Rhizoma gastrodiae pedicel and inflorescence extract as well as preparation method and application thereof

Abstract

The invention provides a pedicel and inflorescence extract of Gastrodia elata Blume, a preparation method of the pedicel and inflorescence extract of Gastrodia elata Blume, and application of the pedicel and inflorescence extract of Gastrodia elata Blume in the fields of cosmetics and medicines. Belongs to the technical field of cosmetics and medicines. The gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7 and ZTM-8 have the effect of promoting collagen secretion, and the ZTM-5 and the ZTM-6 have the activity of promoting collagen secretion in human dermis fibroblasts (human dermis fibroblasts, HDFa) at the extremely low concentration (1 mu g / mL), and have the activity of promoting collagen secretion in human dermis fibroblasts (human dermis fibroblasts, human dermis fibroblasts) at the extremely low concentration (1 mu g / mL) at the extremely low concentration (1 mu g / mL) at the extremely low concentration (1 mu g / mL) at the extremely low concentration (1 mu g / mL) at the extremely low concentration (1 mu g / mL). Under the concentration, the compound has no cytotoxicity and can be used for preparing anti-aging drugs or skin care products.

Description

Technical Field

[0001] The present invention belongs to the field of cosmetics and pharmaceutical technology, and specifically relates to the field of botanical medicine technology, and more specifically, to extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7 and ZTM-8 of Gastrodia elata pedicels and inflorescences, their pharmaceutical compositions, their preparation methods, and their applications in the preparation of drugs and cosmetics. Background Art

[0002] Collagen is a major structural protein in the extracellular matrix of mammals. It is widely present in skin, bones, muscles and other tissues. It is mainly involved in physiological and biochemical behaviors such as cell proliferation, differentiation, migration and signal transduction, and plays a supporting, repairing and protecting role for tissue cells [Ye Tao, Xiang Qi, Yang Yan, Huang Yadong. Research progress on the development and application of collagen. Journal of Biotechnology, 2023, 39(3): 942–960]. Among them, type I collagen is the most abundant protein in human skin, accounting for about 70% of the dry weight of skin. Type I collagen can increase skin elasticity and delay skin aging [Zhang Yuhan, Zhou Jingxuan, Zhang Nanhai, Zhao Liang, Zhang Liebing, Zhou Feng. Research progress on collagen peptides and elastin peptides to improve skin photoaging. Food Research and Development, 2023, 44(11): 208–215]. In addition, on the wound surface, type I collagen promotes wound healing by providing structural support for cell attachment and migration. However, animal-derived type I collagen may be immunogenic, and problems related to the processing technology, degradation rate and sterilization process of type I collagen limit its application [Cheng Y., Li Y., Huang S., Yu F., Bei Y., Zhang Y., Tang J., Huang Y., Xiang Q. Hybrid freeze-dried dressings composed of epidermal growth factor and recombinant human-like collagen enhance cutaneous wound healing in rats. Front. Bioeng. Biotechnol., 2020, 8: 742]. Therefore, screening plant extracts and natural products with the activity of promoting collagen secretion is a current research hotspot.

[0003] The Chinese herbal medicine Gastrodia elata is the dried tuber of the orchid plant Gastrodia elata Blume. It is a precious Chinese herbal medicine in my country and has a good therapeutic effect on nervous system diseases [Duan Xiaoyan, Yan Hui, Chen Yifang, Luo Jifeng, Ding Peichao, Tian Menghua, Wang Yuehu. Study on the chemical composition of the dichloromethane extract of Gastrodia elata Blume and its cholinesterase inhibitory activity. Chinese Herbal Medicine, 2022, 53(23): 7324–7330]. Studies on the pedicel and inflorescence of Gastrodia elata are relatively rare. There are no reports on the extracts of Gastrodia elata pedicel and inflorescence ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, and ZTM-8, nor on their activity, let alone their activity in promoting collagen secretion. Summary of the invention

[0004] In view of the above-mentioned deficiencies in the prior art, the present invention provides extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7 and ZTM-8 of Gastrodia elata pedicels and inflorescences, pharmaceutical compositions containing the extracts, cosmetics containing the extracts, preparation methods thereof, and uses thereof in the preparation of medicines and cosmetics.

