for 10 min until uniform. Use an automatic capsule filling machine to fill into size 00 hard gelatin capsules at a filling weight of 0.6 g / capsule, obtaining the NMN composite anti-aging formulation for activating the SIRT1 pathway of the present invention.

[0061] This embodiment was prepared using an upper limit ratio of active ingredients, with process parameters adjusted accordingly as the raw material dosage increased. Material loss during the entire preparation process complied with pharmaceutical manufacturing specifications, and the test indicators of all intermediate products were superior to the preset quality standards: Dissolution testing of the complex inclusion compound powder showed a 15 min dissolution rate of 93.1% for trans-resveratrol and 91.5% for quercetin, an increase of over 82% compared to the un-included raw materials. The moisture content of the sustained-release pellets after drying was 2.3%, with uniform particle size distribution, and the proportion of pellets in the 20-40 mesh range was 98.6%. The dissolution rate of the enteric-coated sustained-release pellets in pH 1.2 hydrochloric acid medium after standing for 2 h was <2.5%, and the cumulative dissolution rate in pH 6.8 phosphate buffer medium after shaking for 4 h reached 90.2%, superior to the design requirements for enteric sustained-release. The 15 min dissolution rate of the immediate-release granules in pH 6.8 phosphate buffer medium reached 96.1%, demonstrating a significant rapid release effect. In vitro cell experiment tests showed that the formulation of this embodiment increased the activation rate of the SIRT1 pathway by 5.5 times compared to the blank group, the nicotinamide clearance rate reached 98.8%, and the intracellular NAD+ level increased rapidly in a short time and was maintained for over 48 h, with outstanding short-term activation effect, suitable for rapid intervention in aging-related physiological function decline. The formulation had a smooth appearance, with only a small amount of pellets exhibiting slight adhesion, which did not affect the dissolution effect or usability.

[0062] Comparative Example

[0063] This comparative example is a traditional conventional compounded anti-aging formulation, without adding trimethylglycine and without adopting the time-sequential synergistic release biphasic design, forming a contrast with the present invention. The active ingredients total 100 parts by weight, with the specific composition being: P-nicotinamide mononucleotide 60 parts, trans-resveratrol 20 parts, quercetin 10 parts, piperine 10 parts.

[0064] The excipients total 176 parts by weight, with the specific composition being: hydroxypropyl- P-cyclodextrin 200 parts, microcrystalline cellulose 50 parts, magnesium stearate 3 parts; no sustained-release material, enteric coating material, or disintegrant; purified water is a process solvent and its amount is not counted in the total formula.

[0065] The specific preparation method is as follows:

[0066] Simple inclusion compound preparation: Put the prescribed amounts of trans-resveratrol, quercetin, piperine, and hydroxypropyl- P-cyclodextrin into a glass mortar, grind and mix for 10 min until preliminarily uniform, add a small amount of purified water and continue grinding until it forms a paste. Dry the paste in a hot air drying oven at 40 °C. After drying, grind the material using a universal grinder, pass through an 80-mesh standard sieve to obtain a simple inclusion compound powder, collect for later use.

[0067] Total mixing and forming: Put the above simple inclusion compound powder, all of the P-nicotinamide mononucleotide, and microcrystalline cellulose into a trough mixer, mix for 10 min until roughly uniform. Add magnesium stearate and mix again for 5 min. Use a semi-automatic capsule filling machine to fill into size 00 hard gelatin capsules at a filling weight of 0.6 g / capsule, obtaining the traditional conventional compounded anti-aging formulation.

