Retinoic acid derivative as well as preparation method and application thereof

Abstract

The invention discloses a retinoic acid derivative as well as a preparation method and application thereof, and belongs to the technical field of cosmetic raw materials. The derivative is raspberry ketone retinoate, and the structural formula of the derivative is shown in the specification.

Description

Technical Field

[0001] This invention relates to the field of fine chemical and cosmetic raw material technology, specifically to a novel retinoic acid derivative, its preparation method, and its application in the preparation of cosmetics. Background Technology

[0002] As consumers' demands for skincare product efficacy continue to rise and dermatology advances, the development of cosmetic raw materials is shifting from a "single-function" approach to a "precise synergy, multi-effect combination" model. Ideal active ingredients not only need to have a clearly defined core function (such as anti-aging), but should also be able to simultaneously address multiple related skin problems (such as oxidative damage, inflammatory sensitivity, etc.) to achieve more comprehensive skincare results and a better user experience.

[0003] Retinoic acid is considered the "gold standard" ingredient in the anti-aging field. By activating retinoic acid receptors (RAR / RXR) in the cell nucleus, it effectively promotes normal differentiation of keratinocytes, stimulates fibroblasts to synthesize collagen, and inhibits the overexpression of matrix metalloproteinases (MMPs), thus demonstrating outstanding performance in anti-wrinkle and skin texture improvement. However, its strong irritant properties severely limit its widespread application. Common side effects include dry skin, redness, stinging, and peeling, making it intolerable for most people with sensitive skin.

[0004] To reduce the irritation of retinoic acid, existing technologies mainly employ two strategies: one is physical compounding, such as simply mixing retinoic acid with soothing and repairing ingredients like ceramides and panthenol; the other is chemical modification, such as preparing retinol or its ester derivatives (e.g., retinyl palmitate). However, physical compounding suffers from unstable formulation systems and potential interactions between ingredients; while simple esterification modification, although effective in reducing irritation, often results in a significant reduction in bioactivity and fails to integrate other effects such as antioxidant and soothing properties.

[0005] Raspberry ketone is an aromatic ketone compound derived from natural raspberries. Studies have shown that it possesses good antioxidant activity, effectively scavenging free radicals and exhibiting potential anti-inflammatory and soothing properties. However, raspberry ketone alone has limited efficacy in combating skin aging and is unlikely to achieve structural anti-wrinkle and firming effects.

[0006] Therefore, developing a novel single-molecule compound that can retain the core anti-aging activity of retinoic acid, while also possessing excellent antioxidant and soothing properties and significantly reduced irritation has become a pressing technical problem to be solved in this field. Summary of the Invention

[0007] In view of the shortcomings of the prior art, the primary objective of this invention is to provide a retinoid derivative with a novel structure, low irritation and multiple effects.

[0008] Another object of the present invention is to provide a method for preparing the above-mentioned retinoic acid derivative, which uses readily available raw materials, has simple steps, and is suitable for large-scale production.

[0009] Another object of the present invention is to provide the application of the above-mentioned retinoic acid derivatives in the preparation of cosmetics with anti-aging, antioxidant and soothing effects.

[0010] The above-mentioned technical problems of the present invention are mainly solved by the following technical solution: a retinoic acid derivative, characterized in that the retinoic acid derivative is raspberry ketone retinoic acid ester, its tautomer, or a pharmaceutically acceptable salt, prodrug, hydrate, or solvate thereof, and its core structure is .

[0011] A method for preparing retinoic acid derivatives, characterized by comprising the following steps:

[0012] S1. Dissolve retinoic acid and raspberry ketone in an organic solvent, add a condensing agent, and stir the reaction at room temperature for 12-48 hours.

[0013] S2. Monitor the reaction progress. After the raspberry ketone reacts completely, quench the reaction. Then, extract, dry, concentrate and purify to obtain the raspberry ketone retinate.

[0014] In the aforementioned method for preparing retinoic acid derivatives, preferably, the amount of raspberry ketone fed is 1.0-2.0 equivalents of retinoic acid.

[0015] In the aforementioned method for preparing retinoic acid derivatives, preferably, the amount of condensing agent added is 1.0-3.0 equivalents of retinoic acid.

[0016] In the aforementioned method for preparing retinoic acid derivatives, preferably, the condensing agent is selected from one or more of dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), O-benzotriazole-tetramethylurea hexafluorophosphate (HBTU), O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), and benzotriazole-1-yl-oxytripyrrolylphosphine hexafluorophosphate (PyBOP).

