Gastrodin and preparation method and application thereof

Abstract

The present application provides gastrodia elata (Blume) alkaloids PFT201A, PFT201B and PFT203 and a preparation method thereof, and application thereof in the fields of cosmetics and medicine. Gastrodia elata Gastrodia elata (Blume) alkaloids PFT201A, PFT201B and PFT203 significantly promote the secretion of collagen type II in human dermal fibroblasts-adult (HDFa) at an extremely low concentration (0.1 μ M), and have no cytotoxicity at the concentration. These compounds can be used for preparing anti-aging drugs or skin care products.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of cosmetics and medicine, in particular to the field of botanical medicine, more particularly to Gastrodia alkaloids PFT201A, PFT201B and PFT203, pharmaceutical compositions thereof, methods of preparation, and uses thereof in the manufacture of medicaments and in the manufacture of cosmetics. BACKGROUND

[0002] Collagen is a kind of main structural protein in the extracellular matrix of mammals, widely exists in skin, bone, muscle and other tissues, mainly participates in physiological and biochemical behaviors such as cell proliferation, differentiation, migration and signal transmission, and plays a supporting, repairing and protecting role for tissue cells and the like [Ye T, Xiang Q, Yang Y, Huang Y D. Research progress on development and application of collagen. Journal of Bioengineering, 2023, 39(3): 942-960]. Among them, collagen I is the most abundant protein in human skin, accounting for about 70% of the dry weight of the skin. Collagen I can increase skin elasticity and delay skin aging [Zhang Y H, Zhou J X, Zhang N H, Zhao L, Zhang L B, Zhou F. Research progress on collagen peptide and elastin peptide for improving photoaging skin. Food Research and Development, 2023, 44(11): 208-215]. In addition, on the wound surface, collagen I promotes wound healing by providing structural support for cell attachment and migration. However, animal-derived collagen I may have immunogenicity, and problems related to processing technology, degradation rate and disinfection process of collagen I 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 of plant extracts and natural products with collagen secretion promoting activity is a current research hotspot.

[0003] Traditional Chinese medicine Gastrodia elata Blume is the dried tuber of Gastrodia elata Blume of Orchidaceae, which is a Chinese medicinal material and has good therapeutic effect on nervous system diseases [Duan X Y, Yan H, Chen Y F, Luo J F, Ding P C, Tian M H, Wang Y H. Chemical constituents of dichloromethane extract of Gastrodia elata and their cholinesterase inhibitory activities. Zhongcaoyao, 2022, 53(23): 7324-7330]. The research on Gastrodia elata alkaloids is relatively rare. So far, there is no report on Gastrodia elata alkaloids PFT201A, PFT201B and PFT203 in the prior art, and there is no report on their collagen secretion promoting activity. SUMMARY

[0004] The purpose of the present application is to provide Gastrodia elata alkaloids PFT201A, PFT201B and PFT203, a pharmaceutical composition containing the same, a preparation method thereof, and the use thereof in the preparation of drugs and cosmetics, in view of the above-mentioned deficiencies in the prior art.

[0005] In order to achieve the above-mentioned purpose of the present application, the present application provides the following technical solution:

[0006] Gastrodia elata alkaloids PFT201A, PFT201B and PFT203 as shown in the following structural formula,

[0007]

[0008] A preparation method of Gastrodia elata alkaloids PFT201A, PFT201B and PFT203, the method comprising the following steps:

