A sodium hyaluronate and a composition thereof

Abstract

The present application provides a kind of retinyl hyaluronate sodium and its composition applied in the field of cosmetics and medicine.The retinyl hyaluronate sodium of the present application is characterized by all-trans retinoic acid and 13-cis retinoic acid esterification with sodium hyaluronate in a ratio of 1.5-3.0:1 (molar ratio), and the esterification degree is 1%-12%;The composition of the present application is characterized by retinyl hyaluronate sodium and polyol compound in a ratio of 1:1-10 (mass ratio).The retinyl hyaluronate sodium of the present application has significant effect in preventing and treating acne, and the composition is beneficial to improve the stability of retinyl hyaluronate sodium, prolong the shelf life of retinyl hyaluronate sodium raw material product.The retinyl hyaluronate sodium and its composition of the present application can be used for preparing whitening, anti-aging, oil control and acne-removing cosmetics and medical products.

Description

Technical Field

[0001] This invention relates to sodium retinyl hyaluronic acid and its compositions for use in cosmetics and pharmaceuticals. Background Technology

[0002] Both retinoic acid (TR) and isotretinoin (ITR) are vitamin A derivatives. Retinoic acid, also known as retinoic acid or all-trans retinoic acid, has the chemical name 3,7-dimethyl-9-(2,6,6-trimethylcyclohexene)-2,4,6,8-all-trans nonanotetraene. Isotretinoin, an isomer of retinoic acid, is also known as 13-cis-retinoic acid, with the chemical name 3,7-dimethyl-9-(2,6,6-trimethyl-1-cyclohexenyl)-2cis-4trans-6trans-8trans-nonanotetraenoic acid. Retinoic acid promotes skin tissue regeneration, exfoliation, eliminates acne lesions, promotes lesion healing, reduces melanin formation, and has anti-inflammatory effects. Isotretinoin can inhibit sebum secretion, shrink sebaceous gland tissue, and inhibit the growth and reproduction of acne bacteria. Both can be used topically to treat acne and pimples.

[0003] Hyaluronic acid (HA), also known as glass acid or hyaluronic acid, is one of the main components of the extracellular matrix of skin cells. It is a chain-like polyanionic mucopolysaccharide composed of repeating (1-β-4)D-glucuronic acid and (1-β-3)N-acetyl-D-glucosamine disaccharide units. HA is considered an ideal natural moisturizing factor when applied to the skin surface. Small-molecule HA also regulates epidermal cell differentiation and migration, and scavenge free radicals. Therefore, in addition to moisturizing effects, it also has skin nourishing, sun protection and repair, lubrication, film-forming, and thickening effects. Retinoic acid and isotretinoin are both insoluble in water, have poor stability, and both can cause skin irritation symptoms such as burning, erythema, and desquamation when applied topically. To overcome these shortcomings, a series of esterified derivatives have been developed in recent years, including the direct esterification of HA with retinoic acid [Huerta- G. et al. [Carbohydrate Polymers (2019) 231(7): 115733 and CN117964796] or through small molecule bridging esterification (CN116284495) prepared all-trans retinoic acid derivatives, which improved the water solubility and stability of retinoic acid to a certain extent, while reducing the skin irritation caused by retinoic acid. Previous studies have shown that the occurrence of acne is closely related to excessive sebum secretion and Propionibacterium acnes, while the role of retinoic acid in controlling oil production is limited. The role of all-trans retinoic acid (HA) derivatives in the prevention and treatment of acne needs to be strengthened, and the stability of its raw materials also needs to be further improved.

[0004] This invention provides a sodium retinyl hyaluronic acid derivative, which utilizes all-trans retinoic acid (retinoic acid) and 13-cis-retinoic acid (isotretinoin) to simultaneously esterify sodium hyaluronate, thereby enhancing its efficacy in preventing and treating acne. Combining the derivative of this invention with a polyol compound can further improve its stability and extend the shelf life of the raw material product. The derivative and composition of this invention can be widely used in the preparation of cosmetics and medical products with whitening, anti-aging, oil-controlling, and acne-removing effects. Summary of the Invention

[0005] The purpose of this invention is to provide a sodium retinyl hyaluronic acid derivative and its composition with oil-controlling, acne-removing, whitening and anti-aging effects, as well as their applications in the cosmetics and pharmaceutical fields.

[0006] The sodium retinyl hyaluronic acid of the present invention has a structure represented by the following general formula:

[0007]

[0008] in,

[0009] a)a:c=1.5~3.0:1(mol / mol);

[0010] and b) (a+c) / (a+b+c)=1 to 12% (mol / mol).