[0017] In the aforementioned method for preparing retinoic acid derivatives, preferably, the organic solvent is selected from one or more of petroleum ether, ethyl acetate, dichloromethane, tetrahydrofuran, diethyl ether, acetone, toluene, N,N-dimethylformamide (DMF), and dimethyl sulfoxide (DMSO).

[0018] An application of the above-mentioned retinoic acid derivative is characterized in that it is used to prepare cosmetics, said cosmetics having at least one of the following effects: anti-aging, anti-wrinkle, antioxidant, or skin-soothing.

[0019] The aforementioned retinoic acid derivatives are further used to prepare formulations that promote skin collagen synthesis and / or inhibit matrix metalloproteinase expression.

[0020] The aforementioned retinoic acid derivatives are further used to prepare formulations that inhibit the expression of the TRPV1 gene in skin keratinocytes.

[0021] The aforementioned retinoic acid derivatives are further used in the preparation of agents that scavenge reactive oxygen species in skin cells.

[0022] Furthermore, this technical solution provides a cosmetic composition comprising a safe and effective amount of the aforementioned retinoic acid derivative and a cosmetically acceptable carrier or excipient. The dosage form of the cosmetic composition can be any form conventional in the art, such as, but not limited to, serums, lotions, creams, gels, masks, eye creams, or skin oils.

[0023] Compared with the prior art, the technical solution provided by the present invention has the following significant effects:

[0024] 1. Innovative Structure and Integrated Efficacy: This invention creatively combines retinoic acid and raspberry ketone through a covalent ester bond to form a single molecule. This design ingeniously integrates the powerful anti-aging mechanism of retinoic acid with the antioxidant and soothing potential of raspberry ketone, achieving a synergistic effect of multiple benefits—anti-aging, antioxidant, and soothing—at the molecular level, thus solving the pain points of existing raw materials having single efficacy and poor synergistic effects in compound formulations.

[0025] 2. Significantly Reduced Irritation: Esterification modification masks the free carboxyl group of retinoic acid, effectively reducing its direct irritation to the skin. In vitro cytotoxicity experiments show that the cellular safety of the derivative of this invention is significantly better than that of retinoic acid itself within the effective concentration range, and it is expected to be applicable to a wider range of skin types (including sensitive skin).

[0026] 3. Preservation and Enhancement of Bioactivity: Experiments have shown that the raspberry ketone retinoic acid ester of this invention can significantly promote collagen synthesis and inhibit the expression of collagen-degrading enzymes even at low concentrations, thus preserving and even enhancing its anti-aging and anti-wrinkle activities. Simultaneously, this derivative exhibits excellent free radical scavenging ability and inhibition of inflammation-related channels (such as TRPV1) expression, confirming its design intent.

[0027] 4. Raw materials are readily available and preparation is simple: The starting materials required for synthesis, retinoic acid and raspberry ketone, are both commercially available bulk chemicals or natural product extracts, with stable sources. The synthesis method adopts a conventional esterification reaction, with mild conditions, simple operation, and convenient post-processing, making it easy to realize industrial production and possessing good market transformation prospects.

[0028] In summary, this invention provides a novel retinoic acid derivative with multiple synergistic effects, offering a new core ingredient option for developing highly effective, gentle, and multifunctional anti-aging cosmetics. Attached Figure Description

[0029] Figure 1 This is a hydrogen nuclear magnetic resonance spectrum result from an embodiment of the present invention.

[0030] Figure 2 This is a carbon NMR spectrum result from an embodiment of the present invention.

[0031] Figure 3 This is a mass spectrometry result diagram from an embodiment of the present invention.

[0032] Figure 4 This is a graph of HDF cytotoxicity test in an embodiment of the present invention.

[0033] Figure 5 This is a test diagram of HDF cell anti-aging and anti-wrinkle effects in an embodiment of the present invention.

[0034] Figure 6 This is a graph of HDF cell antioxidant test in an embodiment of the present invention.

[0035] Figure 7 This is a diagram of HaCaT cytotoxicity test in an embodiment of the present invention.

[0036] Figure 8 This is a diagram of HaCaT cell relaxation test in an embodiment of the present invention. Detailed Implementation

[0037] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to specific experimental data and embodiments. It should be understood that the embodiments described herein are for illustrative purposes only and do not constitute a limitation thereof. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without inventive effort are within the scope of protection of this invention.