[0009] After the dried tuber, peduncle and inflorescence of Gastrodia elata Blume are crushed, they are extracted with 90% ethanol water solution at 70°C for 3h, and then filtered to obtain filtrate and residue. The residue is extracted again for 2 times by the same method. The filtrates are combined and concentrated under reduced pressure to obtain an extract. The extract is mixed with distilled water to form a suspension, and the pH is adjusted to 2-3 by adding HCl. The suspension is extracted with petroleum ether for 5 times. The organic phase is concentrated under reduced pressure at 60°C by a rotary evaporator to obtain a petroleum ether extraction part. After the petroleum ether extraction, the remaining aqueous phase is extracted with an equal volume of ethyl acetate for 5 times. The organic phase is concentrated under reduced pressure at 60°C by a rotary evaporator to obtain an ethyl acetate extraction part. After the ethyl acetate extraction, the remaining aqueous phase is adjusted to pH 9-10 with NaOH aqueous solution, and then extracted with dichloromethane for 5 times. The organic phase is concentrated under reduced pressure at 60°C by a rotary evaporator to obtain a dichloromethane extraction part.

[0010] The dichloromethane extraction part is subjected to silica gel column chromatography with 100-200 mesh, and gradient elution with v / v acetone-petroleum ether 20:1→1:1 to obtain 10 components A, B, C, D, E, F, G, H, I and J.

[0011] Component H was separated by reversed phase C 18 Silica gel column chromatography, gradient elution with methanol-water (5%→100%, v / v), the fraction eluted with 30% methanol was compound PFT203; fraction J6-1 eluted with 40% methanol was separated by 300-400 mesh silica gel column chromatography, gradient elution with petroleum ether-ethyl acetate (10:1→0:1, v / v) to obtain J6-1-1; J6-1-1 was separated by Sephadex LH-20 column chromatography using methanol as eluent to obtain J6-1-1-1; J6-1-1-1 was purified by HPLC to obtain PFT201; PFT201 was separated by HPLC to obtain compounds PFT201A and PFT201B.

[0012] or, Component J was separated by reversed phase C 18 Silica gel column chromatography, gradient elution with methanol-water (5%→100%, v / v), the fraction eluted with 30% methanol was compound PFT203; fraction J6-1 eluted with 40% methanol was separated by 300-400 mesh silica gel column chromatography, gradient elution with petroleum ether-ethyl acetate (10:1→0:1, v / v) to obtain J6-1-1; J6-1-1 was separated by Sephadex LH-20 column chromatography using methanol as eluent to obtain J6-1-1-1; J6-1-1-1 was purified by HPLC to obtain PFT201; PFT201 was separated by HPLC to obtain compounds PFT201A and PFT201B.

[0013] Use of Gastrodia alkaloids PFT201A, PFT201B and PFT203 in the preparation of anti-aging drugs.

[0014] Use of Gastrodia alkaloids PFT201A, PFT201B and PFT203 in the preparation of collagen secretion promoting active agents.

[0015] Pharmaceutical composition, consisting of any one or any combination of Gastrodia alkaloids PFT201A, PFT201B and PFT203 and a pharmaceutically acceptable carrier.

[0016] Use of the pharmaceutical composition in the preparation of anti-aging drugs, in the preparation of collagen secretion promoting active agents.

[0017] The preparation method of the pharmaceutical composition, the method comprising the following steps: using the above preparation method to prepare Gastrodia alkaloids PFT201A, PFT201B and PFT203 first, taking at least one of them, adding a pharmaceutically acceptable carrier.

[0018] Use of Gastrodia Elata Alkaloids PFT201A, PFT201B and PFT203 in the Preparation of Cosmetics.

[0019] Cosmetics containing at least one of Gastrodia Elata Alkaloids PFT201A, PFT201B and PFT203 as effective components.

[0020] The preparation method of the cosmetics comprises the following steps: first preparing Gastrodia Elata Alkaloids PFT201A, PFT201B and PFT203 by the above-mentioned preparation method, and then adding at least one of the three alkaloids and common cosmetic auxiliaries.

