Oil control composition as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of skin care products, and particularly relates to an oil control composition as well as a preparation method and application thereof. The oil control composition is prepared by compounding a dracocephalum integrifolium extract and a callicarpa nudiflora extract. The dracocephalum chinense extract is extracted through a deep-eutectic solvent in cooperation with an ultrasonic-microwave process, and the callicarpa nudiflora extract is extracted through an enzymolysis-supercritical CO2 microencapsulation process. Oil control is realized through a double-target mechanism, including blocking of SREBP-1 nuclear translocation, down-regulation of PPAR gamma and the like, in cooperation with the two. The composition can be applied to oil-control cosmetics such as essence and emulsion, the process is efficient and stable, and a safe and multi-effect solution is provided for oily skin care.

Description

Technical Field

[0001] The present invention belongs to the technical field of skin care products, and particularly relates to an oil-control composition, a preparation method thereof, and an application thereof. Background Art

[0002] The over-activity of sebaceous glands and abnormal lipid metabolism are the core mechanisms of oily skin and acne pathogenesis, and their regulation involves the interaction of multiple signaling pathways. Research has confirmed that the mTORC1 pathway drives the expression of fatty acid synthase (FAS) and acetyl-CoA carboxylase (ACC) by activating the downstream transcription factor SREBP-1, promoting de novo fatty acid synthesis; the PPARγ pathway enhances lipid storage and secretion capacity by upregulating the expression of lipid droplet formation proteins (such as PLIN2). In addition, androgen signaling (such as the DHT-AR axis) further amplifies the activity of key lipid metabolism enzymes (such as DGAT1, SCD1) by activating the PI3K / AKT / mTOR cascade reaction, forming a positive feedback loop of "synthesis-storage-secretion".

[0003] Existing intervention methods mostly focus on single-target inhibition (such as blocking 5α-reductase to reduce DHT production, or antagonizing mTOR phosphorylation). For example, CN112569152A discloses an oil-control and anti-inflammatory plant combination extract, which is prepared from the following raw materials in parts by mass: 15-45 parts of olive leaf, 15-45 parts of swertia mileensis, 15-45 parts of glossy privet fruit, 4-25 parts of coptis chinensis, 4-25 parts of angelica sinensis, and 4-25 parts of liquorice. It uses hydrophilic acids (such as oleanolic acid, maslinic acid, arbutinolic acid) in the combination extract to inhibit 5α-reductase and thus achieve the oil-control effect. However, such strategies have limitations: excessive inhibition of mTOR may interfere with normal cell metabolic homeostasis, and simply blocking androgen signaling cannot comprehensively regulate the lipid synthesis network.

[0004] In the prior art, natural plant extracts are limited by problems such as component synergy, process complexity, and safety, and it is difficult to meet consumers' demands for high efficiency, mildness, and multi-effect integration. CN118948725A discloses an oil-control composition that relies on flavonoid components in hypericum perforatum extract to inhibit the p-Akt / pErk 1 / 2 pathway. However, its photosensitive component hypericin generates free radicals under ultraviolet irradiation, resulting in skin cell damage; at the same time, hypericum perforatum extract may activate inflammatory factors such as TNF-α, triggering contact dermatitis.

[0005] CN118453484A discloses an oil-control plant composition, comprising: 1-4 parts of Artemisia annua extract; 0.5-1 part of Coptis chinensis extract; 1-3 parts of Scutellaria baicalensis extract; 3-5 parts of white tea extract; 3-7 parts of lophatherum gracile extract; 0.1-1 part of Hamamelis virginiana callus tissue extract; and 6-10 parts of pumpkin seed oil. However, berberine in the Scutellaria baicalensis extract may irritate the skin, causing dryness and peeling; tannins are rich in the white tea extract, lophatherum gracile extract and Hamamelis virginiana callus tissue extract. Although its astringency can reduce sebum secretion in the short term, excessive astringency may inhibit the normal metabolism of the skin, damage the skin barrier, resulting in dryness and tightness; pumpkin seed oil may cause the product to be sticky and affect the use experience.

[0006] In addition, some synthetic drugs (such as retinoids) are prone to cause skin barrier damage due to their non-selective effects. Therefore, developing a technical solution that can synergistically regulate multiple pathway nodes (such as simultaneously intervening in the mTOR / SREBP-1, PPARγ and androgen signals) and has safety has become an important direction for optimizing the regulation of sebaceous gland function. Summary of the Invention

[0007] The present invention aims to re-screen natural plant combinations, construct a more concise, safe and multi-target oil-control system, and optimize the extraction process at the same time, so as to solve the core pain points of the existing technology and promote the scientific and precise development of natural skin care products.

[0008] Dracocephalum integrifolium, also known as Dracocephalum integrifolium and Dracocephalum integrifolium, is the dried above-ground part of Dracocephalum integrifolium Bunge of the genus Dracocephalum in the Lamiaceae family. It is widely distributed in the mountain slopes, forest edges and river valleys in the northwest, north and northeast of China. It has strong adaptability, rich resource reserves and is easy to be cultivated on a large scale. Its stem is erect, 20-50 cm high, quadrangular, much branched, densely covered with short pubescence; the leaves are opposite, entire and toothless, lanceolate to ovate-lanceolate, 2-5 cm long, 0.5-1.5 cm wide, dark green on the surface, gray-green on the back, with glandular dots; the verticillate inflorescence is terminal, the corolla is blue-violet, bilabiate, the upper lip is erect, and the lower lip is 3-lobed. It is brittle and easy to break, with a faint fragrance and a slightly pungent smell, bitter and sweet in taste.

[0009] Dracocephalum integrifolium is rich in active ingredients such as flavonoids (such as luteolin, apigenin, kaempferol-7-O-glucoside), triterpenoids (such as oleanolic acid, ursolic acid), volatile oils (containing limonene, menthone, linalool), phenolic acids (rosmarinic acid, caffeic acid) and phenyl ethanol glycosides (verbascoside, isoverbascoside), etc. Its extract has a history of safe application for hundreds of years in traditional folk medicine, and there are no records of toxicity or adverse reactions. Modern toxicological studies have also confirmed that its LD50 value is significantly higher than the conventional medicinal dose, and there is no risk of mutagenicity or teratogenicity, meeting the requirements of the Chinese Pharmacopoeia and the safety standards of cosmetic raw materials. Traditional medicine believes that it has the effects of clearing heat and detoxifying, relieving cough and asthma, and anti-inflammatory and analgesic, and is often used for the adjuvant treatment of exogenous cough, bronchitis and skin inflammation.