[0038] This embodiment describes a retinoic acid derivative, raspberry ketone retinoic acid ester, whose core structure is as follows: .

[0039] This embodiment describes a method for preparing a retinoic acid derivative, with the following steps:

[0040] S1. In a synthetic vessel, protected from light, dissolve 10 mmol of raspberry ketone and 10 mmol of retinoic acid in 30 mL of anhydrous dichloromethane and stir until the solids are completely dissolved. After cooling, slowly add 12.5 mmol of dicyclohexylcarbodiimide (DCC) while stirring, and stir the reaction at room temperature for 24 hours.

[0041] S2. The reaction process was monitored by TLC. After the raspberry ketone raw material was completely reacted, 30 mL of ethyl acetate was added to quench the reaction. The reaction solution was concentrated to about 20 mL. The solid was filtered through diatomaceous earth. The organic phase was washed twice with saturated sodium chloride aqueous solution, extracted with ethyl acetate, collected, dried with anhydrous sodium sulfate, concentrated, and then purified by column chromatography to finally obtain the target retinic acid derivative. The column chromatography developing solvent was: petroleum ether: ethyl acetate = 3:1 (v / v).

[0042] The yield of raspberry ketone retinate obtained above was 62%, HRMS(ESI) calculated for [C30H38O3+Na+]: 469.2713, found: 469.2711.

[0043] The NMR results of the prepared raspberry ketone retinate are as follows: Figure 1 , Figure 2 As shown:

[0044] Analysis of 1H NMR spectrum: 1 H NMR (400 MHz, CDCl3) δ 7.34 (d, J = 8.8 Hz, 2H), 7.17(t, J = 8.4 Hz, 1H), 6.54-6.44 (m, 2H), 6.35-6.29 (m, 2H), 6.13 (s, 1H), 3.04(t, J = 7.2 Hz, 2H), 2.90 (t, J =7.6 Hz, 2H), 2.55 (s, 3H), 2.29 (s, 3H), 2.20-2.17 (m, 5H), 1.87 (s, 3H), 1.80-1.76 (m, 3H), 1.64-1.61 (m, 2H), 1.19(s, 6H).

[0045] Analysis of carbon NMR spectrum: 13 C NMR (101 MHz, CDCl3)δ 207.94, 165.64, 155.51,149.11, 140.41, 138.25, 137.71, 137.23, 134.81, 132.01, 130.23, 129.46,129.21, 129.15, 121.81, 117.24, 45.19, 39.66, 34.32, 33.17, 30.13, 29.14,29.02, 21.81, 19.26, 14.11, 13.01.

[0046] The above spectral data are in perfect agreement with the structure of the target compound, raspberry ketone retinate, confirming its structure. Figure 3 As shown.

[0047] The application of a retinoic acid derivative in the preparation of cosmetics, with an addition amount of 0.1-1.0 μM.

[0048] Application of a retinoic acid derivative in the preparation of cosmetics with anti-aging, anti-wrinkle, antioxidant, or skin-soothing effects.

[0049] The use of a retinoic acid derivative in cosmetics also includes tautomers of the retinoic acid derivative, pharmaceutically acceptable salts, prodrugs, hydrates, or solvates of the retinoic acid derivative or its tautomers.

[0050] Verification experiment:

[0051] Experimental reagents: Phosphate-buffered saline (PBS, Gibco), high-glucose DMEM medium (Gibco), low-glucose DMEM medium (Gibco), 0.25% trypsin solution (Gibco), antibiotic penicillin-streptomycin (Gibco), fetal bovine serum (FBS, Gibco), CCK8 reagent (Beyotime), dimethyl sulfoxide (DMSO) (Sinopharm), reactive oxygen species detection kit (Beyotime); TB Green® Fast qPCR Mix (TAKARA), PrimeScript II 1st Stand cDNA Synthesis Kit (TAKARA), RNA Extraction Kit (TAKARA).

[0052] Experimental materials: Human immortalized keratinocytes (HaCaT cells), provided by Cybio (Shanghai) Biotechnology Co., Ltd.; human primary fibroblasts (HDF cells), provided by Boxi Biotechnology Co., Ltd.