[0021] In the present application, when the Gastrodia Elata Alkaloids PFT201A, PFT201B and PFT203 or their combinations are used in the preparation of medicines, the content of the compounds or their combinations in the medicines is preferably 0.1-99%; and in the medicine combinations, the content of any one of the active ingredients or any combination thereof in the medicine combinations is preferably 0.5-90%. The medicine combinations of the present application are preferably used in the form of unit body weight dosage. In the present application, the prepared medicines are preferably administered in two forms of injection (intravenous injection, intramuscular injection) and oral administration. The medicine preparations include tablets, capsules, nasal sprays, pills, dripping pills, etc., but are not limited to the above preparations.

[0022] In the present application, when the Gastrodia Elata Alkaloids PFT201A, PFT201B and PFT203 are used in the preparation of cosmetics, the preparations of the cosmetics are not limited, and can be prepared by the conventional preparation method of cosmetics.

[0023] Compared with the prior art, the present application has the following advantages:

[0024] 1. The present application provides compounds PFT201A, PFT201B and PFT203, which have the effect of promoting collagen secretion at very low concentrations (0.1 μM).

[0025] 2. The compounds PFT201A, PFT201B and PFT203 have no toxicity to adult dermal fibroblasts at the concentration at which they exert the pharmacological effect.

[0026] 3. The extraction raw material of the compounds is the waste (rhizomes, peduncles and inflorescences) after the collection of seeds of Gastrodia Elata, which is easy to obtain and low in cost. BRIEF DESCRIPTION OF DRAWINGS

[0027] Figure 1 The figure is a schematic diagram of the chemical structural formula of the compounds PFT201A, PFT201B and PFT203 of the present application.

[0028] Figure 2 ECD spectra of the experiments of the present application and calculated ECD spectra of the (S)-isomer and (R)-isomer. DETAILED DESCRIPTION

[0029] The essential content of the present application is further illustrated below with the embodiments of the present application in conjunction with the accompanying drawings, but the present application is not limited to the embodiments.

[0030] Example 1

[0031] Preparation of Gastrodia alkaloids PFT201A, PFT201B and PFT203:

[0032] The dried Gastrodia tubers, peduncles and inflorescences (21.4 kg) were ground together and extracted with 80 L of 90% aqueous ethanol at 70°C for 3 h. The filtrate and residue were obtained by filtration, and the residue was extracted again twice by the same method. The filtrates were combined and concentrated under reduced pressure to obtain an extract 6.6 kg. The extract was mixed with distilled water (6.8 L) to form a suspension, and 0.1% HC1 (8 L) was added to adjust the pH to 2-3. The mixture was extracted with an equal volume of petroleum ether 5 times. The organic phase was concentrated under reduced pressure at 60°C using a rotary evaporator to obtain the petroleum ether extraction fraction (142.2 g). After the petroleum ether extraction, the remaining aqueous phase was extracted with an equal volume of ethyl acetate 5 times. The organic phase was concentrated under reduced pressure at 60°C using a rotary evaporator to obtain the ethyl acetate extraction fraction (104.4 g). After the ethyl acetate extraction, the remaining aqueous phase was adjusted to pH 9-10 with 5% NaOH aqueous solution (4 L) and extracted with an equal volume of dichloromethane 5 times. The organic phase was concentrated under reduced pressure at 60°C using a rotary evaporator to obtain the dichloromethane extraction fraction (25.6 g).

[0033] The dichloromethane extraction fraction was subjected to gradient elution column chromatography on a 100-200 mesh silica gel column (acetone-petroleum ether, 20:1→1:1, v / v) to obtain 10 fractions, i.e., fractions A (449.2 mg), B (103.0 mg), C (336.1 mg), D (57.5 mg), E (962.8 mg), F (422.7 mg), G (3.9 g), H (1.6 g), I (1.4 g) and J (10.2 g).