[0005] In order to achieve the above-mentioned object of the present invention, the present invention provides the following technical solutions:

[0006] Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, ZTM-8 are prepared by the following method:

[0007] After the dried Gastrodia elata pedicels and inflorescences were crushed, the powder was taken, water was added, and the mixture was heated to 60°C, and ultrasonic-assisted extraction was performed for 1 hour. The extract was filtered, and the filtrate was concentrated to obtain ZTM-2;

[0008] After the dried pedicels and inflorescences of Gastrodia elata are crushed, the powder is taken, 95% ethanol is added, the mixture is heated to 60°C, ultrasonic-assisted extraction is performed for 30 minutes, the mixture is filtered to obtain a filtrate, and the extract is concentrated to obtain an extract, the extract is dissolved in water to form a suspension, and the mixture is extracted three times with an equal volume of petroleum ether, the organic phase is subjected to reduced pressure distillation to recover the solvent, and a petroleum ether extraction portion is obtained. The aqueous phase after the petroleum ether extraction is adjusted to a pH value of 2 with 1.5% HCl, extracted three times with an equal volume of ethyl acetate, and the organic phase is subjected to reduced pressure distillation to recover the solvent, and an ethyl acetate extraction portion is obtained; the aqueous phase after the ethyl acetate extraction is adjusted to a pH value of 9 with 5% NaOH, extracted three times with an equal volume of dichloromethane, and the organic phase is subjected to reduced pressure distillation to recover the solvent, and ZTM-4 is obtained;

[0009] The dried Gastrodia elata pedicel and inflorescence are crushed, and the powder is taken, 95% ethanol is added, and the mixture is heated to 60°C, and ultrasonic-assisted extraction is performed for 30 minutes. The mixture is filtered to obtain a filtrate, and the filtrate is concentrated under reduced pressure to obtain an extract, and water is added to the extract to prepare a suspension, and the suspension is subjected to column chromatography separation using a D101 macroporous resin, and eluted with 6 column volumes of distilled water at a flow rate of 0.5 mL / min, and the eluate is collected, and the mixture is concentrated under reduced pressure to obtain component ZTM-5; eluted with 6 column volumes of 10% ethanol at a flow rate of 0.5 mL / min, and the eluate is collected, and the mixture is concentrated under reduced pressure to obtain ZTM-6; eluted with 6 column volumes of 50% ethanol at a flow rate of 0.5 mL / min, and the eluate is collected, and the mixture is concentrated under reduced pressure to obtain ZTM-7; eluted with 6 column volumes of 90% ethanol at a flow rate of 0.5 mL / min, and the eluate is collected, and the mixture is concentrated under reduced pressure to obtain ZTM-8.

[0010] The present invention also provides a method for preparing Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7 and ZTM-8, which comprises the following steps:

[0011] After the dried Gastrodia elata pedicels and inflorescences were crushed, the powder was taken, water was added, and the mixture was heated to 60°C, and ultrasonic-assisted extraction was performed for 1 hour. The extract was filtered, and the filtrate was concentrated to obtain ZTM-2;

[0012] After the dried pedicels and inflorescences of Gastrodia elata are crushed, the powder is taken, 95% ethanol is added, the mixture is heated to 60°C, ultrasonic-assisted extraction is performed for 30 minutes, the mixture is filtered to obtain a filtrate, and the extract is concentrated to obtain an extract, the extract is dissolved in water to form a suspension, and the mixture is extracted three times with an equal volume of petroleum ether, the organic phase is subjected to reduced pressure distillation to recover the solvent, and a petroleum ether extraction portion is obtained. The aqueous phase after the petroleum ether extraction is adjusted to a pH value of 2 with 1.5% HCl, extracted three times with an equal volume of ethyl acetate, and the organic phase is subjected to reduced pressure distillation to recover the solvent, and an ethyl acetate extraction portion is obtained; the aqueous phase after the ethyl acetate extraction is adjusted to a pH value of 9 with 5% NaOH, extracted three times with an equal volume of dichloromethane, and the organic phase is subjected to reduced pressure distillation to recover the solvent, and ZTM-4 is obtained;

[0013] The dried Gastrodia elata pedicel and inflorescence are crushed, and the powder is taken, 95% ethanol is added, and the mixture is heated to 60°C, and ultrasonic-assisted extraction is performed for 30 minutes. The mixture is filtered to obtain a filtrate, and the filtrate is concentrated under reduced pressure to obtain an extract, and water is added to the extract to prepare a suspension, and the suspension is subjected to column chromatography separation using a D101 macroporous resin, and eluted with 6 column volumes of distilled water at a flow rate of 0.5 mL / min, and the eluate is collected, and the mixture is concentrated under reduced pressure to obtain component ZTM-5; eluted with 6 column volumes of 10% ethanol at a flow rate of 0.5 mL / min, and the eluate is collected, and the mixture is concentrated under reduced pressure to obtain ZTM-6; eluted with 6 column volumes of 50% ethanol at a flow rate of 0.5 mL / min, and the eluate is collected, and the mixture is concentrated under reduced pressure to obtain ZTM-7; eluted with 6 column volumes of 90% ethanol at a flow rate of 0.5 mL / min, and the eluate is collected, and the mixture is concentrated under reduced pressure to obtain ZTM-8.

