A spermidine composition having an anti-aging effect

Abstract

The application discloses a spermidine composition with an anti-aging effect and a preparation method thereof, and relates to the technical field of medical cosmetics. The spermidine composition with the anti-aging effect is composed of the following components in mass parts: 5-15 parts of spermidine, 3-8 parts of fucoxanthin, 1-4 parts of a SASP inhibitor, 10-20 parts of nicotinamide, 2-6 parts of sodium hyaluronate, 12-25 parts of soybean lecithin and 3-5 parts of ergothioneine. The SASP inhibitor is a compound with a specific structure. The composition can synergistically act on multiple pathways such as cell autophagy, oxidative stress and aging-related secretory phenotype by compounding spermidine, fucoxanthin and the specific SASP inhibitor, and compared with a compounding scheme of single components or other alternative components, the composition shows a more optimal comprehensive effect in protecting skin fibroblasts and delaying cell aging.

Description

Technical Field

[0001] This invention relates to the field of pharmaceutical and cosmetic technology, specifically to a spermidine composition with anti-aging effects. Background Technology

[0002] In current technologies, utilizing natural active ingredients for anti-aging intervention has become an important research direction in the pharmaceutical and cosmetic fields. Spermine, as a naturally occurring polyamine, has been proven to have the potential to induce autophagy and maintain cellular homeostasis, thus exhibiting anti-aging activity. To enhance the effect of single ingredients, researchers often attempt to combine them with other active substances. Fucoxanthin, due to its antioxidant properties, is also used in anti-aging formulations; theoretically, the combination of the two can enhance protective effects through multi-pathway regulation of cells. However, existing spermidine complex compositions still face significant challenges in achieving synergistic anti-aging effects. Common formulations often only involve simple physical mixing, lacking effective synergistic effects between components. This results in the scavenging of age-related secretory phenotypic factors and the comprehensive protection of skin fibroblasts failing to meet expectations. In particular, how to fully leverage the synergistic effects of spermidine and fucoxanthin, the two core active ingredients, in a stable system to protect cells and combat aging remains a problem that current technologies have not yet adequately solved. On the other hand, there are also bottlenecks in the formulation process. The physicochemical properties of components such as spermidine and fucoxanthin differ significantly, and simple mixing cannot guarantee the long-term stability of the system, easily leading to problems such as layering and activity degradation, directly affecting the product's bioavailability and final efficacy. Conventional emulsification processes may not achieve high uniformity and stability of the carrier, making it difficult to ensure efficient delivery of active ingredients and their action on the target. Therefore, developing a spermidine composition that maximizes the synergistic anti-aging effect of each component and possesses good stability is of great significance for meeting practical application needs. Summary of the Invention