[0034] Fraction H was subjected to reverse phase (RP) HPLC (YMC Pack ODS-A, 250 mm x 20 mm, 5 μm) to obtain fractions H1 (0.6 g) and H2 (0.4 g). 18Sephadex LH-20 column chromatography using methanol as eluent to give H1-1-1-3-1 (40.1 mg); H1-1-1-3-1 was separated by HPLC (Agilent Zorbax SB-C 18 column, φ 6.4 x 250 mm, 5 μm; MeOH-H2O, 40:60, 2 mL / min) to give PFT201 (14.2 mg, t R = 29.0 min); PFT201 was separated by HPLC (Shiseido Chiral CD-Ph column, φ 4.6 x 250 mm, 5 μm; MeOH-H2O, 60:40, 1 mL / min) to give compound PFT201A (6.3 mg, t R = 9.6 min) and PFT201B (3.9 mg, t R = 11.3 min).

[0035] Component J was separated by reverse phase (RP) C 18 silica gel column chromatography (methanol-water, 5%→100%, v / v) to give compound PFT203 (30.0 mg) at 30% methanol elution fraction. The 40% methanol elution fraction (J6-1, 209.6 mg) was separated by 300-400 mesh silica gel column chromatography (petroleum ether-ethyl acetate, 10:1→0:1, v / v) to give J6-1-1 (39.0 mg); J6-1-1 was separated by Sephadex LH-20 column chromatography using methanol as eluent to give J6-1-1-1 (16.0 mg); J6-1-1-1 was purified by HPLC (Welch Ultimate AQ-C 18 column, φ 7.8 x 250 mm, 5 μm; MeOH-H2O, 55:45, 2 mL / min) to give PFT201 (10.0 mg, t R = 21.6 min).

[0036] Example 2

[0037] PFT201A, PFT201B and PFT203 Figure 1 Spectral data and structure identification of PFT201A, PFT201B and PFT203

[0038] Compound PFT201A, Chinese name: (+)-(S)-4-(2-hydroxy-2-quinolin-4- ylethyl)phenol, English name: (+)-(S)-4-(2-Hydroxy-2-quinolin-4-ylethyl)phenol, white solid, molecular formula C 17 H 15 NO2; [a] 2 D 5 +17(c 0.31, MeOH); ECD(c 0.008, MeOH) λ max (Δε) 289 (-0.11), 233 (5.50), 203 (-2.05) nm; 1 H NMR (500 MHz, chloroform-d) δ H 8.85 (1H, d, J = 4.5 Hz, H-2), 8.16 (1H, br d, J = 8.5 Hz, H-8), 8.08 (1H, br d, J = 8.5 Hz, H-5), 7.72 (1H, m, H-7), 7.60 (1H, m, H-6), 7.51 (1H, d, J = 4.5 Hz, H-3), 7.00 (2H, br d, J = 8.5 Hz, H-2', 6'), 6.77 (2H, br d, J = 8.5 Hz, H-3', 5'), 5.66 (1H, dd, J = 8.1, 3.8 Hz, H-8'), 3.24 (1H, dd, J = 14.1, 3.8 Hz, H-7'a), 2.96 (1H, dd, J = 14.1, 8.1 Hz, H-7'b); 13 CNMR (126 MHz, chloroform-d) δ C 155.4 (C, C-4'), 150.0 (CH, C-2), 149.6 (C, C-4), 147.7 (C, C-8a), 130.7 (CH, C-2', 6'), 130.0 (CH, C-8), 129.3 (CH, C-7), 128.4 (C, C-1'), 126.8 (CH, C-6), 125.4 (C, C-4a), 122.9 (CH, C-5), 117.8 (CH, C-3), 115.6 (CH, C-3', 5'), 71.0 (CH, C-8'), 43.9 (CH2, C-7'); ESIMS m / z 266 [M+H] +Its NMR data are reported for the first time and its absolute configuration was determined by ECD calculation Figure 2 ).