[0014] The present invention provides a pharmaceutical composition, which is composed of any one of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, and ZTM-8 or any combination thereof and a pharmaceutically acceptable carrier.

[0015] The present invention also provides a method for preparing the pharmaceutical composition, firstly preparing Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, and ZTM-8, then taking one of them or any combination thereof and adding a pharmaceutically acceptable carrier.

[0016] At the same time, the invention provides the use of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7 and ZTM-8 in the preparation of anti-aging drugs, antibacterial drugs and anti-inflammatory drugs.

[0017] In addition, the present invention provides a cosmetic comprising any one of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, and ZTM-8 or any combination thereof as an active ingredient, and conventional auxiliary materials in the cosmetics field.

[0018] The present invention also provides a method for preparing the cosmetics, firstly preparing Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7 and ZTM-8, then taking one of them or any combination thereof and adding conventional cosmetic excipients.

[0019] The invention also provides the application of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7 and ZTM-8 in the preparation of cosmetics.

[0020] In the present invention, when the extract or its composition is used to prepare a drug, the content of the extract or its composition in the drug is preferably 0.1-99%; in the pharmaceutical composition, the content of any one of the active ingredient extracts or any combination thereof in the pharmaceutical composition is preferably 0.5-90%. The pharmaceutical composition of the present invention is preferably used in the form of a dosage per unit body weight. In the present invention, the prepared drug can preferably be administered in two forms: injection (intravenous injection, intramuscular injection) and oral administration. The pharmaceutical preparations include tablets, capsules, nasal sprays, pills, dripping pills, etc., including but not limited to the above-mentioned preparations.

[0021] In the present invention, when the extract is used to prepare cosmetics, the preparation of the cosmetics is not limited and can be prepared by conventional preparation methods of cosmetics.

[0022] Compared with the prior art, the present invention has the following advantages:

[0023] 1. The present invention provides Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7 and ZTM-8, which have the effect of promoting collagen secretion, among which ZTM-5 and ZTM-6 have the effect of promoting collagen secretion at relatively low concentrations.

[0024] 2. The extracts of Gastrodia elata pedicel and inflorescence, ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, and ZTM-8, have no toxicity to adult dermal fibroblasts at the concentrations at which they exert their efficacy.

[0025] 3. The extracts of Gastrodia elata pedicel and inflorescence ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, and ZTM-8 also have antioxidant and anti-inflammatory activities, among which ZTM-6 has significant antioxidant and anti-inflammatory activities.

[0026] 4. The present invention provides a method for preparing Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7 and ZTM-8, which has readily available raw materials, simple process, convenient operation and low cost. DETAILED DESCRIPTION

[0027] The following embodiments of the present invention are used to further illustrate the essential content of the present invention, but the present invention is not limited thereto.

[0028] Example 1

[0029] Preparation of Gastrodia elata pedicel and inflorescence extracts ZTM-2 and ZTM-4:

[0030] After seed harvesting, Gastrodia elata pedicels and inflorescences were collected from the Xiaocaoba Experimental Base in Yiliang County, Zhaotong Gastrodia Research Institute in July 2023.

[0031] After the dried Gastrodia elata pedicels and inflorescences were crushed, 20 g of the powder was taken, 400 mL of water was added, and the mixture was heated to 60° C., ultrasonic-assisted extraction was performed for 1 h, the extract was filtered, and the filtrate was concentrated to obtain ZTM-2 (3.5 g).