[0011] The present invention relates to a composition containing sodium retinyl hyaluronic acid, characterized in that sodium retinyl hyaluronic acid and a polyol compound are composed in a ratio of 1:1 to 10 (by mass).

[0012] The present invention relates to a composition containing sodium retinyl hyaluronic acid, characterized in that the structure of the sodium retinyl hyaluronic acid is represented by the following general formula:

[0013]

[0014] in,

[0015] a)a:c=1.5~3.0:1(mol / mol);

[0016] and b) (a+c) / (a+b+c)=1 to 12% (mol / mol).

[0017] The ratio of a to c is the molar ratio obtained by converting the amounts of retinoic acid and isotretinoin grafted onto the hyaluronic acid molecular backbone determined by high performance liquid chromatography (HPLC). The ratio ranges from 1.5 to 3.0:1, preferably from 2.0 to 2.5:1.

[0018] The (a+c) / (a+b+c) ratio represents the ratio of retinoic acid and isotretinoin molecules grafted onto the hyaluronic acid molecular backbone to the hyaluronic acid disaccharide molecules, i.e., the grafting rate of the retinyl hyaluronic acid sodium of this invention, also known as the degree of substitution or esterification. The number of retinoic acid and isotretinoin molecules (a+c) is obtained by hydrolyzing the retinyl hyaluronic acid sodium and determining their contents using high-performance liquid chromatography (HPLC), followed by conversion. The number of hyaluronic acid disaccharide molecules (a+b+c) is obtained by determining the glucuronic acid content using the carbazole method and then converting. This ratio ranges from 1% to 12%, preferably from 4.0% to 8.0%.

[0019] This invention relates to sodium retinyl hyaluronic acid and compositions containing sodium retinyl hyaluronic acid, wherein the average molecular weight of the sodium retinyl hyaluronic acid is 5 kDa to 500 kDa, preferably 8 kDa to 300 kDa, and more preferably 10 kDa to 100 kDa. The average molecular weight is the weight-average molecular weight obtained by gel permeation chromatography (GPC) and / or laser scattering-gel permeation chromatography coupled with laser scattering-gel permeation chromatography (LLS-GPC).

[0020] The present invention relates to a composition containing sodium retinyl hyaluronic acid, characterized in that the ratio of sodium retinyl hyaluronic acid to a polyol compound ranges from 1:1 to 10 (mass ratio), preferably from 1:3 to 5.

[0021] The present invention relates to a composition containing sodium retinyl hyaluronic acid, characterized in that the polyol compound is selected from one or more of glycerol, 1,2-propanediol, polyethylene glycol, and polyglycerol. Preferably, it is a combination of one or more of glycerol, polyglycerol 3, and / or polyglycerol 10, more preferably polyglycerol 10.

[0022] The sodium retinyl hyaluronic acid with the technical features of this invention is produced by simultaneously esterifying sodium hyaluronate with all-trans retinoic acid and 13-cis retinoic acid (isotretinoin). When applied topically to the skin surface, in addition to the exfoliating, anti-inflammatory, acne-eliminating, repairing, whitening, and anti-aging functions of retinoic acid, it also has the effect of inhibiting sebum secretion and acne bacteria growth of 13-cis retinoic acid (isotretinoin), thus enhancing its efficacy in preventing and treating acne.

[0023] The composition containing sodium retinyl hyaluronic acid, which has the technical features of the present invention, is a complex formed by uniformly mixing sodium retinyl hyaluronic acid with a polyol compound. This complex can further improve the stability of sodium retinyl hyaluronic acid and extend the shelf life of its raw material products.

[0024] The sodium retinyl hyaluronic acid and its compositions, possessing the technical features of this invention, exhibit good water solubility and mild skin irritation, and show significant inhibitory effects on 5α-reductase, effectively controlling oil production. They are particularly effective in preventing and treating acne vulgaris and improving the skin's oil-water balance. They can be widely used in the preparation of topical skin products with whitening, anti-aging, oil-controlling, and acne-removing effects, including cosmetics and medical products.

[0025] The sodium retinyl hyaluronic acid and its composition of the present invention are combined with commonly used cosmetic ingredients to formulate various forms such as solutions, gels, ointments, creams, or lotions, including balancing moisturizing lotions, balancing moisturizing sprays, balancing whitening masks, oil-controlling toners, whitening lotions, softening lotions, exfoliating gels, acne-removing gels, balancing moisturizing and whitening emulsions, face creams, body creams, hand creams, foot creams, repairing essences, moisturizing creams, repairing lotions, acne scar creams, scalp care solutions, etc.

