Water-soluble saffron crocus extract composition and application thereof

Abstract

The invention belongs to the field of medicinal chemistry and / or cosmetics, and particularly relates to a water-soluble saffron crocus extract composition and application thereof. The water-soluble saffron crocus extract composition is prepared through the following steps that saffron crocus is smashed, a solvent is added for ultrasonic cold extraction, then centrifugal filtration is conducted, supernate is taken to serve as the water-soluble saffron crocus extract composition, the solvent is a mixture of deionized water and low alcohol, and the weight ratio of saffron crocus to the solvent is 1: 350-1: 0.8. The composition provided by the invention can inhibit the expression of 11 beta-HSD1, so that the conversion of cordeson to cortisol can be weakened, and skin problems caused by imbalance of cortisol can be prevented and / or relieved.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceutical chemistry and / or cosmetics, and in particular, relates to a water-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 a water-soluble saffron extract composition can inhibit the expression of 11β-hydroxysteroid dehydrogenase (11β-HSD1), and a water-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 the present invention was completed on this basis.

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

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

[0011] Still another object of the present invention is to provide a water-soluble saffron extract composition-culture medium mixture.

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

[0013] The saffron is crushed, a solvent is added for ultrasonic cold extraction, and then centrifugation is performed to obtain a supernatant as a water-soluble saffron extract composition.

[0014] Wherein, the solvent is a mixture of deionized water and lower alcohol,

[0015] Among them, the weight ratio of saffron to solvent is 1:350 to 1:0.8.

[0016] According to one embodiment of the present invention, the lower alcohol is butanediol, glycerol or pentanediol.

[0017] According to one embodiment of the present invention, the weight ratio of the deionized water to the lower alcohol is 1:1 to 50:1, preferably 10:1 to 40:1, and more preferably 20:1 to 35:1.

[0018] The content of the water-soluble saffron extract as an active substance is 0.1-25% by mass, preferably 3.5% by mass.

[0019] In some embodiments, a common grinder or a wall-breaking grinder can be used to grind saffron until it can pass through a 200-mesh sieve.

[0020] According to another aspect of the present invention, it provides use of the water-soluble saffron extract composition in the preparation of medicines and / or cosmetics.

[0021] Preferably, the medicament is for treating a disease or condition associated with 11β-HSD1 expression.

[0022] Preferably, the disease or condition is low mood or depression.

[0023] Preferably, the cosmetic is a cosmetic for improving skin problems caused by negative emotions.

[0024] According to another aspect of the present invention, there is provided a pharmaceutical composition comprising the water-soluble saffron extract composition described above, and pharmaceutically acceptable excipients.

[0025] Wherein, the pharmaceutical composition contains a water-soluble saffron extract composition in a concentration of 1%-5% based on the weight of the pharmaceutical composition.

[0026] According to another aspect of the present invention, a water-soluble saffron extract is provided, wherein the extract is prepared by the following method:

[0027] The saffron is crushed, a solvent is added for ultrasonic cold extraction, and then centrifugation is performed, and the supernatant is taken for vacuum distillation to dryness to obtain a water-soluble saffron extract.

[0028] Wherein, the solvent is a mixture of deionized water and lower alcohol,

[0029] Among them, the weight ratio of saffron to solvent is 1:350 to 1:0.8.

[0030] Beneficial Effects

[0031] The inventors of the present application have found that the water-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 pharmaceutical compositions, the water-soluble saffron extract composition has a large down-regulation rate of 11β-HSD1 at a specific content, and when the content is further increased, its down-regulation rate of 11β-HSD1 is reduced, and the stability of the water-soluble saffron extract composition at a specific content is better. DETAILED DESCRIPTION

[0032] The present invention is described in detail below with reference to specific embodiments.

[0033] 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.

[0034] Test system

[0035] The cells used in this test are normal human epidermal keratinocytes, provided by Beina Biotechnology.

