Oil-soluble saffron extract composition and use thereof

Abstract

The present invention belongs to the field of pharmaceutical chemistry and / or cosmetics, and in particular to an oil-soluble saffron extract composition and its use. The oil-soluble saffron extract composition of the present invention is prepared by the following method: saffron is crushed, vegetable oil is added for ultrasonic cold extraction, and then centrifugal filtration is performed to obtain the supernatant, wherein the content ratio of saffron to vegetable oil is 1:4 to 1:999 by mass. The composition of the present invention can inhibit the expression of 11β-HSD1, thereby reducing the conversion of cortisone to cortisol, and preventing and / or alleviating skin problems caused by cortisol imbalance.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry and / or cosmetics, and in particular, relates to an oil-soluble saffron extract composition having the function of an 11β-HSD1 expression inhibitor, and use thereof in preparing medicines and / or cosmetics. Background Art

[0002] With the improvement of living standards, consumers are becoming more concerned about their inner feelings. Consumers are also becoming more aware of the correlation between their emotions and their skin. When you feel happy and relaxed for a long time, your skin will usually have a healthy and bright luster. However, when you are anxious, stressed or depressed for a long time, your skin may have various problems. For example, long-term stress may slow down the metabolism of the skin, making the skin dark and rough, and even cause skin problems such as acne and blackheads.

[0003] Therefore, the existing technology has a method of using fragrance or perfume to relieve emotional problems such as tension, anxiety, and stress, so as to maintain the skin. However, using fragrance or perfume to affect emotions and regulate the state of the skin is still an indirect means. Moreover, these means only treat the symptoms and not the root cause. There is no direct and fundamental solution to this problem. In addition, this kind of emotional regulation is a sensory indicator, and it is difficult to quantify it with specific values ​​or indicators.

[0004] The latest research has found that when the human body perceives negative emotions such as tension, anxiety, and isolation, the hypothalamus-pituitary-adrenal (HPA) axis in the human body is activated, further affecting other physiological systems in the body to resist stress. In the regulation process, cortisol (hydrocortisone) is the relevant response glucocorticoid and is regarded as a "stress hormone" and "emotional factor."

[0005] The secretion of cortisol in response can increase blood sugar and heart rate to cope with stress. However, excessive expression of cortisol can also affect the body's normal level. For example, long-term exposure to stress can cause cortisol to cause stress obesity, stress hair loss, acne or eczema, expression lines, etc. This explains the connection between emotions / stress and skin problems from one aspect.

[0006] An imbalance in cortisol levels can have adverse effects on people of all ages. Only by actively establishing a feedback mechanism can the body return to a normal physiological state, thereby eliminating the consequences of cortisol and restoring homeostasis. However, as the most important glucocorticoid in the human body, cortisol has a series of important physiological functions. Therefore, how to locally reduce the skin problems caused by excessive conversion of cortisol to cortisol in the skin is still an urgent problem to be solved.

[0007] In plant extracts, the oil-soluble components and water-soluble components are not exactly the same, so the activity cannot be easily predicted for different extracts. Summary of the invention

[0008] The inventors of the present application unexpectedly discovered that an oil-soluble saffron extract composition can inhibit the expression of 11β-hydroxysteroid dehydrogenase (11β-HSD1), and that an oil-soluble saffron extract composition at a specific concentration can more effectively inhibit the expression of 11β-HSD1, thereby weakening the conversion of cortisone to cortisol, and has better stability, thereby being able to stably prevent and / or alleviate skin problems caused by cortisol imbalance, and on this basis, the present invention was completed.

[0009] An object of the present invention is to provide an oil-soluble saffron extract composition.

[0010] Another object of the present invention is to provide use of the oil-soluble saffron extract composition in the preparation of medicines and / or cosmetics.

[0011] Another object of the present invention is to provide an oil-soluble saffron extract composition-culture medium mixture.

[0012] According to one aspect of the present invention, an oil-soluble saffron extract composition is provided, wherein the extract composition is prepared by the following method:

[0013] The saffron was crushed, a certain amount of vegetable oil was added for ultrasonic cold extraction, and then the supernatant was obtained by centrifugal filtration.

[0014] The content ratio of saffron to vegetable oil is 1:4 to 1:999 by mass, preferably 1:4.

[0015] According to one embodiment of the present invention, the vegetable oil is sunflower seed oil.

[0016] According to one embodiment of the present invention, saffron is crushed by a common grinder or a wall-breaking grinder until it can pass through a 200-mesh sieve.

[0017] According to another aspect of the present invention, it provides use of the oil-soluble saffron extract composition in the preparation of medicines and / or cosmetics.

[0018] Preferably, the medicament is for treating a disease or condition associated with 11β-HSD1 expression.

[0019] Preferably, the disease or condition is low mood or depression.

[0020] Preferably, the cosmetic is a cosmetic for improving skin problems caused by negative emotions.

[0021] According to another aspect of the present invention, there is provided a pharmaceutical composition comprising the oil-soluble saffron extract composition described above, and pharmaceutically acceptable excipients.

