Retinoyl sodium hyaluronate and composition thereof

Abstract

The invention provides retinyl sodium hyaluronate and a composition thereof, which are applied to the fields of cosmetics and medicines. The retinoyl sodium hyaluronate is characterized in that all-trans retinoic acid and 13-cis-retinoic acid are esterified with sodium hyaluronate according to a molar ratio of (1.5-3.0): 1, and the esterification degree is 1%-12%; the composition disclosed by the invention is characterized by comprising retinyl sodium hyaluronate and a polyhydric alcohol compound in a mass ratio of 1: (1-10). The sodium retinoyl hyaluronate has a remarkable effect of preventing and treating acne, and the composition is beneficial to improving the stability of the sodium retinoyl hyaluronate and prolonging the validity period of a raw material product of the sodium retinoyl hyaluronate. The retinyl sodium hyaluronate and the composition thereof can be used for preparing cosmetics and medical products for whitening, resisting aging, controlling oil and removing acnes.

Description

Technical Field

[0001] The present invention relates to sodium retinoyl hyaluronate and a composition thereof which are applied in the fields of cosmetics and medicine. Background Art

[0002] Tretinoin (TR) and isotretinoin (ITR) are both vitamin A derivatives. Tretinoin is also known as retinoic acid and all-trans retinoic acid, and its chemical name is 3,7-dimethyl-9-(2,6,6-trimethylcyclohexene)-2,4,6,8-all-trans nonatetraene. Isotretinoin is an isomer of retinoic acid, also known as 13-cis retinoic acid, and its chemical name is 3,7-dimethyl-9-(2,6,6-trimethyl-1-cyclohexenyl)-2-cis-4-trans-6-trans-8-nonatetraene. Tretinoin has the effects of promoting skin tissue proliferation, promoting the shedding of the stratum corneum, eliminating acne lesions, promoting the healing of skin lesions, reducing melanin formation, and anti-inflammatory. Isotretinoin can inhibit sebum secretion, reduce the proliferation of sebaceous gland tissue, and inhibit the growth and reproduction of acne bacteria. Both can be used externally to treat acne and blackheads.

[0003] Hyaluronic acid (HA), also known as glassy acid and hyaluronic acid, is one of the main components of the extracellular matrix of the skin. It is a chain polyanionic mucopolysaccharide composed of repeated connections of (1-β-4) D-glucuronic acid and (1-β-3) N-acetyl-D-aminoglucose disaccharide units. HA is considered to be an ideal natural moisturizing factor when applied to the skin surface. Small molecule HA also has the functions of regulating the differentiation and migration of epidermal cells and scavenging free radicals. Therefore, in addition to moisturizing effects, it also has skin nutrition, sunscreen repair, lubrication, film formation and thickening effects. Both retinoic acid and isotretinoin are insoluble in water and have poor stability. Both of them can cause skin irritation symptoms when used externally, such as burning sensation, erythema and desquamation. In order to overcome these defects, a series of esterified derivatives have been developed in recent years, among which HA is directly esterified with retinoic acid [Huerta- G. et al. Carbohydrate Polymers (2019) 231(7): 115733 and CN117964796] or prepared all-trans retinoic acid derivatives by small molecule bridging esterification (CN116284495), which improved the water solubility and stability of retinoic acid to a certain extent, and reduced the irritation of retinoic acid to the skin. Studies have shown that the occurrence of acne is closely related to excessive sebum secretion and Proteobacterium acnes, while retinoic acid has limited effect on skin oil control. 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 material products also needs to be further improved.

[0004] The present invention provides a sodium retinoyl hyaluronate derivative, which uses all-trans retinoic acid (retinoic acid) and 13-cis retinoic acid (isoretinoic acid) to simultaneously form an ester with sodium hyaluronate to enhance its efficacy in preventing and treating acne. The derivative of the present invention and a polyol compound are combined to form a complex, which can further improve its stability and extend the shelf life of its raw material product. The derivative and composition of the present invention can be widely used in the preparation of cosmetics and medical products with whitening, anti-aging, oil control and acne removal effects. Summary of the invention

[0005] The purpose of the present invention is to provide a sodium retinoyl hyaluronate derivative having oil control, acne removal, whitening and anti-aging effects and a composition thereof, as well as their application in the fields of cosmetics and medicine.