[0053] Experiment 1 HDF Cytotoxicity Test

[0054] Log-phase human primary fibroblasts were seeded into 96-well plates at a density of 8 × 10⁶ cells / well. 3 Cells were cultured at concentrations of 100 μL / mL until they adhered to the culture vessel and reached a density of 80-90%. Then, they were divided into groups and administered the drug at concentration gradients of 0 μM, 0.1 μM, 0.5 μM, 1 μM, 2.5 μM, 5 μM, and 10 μM. After culturing in a CO2 incubator for 24 h, 100 μL of CCK8 reagent was added to each well. After incubation at 37°C for 1 h, the absorbance was measured at 450 nm using a microplate reader to calculate cell viability. Relative cell viability (%) = (sample absorbance - absorbance of the zero-adjustment well) / (absorbance of the blank group - absorbance of the zero-adjustment well) * 100%. The results are shown in Table 1.

[0055]

[0056] From Table 1 and Appendix Figure 4 It can be seen that when the concentration of raspberry ketone retinate obtained in the examples is above 2.5 μM, the cell viability of HDF cells decreases significantly.

[0057] Experiment 2: HDF Cell Anti-aging and Anti-wrinkle Test

[0058] Human primary fibroblasts (HDF) were seeded in 6-well plates at a density of 2.5 × 10⁻⁶. 5 When the cell plate density reached 40%–60%, cells were divided into groups and administered the drug. 2 mL of culture medium was added to each well in the blank control group and model group, while 2 mL of raspberry ketone retinate containing 1 μM was added to each well in the sample group. After culturing in a CO2 incubator for 24 h, the well plates were removed, the culture medium was discarded, and an appropriate amount of PBS was added to cover the cell surface. The model group and sample group were then exposed to UVA irradiation at a dose of 10 J. After irradiation, the cells were cultured in a CO2 incubator for another 24 h. Total RNA was extracted from each group of cells, and the expression levels of COL1, COL3, MMP3, and p21 genes in the cells were detected according to the real-time quantitative PCR instructions. The internal reference gene was β-actin, and its sequence is shown in Table 2.

[0059] Note: All sequences are from humans, Latin name (Homo sapiens), and sequence type is RNA.

[0060] Transcription experiments were performed according to the instructions of the corresponding kits (TAKARA, Kyoto, Japan). Primers were ordered from Qingke Biotechnology Co., Ltd. (internal control gene was -actin).

[0061]

[0062] The results are shown in Table 3.

[0063]

[0064] From Table 3 and Appendix Figure 5 As can be seen, compared with the NC group irradiated by UVA, the raspberry ketone retinoic acid ester obtained in the examples significantly increased the relative expression of COL1 and COL3 mRNA, with increase rates of 83.33% and 74.19%, respectively; and significantly decreased the relative expression of MMP3 and p21 mRNA, with decrease rates of 78.64% and 21.23%, respectively. This indicates that raspberry ketone retinoic acid ester has anti-aging and anti-wrinkle effects.

[0065] Experiment 3: HDF Cell Antioxidant Test

[0066] After UVA modeling of human primary fibroblasts in Experiment 2 above, the old culture medium was discarded, and the cells were washed three times with HBSS. After washing, the HBSS in the wells was aspirated and discarded. An appropriate amount of DCFH-DA probe was added to each well at a 1:1000 dilution, and the cells were incubated in a CO2 incubator for 30 min. Then, the cells were washed three times with HBSS, and complete culture medium was added. Cells were photographed under a 63× oil immersion lens using a Leica laser confocal microscope.

[0067] From the appendix Figure 6 It can be seen that 10 J / cm 2 Under UVA induction, the ROS fluorescence intensity generated by the model group NC was significantly higher than that of the blank control group BC, indicating that the modeling system effectively promoted ROS production. Compared with the model group NC, the ROS fluorescence intensity generated by 1 μM raspberry ketone retinoic acid ester was significantly reduced, indicating that raspberry ketone retinoic acid ester has an antioxidant effect.

[0068] Experiment 4 HaCaT Cytotoxicity Test

[0069] Log-phase human immortalized keratinocytes were seeded into 96-well plates at a density of 1 × 10⁻⁶. 4 Cells were cultured at concentrations of 100 μL / mL until they adhered to the culture vessel and reached a density of 80-90%. Then, they were grouped and administered the drug. The sample concentration gradients were 0 μM, 0.1 μM, 0.5 μM, 1 μM, 2.5 μM, 5 μM, and 10 μM. After culturing in a CO2 incubator for 24 h, 100 μL of CCK8 reagent was added to each well, and the cells were incubated at 37°C for 1 h. The absorbance was then measured at 450 nm using a microplate reader to calculate cell viability. Relative cell viability (%) = (sample absorbance - absorbance of the zero-adjustment well) / (absorbance of the blank group - absorbance of the zero-adjustment well) * 100%. The results are shown in Table 4.