[0022] Wherein, the pharmaceutical composition contains the oil-soluble saffron extract composition in a concentration of 0.5%-2% by weight of the pharmaceutical composition.

[0023] Beneficial Effects

[0024] The inventors of the present application have found that the oil-soluble saffron extract composition of the present invention can inhibit the expression of 11β-HSD1, thereby reducing the conversion of cortisol to cortisol, and preventing and / or alleviating skin problems caused by cortisol imbalance. Moreover, the inventors have unexpectedly found that for the oil-soluble saffron extract composition, the oil-soluble saffron extract composition has a large down-regulation rate of 11β-HSD1 at a specific concentration, and when the content is further increased, its down-regulation rate of 11β-HSD1 is reduced, and the oil-soluble saffron extract composition at a specific content has better stability. DETAILED DESCRIPTION

[0025] The present invention is described in detail below with reference to specific embodiments.

[0026] The following examples are merely listed as examples of embodiments of the present invention and do not constitute any limitation to the present invention. Those skilled in the art will appreciate that modifications within the scope of the spirit and concept of the present invention fall within the protection scope of the present invention.

[0027] Test system

[0028] The cells used in this test are normal human epidermal keratinocytes, provided by Beina Biotechnology.

[0029] Main reagents

[0030] EMEM complete medium (Beina Biotechnology), PBS (Solarbio), TRIzol (Invitrogen), reverse transcription kit (Nearshore Protein), fluorescent dye (Nearshore Protein)

[0031] Main Equipment

[0032] Ultrafine grinder (Guangzhou Leimai ZD-6L), refrigerated centrifuge (Xinao, GENIUES 6K-R), ultrasonic oscillator (KQ3200DE), CO2 incubator (Thermo, 150I), clean bench (Suzhou Antai, SW-CJ-1F), gradient PCR instrument (eppendorf, mastercycler pro s), fluorescence quantitative PCR instrument (BioRad, CFX-96).

[0033] Unless otherwise specified, other reagents and materials used in the present invention are reagents and materials that are conventionally available (commercially available) in the art, and the methods and equipment used are methods and equipment conventionally used in the art.

[0034] The invention of this application unexpectedly found that the oil-soluble saffron extract composition of the present invention can inhibit the expression of 11β-HSD1, thereby reducing the conversion of cortisol to cortisol, and preventing and / or alleviating skin problems caused by cortisol imbalance. Among them, the key indicator 11β-HSD1 is related to the amount of cortisol and can be directly quantitatively detected.

[0035] Preparation Example 1: Oil-soluble saffron extract composition

[0036] The oil-soluble saffron extract composition used herein was prepared according to the following method.

[0037] 20 parts of saffron were finely crushed for 20 minutes using a Guangzhou Raymai ZD-6L ultrafine grinder under vibration, and 80 parts of sunflower seed oil were added for ultrasonic cold extraction for 15-75 minutes, with an extraction temperature of 45°C, an ultrasonic power of 30-150W, and an ultrasonic frequency of 40kHz. Then, centrifugal filtration was performed at 3000 rpm, and the supernatant was taken, which was an oil-soluble saffron extract composition.

[0038] Preparation Example 2:

[0039] An oil-soluble saffron extract composition was obtained in the same manner as in Preparation Example 1, except that 19980 parts of sunflower seed oil was used instead of 80 parts of sunflower seed oil, that is, the content of saffron added was 0.1%.

[0040] Preparation Example 3:

[0041] An oil-soluble saffron extract composition was obtained in the same manner as in Preparation Example 1, except that 1980 parts of sunflower seed oil was used instead of 80 parts of sunflower seed oil, that is, the content of saffron added was 1%.

[0042] Preparation Example 4:

[0043] An oil-soluble saffron extract composition was obtained in the same manner as in Preparation Example 1, except that 180 parts of sunflower seed oil was used instead of 80 parts of sunflower seed oil, that is, the content of saffron added was 10%.

[0044] Comparative Preparation Example 1:

[0045] An oil-soluble saffron extract composition was obtained in the same manner as in Preparation Example 1, except that 46.67 parts of sunflower seed oil was used instead of 80 parts of sunflower seed oil, that is, the content of saffron added was 30%.

[0046] Comparative Preparation Example 2:

[0047] An oil-soluble saffron extract composition was obtained in the same manner as in Preparation Example 1, except that 60 parts of sunflower seed oil was used instead of 80 parts of sunflower seed oil, that is, the content of saffron added was 25%.

[0048] Preparation Example 5: Composition-medium mixtures of different concentrations

[0049] The oil-soluble saffron extract composition prepared in Preparation Example 1 was taken, mixed with DMSO to prepare a concentrated stock solution, added to EMEM complete medium, and dispersed under vortex conditions for 3 minutes to obtain composition-medium mixtures 1 to composition-medium mixtures 7 with concentrations of 10%, 5%, 2%, 1%, 0.5%, 0.1%, and 0.01% of the oil-soluble saffron extract composition, respectively.