[0036] Main reagents

[0037] EMEM complete medium (Beina Biotechnology), PBS (Solarbio), TRIzol (Invitrogen), reverse transcription kit (Nearshore Protein), fluorescent dye (Nearshore Protein)

[0038] Main Equipment

[0039] Ultrafine pulverizer (Guangzhou Leimai ZD-6L), refrigerated centrifuge (Xinao, GENIUES 6K-R), ultrasonic oscillator (KQ3200DE), CO 2 Incubator (Thermo, 150I), clean bench (Suzhou Antai, SW-CJ-1F), gradient PCR instrument (eppendorf, mastercycler pro s), and fluorescence quantitative PCR instrument (BioRad, CFX-96).

[0040] Unless otherwise specified, the 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.

[0041] The invention of the present application unexpectedly found that the water-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.

[0042] Preparation Example 1: Water-soluble saffron extract composition

[0043] The water-soluble saffron extract composition used herein was prepared according to the following method.

[0044] 10.7 parts of saffron were finely ground under vibration using a Guangzhou Leimai ZD-6L ultrafine grinder for 20 minutes, 93.5 parts of deionized water and 3 parts of butanediol were added for ultrasonic cold extraction, and then centrifuged and filtered. The supernatant was taken, and the solute content of some samples was tested. It was found that the water-soluble saffron extract content was 3.5%, water 93.5%, and butanediol 3%. The supernatant was the water-soluble saffron extract composition.

[0045] The solute content detection method is as follows: weigh 2 grams of extract (accurate to 0.0001 grams) in a dried constant-weight beaker, dry it in a constant-temperature drying oven at 105±1℃ for 3 hours, take it out and put it in a desiccator to cool to room temperature, and weigh it (accurate to 0.0001 grams).

[0046] Preparation Example 2:

[0047] A water-soluble saffron extract composition was obtained in the same manner as in Preparation Example 1, except that the ratios of deionized water, butylene glycol, and saffron were adjusted to 96.9 parts, 3 parts, and 0.31 parts, respectively. As a result, the content of the water-soluble saffron extract was 0.1%, water 96.9%, and butylene glycol 3%.

[0048] Preparation Example 3:

[0049] A water-soluble saffron extract composition was obtained in the same manner as in Preparation Example 1, except that the ratios of deionized water, butylene glycol, and saffron were adjusted to 96 parts, 3 parts, and 3.1 parts, respectively. As a result, the content of the water-soluble saffron extract was 1%, water 96%, and butylene glycol 3%.

[0050] Preparation Example 4:

[0051] A water-soluble saffron extract composition was obtained in the same manner as Preparation Example 1, except that the ratios of deionized water, butylene glycol, and saffron were adjusted to 87 parts, 3 parts, and 30.6 parts, respectively, wherein the content of the water-soluble saffron extract was 10%, water 87%, and butylene glycol 3%.

[0052] Preparation Example 5:

[0053] A water-soluble saffron extract composition was obtained in the same manner as Preparation Example 1, except that the ratios of deionized water, butylene glycol, and saffron were adjusted to 77 parts, 3 parts, and 61 parts, respectively, wherein the content of the water-soluble saffron extract was 20%, water 77%, and butylene glycol 3%.

[0054] Preparation Example 6:

[0055] A water-soluble saffron extract composition was obtained in the same manner as Preparation Example 1, except that the ratios of deionized water, butylene glycol, and saffron were adjusted to 72 parts, 3 parts, and 76.5 parts, respectively, wherein the content of the water-soluble saffron extract was 25%, water 72%, and butylene glycol 3%.

[0056] Comparative Preparation Example 1:

[0057] A water-soluble saffron extract composition was obtained in the same manner as Preparation Example 1, except that the ratios of deionized water, butylene glycol, and saffron were adjusted to 67 parts, 3 parts, and 91.7 parts, respectively, wherein the content of the water-soluble saffron extract was 30%, water 67%, and butylene glycol 3%.

[0058] Preparation Example 7: Using different ratios of water-soluble saffron extract compositions to obtain composition-medium mixtures of different concentrations

[0059] The water-soluble saffron extract composition prepared in Preparation Example 1 was taken, mixed with EMEM complete medium, and dispersed under vortex conditions for 3 minutes to obtain composition-medium mixtures 1 to composition-medium mixtures 6 with concentrations of water-soluble saffron extract compositions of 20%, 10%, 5%, 2.5%, 1%, and 0.1%, respectively.