[0006] The structure of the sodium retinoyl hyaluronate of the present invention is 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 composition containing sodium retinoyl hyaluronate of the present invention is characterized in that the sodium retinoyl hyaluronate and the polyol compound are composed in a ratio of 1:1 to 10 (mass ratio).

[0012] The composition containing sodium retinoyl hyaluronate of the present invention is characterized in that the structure of the sodium retinoyl hyaluronate 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 amount of retinoic acid and isotretinoin grafted onto the hyaluronic acid molecular skeleton measured by high performance liquid chromatography (HPLC), and the ratio ranges from 1.5 to 3.0:1, preferably 2.0 to 2.5:1.

[0018] The meaning of (a+c) / (a+b+c) is the ratio of the retinoic acid and isotretinoic acid molecules grafted on the hyaluronic acid molecular skeleton to the hyaluronic acid disaccharide molecules, that is, the grafting rate of the sodium retinoyl hyaluronate of the present invention, also known as the degree of substitution or degree of esterification. The number of retinoic acid and isotretinoic acid molecules (a+c) is obtained by converting the content of the two molecules after hydrolyzing the sodium retinoyl hyaluronate using high performance liquid chromatography (HPLC), and the number of hyaluronic acid disaccharide molecules (a+b+c) is obtained by converting the content of glucuronic acid after measuring the content of glucuronic acid using the carbazole method. The ratio ranges from 1% to 12%, preferably from 4.0% to 8.0%.

[0019] The sodium retinoyl hyaluronate and the composition containing the sodium retinoyl hyaluronate of the present invention have an average molecular weight of 5 kDa to 500 kDa, preferably an average molecular weight of 8 kDa to 300 kDa, and more preferably an average molecular weight of 10 kDa to 100 kDa. The average molecular weight is a weight average molecular weight measured by gel permeation chromatography (GPC) and / or laser scattering-gel permeation chromatography (LLS-GPC).

[0020] The composition containing sodium retinoyl hyaluronate of the present invention is characterized in that the ratio of sodium retinoyl hyaluronate to the polyol compound is in the range of 1:1 to 10 (mass ratio), preferably 1:3 to 5.

[0021] The composition containing sodium retinoyl hyaluronate of the present invention is characterized in that the polyol compound is selected from one or a combination of two or more of glycerol, 1.2-propylene glycol, polyethylene glycol and polyglycerol. Among them, one or a combination of two or more of glycerol, polyglycerol 3 and / or polyglycerol 10 is preferred, and polyglycerol 10 is more preferred.

[0022] The sodium retinoyl hyaluronate with the technical features of the present invention adopts all-trans retinoic acid and 13-cis retinoic acid (isotretinoin) to simultaneously form an ester with sodium hyaluronate. When applied to the skin surface, in addition to the functions of retinoic acid in exfoliating, anti-inflammatory, eliminating acne, repairing, whitening and anti-aging, it also has the function of 13-cis retinoic acid (isotretinoin) in inhibiting sebum secretion and the growth of acne bacteria, thereby enhancing its efficacy in preventing and treating acne.

[0023] The composition containing sodium retinoyl hyaluronate having the technical features of the present invention, and the complex formed by uniformly mixing sodium retinoyl hyaluronate and a polyol compound, can further improve the stability of sodium retinoyl hyaluronate and extend the shelf life of its raw material product.

[0024] The sodium retinoyl hyaluronate and its composition with the technical features of the present invention, in addition to having good water solubility and relatively mild skin irritation, have a significant inhibitory effect on 5α-reductase, can effectively control oil, and are particularly effective in preventing and treating acne vulgaris and improving the water-oil balance of the skin. It can be widely used in the preparation of skin external-use products with whitening, anti-aging, oil control and acne removal effects, including cosmetics and medical products.