[0026] The sodium xanthoyl hyaluronic acid and its composition of the present invention are combined with commonly used excipients in the pharmaceutical field to formulate various dosage forms such as solutions, gels, and creams, for example, acne creams, acne gels, and acne lotions. Detailed Implementation

[0027] The following examples are provided to better illustrate the present invention and are not intended to limit the invention.

[0028] Example 1: Sodium Retinyl Hyaluronic Acid

[0029] Hyaluronic acid, retinoic acid, and / or isotretinoin are dissolved in dimethylformamide or dimethyl sulfoxide, and an appropriate amount of carbonyl diimidazole is added. After reacting by heating in the dark, saturated sodium chloride solution is added for dilution, followed by ethanol for precipitation and washing. The precipitate is collected after filtration and dried under reduced pressure to obtain a pale yellow to yellow granular powder, which is the sodium retinyl hyaluronic acid of the present invention (samples 1-1 to 1-12).

[0030] Table 1 Sodium Retinyl Hyaluronic Acid

[0031]

[0032] Retinoic acid [C] TR [% (g / g)] and isotretinoin [C ITR Method for determining the content of [% (g / g)]:

[0033] The sodium retinyl hyaluronic acid of this invention was hydrolyzed with an appropriate amount of alkaline solution and then determined by HPLC. Retinoic acid and isotretinoin were used as reference standards. An octadecylsilane-bonded silica gel column and a UV detector were used. The mobile phase was methanol:2% acetic acid = 81:19. The retinyl hyaluronic acid concentration was calculated using the external standard method based on peak area. TR (g / g)] and the content of isotretinoin [C ITR (g / g)]

[0034] Method for determining glucuronic acid [T / % (g / g)] content:

[0035] According to the literature [Bitter T, Muir H MA modified uronic acid carbarbazolereationl J J. Anal Biochem. 1962, 4: 330-333.], the glucuronic acid content T / % (g / g) of the sample was determined by the modified carbazole method.

[0036] Calculation of retinoic acid / isotretinoin (mol / mol): Since they have the same molecular weight, the ratio of the two is...

[0037] K = C TR / C ITR .

[0038] Calculation of degree of substitution (%) (mol / mol):

[0039] DS = (C TR +C ITR )×194.14 / T / 300.44=0.646×(C TR +C ITR ) / T

[0040] Example 2: Composition containing sodium retinyl hyaluronic acid

[0041] The sodium retinyl hyaluronic acid of the present invention was mixed with a polyol compound in a certain proportion to obtain the composition samples 2-1 to 2-10 of the present invention (Table 2).

[0042] Table 2. Components and proportions of different formulations

[0043]

[0044] Example 3 Stability Test

[0045] Samples 1-7 from Example 1 of this invention were dispensed into sealed aluminum-plastic bags and brown screw-top glass sample bottles. Additionally, polyol compounds were added to samples 1-7 from Example 1, and after mixing, the mixtures were dispensed into brown screw-top glass sample bottles as samples for stability testing (Table 3). Stability was investigated by placing the samples in different temperature environments and taking samples at different times. The sum of the contents of free retinoic acid and isotretinoin bound to HA molecules, Ci = C, was measured. TR +C ITR(%). The results showed that there was no significant increase in free retinoic acid and isotretinoin after different conditions, and the ratio (K value) of retinoic acid and isotretinoin did not change significantly; the change rate of retinoic acid and isotretinoin content in samples taken under different conditions and at different times was R = (Ci-C0) / C0*100%, where C0 is the sum of retinoic acid and isotretinoin content in the sample taken on day 0. The results are shown in Tables 4 and 5.

[0046] Table 3. Samples of sodium retinyl hyaluronic acid and different ratio compositions.

[0047]

[0048] Table 4. Results of stability test change rate (R value) under low temperature conditions*

[0049]

[0050] * "-" indicates a decrease in content.

[0051] Table 5. Results of R-value determination in stability tests under high temperature conditions*

[0052]

[0053] * "-" indicates a decrease in content.

[0054] The stability test results under low temperature conditions show that lower storage temperature is beneficial to the stability of raw materials. After nitrogen protection in glass bottle packaging, the stability of raw materials is significantly improved, indicating that isolating them from air is beneficial to the stability of raw materials.