[0010] Callicarpa nudiflora, also known as purple pearl grass and naked flower purple pearl grass, is the dried leaves and tender branches of Callicarpa nudiflora Hook.&Arn. of the genus Callicarpa in the Verbenaceae family. It is widely distributed in the subtropical and tropical regions of Guangdong, Guangxi, Yunnan, Fujian and other provinces in China, such as hillside shrubs, forest edges by streams and hilly areas. The wild resources are abundant and the artificial cultivation technology is mature, with the advantage of sustainable utilization. Its plants are deciduous shrubs, 1-3 meters high, with quadrangular branches densely covered with grayish-white stellate villi; the leaves are opposite, ovate-lanceolate to elliptic, 5-15 cm long and 2-6 cm wide, with fine serrations on the edge, dark green on the surface and densely covered with grayish-white pubescence on the back; the cymes are axillary, the corolla is purple, 4-lobed; the fruits are spherical and bright purple when mature. It is light and brittle, with a slightly fragrant smell and a bitter and astringent taste that turns sweet aftertaste.

[0011] Callicarpa nudiflora is rich in active ingredients such as flavonoids (such as luteolin, apigenin, quercetin-3-O-glucoside), triterpenoid saponins (such as callicarpa nudiflora saponins A, B, C), phenyl ethanol glycosides (verbascoside, isoverbascoside), volatile oils (containing α-pinene, β-caryophyllene, eucalyptol) and tannins, polyphenols, etc. Its extract has been used in folk medicine in the Lingnan region for thousands of years, and is often used for traumatic hemostasis, burn repair and skin infection treatment. There are no records of its toxic reactions in ancient herbal classics. Modern toxicological evaluations show that its acute oral LD50 > 5000 mg / kg (rats), and it has no skin irritation, sensitization and genetic toxicity, meeting the requirements of the Chinese Pharmacopoeia and international safety specifications for cosmetic raw materials (such as ECHA, CIR).

[0012] In order to achieve the above object, the present invention provides the following technical solutions:

[0013] An oil-control composition is composed of Dracocephalum integrifolium extract and Callicarpa nudiflora extract, wherein:

[0014] The mass ratio of the Dracocephalum integrifolium extract to the Callicarpa nudiflora extract is 1:(0.5~2);

[0015] The preparation method of the whole-leaf blue orchid extract comprises the following steps:

[0016] S11: crushing the dried whole-leaf blue orchid plant into 50-70 meshes to obtain a whole-leaf blue orchid pre-treated matrix;

[0017] S12: mixing the pretreated matrix obtained in step S11 with a solvent to obtain a whole-leaf blue indigo mixed system, wherein: the solvent is a mixture of a choline chloride-lactic acid low eutectic solvent and ethanol, the concentration of choline chloride in the solvent is 0.1-1.0 mol / L, the molar ratio of choline chloride to lactic acid is 1:2, and the mass volume ratio of the pretreated matrix to the solvent is 1 g: (10-50) ml;

[0018] S13: subjecting the whole-leaf cyanidin mixed system obtained in step S12 to ultrasonic wall-breaking treatment at a temperature of 45-55° C., an ultrasonic power of 250-350 W, a frequency of 35-45 kHz, and a treatment time of 10-20 min to obtain a whole-leaf cyanidin ultrasonic-treated liquid;

[0019] S14: subjecting the whole-leaf cyanus ultrasonic treatment liquid obtained in step S13 to microwave enhanced mass transfer treatment, and performing microwave-assisted extraction at 65-75° C., a microwave power of 550-650 W, and an extraction time of 8-12 min to obtain a whole-leaf cyanus microwave extract;

[0020] S15: centrifuging the whole-leaf cyanidin microwave extract obtained in step S14 at a speed of 7500-8500 rpm for 10-20 min to obtain a whole-leaf cyanidin supernatant;

[0021] S16: Concentrating the whole-leaf cyanidin supernatant obtained in step S15 under reduced pressure to obtain an extract, and freeze-drying to obtain a whole-leaf cyanidin active crude extract;

[0022] The preparation method of the Callicarpa nudiflora extract comprises the following steps:

[0023] S21: Quick-freeze the leaves of Callicarpa nudiflora with liquid nitrogen and then ball-mill them to 60-100 meshes to obtain Callicarpa nudiflora powder;

[0024] S22: adding a citric acid buffer having a pH value of 4.0 to 5.0 to the Callicarpa nudiflora powder obtained in step S21, wherein the citric acid buffer contains 1.0 to 2.0 wt % of cellulase, and performing enzymolysis at 40 to 50° C. for 60 to 120 min to obtain an enzymatic hydrolyzate of Callicarpa nudiflora;

[0025] S23: After the enzyme is inactivated, the enzymatic hydrolyzate of Callicarpa nudiflora obtained in step S22 is introduced into a supercritical CO2 system, and dynamic countercurrent extraction is performed at 20-30 MPa and 50-60° C. for 1.5-2.5 h to obtain a Callicarpa nudiflora separation extract;

[0026] S24: The extract of Callicarpa nudiflora obtained in step S23 is included with hydroxypropyl-β-cyclodextrin at a mass ratio of 1:(3-5), and then spray-dried to obtain the microencapsulated extract of Callicarpa nudiflora.

[0027] The pH value of the solvent described above is generally 3.5-5.5.

[0028] Preferably, the mass ratio of the extract of Dracocephalum integrifolium to the extract of Callicarpa nudiflora is 1:1.

[0029] Furthermore, the total concentration of the crude drug of the extract of Dracocephalum integrifolium and the crude drug of the extract of Callicarpa nudiflora in the oil-control composition is 0.01-10 wt%. Most preferably 0.1-1 wt%, so as to ensure both biological safety and effectively improve the oil-control effect.

[0030] Preferably, the ultrasonic power / microwave power is 1:2.

[0031] Preferably, in step S12, the concentration of choline chloride is 0.5 mol / L, the concentration of lactic acid is 1.0 mol / L, and the mass-volume ratio of the pretreatment matrix to the solvent is 1 g:30 ml.