[0025] The sodium retinoyl hyaluronate and the composition thereof of the present invention are formulated with ingredients commonly used in cosmetics to be prepared into various forms such as solutions, gels, ointments, creams or lotions, for example, balancing moisturizing lotion, balancing moisturizing spray, balancing whitening facial mask, oil-control toner, whitening lotion, softening lotion, exfoliating gel, anti-acne gel, balancing moisturizing whitening lotion, face cream, body cream, hand cream, foot cream, repair essence, moisturizing cream, repair lotion, acne scar cream, scalp care lotion, etc.

[0026] The sodium yellow hyaluronate and the composition thereof of the present invention are formulated with auxiliary materials commonly used in the medical field to form various dosage forms such as solutions, gels, creams, etc., such as acne creams, acne gels, acne lotions, etc. DETAILED DESCRIPTION

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

[0028] Example 1 Sodium retinoyl hyaluronate

[0029] Dissolve hyaluronic acid, retinoic acid and / or isotretinoin in dimethylformamide or dimethyl sulfoxide, add an appropriate amount of carbonyldiimidazole, heat in the dark for reaction, add saturated sodium chloride solution for dilution, add ethanol for precipitation and washing, collect the precipitate after filtration, and dry under reduced pressure to obtain a light yellow to yellow granular powder, which is the sodium retinoyl hyaluronate of the present invention (samples 1-1 to 1-12).

[0030] Table 1 Sodium retinoyl hyaluronate

[0031]

[0032] Retinoic acid [C TR % (g / g)] and isotretinoin [C ITR % (g / g)] content determination method:

[0033] The sodium retinoyl hyaluronate of the present invention was taken, and after being hydrolyzed by adding an appropriate amount of alkaline solution, it was determined by HPLC. Retinoic acid and isotretinoin were used as reference substances, an octadecylsilane bonded silica gel chromatographic column and an ultraviolet detector were used, and the mobile phase was methanol: 2% acetic acid = 81:19. Retinoic acid [C TR (g / g)] and the content of isotretinoin [C ITR (g / g)]

[0034] Glucuronic acid [T / % (g / g)] content determination method:

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

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

[0037] K=C TR / C ITR .

[0038] Calculation of substitution degree / % (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 retinoyl hyaluronate

[0041] The sodium retinoyl hyaluronate of the present invention and the polyol compound are uniformly mixed according to a certain proportion to obtain the composition samples 2-1 to 2-10 of the present invention (Table 2).

[0042] Table 2 Ingredients and proportions of different formula compositions

[0043]

[0044] Example 3 Stability Test

[0045] Samples 1-7 in Example 1 of the present invention were divided into sealed aluminum-plastic bags and brown spiral-mouthed glass sample bottles. Polyol compounds were added to samples 1-7 in Example 1, mixed and divided into brown spiral-mouthed glass sample bottles as samples for stability test (Table 3). They were placed in different temperature environments for stability investigation. Samples were taken at different times to detect the sum of the contents of free retinoic acid and isotretinoin bound to HA molecules in the samples. Ci=C TR +C ITR (%). The results showed that there was no significant increase in free retinoic acid and isotretinoin after different conditions were investigated, and the ratio of retinoic acid to isotretinoin (K value) did not change significantly; the change rate of retinoic acid and isotretinoin content in different conditions and different time samples was R = (Ci-C0 ) / C 0 *100%, of which C 0 is the sum of the contents of retinoic acid and isotretinoin in the 0-day sample. The results are shown in Tables 4 and 5.