[0003] This invention addresses the problems existing in the prior art by providing a spermidine composition with significant synergistic effects, high system stability, excellent bioavailability, enhanced cell protection, improved anti-aging efficacy, and stable applicability in skin care and anti-aging products. To achieve an anti-aging spermidine composition, comprising a SASP inhibitor as shown in Formula 1: Formula 1: ; R1 in Formula 1 is selected from any one of methyl, methoxy, ethyl, phenyl, isopropyl, and butyl. Furthermore, R1 is selected from any one of methyl, methoxy, ethyl, and phenyl. Furthermore, the SASP inhibitor is any one of the compounds shown in the following structures: ; . A spermidine composition with anti-aging effects is composed of the following components in parts by weight: 5-15 parts spermidine, 3-8 parts fucoxanthin, 1-4 parts SASP inhibitor, 10-20 parts nicotinamide, 2-6 parts sodium hyaluronate, 12-25 parts soybean lecithin, and 3-5 parts ergothioneine. Furthermore, the mass ratio of spermidine to fucoxanthin is 2-3:1. Furthermore, the purity of the spermidine is 99.99%; The purity of the fucoxanthin is 99.99%. Furthermore, the soybean lecithin is high-purity hydrogenated soybean lecithin, wherein the purity of phosphatidylcholine is 95%. A method for preparing a spermidine composition with anti-aging effects includes the following steps: S1. Dissolve the soybean lecithin and SASP inhibitor in anhydrous ethanol, and stir at a constant temperature of 45-55℃ to form a clear lipid solution; S2. The spermidine, fucoxanthin, nicotinamide, sodium hyaluronate and ergothioneine are dispersed in deionized water and premixed at a pressure of 20-40 MPa to obtain a mixed aqueous phase; S3. The lipid solution obtained in step S1 is added dropwise to the mixed aqueous phase obtained in step S2 at a rate of 5-10 ml per minute, while a high shear disperser with a shear force of 10000-15000 r / min is turned on for emulsification. S4. The emulsified material is fed into a high-pressure homogenizer and homogenized 3-5 times at a pressure of 80-120MPa. Then, residual ethanol is removed using a vacuum rotary evaporator to obtain the composition. Furthermore, the premixing process in step S2 is carried out under nitrogen gas protection; In step S4, the condensation temperature of the vacuum rotary evaporator is controlled between -5 and 0°C. The application of a spermidine composition with anti-aging effects in the preparation of a drug for clearing aging-related secretory phenotypic factors and protecting skin fibroblasts from damage. In the synergistic anti-aging system composed of spermidine and fucoxanthin, SASP inhibitors play a crucial regulatory role. Speridine primarily promotes autophagy and maintains homeostasis, while fucoxanthin focuses on providing antioxidant defense. The introduction of SASP inhibitors specifically regulates the abnormally active inflammatory signaling network in senescent cells. Their molecules may influence the production and release of pro-inflammatory factors through interactions with related proteins in the pathway, thereby helping to alleviate the chronic low-grade inflammatory microenvironment induced by age-related secretory phenotypes. This regulation of inflammatory pathways, combined with the autophagy-promoting effects of spermidine and the antioxidant effects of fucoxanthin, forms a multi-dimensional protective network at the cellular level, jointly enhancing the protection of target cells such as skin fibroblasts and synergistically delaying the cellular aging process. The structural characteristics of this SASP inhibitor also offer certain benefits during the construction and stabilization of the composition. Its molecules possess both hydrophilic and hydrophobic regions, which helps achieve a more balanced distribution in complex systems containing spermidine, fucoxanthin, and other components, reducing the tendency for phase separation due to differences in properties. During preparation steps such as emulsification or homogenization, its presence may have a mild influence on the interfacial properties of the two phases, contributing to the formation and maintenance of a more uniform and delicate emulsion or dispersion structure, which is beneficial for the long-term physical stability of the active ingredients. Simultaneously, its presence in the formulation system, through intermolecular interactions, provides a buffering effect on core active substances such as spermidine and fucoxanthin, reducing the direct impact of external environmental factors on the active ingredients and supporting the maintenance of the composition's biological efficacy during storage. Therefore, the mechanism of action of this SASP inhibitor lies in the following: From a biological activity perspective, it modulates specific aging-related inflammatory pathways, compensating for the key links in the anti-aging spectrum of spermidine and fucoxanthin, achieving synergistic effects across multiple targets and pathways, and enhancing the overall ability of the composition to protect cells and resist aging; from a physicochemical perspective, its molecular characteristics help improve the homogeneity and stability of the entire composition system. Combined with the core efficacy of spermidine and fucoxanthin, it forms an important foundation for the significant synergistic effect and good application stability of this anti-aging composition. Compared with the prior art, the beneficial effects of the present invention are: 1. Achieving multi-target synergistic anti-aging effects: Addressing the lack of effective synergy in existing technologies with simple compounding, this approach introduces a SASP inhibitor with a specific structure. This inhibitor complements the autophagy-promoting effect of spermidine and the antioxidant effect of fucoxanthin, synergistically regulating cell autophagy, redox balance, and aging-related secretory phenotypes, thereby providing a more comprehensive intervention in the skin cell aging process. 2. Improve the stability and bioavailability of the composition system: In response to the problems of system instability and easy degradation of activity caused by large differences in the physicochemical properties of components in the existing technology, a uniform and stable carrier system is formed by optimizing the component ratio and adopting a specific high-pressure emulsification and homogenization preparation process, which is conducive to the retention and efficient delivery of active ingredients. 3. Enhanced comprehensive protective efficacy against skin fibroblasts: Compared with the unsatisfactory protective effects mentioned in the background art, this composition, through the above-mentioned multi-component synergy and system optimization, shows a better ability to maintain cell viability, promote the expression of autophagy markers and inhibit the secretion of aging-related inflammatory factors in cell models, providing a more effective solution for skin anti-aging applications. Attached Figure Description