[0032] According to the specific embodiments of the present invention, in step S16, the parameters of the vacuum concentration are a vacuum degree of -0.08 to -0.1 MPa and a concentration temperature of 40-60 °C.

[0033] According to the specific embodiments of the present invention, in step S16, the freeze-drying includes pre-freezing at -40 to -50 °C for 2-4 h, then sublimation drying under the conditions of 10-30 Pa and -20 to 0 °C, and finally analytical drying at 25-35 °C until the water content ≤ 3%.

[0034] Preferably, in step S22, the addition amount of the cellulase is 1.5 wt%, the pH of the citrate buffer solution is 4.5, and the enzymolysis time is 90 min; in step S23, the enzymolyzed solution of Callicarpa nudiflora is introduced into the CO2 supercritical system within 30 min after inactivating the enzyme, and the supercritical parameters are 25 MPa and 55 °C, and the extraction time is 2 h.

[0035] According to the specific embodiments of the present invention, in step S24, the parameters of the spray drying are an inlet air temperature of 160-180 °C and an outlet air temperature of 80-90 °C.

[0036] The present invention also provides the application of the oil-control composition described above in the preparation of oil-control cosmetics. By way of example, the oil-control cosmetics are selected from essence, emulsion, facial mask or facial cleansing foam.

[0037] The total crude drug concentration of Dracocephalum integrifolium extract and Callicarpa nudiflora extract in the oil-control cosmetic is 0.01% - 10 wt%, preferably 0.1 - 5.0 wt%, and most preferably 0.1 - 1 wt%.

[0038] Compared with the prior art, the present invention has the following advantages:

[0039] The composition of the present invention is composed of Dracocephalum integrifolium extract and Callicarpa nudiflora extract in combination. The two work together through a dual-target mechanism (blocking the nuclear translocation of SREBP-1 and down-regulating PPARγ) to achieve oil control. This composition can be applied to oil-control cosmetic and pharmaceutical products such as essence and lotion. The process is efficient and stable, providing a safe and multi-effective solution for oily skin care. The present invention breaks through the technical limitations of traditional mixed extraction by establishing a precise matching mechanism between plant characteristics and process characteristics, and provides an innovative solution for the collaborative development of multi-component natural products while maintaining the safety of the extract. Detailed implementation manners

[0040] Combined with the following specific embodiments, the present invention is further described in detail. The protection scope of the present invention is not limited to the following embodiments. Without departing from the spirit and scope of the inventive concept, changes and advantages that can be conceived by those skilled in the art are included in the present invention, and the appended claims are used as the protection scope. The processes, conditions, reagents, experimental methods, etc. for implementing the present invention, except for the specifically mentioned content below, are all common knowledge and well-known common sense in the art, and the present invention has no special limitations. Unless otherwise specified, % in the following embodiments refers to wt%; the ratio of material to liquid refers to the mass-volume ratio of the pretreated matrix to the solvent (g:ml); freeze-drying includes pre-freezing at -40 to -50 °C for 2 to 4 h, then sublimation drying under the conditions of 10 to 30 Pa and -20 to 0 °C, and finally analytical drying at 25 to 35 °C until the water content ≤ 3%.

[0041] Example 1

[0042] Preparation of Dracocephalum integrifolium extract:

[0043] S1: Crush the dried Dracocephalum integrifolium plants to 60 mesh, weigh 10 g of the powder to obtain the pretreated matrix of Dracocephalum integrifolium;

[0044] S2: Mix the pretreated matrix obtained in step S1 with the solvent to obtain a mixed system of Dracocephalum integrifolium, wherein: the solvent is a mixture of choline chloride-lactic acid deep eutectic solvent and ethanol, and the molar concentrations of choline chloride and lactic acid in the solvent are 0.5 mol / L and 1.0 mol / L respectively; the ratio of material to liquid is 1:30;

[0045] S3: Subject the Dracocephalum integrifolium mixture system obtained in step S2 to ultrasonic cell wall breaking treatment at a temperature of 50°C, an ultrasonic power of 300 W, a frequency of 40 kHz, and a treatment time of 15 min to obtain a Dracocephalum integrifolium ultrasonic treatment solution;

[0046] S4: Subject the Dracocephalum integrifolium ultrasonic treatment solution obtained in step S3 to microwave-assisted mass transfer treatment, perform microwave-assisted extraction at 70°C, with a microwave power of 600 W and an extraction time of 10 min to obtain a Dracocephalum integrifolium microwave extraction solution;

[0047] S5: Centrifuge the Dracocephalum integrifolium microwave extraction solution obtained in step S4 at a rotational speed of 8000 rpm for 15 min to obtain a Dracocephalum integrifolium supernatant;

[0048] S6: Concentrate the Dracocephalum integrifolium supernatant obtained in step S5 under reduced pressure to an extract, with a vacuum degree of -0.09 MPa and a concentration temperature of 50°C, and obtain a crude extract of Dracocephalum integrifolium active ingredients after freeze-drying.

[0049] Example 2

[0050] Preparation of Dracocephalum integrifolium extract:

[0051] S1: Grind the dried Dracocephalum integrifolium plants to 50 mesh, weigh 15.0 g of the powder to obtain a pretreated matrix of Dracocephalum integrifolium;

[0052] S2: Mix the pretreated matrix obtained in step S1 with a solvent to obtain a Dracocephalum integrifolium mixture system, wherein: the solvent is a mixture of choline chloride-lactic acid deep eutectic solvent and ethanol, and the molar concentrations of choline chloride and lactic acid in the solvent are 0.1 mol / L and 0.2 mol / L respectively; the solid-liquid ratio is 1:10;

[0053] S3: Subject the Dracocephalum integrifolium mixture system obtained in step S2 to ultrasonic cell wall breaking treatment at a temperature of 45°C, an ultrasonic power of 350 W, a frequency of 35 kHz, and a treatment time of 20 min to obtain a Dracocephalum integrifolium ultrasonic treatment solution;

[0054] S4: Subject the Dracocephalum integrifolium ultrasonic treatment solution obtained in step S3 to microwave-assisted mass transfer treatment, perform microwave-assisted extraction at 65°C, with a microwave power of 650 W and an extraction time of 12 min to obtain a Dracocephalum integrifolium microwave extraction solution;

[0055] S5: Centrifuge the Dracocephalum integrifolium microwave extraction solution obtained in step S4 at a rotational speed of 7500 rpm for 20 min to obtain a Dracocephalum integrifolium supernatant;

[0056] S6: Concentrate the Dracocephalum integrifolium supernatant obtained in step S5 under reduced pressure to an extract, with a vacuum degree of -0.08 MPa and a concentration temperature of 60°C, and obtain a crude extract of Dracocephalum integrifolium active ingredients after freeze-drying.