[0050] Example 1: Testing of the oil-soluble saffron extract composition-medium mixture based on normal human epidermal keratinocytes

[0051] Based on normal human epidermal keratinocytes, the qRT-PCR method was used to detect the changes in the expression level of 11β-HSD1 (HSD1) gene in normal human epidermal keratinocytes after the oil-soluble saffron extract composition-culture medium mixture was detected.

[0052] The specific method is as follows:

[0053] 1) Cell inoculation: After the cells are revived, when the plating rate reaches about 60%, the cells are inoculated into a 6-well plate and incubated overnight in a CO2 incubator (37°C, 5% CO2).

[0054] 2) Liquid preparation: Prepare the working solution of the test substance according to the test group.

[0055] 3) Administration: According to the test grouping, when the cell plating rate in the 6-well plate reaches 40% to 60%, group administration is carried out, and 3 replicate wells are set up for each group. 2 mL of culture medium is added to each well of the blank control group, and 2 mL of culture medium containing the corresponding concentration of the sample to be tested is added to each well of the sample group. After the administration is completed, the 6-well plate is placed in a CO2 incubator (37°C, 5% CO2) and cultured for 24 hours.

[0056] 4) Collecting cells: After culturing for 24 hours, discard the old solution, wash twice with PBS, add 1 mL of TRIzol to each well, pipette and lyse the cells, and collect the samples.

[0057] 5) Gene expression detection: RNA was extracted and reverse transcribed into cDNA, and then fluorescence quantitative PCR was performed. -△△CT Method to calculate the results.

[0058] 6) Calculation of reduction rate: (1-2 -△△CT )×100%

[0059] 7) Statistical analysis of results: GraphPad Prism was used for graphing, and the results were expressed as Mean ± SD. The t-test was used for statistical analysis between the groups. All statistical analyses were two-tailed. P < 0.05 was considered to be significantly different, and P < 0.01 was considered to be extremely significantly different.

[0060] Based on normal human epidermal keratinocytes, compared with the blank control group (see the administration part of 3) above), the oil-soluble saffron extract composition-culture medium mixture was added to the test system at the test concentration, and the resulting downregulation rate of 11β-HSD1 is shown in Table 1 below.

[0061] Table 1: Down-regulation rate of 11β-HSD1 by oil-soluble saffron extract composition-medium mixture

[0062]

[0063] As can be seen from Table 1, at each tested concentration (except 10%), 11β-HSD1 can be significantly down-regulated. Starting from 0.01%, as the concentration increases, at 1%, the down-regulation rate reaches the maximum, and as the concentration increases further, the down-regulation rate decreases.

[0064] Example 2: Stability Test

[0065] Stability test: The oil-soluble saffron extract compositions of Preparation Examples 1-4 and Comparative Preparation Examples 1-2 were stored at -18, 4, 25, 40, and 48°C for 3 months, and the color, odor, and appearance were observed. If there was obvious change in taste, color, turbidity, etc., it was judged that there was a corresponding problem with the stability. The results are shown in Table 2 below.

[0066] Table 2

[0067] Sample name Saffron Content Sunflower seed oil content 3-month stability challenge Preparation Example 2 0.1％ 99.9％ Stablize Preparation Example 3 1％ 99％ Stablize Preparation Example 4 10％ 90％ Stablize Preparation Example 1 20％ 80％ Stablize Comparative Preparation Example 2 25％ 75％ Turbidity appears Comparative Preparation Example 1 30％ 70％ Turbidity appears

[0068] According to the results in Table 2, it can be seen that when the saffron content exceeds 25%, unstable phenomena such as precipitation occur.

Claims

1. An oil-soluble saffron extract composition, the extract composition is prepared by the following method: The saffron was crushed, vegetable oil was added for ultrasonic cold extraction, and then the supernatant was obtained by centrifugal filtration. The content ratio of saffron to vegetable oil is 1:4 to 1:999 by mass.

2. The oil-soluble saffron extract composition according to claim 1, wherein The content ratio of saffron to vegetable oil is 1:4 by mass.

3. The oil-soluble saffron extract composition according to claim 1, wherein The vegetable oil is sunflower seed oil.

4. The oil-soluble saffron extract composition according to claim 1, wherein Use a common grinder or a wall-breaking grinder to grind the saffron until it can pass through a 200-mesh sieve.

5. Use of the oil-soluble saffron extract composition according to claim 1 in the preparation of medicines and / or cosmetics.

6. The use according to claim 5, wherein The drug is used for treating a disease or condition associated with 11β-HSD1 expression.

7. The use according to claim 6, wherein The disease or condition is low mood or depression.

8. The use according to claim 5, wherein The cosmetics are cosmetics used to improve skin problems caused by negative emotions.

9. A pharmaceutical composition comprising the oil-soluble saffron extract composition according to any one of claims 1 to 4, and pharmaceutically acceptable excipients, in, The pharmaceutical composition comprises the oil-soluble saffron extract composition in a concentration of 0.5%-2% based on the weight of the pharmaceutical composition.

Images