[0032] After the dried Gastrodia elata pedicel and inflorescence were crushed, 100 g of the powder was taken, 2000 mL of 95% ethanol was added, heated to 60 ° C, ultrasonic-assisted extraction was performed for 30 min, filtered, and the filtrate was obtained. After concentration, the extract (7.0 g) was obtained. The extract was dissolved in 35 mL of water to form a suspension, extracted 3 times with an equal volume of petroleum ether, and the organic phase was distilled under reduced pressure to recover the solvent to obtain a petroleum ether extraction portion (1.2 g). The aqueous phase after petroleum ether extraction was adjusted to a pH value of about 2 with 1.5% HCl, extracted 3 times with an equal volume of ethyl acetate, and the organic phase was distilled under reduced pressure to recover the solvent to obtain an ethyl acetate extraction portion (1.0 g). The aqueous phase after ethyl acetate extraction was adjusted to a pH value of about 9 with 5% NaOH, extracted 3 times with an equal volume of dichloromethane, and the organic phase was distilled under reduced pressure to recover the solvent to obtain ZTM-4 (14.5 mg).

[0033] Embodiment 2:

[0034] Preparation of Gastrodia elata pedicel and inflorescence extracts ZTM-5, ZTM-6, ZTM-7 and ZTM-8:

[0035] After seed harvesting, Gastrodia elata pedicels and inflorescences were collected from the Xiaocaoba Experimental Base in Yiliang County, Zhaotong Gastrodia Research Institute in July 2023.

[0036] After the dried Gastrodia elata pedicel and inflorescence were crushed, 100 g of the powder was taken, 2000 mL of 95% ethanol was added, heated to 60°C, ultrasonic-assisted extraction was performed for 30 min, filtered, and a filtrate was obtained. The filtrate was concentrated under reduced pressure to obtain an extract (6.7 g), and 100 mL of water was added to the extract to prepare a suspension. The suspension was separated by column chromatography using D101 macroporous resin (50 g). Elution was performed with 6 column volumes of distilled water at a flow rate of 0.5 mL / min, the eluate was collected, and the mixture was concentrated under reduced pressure to obtain component ZTM-5 (4.4 g); elution was performed with 6 column volumes of 10% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and the mixture was concentrated under reduced pressure to obtain component ZTM-6 (0.1 g); elution was performed with 6 column volumes of 50% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and the mixture was concentrated under reduced pressure to obtain component ZTM-7 (0.7 g); elution was performed with 6 column volumes of 90% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and the mixture was concentrated under reduced pressure to obtain component ZTM-8 (0.7 g).

[0037] Example 3

[0038] Extracts promote collagen secretion experiment.

[0039] 1. Cells

[0040] Human dermal fibroblasts-adult (HDFa) were purchased from Beina Biotechnology.

[0041] 2. Reagents

[0042] DMEM (high glucose) medium, PBS, Hank's balanced salt solution (HBSS), penicillin-streptomycin and fetal bovine serum (FBS) were purchased from Hyclone; 0.25% trypsin (containing EDTA) was purchased from ThermoFisher; transforming growth factor beta (TGF-β) was purchased from Peprotech; collagen ELISA kit was purchased from TaKaRa; MTS reagent was purchased from Promega.

[0043] 3. Experimental Methods

[0044] On a 96-well cell culture plate, HDFa cells were mixed with the sample to be tested, and a blank control without extract and a TGF-β positive control were set up; cultured at 37°C, 5% CO2 for 3 days, the cell culture supernatant was collected and stored at -80°C; MTS was added, and the OD value at 490nm was detected by MTS colorimetry; the secretion of collagen was detected according to the method provided in the collagen ELISA kit, and the OD value was measured by a microplate reader, and the detection wavelength was 450nm. The increase rate of collagen secretion was calculated.

[0045] Collagen secretion increase rate (%) = (OD 450 nm / Cell viability / blank well OD 450 nm -1)×100%

[0046] 4. Experimental Results

[0047] The present invention evaluates the activity of 6 Gastrodia elata extracts in promoting the secretion of HDFa collagen, and the test results are shown in Table 1. At a concentration of 10 μg / mL, extracts ZTM-2 and ZTM-4 have a significant promoting effect on collagen secretion, and the increase rates of collagen secretion are 29.3% and 55.2%, respectively. At a concentration of 1 μg / mL, extracts ZTM-5 and ZTM-6 have a significant promoting effect on collagen secretion, and the increase rates of collagen secretion are 31.1% and 40.3%, respectively.

[0048] Table 1. The promoting effect of extracts from Gastrodia elata pedicel and inflorescence on HDFa collagen secretion

[0049]

[0050] Example 4

[0051] Antioxidant test of extracts.

[0052] 1. Experimental Principle

[0053] This experiment uses the DPPH (1,1-diphenyl-2-trinitrophenylhydrazine) method to evaluate the in vitro antioxidant capacity of the samples. DPPH is a stable nitrogen-centered free radical with a strong absorption peak at 515nm. When a free radical scavenger is present, DPPH will combine with the protons provided by the scavenger to form DPPH-H, thereby changing the color of the solution from purple to yellow and reducing the absorbance at 515nm. This is used to screen samples with in vitro antioxidant capacity.