[0055] The stability test results under high temperature conditions show that, compared with 3-7, the absolute values ​​of R in 3-1 to 3-6 and 3-8 are significantly smaller, that is, the degradation of the raw material product is significantly reduced (the change in the degree of substitution is reduced). This indicates that after the sodium retinyl hyaluronic acid of the present invention is formulated with polyol, the influence of air can be effectively isolated under the condition of no nitrogen protection, thereby reducing the degradation of the raw material product and improving its stability.

[0056] Based on the results of high-temperature tests at 40℃ and 60℃, and cold storage at 2-10℃ and 25℃, the composition of sodium retinyl hyaluronic acid with two types of polyglycerols in this invention has relatively good stability (comparable to the sample of sodium retinyl hyaluronic acid alone in a sealed aluminum-plastic bag), while the composition with glycerol and propylene glycol is slightly less stable.

[0057] Example 4: Inhibitory effect of sodium retinyl hyaluronic acid on 5α-reductase

[0058] According to the literature method (Pan Jifei et al. Establishment and application of in vitro evaluation system for 5α-reductase inhibitors. Daily Chemical Industry (Chinese and English), 2023, 53(11):1280-1284), the following were determined: sodium retinyl hyaluronic acid (samples 1-1, 1-2, 1-7, 1-11 and 1-12 of Example 1 of this invention), sodium hyaluronic acid, retinoic acid, isotretinoin and all-trans retinoic acid HA derivative [according to the literature Huerta- The method disclosed in G. et al. Carbohydrate Polymers (2019) 231(7): 115733 was used to prepare retinoic acid with a substitution degree of 7.5% and the relative inhibition rate of 5α-reductase (5AR) was compared in vitro to control oil production.

[0059] The test sample was added to the appropriate solvent and diluted to a solution with a retinoic acid content of 0.15–0.3 mg / ml and an isotretinoin content of 0.1 mg / ml as the test solution. A 0.2 mM dutasteride (DTS) solution was used as the positive control (PC). The relative inhibition rate was determined and calculated.

[0060] The results (Table 6) showed that, except for sodium hyaluronate (HA), all other groups had varying degrees of inhibitory effects on 5α-reductase. Comparing 4-2 and 4-3, and 4-6 and 4-7, there was no significant difference in the inhibitory effects on 5α-reductase, indicating that the mixture of sodium hyaluronate (HA) and retinoic acid or isotretinoin had no effect on the inhibitory effect on 5α-reductase. The relative inhibition rates of 5α-reductase in samples 4-4, 4-5, and 4-6 showed no significant difference, indicating that the inhibitory effect of retinoic acid on 5α-reductase tended to be stable within the range of 0.15–0.30 mg / ml. The inhibitory effect of 4-2 on 5α-reductase was significantly higher than that of samples 4-4, 4-5, and 4-6, indicating that isotretinoin had a more significant inhibitory effect on 5α-reductase compared to retinoic acid.

[0061] The inhibitory effect of the retinyl hyaluronic acid sodium of the present invention on 5α-reductase is higher than that of retinoic acid and all-trans retinoic acid HA derivatives. The inhibitory effect of samples 4-13, 4-8, 4-9, 4-10 and 4-12 on 5α-reductase increases sequentially, indicating that the proportion of grafted isotretinoin in the retinyl hyaluronic acid sodium of the present invention is increased, and the inhibitory effect on 5α-reductase is enhanced. There is no significant difference in the relative inhibition rate of 5α-reductase between samples 4-10 and 4-12, indicating that when the K value is below 1.51, the inhibitory effect of the samples on 5α-reductase tends to be stable. There is no significant difference in the relative inhibition rate of 5α-reductase between samples 4-8 and 4-13, indicating that when the K value is above 2.96 (the proportion of isotretinoin decreases to a certain extent), the inhibitory effect of the samples on 5α-reductase tends to be stable.

[0062] Table 6. Relative inhibition rates of different test samples against 5α-reductase

[0063]

[0064] Example 5: Antioxidant effect of sodium retinyl hyaluronic acid

[0065] Following the method described in the literature [Xu Yanxia, ​​Wang Rongrong, Mou Hongxia, et al. Extraction of polysaccharides from Zhangye yam and study on the antioxidant properties of polysaccharides, Guangdong Chemical Industry, 2022, No. 17], with vitamin C (Vc) as a positive control, 2 ml of different concentrations (100 μg / ml, 200 μg / ml, 500 μg / ml, 800 μg / ml, 1000 μg / ml) of Vc solution were placed in test tubes respectively. First, 2 ml each of FeSO4 solution (6 mmol / L) and H2O2 solution (6 mmol / L) were added, shaken well, and allowed to stand for 10 min. Then, 2 ml of salicylic acid solution (6 mmol / L) was added, shaken well, and allowed to stand for 30 min. The absorbance value A was measured at 510 nm. i The above operation was then repeated by replacing the salicylic acid solution with distilled water, and the reference absorbance A was measured. b Alternatively, the sample solution was replaced with distilled water and the above experiment was repeated to obtain the blank absorbance A0.