[0039] Compound PFT201B, Chinese name (-)-(R)-4-(2-hydroxy-2-quinolin-4- ylethyl)phenol, English name (-)-(R)-4-(2-Hydroxy-2-quinolin-4-ylethyl)phenol, white solid, molecular formula C 17 H 15 NO2; [a] 2 D 5 -25 (c 0.29, MeOH); ECD (c 0.006, MeOH) λ max (Δε) 290 (1.13), 223 (-6.96), 202 (+6.99) nm; 1 H NMR (500 MHz, chloroform-d) δ H 8.85 (1H, d, J = 4.5 Hz, H-2), 8.16 (1H, br d, J = 8.5 Hz, H-8), 8.08 (1H, br d, J = 8.5 Hz, H-5), 7.72 (1H, m, H-7), 7.60 (1H, m, H-6), 7.51 (1H, d, J = 4.5 Hz, H-3), 7.00 (2H, br d, J = 8.5 Hz, H-2', 6'), 6.77 (2H, br d, J = 8.5 Hz, H-3', 5'), 5.66 (1H, dd, J = 8.1, 3.8 Hz, H-8'), 3.24 (1H, dd, J = 14.1, 3.8 Hz, H-7'a), 2.96 (1H, dd, J = 14.1, 8.1 Hz, H-7'b); 13 CNMR (126 MHz, chloroform-d) δ C 155.4 (C, C-4'), 150.0 (CH, C-2), 149.6 (C, C-4), 147.7 (C, C-8a), 130.7 (CH, C-2', 6'), 130.0 (CH, C-8), 129.3 (CH, C-7), 128.4 (C, C-1'), 126.8 (CH, C-6), 125.4 (C, C-4a), 122.9 (CH, C-5), 117.8 (CH, C-3), 115.6 (CH, C-3', 5'), 71.0 (CH, C-8'), 43.9 (CH2, C-7'); ESIMS m / z 266 [M+H] +its NMR data are reported for the first time and its absolute configuration is determined by ECD calculation Figure 2 ).

[0040] Compound PFT203, English name Para-topolin riboside, white needle-like crystals, molecular formula C 17 H 19 N5O5; [a] 2 D 5 -68 (c 0.12, MeOH); 1 H NMR (500 MHz, DMSO-d6) δ H 8.36 (1H, s, H-8), 8.31 (1H, br s, 6-NH), 8.21 (1H, s, H-2), 7.14 (2H, d, J = 8.3 Hz, H-2", 6"), 6.67 (2H, d, J = 8.3 Hz, H-3", 5"), 5.88 (1H, d, J = 6.2 Hz, H-1'), 4.60 (1H, m, H-3'), 4.58 (2H, br s, H-7"), 4.14 (1H, m, H-2'), 3.96 (1H, m, H-4'), 3.67 (1H, m, H-5'a), 3.55 (1H, m, H-5'b); 13 C NMR (126 MHz, DMSO-d6) δ C 156.1 (C, C-6), 154.5 (C, C-1"), 152.4 (CH, C-2), 148.4 (C, C-4), 139.8 (CH, C-8), 130.2 (C, C-4"), 128.5 (CH, C-2", 6"), 119.8 (C, C-5), 115.0 (CH, C-3", 5"), 88.0 (CH, C-1'), 86.0 (CH, C-4'), 73.5 (CH, C-3'), 70.7 (CH, C-2'), 61.7 (CH2, C-5'), 42.4 (CH2, C-7"); ESIMS m / z 396 [M+Na] + , 769 [2M+Na] +The compound NMR data is consistent with the literature data [Huang Z.-B., Wu Z., Chen F.-K., Zou L.-B. The protective effects of phenolic constituents from Gastrodia elata on the cytotoxicity induced by KCl and glutamate. Arch. Pharm. Res., 2006, 29(11), 963-968].

[0041] Example 3

[0042] Gastrodia elata alkaloids PFT201A, PFT201B and PFT203 promote collagen type I secretion experiment.

[0043] 1. Cells

[0044] Human dermal fibroblasts-adult (HDFa) were purchased from North Na Biological Technology Co., Ltd.