[0054] 2. Experimental Reagents

[0055] DPPH and positive control drug water-soluble vitamin E (Trolox) were purchased from Sigma. Trolox was dissolved in anhydrous ethanol at a concentration of 10 mM and stored at -20°C.

[0056] 3. Experimental Methods

[0057] The sample was mixed with DPPH (final concentration of 100 μM) for reaction, 3 replicate wells were set, and a blank control without drug and a Trolox positive control were set at the same time. The OD value was measured by an enzyme marker at 30°C for 1 hour, and the detection wavelength was 515 nm. The antioxidant rate was calculated.

[0058] Antioxidant rate (%) = (1 – sample OD 515 nm / Control OD 515 nm )×100%

[0059] 4. Experimental Results

[0060] The results of DPPH free radical scavenging activity showed that at a concentration of 100 μg / mL, ZTM-6 and ZTM-7 had strong DPPH free radical scavenging effects, and ZTM-4 had a relatively strong DPPH free radical scavenging effect (Table 2).

[0061] Table 2 Antioxidant effects of extracts from Gastrodia elata pedicel and inflorescence in vitro

[0062]

[0063] Example 5

[0064] Anti-inflammatory activity test of extracts.

[0065] 1. Experimental Principle

[0066] Nitric oxide (NO) has a wide range of important biological regulatory functions and plays an important role in inflammation, tumors, and the cardiovascular system. When immune cells are stimulated by microbial endotoxins, inflammatory mediators, etc., a large amount of induced nitric oxide synthase (iNOS) will be generated to produce NO for immune response. Therefore, inhibiting NO production is a direct indicator of the anti-inflammatory activity of the sample. Mouse mononuclear macrophage RAW264.7 was treated with lipopolysaccharide (LPS) to induce the production of nitric oxide synthase, and the sample to be tested was added at the same time. The culture medium was aspirated and the absorbance value was measured at a wavelength of 570nm by the Griess method to detect nitrite (NO2 - ).

[0067] 2. Reagents

[0068] Mouse mononuclear macrophage RAW264.7 was purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences, and DMEM medium and fetal bovine serum were purchased from VivaCell. Griess Reagent, LPS and control drug L-NMMA were purchased from Sigma.

[0069] 3. Experimental Methods

[0070] RAW264.7 cells were inoculated into 96-well plates and stimulated with 1 μg / mL LPS. The samples to be tested were added (the final concentration was diluted 2 times from 50 μg / mL) and the drug-free group and L-NMMA positive drug group were set as controls. After the cells were cultured overnight, the culture medium was taken to detect NO generation and the absorbance was measured at 570 nm. MTS was added to the remaining culture medium to detect cell viability to exclude the toxic effects of the samples on the cells.

[0071] NO production inhibition rate (%) = (OD 570 nm - Sample Group OD 570 nm ) / OD of non-drug treated group 570 nm ×100%

[0072] IC 50 (50%concentration of inhibition) was calculated according to Reed & Muench method.

[0073] 4. Experimental Results

[0074] Extracts ZTM-4, ZTM-6, ZTM-7 and ZTM-8 showed significant inhibitory activity against NO production, with IC 50 The values ​​are shown in Table 3. The inhibitory rates of extracts ZTM-2 and ZTM-5 on NO production were 2.87±2.34% and 11.33±2.90%, respectively, at a concentration of 50 μg / mL.

[0075] Table 3 IC of Gastrodia elata pedicel and inflorescence extracts for inhibiting NO production 50 value

[0076]

[0077] Example 6

[0078] Cleansing milk with extracts of Gastrodia elata peduncle and inflorescence.

[0079] 1. Materials

[0080] The extracts of Gastrodia elata pedicel and inflorescence obtained by the same method as in Example 1 are made of cosmetic grade or food grade raw materials.

[0081] 2. Formulation

[0082] 0.5g of any one or any combination of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, and ZTM-8, glycerin 3.0g, butylene glycol 3.0g, propylene glycol 0.6g, sodium EDTA 0.1g, guar gum 0.3g, zinc dioxide 2.0g, C12-15 alcohol benzoate 4.0g, C12-20 alkyl glucoside 3.0g, C14-22 alcohol 0.5g, cetearyl alcohol 1.2g, paraben 0.2g, sodium stearate 0.3g, dimethicone 0.5g, polysorbate 0.2g, and deionized water 60g.