[0004] Figure 1 The present invention provides a synthetic route for the SASP inhibitor. Detailed Implementation

[0005] The technical solution of the present invention will now be clearly and completely described with reference to the accompanying drawings. Obviously, the described embodiments are merely some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. . In the first step, under nitrogen protection, 20.00 g of compound 3, 10.99 g of raw material 2, and 300 ml of a toluene-water mixture (200 ml toluene and 100 ml water) were added sequentially to the dry reaction system. After purging the air with nitrogen, 1.75 g of 1,1-bis(diphenylphosphine)ferrocene palladium dichloride and 22.03 g of potassium carbonate were added sequentially. After purging the air with nitrogen again, the mixture was stirred until homogeneous, heated to 100 °C, and refluxed for 9 h. After the reaction was completed, the mixture was filtered with diatomaceous earth while hot, the filtrate was collected, allowed to stand, separated, and the organic phase was retained. The organic phase was purified by removing the solvent using a rotary evaporator and purified by silica gel column chromatography using a mixture of petroleum ether and ethyl acetate as the eluent. The eluent was removed by a rotary evaporator to obtain 13.68 g of intermediate 1. In the second step, under nitrogen protection, 13.68 g of intermediate 1, 7.47 g of raw material 3, and 168 mL of a mixed solvent of dioxane and diisopropylamine (112 mL / 56 mL) were added sequentially to the dry reaction system. After stirring evenly, 0.3 g of palladium acetate, 0.7 g of 2-dicyclohexylphosphine-2',4',6'-triisopropylbiphenyl, and 0.3 g of CuI were added sequentially. After stirring evenly, the mixture was heated to 85 °C and refluxed for 6 h. After the reaction was completed, the mixture was filtered after cooling to room temperature. The filter cake was washed three times with tetrahydrofuran. The solvent in the filtrate was removed by a rotary evaporator, and the filtrate was purified by silica gel column chromatography using a mixed solution of petroleum ether and ethyl acetate as the eluent. The eluent was removed by a rotary evaporator to obtain 12.38 g of SASP inhibitor 1. Product structure identification: MS[MS+H] of intermediate 1 + :264; MS[MS+H] of SASP inhibitor 1 + 328. Synthesis Example 2-Synthesis Example 4 In Synthesis Examples 2-4, SASP inhibitor 2-SASP inhibitor 4 were synthesized sequentially, following the same synthesis method as in Synthesis Example 1, except that raw material 1 was replaced, and the other conditions remained the same as in Synthesis Example 1. Specific structures of raw material 1, SASP inhibitor 2-SASP inhibitor 4, and MS [MS+H] + The data is shown in Table 1. Table 1. Structures of raw material 1, SASP inhibitor 2-SASP inhibitor 4, and MS[MS+H] involved in Synthetic Examples 2-4 + data. Example 1 Preparation of a spermidine composition with anti-aging effects: 1. Raw material proportions by weight: 10 parts spermidine; 5 parts of fucoxanthin; Two doses of SASP inhibitor; 15 parts of nicotinamide; 4 parts sodium hyaluronate; 18 parts soybean lecithin; Ergothioneine 3 parts. 2. Preparation method: S1. Dissolve 18 parts of soybean lecithin and 2 parts of SASP inhibitor together in 200 ml of anhydrous ethanol, and stir at 50°C for 30 minutes to form a clear lipid solution. S2. Disperse 10 parts spermidine, 5 parts fucoxanthin, 15 parts nicotinamide, 4 parts sodium hyaluronate and 3 parts ergothioneine in 500 ml of deionized water and premix at 30 MPa for 10 minutes to obtain a mixed aqueous phase; the premixing process is carried out under nitrogen protection. S3. The lipid solution obtained in step S1 is added dropwise to the mixed aqueous phase obtained in step S2 at a rate of 7.5 ml per minute, while a high shear disperser with a shear force of 12500 r / min is turned on for emulsification treatment for 15 minutes. S4. The emulsified material is fed into a high-pressure homogenizer and homogenized 4 times at a pressure of 100 MPa. Then, residual ethanol is removed by a vacuum rotary evaporator and the condensation temperature is controlled at -2℃ to obtain the composition. Examples 2-4 The preparation of a spermidine composition with anti-aging effect is carried out by referring to the preparation method of Example 1, except that the SASP inhibitor is replaced by the SASP inhibitor synthesized in Synthetic Examples 2 to 4 in sequence, and the remaining raw materials, proportions and preparation steps are the same as in Example 1. Comparative Example 1 The preparation of a spermidine composition with anti-aging effects is carried out according to the preparation method of Example 1, without the addition of SASP inhibitors, and the remaining raw material ratios and preparation steps are the same as in Example 1. Comparative Example 2 The preparation of a spermidine composition with anti-aging effects is carried out according to the preparation method of Example 1, except that the SASP inhibitor is replaced with an equal mass of resveratrol (CAS No. 501-36-0), and the remaining raw material ratios and preparation steps are the same as in Example 1. Comparative Example 3 The preparation of a spermidine composition with anti-aging effects is carried out according to the preparation method of Example 1, except that spermidine is not