[0045] 2. Reagents

[0046] DMEM (high sugar) medium, PBS, Hank's balanced salt solution (HBSS), penicillin-streptomycin and fetal bovine serum (FBS) were purchased from Hyclone Co., Ltd.; 0.25% trypsin (containing EDTA) was purchased from Thermo Fisher Scientific; transforming growth factor beta (TGF-β) was purchased from Peprotech; collagen ELISA kit was purchased from TaKaRa; MTS reagent was purchased from Promega.

[0047] 3. Experimental method

[0048] HDFa cells were mixed with the sample to be tested on a 96-well cell culture plate, and a blank control without compound and a TGF-β positive control were set. After 3 days of culture at 37°C and 5% CO2, the cell culture supernatant was collected and stored at -80°C. MTS was added, and the OD value at 490 nm was detected by MTS colorimetric method. The secretion of collagen type I was detected according to the method provided in the collagen ELISA kit, and the OD value was detected by a microplate reader at a wavelength of 450 nm. The increase rate of collagen type I secretion was calculated.

[0049] Collagen type I secretion increase rate (%) = (experimental well OD 450 nmCell survival rate / blank well OD 450 nm -1) x 100%

[0050] 4. Experimental results

[0051] The present application evaluated the activity of three compounds PFT201A, PFT201B and PFT203 of the present application in promoting the secretion of collagen type I of HDFa cells, and the test results are shown in Table 1. At a concentration of 0.1 μM, PFT201A, PFT201B and PFT203 had a significant promoting effect on the secretion of collagen type I, with an increase rate of collagen type I secretion of 49.7%, 31.1% and 55.2% respectively, and PFT201A, PFT201B and PFT203 had no toxicity to HDFa cells at this concentration.

[0052] Table 1. Promoting effect of Gastrodia alkaloids on the secretion of collagen type I of HDFa cells

[0053]

[0054] Example 4:

[0055] Tablet: 10 mg of at least one of compounds PFT201A, PFT201B and PFT203, 180 mg of lactose, 55 mg of starch and 5 mg of magnesium stearate;

[0056] Preparation method: mix at least one of compounds PFT201A, PFT201B and PFT203, lactose and starch, uniformly wet with water, sieve and dry the wet 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.

[0057] Example 5:

[0058] Ampoule: 2 mg of at least one of compounds PFT201A, PFT201B and PFT203, and 10 mg of sodium chloride;

[0059] Preparation method: dissolve at least one of compounds PFT201A, PFT201B and PFT203 and sodium chloride in an appropriate amount of water for injection, filter the resulting solution, and fill the ampoule under sterile conditions.

[0060] Example 6:

[0061] Capsule: 10 mg of at least one of compounds PFT201A, PFT201B and PFT203, 187 mg of lactose and 3 mg of magnesium stearate;

[0062] Preparation: Compound PFT201A, PFT201B, PFT203 at least one is mixed with an auxiliary, sieved, mixed uniformly, the obtained mixture is filled into hard gelatin capsules, each capsule weighs 200 mg, the active ingredient content is 10 mg.

[0063] Example 7:

[0064] Cream formulation (W%) containing at least one of the compounds PFT201A, PFT201B, PFT203:

[0065]

[0066]

[0067] The cosmetic of the above formulation of the present application is prepared by the conventional method of making cosmetics.

[0068] Example 8:

[0069] Emulsion formulation (W%) containing at least one of the compounds PFT201A, PFT201B, PFT203:

[0070]

[0071] The cosmetic of the above formulation of the present application is prepared by the conventional method of making cosmetics.