[0083] 3. Preparation process

[0084] Weigh the formula amount of any one or any combination of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, ZTM-8, and add purified water to completely dissolve; take glycerol, butylene glycol, propylene glycol, sodium EDTA, guar gum, zinc dioxide, sodium stearate, C12-15 benzoate, C12-20 alkyl glucoside, C14-22 alcohol, cetearyl alcohol, add purified water to mix; take paraben, polydimethylsiloxane alcohol, polysorbate, add purified water to mix. Mix the above-prepared solutions, stir well, and can.

[0085] Embodiment 7:

[0086] Cream formula (W%) containing any one of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, ZTM-8 or any combination thereof:

[0087]

[0088] The cosmetics of the present invention are prepared by conventional methods for preparing cosmetics.

[0089] Embodiment 8:

[0090] Emulsion formula (W%) containing any one of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, ZTM-8 or any combination thereof:

[0091]

[0092]

[0093] The cosmetics of the present invention are prepared by conventional methods for preparing cosmetics.

[0094] Embodiment 9:

[0095] Excipients are added in a weight ratio of 1:1 between any one of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, and ZTM-8 or any combination thereof and the excipients, and granulation and tableting are performed.

[0096] Embodiment 10:

[0097] Excipients are added in a weight ratio of 1:2 between any one of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, and ZTM-8 or any combination thereof and the excipients, and granulation and tableting are performed.

[0098] Embodiment 11:

[0099] Any one of the extracts of Gastrodia elata pedicel and inflorescence ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7 and ZTM-8 or any combination thereof is used to prepare capsules according to a conventional capsule preparation method.

[0100] Embodiment 12:

[0101] Tablets: any one of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, ZTM-8 or any combination thereof 10 mg, lactose 180 mg, starch 55 mg, magnesium stearate 5 mg;

[0102] Preparation method: Mix any one of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, ZTM-8 or any combination thereof, lactose and starch, evenly moisten with water, sieve and dry the moistened mixture, sieve again, add magnesium stearate, and then compress the mixture into tablets, each tablet weighing 250 mg and containing 10 mg of the compound.

[0103] Embodiment 13:

[0104] Ampoule: 2 mg of any one or any combination of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, ZTM-8, and 10 mg of sodium chloride;

[0105] Preparation method: dissolve any one of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, ZTM-8 or any combination thereof and sodium chloride in an appropriate amount of water for injection, filter the resulting solution, and fill it into an ampoule bottle under sterile conditions.

[0106] Embodiment 14:

[0107] Capsules: 10 mg of any one or any combination of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, ZTM-8, lactose 187 mg, magnesium stearate 3 mg;

[0108] Preparation method: Mix any one or any combination of Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, ZTM-8 with adjuvants, sieve, mix evenly, and fill the resulting mixture into hard gelatin capsules, each capsule weighing 200 mg and containing 10 mg of active ingredients.

Claims

1. Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, ZTM-8, characterized in that: It is prepared by the following method: After the dried pedicels and inflorescences of Gastrodia elata were crushed, the powder was taken, water was added, and the mixture was heated to 60 °C, and ultrasonic-assisted extraction was performed for 1 h. The extract was filtered, and the filtrate was concentrated to obtain ZTM-2; The dried pedicels and inflorescences of Gastrodia elata were crushed, and the powder was taken, 95% ethanol was added, and the mixture was heated to 60 °C, and ultrasonic-assisted extraction was performed for 30 min. The mixture was filtered to obtain a filtrate, and the extract was concentrated to obtain an extract. The extract was dissolved in water to form a suspension, and the mixture was extracted three times with an equal volume of petroleum ether. The organic phase was subjected to reduced pressure distillation to recover the solvent, and the petroleum ether extraction part was obtained. The aqueous phase after the petroleum ether extraction was adjusted to a pH value of 2 with 1.5% HCl, and extracted three times with an equal volume of ethyl acetate. The organic phase was subjected to reduced pressure distillation to recover the solvent, and the ethyl acetate extraction part was obtained. The aqueous phase after the ethyl acetate extraction was adjusted to a pH value of 9 with 5% NaOH, and extracted three times with an equal volume of dichloromethane. The organic phase was subjected to reduced pressure distillation to recover the solvent, and ZTM-4 was obtained. The dried pedicels and inflorescences of Gastrodia elata were crushed, and the powder was taken, 95% ethanol was added, heated to 60 °C, ultrasonic-assisted extraction was performed for 30 min, and filtered to obtain a filtrate. The filtrate was concentrated under reduced pressure to obtain an extract, and water was added to the extract to prepare a suspension. The suspension was separated by column chromatography using D101 macroporous resin, eluted with 6 column volumes of distilled water at a flow rate of 0.5 mL / min, the eluate was collected, and concentrated under reduced pressure to obtain component ZTM-5; eluted with 6 column volumes of 10% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and concentrated under reduced pressure to obtain ZTM-6; eluted with 6 column volumes of 50% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and concentrated under reduced pressure to obtain ZTM-7; eluted with 6 column volumes of 90% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and concentrated under reduced pressure to obtain ZTM-8.