added, and the proportions of other raw materials and preparation steps are the same as in Example 1. Comparative Example 4 The preparation of a spermidine composition with anti-aging effects is carried out according to the preparation method of Example 1, except that fucoxanthin is not added, and the proportions of other raw materials and preparation steps are the same as in Example 1. Comparative Example 5 The preparation of a spermidine composition with anti-aging effects is carried out according to the preparation method of Example 1, without the addition of ergothioneine, and the remaining raw material ratios and preparation steps are the same as in Example 1. Performance testing: Construction of a senescent cell model: Human skin fibroblasts (HSF) were induced to age using hydrogen peroxide (H2O2) to simulate the senescent state of skin cells caused by oxidative stress. Specifically, HSF cells in logarithmic growth phase were treated with 400 μmol / L H2O2 for 1.5 hours. B1. Sample processing solution preparation: Accurately weigh 1.00 g of the composition samples obtained in each example and comparative example, and disperse them in 50 mL of cell culture medium. After sonication with an ice bath probe for 20 minutes (power 300 W), centrifuge at 4°C and 12000 rpm for 15 minutes, and take the supernatant and filter it through a 0.22 μm sterile filter membrane to obtain the sample processing solution for cell experiments. B2. Senescence model treatment: Replace the senescent model cells treated with H2O2 with the sample treatment solution and continue culturing for 24 hours. B3. Cell viability assay (CCK-8 assay): After treatment, culture medium containing 10% CCK-8 reagent was added to each well of cells, and the cells were incubated at 37°C in the dark for 2 hours. The absorbance value at a wavelength of 450 nm was measured using a microplate reader, and the relative cell viability percentage compared with the normal control group cells that had not been treated with H2O2 was calculated. B4. Senescence-related β-galactosidase (SA-β-gal) activity assay: After treatment, cells were fixed and stained using a commercially available SA-β-gal staining kit. Positive (blue) cells were randomly selected from each field of view under an optical microscope and counted. The percentage of positive cells out of the total cell count was calculated to assess the degree of cellular senescence. B5. Detection of aging-related secretory phenotype (SASP) factors: Collect cell culture supernatant and use an IL-6 ELISA kit to determine the concentration of IL-6 in the supernatant, strictly following the instructions. Table 2 Group Relative cell viability (%) SA-β-gal positive cells (%) IL-6 concentration (pg / mL) Example 1 82.5±3.2 28.4±2.8 65.3±4.1 Example 2 81.8±2.9 29.1±3.1 67.8±3.8 Example 3 83.1±3.5 27.9±2.5 63.5±4.5 Example 4 82.0±3.0 28.7±2.9 66.2±4.0 Comparative Example 1 62.3±4.1 52.6±4.3 158.7±7.2 Comparative Example 2 71.5±3.8 41.2±3.6 112.5±6.0 Comparative Example 3 68.9±3.5 45.8±3.9 125.8±6.5 Comparative Example 4 65.4±4.0 49.3±4.1 145.3±6.9 Comparative Example 5 79.2±3.4 32.5±3.0 80.1±5.2 <![CDATA[H2O2 model group]]> 55.1±4.5 68.5±5.2 205.4±9.8 normal control group 100.0±2.5 5.2±1.1 25.3±2.1 Examples 1-4, containing spermidine, fucoxanthin, and a specific SASP inhibitor, showed the highest relative cell viability, the lowest proportion of SA-β-gal positive cells, and the lowest IL-6 secretion level, indicating a significant protective and delaying effect against H2O2-induced cell senescence. Comparative Example 1 (without the SASP inhibitor) showed significantly worse results than the example groups in all indicators, confirming the necessity of the specific SASP inhibitor in synergistic enhancement. Comparative Example 2 (using resveratrol instead of the SASP inhibitor) was better than Comparative Example 1 but less effective than the example groups, indicating that the specific SASP inhibitor described in this invention has superior synergistic performance. The protective effects of Comparative Example 3 (without spermidine) and Comparative Example 4 (without fucoxanthin) decreased to varying degrees, confirming the key roles of spermidine and fucoxanthin as core active ingredients in protecting cells, resisting oxidative stress, and the aging process. The data for Comparative Example 5 (without ergothioneine) were close to but slightly inferior to the example groups, indicating that ergothioneine has an auxiliary enhancing effect on the anti-aging effect of this system. The data trends in Examples 1-4 are consistent with small fluctuations, indicating that SASP inhibitors with different substitutions can all effectively exert synergistic effects in this system. The data demonstrate that spermidine, fucoxanthin, and specific SASP inhibitors work synergistically to effectively protect skin fibroblasts and resist the aging process through multiple mechanisms, including promoting autophagy, reducing oxidative damage, and inhibiting aging-related inflammatory secretion. Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A spermidine composition with anti-aging effects, characterized in that, The composition contains a SASP inhibitor, which is a compound represented by SASP inhibitor Formula 1. Formula 1: ; R1 in Formula 1 is selected from any one of methyl, methoxy, ethyl, phenyl, isopropyl, and butyl.