Claims

1. A method for preparing Gastrodia alkaloids PFT201A and PFT201B as shown in the following structural formula, ， characterized in that The method comprises the following steps: The dried Gastrodia tubers, peduncles and inflorescences after seed collection are crushed, and then extracted with 90% ethanol aqueous solution at 70 °C for 3 h. After filtration, a filtrate and a residue are obtained. The residue is extracted again twice by the same method. The filtrates are combined, and then concentrated under reduced pressure to obtain an extract. Distilled water is added to the extract to form a suspension, and then HCl is added to adjust the pH to 2-3. The suspension is extracted with petroleum ether for 5 times. The organic phase is concentrated under reduced pressure at 60 °C by a rotary evaporator to obtain a petroleum ether extraction part. After the petroleum ether extraction, the remaining aqueous phase is extracted with an equal volume of ethyl acetate for 5 times. The organic phase is concentrated under reduced pressure at 60 °C by a rotary evaporator to obtain an ethyl acetate extraction part. After the ethyl acetate extraction, the remaining aqueous phase is adjusted to pH 9-10 with NaOH aqueous solution, and then extracted with dichloromethane for 5 times. The organic phase is concentrated under reduced pressure at 60 °C by a rotary evaporator to obtain a dichloromethane extraction part. The dichloromethane extraction part is subjected to silica gel column chromatography with 100-200 mesh, and then eluted with v / v acetone-petroleum ether 20:1→1:1 gradient to obtain 10 components A, B, C, D, E, F, G, H, I and J. Fraction H was subjected to reverse phase C 18 Silica gel column chromatography, methanol-water 10%→100% v / v, to give 10% methanol eluted fraction H1; Fraction H1 was subjected to Sephadex LH-20 column chromatography, using methanol as eluent to give H1-1; H1-1 was subjected to H silica gel column chromatography, petroleum ether-acetone 10:1→3:1 v / v to give H1-1-1; H1-1-1 was subjected to Sephadex LH-20 column chromatography, using methanol as eluent to give H1-1-1-3; H1-1-1-3 was subjected to H silica gel column chromatography, petroleum ether-acetone 10:1→1:1 v / v to give H1-1-1-3-1; H1-1-1-3-1 was subjected to HPLC to give PFT201; PFT201 was subjected to HPLC to give compounds PFT201A and PFT201B; or, by component J via reverse phase C 18 Silica gel column chromatography, gradient elution with methanol-water 5%→100% v / v, 40% methanol eluted fraction J6-1 was separated by silica gel column chromatography with 300~400 mesh, gradient elution with petroleum ether-ethyl acetate 10:1→0:1 v / v to obtain J6-1-1; J6-1-1 was separated by Sephadex LH-20 column chromatography using methanol as eluent to obtain J6-1-1-1; PFT201 was obtained by HPLC purification of J6-1-1-1; PFT201A and PFT201B were obtained by HPLC separation of PFT201. 2.The application of Gastrodia alkaloids PFT201A and PFT201B as shown in the following structural formula in the preparation of anti-aging drugs, 。 3.The application of Gastrodia alkaloids PFT201A and PFT201B as shown in the following structural formula in the preparation of collagen secretion promoting active agents, 。 4.A pharmaceutical composition, which comprises any one or any combination of Gastrodia alkaloids PFT201A and PFT201B as shown in the following structural formula and a pharmaceutically acceptable carrier, 。 5.The application of the pharmaceutical composition of claim 4 in the preparation of anti-aging drugs and collagen secretion promoting active agents.

6. The method of preparing the pharmaceutical composition according to claim 4, characterized by, The method comprises the following steps: first, Gastrodia alkaloids PFT201A and PFT201B are obtained by the preparation method of claim 1, and then at least one of them is added to a pharmaceutically acceptable carrier. 7.The application of Gastrodia alkaloids PFT201A and PFT201B as shown in the following structural formula in the preparation of cosmetics, 。 8.A cosmetic, which comprises at least one of Gastrodia alkaloids PFT201A and PFT201B as shown in the following structural formula as an effective component, 。 9. The method for preparing a cosmetic according to claim 8, characterized in that, The method comprises the following steps: first, Gastrodia alkaloids PFT201A and PFT201B are obtained by the preparation method of claim 1, and then at least one of them is added to a cosmetic auxiliary material commonly used.

Images