[0068] This comparative example is a traditional conventional compounded formulation, without adding trimethylglycine, with active ingredients totaling 100 parts by weight and each component at the upper limit ratio. The preparation process is simple with few types of excipients, and there is no targeted control of process parameters. Material loss during preparation was relatively high, and the test indicators of all intermediate products were far below the standards of the formulation of the present invention: Dissolution testing of the simple inclusion compound powder showed a 15 min dissolution rate of only 45.2% for trans-resveratrol, only 41.8% for quercetin, and only 43.5% for piperine, indicating poor dissolution effect. The formulation had no enteric coating design; after standing in pH 1.2 hydrochloric acid medium for 2 h, the dissolution rate of P-nicotinamide mononucleotide reached 98.5%, easily releasing and degrading rapidly in gastric acid. There was no biphasic design of immediate-release and sustained-release, and the dissolution of components was irregular. The 4 h dissolution rate of trans-resveratrol in pH 6.8 phosphate buffer medium was only 62.3%, with a dissolution time difference of over 3 h compared to P-nicotinamide mononucleotide. In vitro cell experiment tests showed that the formulation of this comparative example increased the activation rate of the SIRT1 pathway by only 1.2 times compared to the blank group, had no nicotinamide scavenging ability, with a nicotinamide accumulation rate in the body reaching 89.5%. The intracellular NAD+ level only increased transiently in the initial stage and rapidly declined within 12 h, failing to achieve continuous activation of the SIRT1 pathway. All efficacy indicators did not meet the basic requirements for an anti-aging formulation. Moreover, the formulation was prone to component degradation and particle caking during storage, and the material loss was more than 15% higher than that of the formulation of the present invention.

[0069]

[0070] From the above table, it can be seen that Embodiments 1, 2, and 3 are all NMN composite anti-aging formulations for activating the SIRT1 pathway that meet the design requirements of the present invention. By adjusting the process parameters according to the differences in active ingredient ratios, all three achieved the biphasic release design of enteric sustained-release and immediate-release. All test indicators at each stage met the relevant quality standards of the pharmaceutical industry, with low material loss during preparation and no process defects or efficacy shortcomings. Among them, Embodiment 1 with the optimal ratio achieved the best balance of core efficacy indicators such as SIRT1 pathway activation rate, nicotinamide clearance rate, NAD+ maintenance time, and formulation process indicators, with excellent storage stability and component dissolution, representing the best mode of the present invention. Embodiment 2 with the lower limit ratio, while achieving core benefits, reduced raw material usage, possessing significant cost advantages and suitable for conventional consumption scenarios. Embodiment 3 with the upper limit ratio has outstanding short-term SIRT1 pathway activation effect, capable of rapidly improving aging-related physiological function decline, suitable for targeted rapid intervention scenarios. The two can adapt to different consumer needs and application scenarios. Comparative Example 1, as a traditional conventional compounded formulation, lacks trimethylglycine for regulating nicotinamide metabolism, does not adopt the time-sequential synergistic release biphasic design, and has a simple preparation process without precise parameter control. Its core efficacy indicators and process indicators are still far below those of the formulation of the present 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, fully demonstrating the innovation of the present invention in active ingredient compounding, formulation structure design, and preparation process control.

[0071] The above descriptions are only preferred embodiments of the present invention and are not intended to limit the present invention in any form. Although the present invention has been disclosed as above with preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art, without departing from the scope of the technical solution of the present invention, can make some changes or modifications to equivalent embodiments with equivalent changes using the above-disclosed technical content. However, any simple modification, equivalent change, and modification made to the above embodiments based on the technical essence of the present invention, without departing from the content of the technical solution of the present invention, still fall within the scope of the technical solution of the present invention.

Claims

CLAIMS1. An NMN composite anti-aging formulation for activating the SIRT1 pathway, characterized in that the formulation consists of active ingredients and pharmaceutically acceptable excipients, the active ingredients totaling 100 parts by weight, comprising the following components:30-60 parts by weight of P-nicotinamide mononucleotide;8-20 parts by weight of trans-resveratrol;2-10 parts by weight of quercetin;0.5-3 parts by weight of piperine; and the balance being trimethylglycine.

2. The NMN composite anti-aging formulation for activating the SIRT1 pathway according to claim 1, characterized in that the trimethylglycine is a natural betaine extract with an extraction purity >99%, wherein the free betaine content is >98.5% and the water content is <0.5%.