2. The spermidine composition with anti-aging effects according to claim 1, characterized in that, R1 is selected from any one of methyl, methoxy, ethyl, and phenyl.

3. The spermidine composition with anti-aging effects according to claim 1, characterized in that, The SASP inhibitor is any one of the compounds shown in the following structures: ； 。 4. The spermidine composition with anti-aging effects according to claim 1, characterized in that, The composition comprises the following components in parts by weight: spermidine 5-15 parts, fucoxanthin 3-8 parts, SASP inhibitor 1-4 parts, nicotinamide 10-20 parts, sodium hyaluronate 2-6 parts, soybean lecithin 12-25 parts, and ergothioneine 3-5 parts.

5. The spermidine composition with anti-aging effects according to claim 4, characterized in that, The mass ratio of spermidine to fucoxanthin is 2-3:

1.

6. The spermidine composition with anti-aging effects according to claim 4, characterized in that, The purity of the spermidine is 99.99%; The purity of the fucoxanthin is 99.99%.

7. The spermidine composition with anti-aging effects according to claim 4, characterized in that, The soybean lecithin is a high-purity hydrogenated soybean lecithin, wherein the phosphatidylcholine purity is 95%.

8. A method for preparing a spermidine composition with anti-aging effects according to any one of claims 4-7, characterized in that, Includes the following steps: S1. Dissolve the soybean lecithin and SASP inhibitor in anhydrous ethanol, and stir at a constant temperature of 45-55℃ to form a clear lipid solution; S2. The spermidine, fucoxanthin, nicotinamide, sodium hyaluronate and ergothioneine are dispersed in deionized water and premixed at a pressure of 20-40 MPa to obtain a mixed aqueous phase; S3. The lipid solution obtained in step S1 is added dropwise to the mixed aqueous phase obtained in step S2 at a rate of 5-10 ml per minute, while a high shear disperser with a shear force of 10000-15000 r / min is turned on for emulsification. S4. The emulsified material is fed into a high-pressure homogenizer and homogenized 3-5 times at a pressure of 80-120MPa. Then, residual ethanol is removed using a vacuum rotary evaporator to obtain the spermidine composition with anti-aging effect.

9. A method for preparing a spermidine composition with anti-aging effects according to claim 8, characterized in that, The premixing process in step S2 is carried out under nitrogen gas protection; In step S4, the condensation temperature of the vacuum rotary evaporator is controlled between -5 and 0°C.

10. The use of a spermidine composition with anti-aging effects according to any one of claims 4-7 in the preparation of a medicament for clearing aging-related secretory phenotypic factors and protecting skin fibroblasts from damage.

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