2. A method for preparing Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7 and ZTM-8, characterized in that: The method comprises the following steps: After the dried Gastrodia elata pedicels and inflorescences were crushed, the powder was taken, water was added, and the mixture was heated to 60 °C, and ultrasonic-assisted extraction was performed for 1 h. The extract was filtered, and the filtrate was concentrated to obtain ZTM-2; The dried pedicels and inflorescences of Gastrodia elata were crushed, and the powder was taken, 95% ethanol was added, and the mixture was heated to 60 °C, and ultrasonic-assisted extraction was performed for 30 min. The mixture was filtered to obtain a filtrate, and the extract was concentrated to obtain an extract. The extract was dissolved in water to form a suspension, and the mixture was extracted three times with an equal volume of petroleum ether. The organic phase was subjected to reduced pressure distillation to recover the solvent, and the petroleum ether extraction part was obtained. The aqueous phase after the petroleum ether extraction was adjusted to a pH value of 2 with 1.5% HCl, and extracted three times with an equal volume of ethyl acetate. The organic phase was subjected to reduced pressure distillation to recover the solvent, and the ethyl acetate extraction part was obtained. The aqueous phase after the ethyl acetate extraction was adjusted to a pH value of 9 with 5% NaOH, and extracted three times with an equal volume of dichloromethane. The organic phase was subjected to reduced pressure distillation to recover the solvent, and ZTM-4 was obtained. The dried pedicels and inflorescences of Gastrodia elata were crushed, and the powder was taken, 95% ethanol was added, heated to 60 °C, ultrasonic-assisted extraction was performed for 30 min, and filtered to obtain a filtrate. The filtrate was concentrated under reduced pressure to obtain an extract, and water was added to the extract to prepare a suspension. The suspension was separated by column chromatography using D101 macroporous resin, eluted with 6 column volumes of distilled water at a flow rate of 0.5 mL / min, the eluate was collected, and concentrated under reduced pressure to obtain component ZTM-5; eluted with 6 column volumes of 10% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and concentrated under reduced pressure to obtain ZTM-6; eluted with 6 column volumes of 50% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and concentrated under reduced pressure to obtain ZTM-7; eluted with 6 column volumes of 90% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and concentrated under reduced pressure to obtain ZTM-8.

3. Use of the Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, and ZTM-8 according to claim 1 in the preparation of anti-aging drugs, antibacterial drugs, and anti-inflammatory drugs.

4. A pharmaceutical composition, consisting of any one of the Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, and ZTM-8 or any combination thereof as described in claim 1 and a pharmaceutically acceptable carrier.

5. Use of the pharmaceutical composition according to claim 4 in the preparation of anti-aging drugs, antibacterial drugs, and anti-inflammatory drugs.