[0057] Example 3

[0058] Preparation of Dracocephalum integrifolium Bunge extract:

[0059] S1: Grind the dried plants of Dracocephalum integrifolium Bunge to 70 mesh, weigh 5.0 g of the powder to obtain the pretreated matrix of Dracocephalum integrifolium Bunge;

[0060] S2: Mix the pretreated matrix obtained in step S1 with a solvent to obtain a mixed system of Dracocephalum integrifolium Bunge, wherein: the solvent is a mixture of choline chloride-lactic acid deep eutectic solvent and ethanol, and the molar concentrations of choline chloride and lactic acid in the solvent are 1.0 mol / L and 2.0 mol / L respectively; the solid-liquid ratio is 1:50;

[0061] S3: Perform ultrasonic cell wall breaking treatment on the mixed system of Dracocephalum integrifolium Bunge obtained in step S2 at a temperature of 55 °C, an ultrasonic power of 250 W, a frequency of 45 kHz, and a treatment time of 10 min to obtain the ultrasonic-treated solution of Dracocephalum integrifolium Bunge;

[0062] S4: Perform microwave-assisted mass transfer treatment on the ultrasonic-treated solution of Dracocephalum integrifolium Bunge obtained in step S3, extract with microwave assistance at 75 °C, a microwave power of 550 W, and an extraction time of 8 min to obtain the microwave extract of Dracocephalum integrifolium Bunge;

[0063] S5: Centrifuge the microwave extract of Dracocephalum integrifolium Bunge obtained in step S4 at a rotation speed of 8500 rpm for 10 min to obtain the supernatant of Dracocephalum integrifolium Bunge;

[0064] S6: Concentrate the supernatant of Dracocephalum integrifolium Bunge obtained in step S5 under reduced pressure to an extract, with a vacuum degree of -0.1 MPa and a concentration temperature of 40 °C, and obtain the crude active extract of Dracocephalum integrifolium Bunge after freeze-drying.

[0065] Example 4

[0066] Preparation of Callicarpa nudiflora Hook. et Arn. extract

[0067] S1: Quick-freeze the leaves of Callicarpa nudiflora Hook. et Arn. in liquid nitrogen and then grind them to 80 mesh, weigh 5.0 g of the Callicarpa nudiflora Hook. et Arn. powder;

[0068] S2: Add a citric acid buffer solution with a pH of 4.5 (containing 1.5% cellulase) to the Callicarpa nudiflora Hook. et Arn. powder obtained in step S1 and enzymatically hydrolyze it at 45 °C for 90 min to obtain the enzymatically hydrolyzed solution of Callicarpa nudiflora Hook. et Arn.;

[0069] S3: Introduce the enzymatically hydrolyzed solution of Callicarpa nudiflora Hook. et Arn. obtained in step S2 into a supercritical CO2 system (25 MPa, 55 °C) for dynamic countercurrent extraction for 2.0 h within 30 min after inactivating the enzyme to obtain the separated and extracted product of Callicarpa nudiflora Hook. et Arn.;

[0070] S4: Incorporate the extract of Callicarpa nudiflora obtained in step S3 with hydroxypropyl-β-cyclodextrin (mass ratio of 1:4), and perform spray drying at 170 °C with an air outlet temperature of 85 °C to obtain the microencapsulated extract of Callicarpa nudiflora.

[0071] Example 5

[0072] Preparation of the extract of Callicarpa nudiflora

[0073] S1: Quick-freeze the leaves of Callicarpa nudiflora in liquid nitrogen and then grind them to 60 mesh, and weigh 7.0 g of the Callicarpa nudiflora powder;

[0074] S2: Add a citric acid buffer solution with pH 4.0 (containing 1.0% cellulase) to the Callicarpa nudiflora powder obtained in step S1, and perform enzymatic hydrolysis at 40 °C for 60 min to obtain the enzymolysis solution of Callicarpa nudiflora;

[0075] S3: Inactivate the enzyme in the enzymolysis solution of Callicarpa nudiflora obtained in step S2 and introduce it into a supercritical CO2 system (20 MPa, 60 °C) for dynamic countercurrent extraction for 2.5 h to obtain the separated and extracted extract of Callicarpa nudiflora;

[0076] S4: Incorporate the separated and extracted extract of Callicarpa nudiflora obtained in step S3 with hydroxypropyl-β-cyclodextrin (mass ratio of 1:3), and perform spray drying at 160 °C with an air outlet temperature of 80 °C to obtain the microencapsulated extract of Callicarpa nudiflora.

[0077] Example 6

[0078] Preparation of the extract of Callicarpa nudiflora

[0079] S1: Quick-freeze the leaves of Callicarpa nudiflora in liquid nitrogen and then grind them to 100 mesh, and weigh 3.0 g of the Callicarpa nudiflora powder;

[0080] S2: Add a citric acid buffer solution with pH 5.0 (containing 2.0% cellulase) to the Callicarpa nudiflora powder obtained in step S1, and perform enzymatic hydrolysis at 50 °C for 120 min to obtain the enzymolysis solution of Callicarpa nudiflora;

[0081] S3: Inactivate the enzyme in the enzymolysis solution of Callicarpa nudiflora obtained in step S2 and introduce it into a supercritical CO2 system (30 MPa, 50 °C) for dynamic countercurrent extraction for 1.5 h to obtain the separated and extracted extract of Callicarpa nudiflora;

[0082] S4: Incorporate the separated and extracted extract of Callicarpa nudiflora obtained in step S3 with hydroxypropyl-β-cyclodextrin (mass ratio of 1:5), and perform spray drying at 180 °C with an air outlet temperature of 90 °C to obtain the microencapsulated extract of Callicarpa nudiflora.