[0046] Table 3 Retinoyl hyaluronate sodium and different ratio composition samples

[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 R value determination results of stability test under high temperature conditions*

[0052]

[0053] *“-” indicates a decrease in content

[0054] The results of the stability test under low temperature conditions show that lowering the storage temperature is beneficial to the stability of the raw material product. After the glass bottle packaging is filled with nitrogen for protection, the stability of the raw material product is significantly improved, indicating that isolating from the air is beneficial to the stability of the raw material product.

[0055] The results of the stability test under high temperature conditions show that compared with 3-7, the absolute values ​​of R of 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 degree of substitution is reduced), indicating that after the sodium retinoyl hyaluronate of the present invention is formulated with a polyol, it can effectively isolate the influence of air in the absence of nitrogen protection, thereby reducing the degradation of the raw material product and improving its stability.

[0056] According to the results of high temperature of 40°C and 60°C, cold storage at 2-10°C, and 25°C, the stability of the composition of sodium retinoyl hyaluronate and two polyglycerols of the present invention is relatively good (equivalent to the sample of sodium retinoyl hyaluronate alone in a sealed aluminum-plastic bag), followed by the composition of glycerol and propylene glycol.

[0057] Example 4 Inhibitory Effect of Sodium Retinoyl Hyaluronate on 5α-Reductase

[0058] According to the literature method (Pan Jifei et al. Establishment and application of in vitro evaluation system of 5α-reductase inhibitors. Daily Chemical Industry (Chinese and English), 2023, 53(11): 1280-1284), the sodium retinoyl hyaluronate of the present invention (samples 1-1, 1-2, 1-7, 1-11 and 1-12 of Example 1 of the present invention), sodium hyaluronate, retinoic acid, isotretinoin and all-trans retinoic acid HA derivatives [according to the literature Huerta- G.et al.Carbohydrate Polymers (2019) 231 (7): 115733 prepared by the disclosed method, retinoic acid substitution degree 7.5%] on the relative inhibition rate of 5α-reductase (5AR), and compared the in vitro oil control efficacy.

[0059] The test sample was added to the corresponding 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, and a 0.2 mM dutasteride (DTS) solution was used as the positive control (PC), and the relative inhibition rate was determined and calculated.

[0060] The results (Table 6) showed that, except for sodium hyaluronate (HA), the other groups had different degrees of inhibitory effect on 5α-reductase; there was no significant difference in the inhibitory effect of 4-2 and 4-3, 4-6 and 4-7 on 5α-reductase, indicating that sodium hyaluronate (HA) mixed with retinoic acid or isotretinoin had no effect on the inhibitory effect of 5α-reductase; there was no significant difference in the relative inhibition rate of 4-4, 4-5 and 4-6 samples on 5α-reductase, indicating that the inhibitory effect of retinoic acid samples 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 improved compared with 4-4, 4-5 and 4-6 samples, indicating that compared with retinoic acid, isotretinoin has a more significant inhibitory effect on 5α-reductase.

[0061] The inhibitory effects of the sodium retinoyl hyaluronate of the present invention on 5α-reductase are higher than those of retinoic acid and all-trans retinoic acid HA derivatives. The inhibitory effects of the test samples 4-13, 4-8, 4-9, 4-10 and 4-12 on 5α-reductase increase successively, indicating that the proportion of grafted isotretinoin in the sodium retinoyl hyaluronate of the present invention increases, 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 lower than 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 higher than 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 rate of different test samples on 5α-reductase

[0063]

[0064] Example 5 Antioxidant Effect of Sodium Retinoyl Hyaluronate

[0065] According to the literature method [Xu Yanxia, ​​Wang Rongrong, Mou Hongxia, et al. Extraction of Zhangye yam polysaccharides and study on the antioxidant properties of polysaccharides, Guangdong Chemical Industry, Issue 17, 2022], vitamin C (Vc) was used as a positive control, 2 ml of Vc solution with different concentrations (100 μg / ml, 200 μg / ml, 500 μg / ml, 800 μg / ml, 1000 μg / ml) were placed in test tubes, and FeSO4 solution (6 mmol / L) and H 2 O 2 2 ml of each solution (6 mmol / L) was shaken and allowed to stand for 10 min, then 2 ml of salicylic acid solution (6 mmol / L) was added, shaken and allowed to stand for 30 min, and the absorbance value A was measured at 510 nm. i Then, the salicylic acid solution was replaced with distilled water and the above operation was repeated to measure the reference absorbance A. b ; Replace the sample solution with distilled water and repeat the above test to obtain the blank absorbance A 0 .