3. The NMN composite anti-aging formulation for activating the SIRT1 pathway according to claim 1, characterized in that the P-nicotinamide mononucleotide is P-configuration crystal form I, with a particle size range of 10-50 pm, purity >99.5%, and a-configuration impurity <0.1%; the trans-resveratrol has a trans-isomer content >98% and a melting point of 254-256°C; the quercetin is anhydrous quercetin aglycone, with a particle size <20 pm, purity >98%, and glycoside impurities <1%; the piperine has a purity >95% and a melting point of 128-130 °C.

4. The NMN composite anti-aging formulation for activating the SIRT1 pathway according to claim 1, characterized in that the formulation is a time-sequential 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 composite anti-aging formulation for activating the SIRT1 pathway according to claim 4, characterized in that the immediate-release phase comprises 10%-20% of the total weight of the P-nicotinamide mononucleotide, along with a filler and a disintegrant, the filler being microcrystalline cellulose and the disintegrant being crospovidone; the enteric-coated sustained-release synchronous phase comprises 80%-90% of the total weight of the P-nicotinamide mononucleotide, all of the trans-resveratrol, all of the quercetin, all of the piperine, all of the trimethylglycine, along with a sustained-release material and an enteric coating material.

6. The NMN composite anti-aging formulation for activating the SIRT1 pathway according to claim 1, characterized in that the pharmaceutically acceptable excipients comprise hydroxypropyl-P-cyclodextrin, the degree of substitution of the hydroxypropyl-P-cyclodextrin is 4-9, and the ratio of its mass to the total mass of trans-resveratrol, quercetin, and piperine is 4-8:1.

7. The NMN composite anti-aging formulation for activating the SIRT1 pathway according to claim 1, characterized in that the pharmaceutically acceptable excipients comprise a sustained-release material and an enteric coating material; the sustained-release material is hypromellose K4M, used in an amount of 3%-8% of the total weight of the enteric-coated sustained-release synchronous phase; the enteric coating material is a mixture of acrylic resin II and acrylic resin III, the weight ratio of the two being 1:1-3, and the coating weight gain is 3%-8% of the total weight of the enteric-coated sustained-release pellets.

8. A preparation of an NMN composite anti-aging formulation for activating the SIRT1 pathway, the method being used for preparing the NMN composite anti-aging formulation for activating the SIRT1 pathway according to any one of claims 1-7, characterized in that the preparation method comprises:SI . Preparation of trans-resveratrol-quercetin-piperine complex inclusioncompound: mixing the prescribed amounts of trans-resveratrol, quercetin, piperine with hydroxypropyl- P-cyclodextrin, adding purified water in an amount of 30%-50% of the total mass of the solid materials, grinding and including at a speed of 300-600 r / min for 20-60 min, vacuum drying at 40-60°C for 8-12 h, passing through an 80-100 mesh sieve to obtain a complex inclusion compound powder;52. Preparation of enteric-coated sustained-release synchronous phase: mixing the complex inclusion compound powder obtained in SI, the prescribed amount of trimethylglycine, 80%-90% of the prescribed amount of P-nicotinamide mononucleotide with hypromellose K4M and microcrystalline cellulose, adding purified water to make a soft material, extruding and spheronizing to prepare sustained-release pellets, with an extrusion speed of 50-80 rpm and a spheronization speed of 600-900 rpm, drying at 40-50°C to a moisture content <3%; coating the sustained-release pellets with a mixed coating solution of acrylic resin II and acrylic resin III, and after coating, curing at 40-45 °C for 2-4 h to obtain enteric-coated sustained-release pellets;53. Preparation of immediate-release phase: uniformly mixing the remaining 10%-20% of the prescribed amount of P-nicotinamide mononucleotide with microcrystalline cellulose and crospovidone, dry granulating and passing through a 20-mesh sieve to obtain immediate-release granules;54. Total mixing and forming: uniformly mixing the enteric-coated sustained-release pellets and the immediate-release granules in a weight ratio of 4-6: 1, and filling into capsules or compressing into tablets to obtain the anti-aging formulation.

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