[0083] Comparative Example 1

[0084] Preparation of the extract of Dracocephalum integrifolium

[0085] S1: Quick-freeze Dracocephalum integrifolium leaves in liquid nitrogen and then mill them to 100 mesh, and weigh 3.0 g of Dracocephalum integrifolium powder;

[0086] S2: Add citric acid buffer solution (containing 2.0% cellulase) with pH 5.0 to the Dracocephalum integrifolium powder obtained in step S1, and enzymatically hydrolyze at 50 °C for 120 min to obtain Dracocephalum integrifolium enzymatic hydrolysate;

[0087] S3: Inactivate the enzyme of the Dracocephalum integrifolium enzymatic hydrolysate obtained in step S2, and introduce it into a supercritical CO2 system (30 MPa, 50 °C) for dynamic countercurrent extraction for 1.5 h within 30 min to obtain Dracocephalum integrifolium separated extract;

[0088] S4: Inclusion complex the Dracocephalum integrifolium separated extract obtained in step S3 with hydroxypropyl-β-cyclodextrin (mass ratio of 1:5), and spray-dry at 180 °C with an air outlet temperature of 90 °C to obtain microencapsulated extract of Dracocephalum integrifolium.

[0089] Comparative Example 2

[0090] Preparation of Callicarpa nudiflora extract:

[0091] S1: Grind the dried Callicarpa nudiflora plants to 60 mesh, and weigh 10 g of powder to obtain the Callicarpa nudiflora pretreatment matrix;

[0092] S2: Mix the pretreatment matrix obtained in step S1 with a solvent to obtain a Callicarpa nudiflora mixed system, where: the solvent is a mixture of choline chloride-lactic acid deep eutectic solvent and ethanol, and the molar concentrations of choline chloride and lactic acid in the solvent are 0.5 mol / L and 1.0 mol / L respectively; the solid-liquid ratio is 1:30;

[0093] S3: Perform ultrasonic cell wall breaking treatment on the Callicarpa nudiflora mixed system obtained in step S2 at a temperature of 50 °C, an ultrasonic power of 300 W, a frequency of 40 kHz, and a treatment time of 15 min to obtain Callicarpa nudiflora ultrasonic treatment solution;

[0094] S4: Perform microwave enhanced mass transfer treatment on the Callicarpa nudiflora ultrasonic treatment solution obtained in step S3, and extract with microwave assistance at 70 °C, a microwave power of 600 W, and an extraction time of 10 min to obtain Callicarpa nudiflora microwave extraction solution;

[0095] S5: Centrifuge the Callicarpa nudiflora microwave extraction solution obtained in step S4 at a rotation speed of 8000 rpm and a centrifugation time of 15 min to obtain Callicarpa nudiflora supernatant;

[0096] S6: Concentrate the Callicarpa nudiflora supernatant obtained in step S5 under reduced pressure to an extract, with a vacuum degree of -0.09 MPa and a concentration temperature of 50 °C, and obtain the crude Callicarpa nudiflora active extract after freeze-drying.

[0097] Comparative Example 3

[0098] Preparation of the mixture of Dracocephalum integrifolium and Callicarpa nudiflora extracts:

[0099] S1: Grind the dry plants of Dracocephalum integrifolium and Callicarpa nudiflora to 60 mesh, weigh 5 g of each powder and mix them to obtain the pretreatment matrix of Dracocephalum integrifolium and Callicarpa nudiflora;

[0100] S2: Mix the pretreatment matrix obtained in step S1 with a solvent to obtain a mixed system of Dracocephalum integrifolium and Callicarpa nudiflora, where: the solvent is a mixture of choline chloride-lactic acid deep eutectic solvent and ethanol, and the molar concentrations of choline chloride and lactic acid in the solvent are 0.5 mol / L and 1.0 mol / L respectively; the solid-liquid ratio is 1:30;

[0101] S3: Perform ultrasonic cell wall breaking treatment on the mixed system of Dracocephalum integrifolium and Callicarpa nudiflora obtained in step S2 at a temperature of 50 °C, an ultrasonic power of 300 W, a frequency of 40 kHz, and a treatment time of 15 min to obtain an ultrasonic treatment solution of Dracocephalum integrifolium and Callicarpa nudiflora;

[0102] S4: Perform microwave-assisted mass transfer treatment on the ultrasonic treatment solution of Dracocephalum integrifolium and Callicarpa nudiflora obtained in step S3, carry out microwave-assisted extraction at 70 °C, with a microwave power of 600 W and an extraction time of 10 min to obtain a microwave extraction solution of Dracocephalum integrifolium and Callicarpa nudiflora;

[0103] S5: Centrifuge the microwave extraction solution of Dracocephalum integrifolium and Callicarpa nudiflora obtained in step S4 at a rotation speed of 8000 rpm and a centrifugation time of 15 min to obtain a supernatant of Dracocephalum integrifolium and Callicarpa nudiflora;

[0104] S6: Concentrate the supernatant of Dracocephalum integrifolium and Callicarpa nudiflora obtained in step S5 under reduced pressure to an extract, with a vacuum degree of -0.09 MPa and a concentration temperature of 50 °C, and obtain the crude active extract of Dracocephalum integrifolium and Callicarpa nudiflora after freeze-drying.

[0105] Comparative Example 4

[0106] Preparation of the mixture of Dracocephalum integrifolium and Callicarpa nudiflora extracts:

[0107] S1: Quick-freeze the leaves of Dracocephalum integrifolium and Callicarpa nudiflora in liquid nitrogen and then ball-mill them to 100 mesh, weigh 1.5 g of each and mix them to obtain the powder of Dracocephalum integrifolium and Callicarpa nudiflora;

[0108] S2: Add a citric acid buffer solution with a pH of 5.0 (containing 2.0% cellulase) to the powder of Dracocephalum integrifolium and Callicarpa nudiflora obtained in step S1, and carry out enzymatic hydrolysis at 50 °C for 120 min to obtain an enzymatic hydrolysis solution of Dracocephalum integrifolium and Callicarpa nudiflora;

[0109] S3: Introduce the enzymatic hydrolysis solution of Dracocephalum integrifolium and Callicarpa nudiflora obtained in step S2 into a supercritical CO2 system (30 MPa, 50 °C) for dynamic countercurrent extraction for 1.5 h after inactivating the enzyme to obtain a separated extract of Dracocephalum integrifolium and Callicarpa nudiflora;

[0110] S4: Incorporate the Dracocephalum integrifolium and Callicarpa nudiflora extract obtained in step S3 with hydroxypropyl-β-cyclodextrin (mass ratio 1:5), and perform spray drying at 180 °C with an air outlet temperature of 90 °C to obtain the microencapsulated extract of Dracocephalum integrifolium and Callicarpa nudiflora.