[0070]

[0071] From Table 4 and Appendix Figure 7 It can be seen that when the concentration of raspberry ketone retinate obtained in the examples is above 5 μM, the cell viability of HaCaT cells decreases significantly.

[0072] Experiment 5: HaCaT cell anti-inflammatory test

[0073] Log-phase human immortalized keratinocytes were seeded into 6-well plates at a density of 4.0 × 10⁻⁶. 5 When the cell density reached 40%–60% at a density of [number] cells / mL, cells were administered to different groups. 2 mL of culture medium was added to each well in the blank control group, 2 mL of culture medium containing 10 μM capsaicin was added to each well in the model group, and 2 mL of culture medium containing 1 μM raspberry ketone retinate and 10 μM capsaicin was added to each well in the sample group. After administration, cells were cultured in a CO2 incubator for 24 h. Total RNA was extracted from each group, and the expression level of the TRPV1 gene in the cells was detected according to the real-time quantitative PCR instructions. The internal reference gene was β-actin, and its sequence is shown in Table 5.

[0074]

[0075] The results are shown in Table 6:

[0076]

[0077] From Table 6 and Appendix Figure 8 As can be seen, compared with the capsaicin-stimulated NC group, raspberry ketone retinoic acid significantly reduced the relative expression of TRPV1 mRNA, with a reduction rate of 63.53%. This indicates that raspberry ketone retinoic acid has an anti-inflammatory effect.

[0078] Conclusion: Combining Table 1-6 and Appendix Figure 1-8 The retinoic acid derivative provided in this application is a novel and easily synthesized compound. Systematic in vitro biological evaluation has demonstrated that this compound exhibits excellent anti-aging and anti-wrinkle effects (promoting collagen and inhibiting degradation), antioxidant effects (scavenging ROS), and soothing effects (inhibiting TRPV1) at low micromolar concentrations, indicating its broad application prospects in cosmetics.

Claims

1. A retinic acid derivative, characterized in that, The retinoic acid derivative is raspberry ketone retinoate, its tautomer, or a pharmaceutically acceptable salt, prodrug, hydrate, or solvate thereof, with the core structure being...

2. A method for preparing the retinoic acid derivative as described in claim 1, characterized in that, Includes the following steps: S1. Dissolve retinoic acid and raspberry ketone in an organic solvent, add a condensing agent, and stir the reaction at room temperature for 12-48 hours. S2. Monitor the reaction progress. After the raspberry ketone reacts completely, quench the reaction. Then, extract, dry, concentrate and purify to obtain the raspberry ketone retinate.

3. The method for preparing retinoic acid derivatives according to claim 2, characterized in that, The amount of raspberry ketone fed is 1.0-2.0 equivalents of retinoic acid.

4. The method for preparing retinoic acid derivatives according to claim 2, characterized in that, The amount of the condensing agent added is 1.0-3.0 equivalents of retinoic acid.

5. The method for preparing retinoic acid derivatives according to claim 2, characterized in that, The condensing agent is selected from one or more of the following: dicyclohexylcarbodiimide (DCC), diisopropylcarbodiimide, 1-ethyl-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDCI), O-benzotriazole-tetramethylurea hexafluorophosphate (HBTU), O-(7-azabenzotriazole-1-yl)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU), and benzotriazole-1-yl-oxytripyrrolylphosphine hexafluorophosphate (PyBOP).

6. The method for preparing retinoic acid derivatives according to claim 2, characterized in that, The organic solvent is selected from one or more of petroleum ether, ethyl acetate, dichloromethane, tetrahydrofuran, diethyl ether, acetone, toluene, N,N-dimethylformamide (DMF), and dimethyl sulfoxide (DMSO).

7. An application of the retinoic acid derivative as described in claim 1, characterized in that, Used to prepare cosmetics, said cosmetics having at least one of the following effects: anti-aging, anti-wrinkle, antioxidant, or skin-soothing.

8. An application of the retinoic acid derivative as described in claim 1, characterized in that, This is used to prepare formulations that promote collagen synthesis in the skin and / or inhibit the expression of matrix metalloproteinases.

9. An application of the retinoic acid derivative as described in claim 1, characterized in that, This is used to prepare a formulation that inhibits the expression of the TRPV1 gene in skin keratinocytes.

10. An application of the retinoic acid derivative as described in claim 1, characterized in that, Used to prepare agents that scavenge reactive oxygen species in skin cells.

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