[0060] Example 1: Test on normal human epidermal keratinocytes containing a water-soluble saffron extract composition-medium mixture

[0061] 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 water-soluble saffron extract composition-culture medium mixture.

[0062] The specific method is as follows:

[0063] 1) Cell inoculation: After the cells are revived, when the plating rate reaches about 60%, inoculate the cells into a 6-well plate and incubate at 4 °C for 1 h. 2 Incubator (37°C, 5% CO 2 ) and incubate overnight.

[0064] 2) Liquid preparation: Prepare the working solution of the test substance according to the test group.

[0065] 3) Drug administration: According to the test grouping, when the cell plating rate in the 6-well plate reaches 40% to 60%, group drug administration is carried out, with 3 replicate wells in 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 drug administration, the 6-well plate is placed in a CO 2 Incubator (37°C, 5% CO 2 ) for 24 h.

[0066] 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 then collect the samples.

[0067] 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.

[0068] 6) Calculation of down adjustment rate: (1-2 -△△CT )×100%

[0069] 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.

[0070] Based on normal human epidermal keratinocytes, compared with the blank control group (see the administration part of 3) above), the water-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.

[0071] Table 1: Down-regulation rate of 11β-HSD1 by water-soluble saffron extract

[0072]

[0073] As can be seen from Table 1, at each tested concentration (except 20%), 11β-HSD1 can be significantly down-regulated. Starting from 0.1%, as the concentration increases, at 2.5%, the down-regulation rate reaches the maximum, and as the concentration increases further, the down-regulation rate decreases.

[0074] Example 2: Stability Test

[0075] Stability test: The water-soluble saffron extract compositions of Preparation Examples 1-6 and Comparative Preparation Example 1 were stored at -18, 4, 25, 40, and 48°C for 3 months, and the color, odor, and appearance indicators were observed. If obvious taste change, color change, turbidity, etc. occur, it is judged that the stability has corresponding problems. The results are shown in Table 2 below.

[0076] Table 2

[0077]

[0078] According to the results in Table 2, it can be seen that when the content of the water-soluble saffron extract exceeds 25%, unstable phenomena such as precipitation occur.

Claims

1. A water-soluble saffron extract composition, the extract composition is prepared by the following method: The saffron is crushed, a solvent is added for ultrasonic cold extraction, and then centrifugation is performed to obtain a supernatant as a water-soluble saffron extract composition. in, The solvent is a mixture of deionized water and lower alcohol, Among them, the weight ratio of saffron to solvent is 1:350 to 1:0.

8.

2. The water-soluble saffron extract composition according to claim 1, wherein The lower alcohol is butanediol, glycerol or pentanediol.

3. The water-soluble saffron extract composition according to claim 1, wherein in, The weight ratio of deionized water to lower alcohol is 1:1 to 50:1, preferably 10:1 to 40:1, and more preferably 20:1 to 35:

1.

4. The water-soluble saffron extract composition according to claim 1, wherein The content of the water-soluble saffron extract is 0.1-25% by mass, preferably 3.5% by mass.

5. The water-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.

6. Use of the water-soluble saffron extract composition according to claim 1 in the preparation of medicines and / or cosmetics, preferably, the medicine is used to treat diseases or conditions related to 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 6, wherein The cosmetics are cosmetics used to improve skin problems caused by negative emotions.

9. A pharmaceutical composition comprising the water-soluble saffron extract composition according to any one of claims 1 to 5, and pharmaceutically acceptable excipients, in, The pharmaceutical composition comprises the water-soluble saffron extract composition in a concentration of 1%-5% based on the weight of the pharmaceutical composition.

10. A water-soluble saffron extract, the extract being prepared by the following method: The saffron is crushed, a solvent is added for ultrasonic cold extraction, and then centrifugation is performed, and the supernatant is taken for vacuum distillation to dryness to obtain a water-soluble saffron extract. in, The solvent is a mixture of deionized water and lower alcohol, Among them, the weight ratio of saffron to solvent is 1:350 to 1:0.8.

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