[0111] Effect evaluation 1: Effects of Dracocephalum integrifolium extract, Callicarpa nudiflora extract, and the mixed extract of Dracocephalum integrifolium and Callicarpa nudiflora on the viability of human sebaceous gland cells.

[0112] Dissolve the Dracocephalum integrifolium extract, Callicarpa nudiflora extract, and the mixed extract of Dracocephalum integrifolium and Callicarpa nudiflora obtained in Examples 1 - 6 and Comparative Examples 1 - 4 in deionized water, filter, and prepare samples. The amount of each sample and its corresponding extract amount and crude drug amount are shown in Table 1.

[0113] Table 1

[0114]

[0115] Dilute the test samples with culture medium under sterile conditions. Uniformly dilute each test sample to the crude drug concentration of the extract (i.e., the mass content of the extract calculated based on the crude drug amount): 0.01%, 0.1%, 1%, 10%.

[0116] Seed SZ95 human sebaceous gland cells into 96-well plates at a density of 7×10 3 cells / well, and culture them in a 37 °C, 5% CO2 cell culture incubator for 24 h. When the cell density is about 50%, treat the cells in the treatment group with 100 μl of the test sample (the crude drug concentrations of the extract are 0.01%, 0.1%, 1%, 10% respectively), and add 100 μl of culture medium to the control group (this culture medium is the same as the above test sample culture medium without adding any test sample). Each group has 3 replicate wells. 48 h after adding the samples, detect the cell viability by the CCK-8 method, and calculate the cell viability according to the following formula.

[0117] Cell viability = OD value of the treatment group / OD value of the control group × 100%.

[0118] The experimental results are shown in Table 2.

[0119] Table 2

[0120]

[0121]

[0122] According to the experimental data and process comparative analysis, the Dracocephalum integrifolium Bunge / Callicarpa nudiflora Hook. et Arn. extract in the embodiments of the present invention can maintain or improve the viability of SZ95 human sebaceous gland cells at the crude drug concentrations of 0.01%, 0.1%, 1%, and 10%: the cell survival rate of Example 1 (microwave-DES extraction) reached 107.9% at a concentration of 0.1% (7.9% higher than that of the control group), and the activity remained 94.6% at a concentration of 10%. The cell survival rate of Example 4 (enzymolysis-supercritical coupling) was as high as 109.3% at a concentration of 0.1%, and 92.7% was maintained at a concentration of 10%, indicating that the composite process can selectively enrich active ingredients. The survival rates of Comparative Examples 1-2 dropped sharply to 70.6% and 68.1% (inhibition rate ≥ 31.9%) at a concentration of 10%, the survival rate of Comparative Example 3 was 77.0% (inhibition rate 23.0%) at a concentration of 10%, and the survival rate of Comparative Example 4 was 89.6% (inhibition rate 10.4%) at a concentration of 0.01%. The survival rates of Comparative Examples 1-4 were all inferior to those of Examples 1-6. The experimental data show that the exclusive extraction process of Dracocephalum integrifolium Bunge and Callicarpa nudiflora Hook. et Arn. proposed by the present technology has significant matrix adaptability and technical synergy. When the cross-extraction method is adopted, the cytotoxicity of the two plants is significantly enhanced due to process mismatch. It is speculated that the reason is that Dracocephalum integrifolium Bunge cannot tolerate the biological enzymolysis step in the heterologous process, and harmful components in Callicarpa nudiflora Hook. et Arn. are abnormally transformed in a non-exclusive solvent system. The exclusive process realizes the safe release of active ingredients through the adaptation of physical cell wall breaking and low-polarity solvents, and the coupling of biological enzymolysis and supercritical targeted extraction. In addition, when mixed extraction is carried out, negative interactions will occur in the key steps of the two processes. For example, the acidic environment activates the residual enzyme activity or causes component oxidation, further verifying the technical necessity of step-by-step independent extraction in reverse. In summary, the present invention breaks through the technical limitations of traditional mixed extraction by establishing a precise matching mechanism between plant characteristics and process characteristics, and provides an innovative solution for the collaborative development of multi-component natural products while maintaining the safety of the extract.

[0123] Effect evaluation 2: Inhibition of lipid production in SZ95 human sebaceous gland cells by Dracocephalum integrifolium Bunge extract, Callicarpa nudiflora Hook. et Arn. extract, and the mixture of Dracocephalum integrifolium Bunge and Callicarpa nudiflora Hook. et Arn. extract

[0124] Under aseptic conditions, dilute the test samples with the culture medium, and uniformly dilute each test sample to the crude drug concentration of the extract: 0.01%, 0.025%, 0.05%, 0.1%.

[0125] Seed SZ95 human sebaceous gland cells at 7×10 3Inoculate at a density of cells per well in a 96-well plate and culture in a 37°C, 5% CO₂ cell incubator for 24 h. When the cell density reaches about 50%, add 100 μl of medium to the control group, add 100 μl of medium containing 10 μg / ml insulin (Ins) and 1 μM T0901317 to the model group to stimulate cell lipid production, and add 100 μl of medium containing the sample to be tested (crude drug concentration of the extract: 0.01%, 0.025%, 0.05%, 0.1%), 10 μg / ml Ins and 1 μM T0901317 to the dosing group to treat the cells. Each group is set up with 3 replicate wells. After adding the samples for 48 h, detect the inhibition rate of cell sebum production by the Nile red method and calculate the inhibition rate of cell sebum production according to the following formula.