6. The method for preparing the pharmaceutical composition according to claim 4, characterized in that: The method comprises the following steps: After the dried pedicels and inflorescences of Gastrodia elata were crushed, the powder was taken, water was added, and the mixture was heated to 60 °C, and ultrasonic-assisted extraction was performed for 1 h. The extract was filtered, and the filtrate was concentrated to obtain ZTM-2; The dried pedicels and inflorescences of Gastrodia elata were crushed, and the powder was taken, 95% ethanol was added, and the mixture was heated to 60 °C, and ultrasonic-assisted extraction was performed for 30 min. The mixture was filtered to obtain a filtrate, and the extract was concentrated to obtain an extract. The extract was dissolved in water to form a suspension, and the mixture was extracted three times with an equal volume of petroleum ether. The organic phase was subjected to reduced pressure distillation to recover the solvent, and the petroleum ether extraction part was obtained. The aqueous phase after the petroleum ether extraction was adjusted to a pH value of 2 with 1.5% HCl, and extracted three times with an equal volume of ethyl acetate. The organic phase was subjected to reduced pressure distillation to recover the solvent, and the ethyl acetate extraction part was obtained. The aqueous phase after the ethyl acetate extraction was adjusted to a pH value of 9 with 5% NaOH, and extracted three times with an equal volume of dichloromethane. The organic phase was subjected to reduced pressure distillation to recover the solvent, and ZTM-4 was obtained. After the dried pedicels and inflorescences of Gastrodia elata were crushed, the powder was taken, 95% ethanol was added, and the mixture was heated to 60 °C. Ultrasonic-assisted extraction was performed for 30 min, and the mixture was filtered to obtain a filtrate. The filtrate was concentrated under reduced pressure to obtain an extract. Water was added to the extract to prepare a suspension. The suspension was separated by column chromatography using D101 macroporous resin, and eluted with 6 column volumes of distilled water at a flow rate of 0.5 mL / min. The eluate was collected and concentrated under reduced pressure to obtain component ZTM-5; eluted with 6 column volumes of 10% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and the mixture was concentrated under reduced pressure to obtain ZTM-6; eluted with 6 column volumes of 50% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and the mixture was concentrated under reduced pressure to obtain ZTM-7; eluted with 6 column volumes of 90% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and the mixture was concentrated under reduced pressure to obtain ZTM-8; Then, any one of the extracts of Gastrodia elata pedicel and inflorescence ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, and ZTM-8 or any combination thereof is added to a pharmaceutically acceptable carrier.

7. A cosmetic comprising any one of the extracts of Gastrodia elata pedicel and inflorescence ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7, and ZTM-8 as claimed in claim 1 or any combination thereof as an active ingredient, and conventional cosmetic excipients.

8. Use of the Gastrodia elata pedicel and inflorescence extracts ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7 and ZTM-8 according to claim 1 in the preparation of cosmetics.

9. The method for preparing the cosmetics according to claim 6, characterized in that: The method comprises the following steps: After the dried Gastrodia elata pedicels and inflorescences were crushed, the powder was taken, water was added, and the mixture was heated to 60 °C, and ultrasonic-assisted extraction was performed for 1 h. The extract was filtered, and the filtrate was concentrated to obtain ZTM-2; The dried pedicels and inflorescences of Gastrodia elata were crushed, and the powder was taken, 95% ethanol was added, and the mixture was heated to 60 °C, and ultrasonic-assisted extraction was performed for 30 min. The mixture was filtered to obtain a filtrate, and the extract was concentrated to obtain an extract. The extract was dissolved in water to form a suspension, and the mixture was extracted three times with an equal volume of petroleum ether. The organic phase was subjected to reduced pressure distillation to recover the solvent, and the petroleum ether extraction part was obtained. The aqueous phase after the petroleum ether extraction was adjusted to a pH value of 2 with 1.5% HCl, and extracted three times with an equal volume of ethyl acetate. The organic phase was subjected to reduced pressure distillation to recover the solvent, and the ethyl acetate extraction part was obtained. The aqueous phase after the ethyl acetate extraction was adjusted to a pH value of 9 with 5% NaOH, and extracted three times with an equal volume of dichloromethane. The organic phase was subjected to reduced pressure distillation to recover the solvent, and ZTM-4 was obtained. After the dried pedicels and inflorescences of Gastrodia elata were crushed, the powder was taken, 95% ethanol was added, and the mixture was heated to 60 °C. Ultrasonic-assisted extraction was performed for 30 min, and the mixture was filtered to obtain a filtrate. The filtrate was concentrated under reduced pressure to obtain an extract. Water was added to the extract to prepare a suspension. The suspension was separated by column chromatography using D101 macroporous resin, and eluted with 6 column volumes of distilled water at a flow rate of 0.5 mL / min. The eluate was collected and concentrated under reduced pressure to obtain component ZTM-5; eluted with 6 column volumes of 10% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and the mixture was concentrated under reduced pressure to obtain ZTM-6; eluted with 6 column volumes of 50% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and the mixture was concentrated under reduced pressure to obtain ZTM-7; eluted with 6 column volumes of 90% ethanol at a flow rate of 0.5 mL / min, the eluate was collected, and the mixture was concentrated under reduced pressure to obtain ZTM-8; Then, any one of the extracts of Gastrodia elata pedicel and inflorescence ZTM-2, ZTM-4, ZTM-5, ZTM-6, ZTM-7 and ZTM-8 or any combination thereof is added to conventional cosmetic excipients.

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