[0066] The ·OH scavenging rate is calculated using equation (1).

[0067]

[0068] The results are shown in Table 7. A log-linear model was used to fit the relationship between vitamin C concentration (μg / ml) and the corresponding clearance rate (%), and the regression equation was calculated as y = 39.726lnx - 170.5, r 2 =0.9928, where x is the concentration of Vc (μg / ml) and y is the corresponding ·OH scavenging rate (%).

[0069] Following the above operating steps, sodium hyaluronate (average molecular weight 7.9 kDa) and the retinyl hyaluronate of the present invention (samples 1-1, 1-2, 1-7, 1-11 and 1-12 of Example 1 of the present invention) and all-trans retinoic acid HA derivatives [according to Huerta- The method disclosed in G. et al. Carbohydrate Polymers (2019) 231(7): 115733 was used to prepare the sample with a retinoic acid substitution degree of 7.5%. The ·OH scavenging rate of the sample was compared with the in vitro antioxidant activity.

[0070] The results (Table 7) show that, compared with sodium hyaluronate (5-1) of similar average molecular weight, the ·OH scavenging rates of the retinyl hyaluronate (5-2 to 5-6) and all-trans retinoic acid HA derivative (5-7) samples of the present invention are significantly improved, and both have significant antioxidant effects. Compared with all-trans retinoic acid HA derivative (5-7), the ·OH scavenging rates of the retinyl hyaluronate (5-3, 5-4, and 5-5) samples of the present invention are not significantly different, indicating that their antioxidant effects are comparable. The ·OH scavenging rates of 5-2, 5-3, 5-4, 5-5, and 5-6 increase sequentially, indicating that the proportion of grafted retinoic acid in the retinyl hyaluronate of the present invention is increased, and the antioxidant effect is enhanced. Compared with 5-3, the ·OH scavenging rates of 5-2 and 5-3 samples are significantly different, indicating that when the K value is lower than 1.51, the antioxidant effect of the samples decreases significantly.

[0071] According to the regression equation for the antioxidant properties of vitamin C, the antioxidant properties of the sodium retinyl hyaluronic acid (samples 1-11 in Example 1) at 5 mg / ml are comparable to those of the vitamin C solution at 223 μg / ml.

[0072] Table 7 ·OH scavenging rate of different samples

[0073]

Claims

1. A sodium retinyl hyaluronic acid, the structure of which is represented by the following general formula: （1） in, a) The molar ratio of a to c is 1.5~3.0 : 1; The molar ratio of (a+c) to (a+b+c) is 1~12%.

2. The sodium retinyl hyaluronic acid according to claim 1, characterized in that, The average molecular weight of the retinyl hyaluronic acid sodium is 5kDa~500kDa.

3. A composition containing sodium retinyl hyaluronic acid as described in claim 1, characterized in that... It consists of sodium retinyl hyaluronic acid and polyol compounds in a mass ratio of 1:1 to 10.

4. The composition of claim 3, wherein the average molecular weight of the sodium retinyl hyaluronic acid is 5 kDa to 500 kDa.

5. The composition according to claim 3, characterized in that, The polyol compound is selected from one or more of glycerol, 1,2-propanediol, polyethylene glycol, and polyglycerol.

6. The composition according to claim 3, characterized in that, The polyol compound is polyglycerol 3 and / or polyglycerol 10.

7. The composition according to claim 3, characterized in that, The polyol compound mentioned is glycerol.

8. The use of sodium retinyl hyaluronic acid as described in claim 1 in the preparation of cosmetics for whitening, anti-aging, oil control and acne removal.

9. The use of the composition of claim 3 in the preparation of cosmetics for whitening, anti-aging, oil control and acne treatment.

10. The use of sodium retinyl hyaluronic acid as described in claim 1 in the preparation of acne creams, acne gels, and acne lotions.

11. The use of the composition of claim 3 in the preparation of acne cream, acne gel, and acne lotion.

Images