[0126]

[0127] Among them, the sample to be tested in dosing group A is the Dracocephalum integrifolium Bunge extract diluted in Example 1, and the crude drug concentrations of the extract are: 0.01%, 0.025%, 0.05%, 0.1%; the sample to be tested in dosing group B is the Callicarpa nudiflora Hook. extract diluted in Example 4, and the crude drug concentrations of the extract are: 0.01%, 0.025%, 0.05%, 0.1%; dosing group C: includes 4 groups, and the sample to be tested is a mixture of Dracocephalum integrifolium Bunge extract (diluted in Example 1, crude drug concentrations of the extract are 0.01%, 0.025%, 0.05%, 0.1%) and Callicarpa nudiflora Hook. extract (diluted in Example 4, corresponding concentrations) mixed at a mass ratio of 1:0.5; dosing group D: includes 4 groups, and the sample to be tested is a mixture of Dracocephalum integrifolium Bunge extract (diluted in Example 1, crude drug concentrations of the extract are 0.01%, 0.025%, 0.05%, 0.1%) and Callicarpa nudiflora Hook. extract (diluted in Example 4, corresponding concentrations) mixed at a mass ratio of 1:1; dosing group E: includes 4 groups, and the sample to be tested is a mixture of Dracocephalum integrifolium Bunge extract (diluted in Example 1, crude drug concentrations of the extract are 0.01%, 0.025%, 0.05%, 0.1%) and Callicarpa nudiflora Hook. extract (diluted in Example 4, corresponding concentrations) mixed at a mass ratio of 1:2.

[0128] The experimental results are shown in Table 3.

[0129] Table 3

[0130]

[0131] As can be seen from Table 3, according to the experimental data, the compound combinations of Dracocephalum integrifolium Bunge extract (Example 1) and Callicarpa nudiflora Hook. extract (Example 4) show synergistic effects at different mass ratios, and the synergistic effect is the strongest in the 1:1 mass ratio group.

[0132]

[0133] Among them, the highest monomer inhibition rate refers to the highest value of the cell sebum production inhibition rate in dosing group A and dosing group B; the combined inhibition rate is the cell sebum production inhibition rate in dosing group C, dosing group D or dosing group E.

[0134] At a concentration of 0.1%, the synergistic ratio of the 1:1 mass ratio is significantly higher than other ratios (36.2% vs 20.3%, 9.0%), confirming its optimal synergistic ratio; while the 1:2 combination shows weakened synergism due to the ratio imbalance, further verifying that the mass ratio of 1:1 is the best synergistic ratio of the active ingredients.

[0135] Effect evaluation 3: Effects of Dracocephalum integrifolium Bunge extract, Callicarpa nudiflora Hook. et Arn. extract and the mixture of Dracocephalum integrifolium Bunge and Callicarpa nudiflora Hook. et Arn. extract on lipid metabolism-related genes in SZ95 human sebaceous gland cells

[0136] Under sterile conditions, the test samples were diluted with the culture medium, and each test sample was uniformly diluted to the crude drug concentration of the extract: 0.025%, 0.05%, 0.1%.

[0137] SZ95 cells were cultured in Sebomed basal medium, placed in an incubator at 37°C and 5% CO2. When the cells were fused to 80%, cell passage inoculation was carried out. The cells were digested with trypsin to detach them from the culture flask, and the cells were inoculated in 6-well plates, with about 2×10 5 cells in each well. After culturing for 24 h, the control group was added with 100 μl of culture medium and cultured for 24 h, the model group was added with 100 μl of culture medium containing 10 μg / ml Ins and 1 μM T0901317 and cultured for 24 h, and the dosing groups were added with 100 μl of culture medium containing 10 μg / ml Ins, 1 μM T0901317 and several samples at different concentrations and cultured for 24 h. Total RNA in the cells was extracted, reverse transcribed into cDNA after removing gDNA, and qPCR experiments were carried out. A 20 μL system was used with a 96-well plate, and the internal reference gene and target gene of each component were each made in 3 replicates. The grouping of dosing groups A-D refers to effect evaluation 2. The relative gene expression level was calculated according to the following formula:

[0138] Relative gene expression level = 2 -ΔΔCt .

[0139] ΔCt = Ct value of the target gene - Ct value of the internal reference gene of the corresponding sample

[0140] ΔΔCt = ΔCt value of the model group or dosing group - average value of ΔCt of the control group

[0141] The results of the effects of the samples on the expression of FAS in Ins / T0901317-induced SZ95 cells at the mRNA level are shown in Table 4.

[0142] Table 4

[0143]

[0144]

[0145] The results of the effect of the sample on the expression of ACC in SZ95 cells induced by Ins / T0901317 at the mRNA level are shown in Table 5 as follows.

[0146] Table 5

[0147]

[0148]

[0149] The results of the effect of the sample on the expression of SREBP-1 in SZ95 cells at the mRNA level are shown in Table 6 as follows.

[0150] Table 6

[0151]

[0152]

[0153] The results of the effect of the sample on the expression of PPARγ in SZ95 cells induced by Ins / T0901317 at the mRNA level are shown in Table 7 as follows.

[0154] Table 7

[0155]

[0156] As can be seen from Tables 4, 5, 6, and 7, both Dracocephalum integrifolium Bunge extract (Example 1) and Callicarpa nudiflora Hook. extract (Example 2) can significantly inhibit the expression of lipid synthesis-related genes SREBP-1, PPARγ, FAS, and ACC, and both have a strict dose-effect relationship (0.1% > 0.05% > 0.025%). When the Dracocephalum integrifolium Bunge extract and Callicarpa nudiflora Hook. extract are compounded in different ratios, Example 1:Example 2 = 1:(0.5 - 2), the inhibitory effect on lipid synthesis-related genes SREBP-1, PPARγ, FAS, and ACC is significantly improved, and the inhibitory effect on lipid synthesis genes is most significantly improved after compounding at 1:1.

[0157] FAS gene: Synergy rate +27.41% (inhibitory rate of the compound group 40.92% vs. the best monomer inhibitory rate 32.12%);

[0158] ACC gene: Synergy rate +31.44% (inhibitory rate of the compound group 68.26% vs. the best monomer inhibitory rate 51.93%);

[0159] SREBP-1 gene: Synergy rate +19.84% (inhibition rate of the compound group 92.46% vs. the best monomer inhibition rate 77.15%);

[0160] PPARγ gene: Synergy rate +38.17% (inhibition rate of the compound group 58.87% vs. the best monomer inhibition rate 42.61%);

[0161] Comprehensive synergy effect index (CSI) = (27.41% + 31.44% + 19.84% + 38.17%) / 4 ≈ 29.31%.