[0066] The OH removal rate was calculated by formula (1).

[0067]

[0068] The results are shown in Table 7. The Vc concentration (μg / ml) and the corresponding clearance rate (%) were fitted according to the logarithmic linear model, and the regression equation was calculated as y=39.726lnx-170.5, r 2 =0.9928, where x is the Vc concentration (μg / ml) and y is the corresponding ·OH clearance rate (%).

[0069] According to the above operation steps, sodium hyaluronate (average molecular weight 7.9 kDa) and sodium retinoyl 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 samples were prepared by the method disclosed in G.et al.Carbohydrate Polymers (2019) 231(7): 115733, and the ·OH clearance rate of the samples [retinoic acid substitution degree 7.5%] was compared for in vitro antioxidant effects.

[0070] The results (Table 7) show that compared with sodium hyaluronate (5-1) with equivalent average molecular weight, the ·OH clearance rates of the samples of sodium retinoyl hyaluronate (5-2 to 5-6) and the all-trans retinoic acid HA derivative (5-7) of the present invention are significantly improved, and both have significant antioxidant effects; compared with the all-trans retinoic acid HA derivative (5-7), the ·OH clearance rates of the samples of sodium retinoyl hyaluronate (5-3, 5-4 and 5-5) of the present invention are not significantly different, indicating that their antioxidant effects are equivalent; the ·OH clearance rates of 5-2, 5-3, 5-4, 5-5 and 5-6 increase in sequence, indicating that the proportion of grafted retinoic acid in the sodium retinoyl hyaluronate of the present invention increases and the antioxidant effect is enhanced; compared with 5-2 and 5-3, the ·OH clearance rates of the 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 of the antioxidant performance of Vc, the antioxidant performance of 5 mg / ml of the sodium retinoyl hyaluronate (Sample 1-11 of Example 1) of the present invention is equivalent to that of 223 μg / ml of Vc solution.

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

[0073]

Claims

1. A sodium retinoyl hyaluronate, the structure of which is represented by the following general formula: in, a)a:c=1.5~3.0:1(mol / mol); and b) (a+c) / (a+b+c)=1 to 12% (mol / mol).

2. The sodium retinoyl hyaluronate according to claim 1, characterized in that The average molecular weight of the sodium retinoyl hyaluronate is 5kDa to 500kDa.

3. A composition containing sodium retinoyl hyaluronate, characterized in that: The sodium retinoyl hyaluronate and the polyol compound are composed in a ratio of 1:1 to 10 (mass ratio).

4. The composition according to claim 3, characterized in that The structure of the sodium retinoyl hyaluronate is represented by the following general formula: in, a)a:c=1.5~3.0:1(mol / mol); and b) (a+c) / (a+b+c)=1 to 12% (mol / mol).

5. The composition of claim 3, wherein the average molecular weight of the sodium retinoyl hyaluronate is 5 kDa to 500 kDa.

6. The composition according to claim 3, characterized in that The polyol compound is selected from one or a combination of two or more of glycerol, 1.2-propylene glycol, polyethylene glycol and polyglycerol.

7. The composition of claim 3, characterized in that The polyol compound is polyglycerol 3 and / or polyglycerol 10.

8. The composition of claim 3, characterized in that The polyol compound is glycerol.

9. The sodium retinoyl hyaluronate according to claim 1 is used for preparing cosmetics and medical products for whitening, anti-aging, oil control and acne removal.

10. The composition of claim 3 is used for preparing cosmetics and medical products for whitening, anti-aging, oil control and acne removal.

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