[0162] Among them, the inhibition rate = (relative gene expression level in the model group - relative gene expression level in the drug administration group) / relative gene expression level in the model group × 100%;

[0163] Best monomer inhibition rate: The highest inhibition rate value in Examples 1 and 4 at the same concentration;

[0164] Inhibition rate of the compound group: The highest inhibition rate value in the drug administration group C-E at the same concentration.

[0165] Synergy rate = (inhibition rate of the compound group - inhibition rate of the best monomer group) / inhibition rate of the best monomer group × 100%

[0166] The comprehensive synergy effect index (CSI = 29.21%) indicates that the inhibition of sebum gland cell lipid synthesis has a better effect after the extract of Dracocephalum integrifolium and the extract of Callicarpa nudiflora are compounded in different ratios. This compound system shows significant technical advantages in lipid metabolism regulation through target complementary synergy and dual enhancement of process-component. Its optimization of the "breaking-wall-extraction" process and design of "multi-target-low-toxicity" components break through the efficiency bottleneck of traditional single-agent preparations, providing theoretical and practical basis for the development of oil-control active ingredients with both high efficiency and safety.

Claims

1. An oil control composition, characterized in that: It is composed of whole leaf cymbidium extract and callicarpa nudiflora extract, including: The mass ratio of the whole leaf blue cymbidium extract to the callicarpa nudiflora extract is 1:(0.5-:2); The preparation method of the whole-leaf blue orchid extract comprises the following steps: S11: crushing the dried whole-leaf blue orchid plant into 50-70 meshes to obtain a whole-leaf blue orchid pre-treated matrix; S12: mixing the pretreated matrix obtained in step S11 with a solvent to obtain a whole-leaf blue orchid mixed system, wherein: the solvent is a mixture of a choline chloride-lactic acid low eutectic solvent and ethanol, the concentration of choline chloride in the solvent is 0.1-1.0 mol / L, the molar ratio of choline chloride to lactic acid is 1:2, and the pH value of the solvent is 3.5-5.5; the mass volume ratio of the pretreated matrix to the solvent is 1 g: (10-50) ml; S13: subjecting the whole-leaf cyanidin mixed system obtained in step S12 to ultrasonic wall-breaking treatment at a temperature of 45-55° C., an ultrasonic power of 250-350 W, a frequency of 35-45 kHz, and a treatment time of 10-20 min to obtain a whole-leaf cyanidin ultrasonic-treated liquid; S14: subjecting the whole-leaf cyanus ultrasonic treatment liquid obtained in step S13 to microwave enhanced mass transfer treatment, and performing microwave-assisted extraction at 65-75° C., a microwave power of 550-650 W, and an extraction time of 8-12 min to obtain a whole-leaf cyanus microwave extract; S15: centrifuging the whole-leaf cyanidin microwave extract obtained in step S14 at a speed of 7500-8500 rpm for 10-20 min to obtain a whole-leaf cyanidin supernatant; S16: Concentrating the whole-leaf cyanidin supernatant obtained in step S15 under reduced pressure to obtain an extract, and freeze-drying to obtain a whole-leaf cyanidin active crude extract; The preparation method of the Callicarpa nudiflora extract comprises the following steps: S21: Quick-freeze the leaves of Callicarpa nudiflora with liquid nitrogen and then ball-mill them to 60-100 meshes to obtain Callicarpa nudiflora powder; S22: adding a citric acid buffer having a pH value of 4.0 to 5.0 to the Callicarpa nudiflora powder obtained in step S21, wherein the citric acid buffer contains 1.0 to 2.0 wt % of cellulase, and performing enzymolysis at 40 to 50° C. for 60 to 120 min to obtain an enzymatic hydrolyzate of Callicarpa nudiflora; S23: After the enzyme is inactivated, the enzymatic hydrolyzate of Callicarpa nudiflora obtained in step S22 is introduced into a supercritical CO2 system, and dynamic countercurrent extraction is performed at 20-30 MPa and 50-60° C. for 1.5-2.5 h to obtain a Callicarpa nudiflora separation extract; S24: The Callicarpa nudiflora isolated extract obtained in step S23 is included with hydroxypropyl-β-cyclodextrin in a mass ratio of 1:(3-:5), and spray-dried to obtain a Callicarpa nudiflora microencapsulated extract.

2. The oil control composition according to claim 1, characterized in that The total concentration of the crude drug of the whole-leaf cycad extract and the crude drug of the callicarpa nudiflora extract in the oil-control composition is 0.01-10 wt %.

3. The oil control composition according to claim 1, characterized in that In step S12, the concentration of choline chloride is 0.5 mol / L, the concentration of lactic acid is 1.0 mol / L, and the mass volume ratio of the pretreated matrix to the solvent is 1 g:30 ml.

4. The oil control composition according to claim 1, characterized in that In step S16, the parameters of the reduced pressure concentration are a vacuum degree of -0.08 to -0.1 MPa and a concentration temperature of 40 to 60°C.

5. The oil control composition according to claim 1, characterized in that In step S16, the freeze drying includes pre-freezing at -40 to -50°C for 2 to 4 hours, sublimation drying at 10 to 30 Pa and -20 to 0°C, and finally desorption drying at 25 to 35°C to a water content of ≤3%.

6. The oil control composition according to claim 1, characterized in that: In step S22, the amount of cellulase added is 1.5wt%, the pH of the citric acid buffer is 4.5, and the enzymatic hydrolysis time is 90min; in step S23, the enzymatic hydrolyzate of Callicarpa nudiflora is introduced into a CO2 supercritical system within 30min after the enzyme is inactivated, the supercritical parameters are 25MPa, 55°C, and the extraction time is 2h.

7. The oil control composition according to claim 1, characterized in that The spray drying parameters in step S24 are 160-180°C for inlet air and 80-90°C for outlet air.

8. Use of the oil control composition according to any one of claims 1 to 7 in the preparation of oil control cosmetics.

9. The use according to claim 8, characterized in that: The oil-control cosmetic is selected from essence, lotion, facial mask or cleansing foam.

10. The use according to claim 8, characterized in that: The sum of the crude drug concentrations of the whole-leaf cymbidium extract and the callicarpa nudiflora extract in the oil-control cosmetic is 0.01% to 10wt%.

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