Application of Paris polyphylla extract in the preparation of cosmetics and pharmaceuticals

Abstract

The present application provides a paridis Paris rugosa The present application provides a preparation method of paridis μg The present application provides a preparation method of paridis μg The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation method of paridis The present application provides a preparation

Description

Technical Field

[0001] This invention belongs to the fields of cosmetics and pharmaceutical technology, specifically relating to the field of herbal medicine technology, and more specifically, to the extracts PFVB6 and PFEA6 of Paris polyphylla, which have collagen-promoting activity, as well as the preparation method and application of the extracts PFVB6 and PFEA6 of Paris polyphylla. Background Technology

[0002] With the improvement of living standards and the development of medical technology, while meeting basic living needs, delaying aging has become a new pursuit for people. Skin aging is the result of various factors working together, mainly divided into two types: one is endogenous aging, mainly influenced by genetic factors, which is a natural aging process; the other is exogenous aging caused by external environmental factors such as ultraviolet radiation, smoking, gravity, and chemicals. Maintaining youthful skin, solving skin problems, and restoring skin health have become new research hotspots. 75% of the skin is composed of collagen, and skin growth, repair, and nutrition all depend on collagen. The two key aspects of healthy skin—moisturizing and anti-wrinkle—are both related to collagen. The effects of anti-aging active substances include scavenging free radicals, increasing cell proliferation rate, slowing down the degradation rate of the extracellular matrix, and replenishing collagen in the skin, restoring and rebuilding the collagen layer. Currently, common methods of supplementing collagen include oral intake of collagen-rich foods, subcutaneous injections, and the use of collagen-rich skincare products, but their effects are limited, the time required is long, they are prone to causing allergies and infections, and the absorption effect is limited. Adding active ingredients that stimulate the skin's own collagen synthesis to skincare products or medications to replenish collagen has become a new focus in anti-aging drug and skincare product research and development. Therefore, finding natural anti-aging products that promote the skin's own collagen secretion has become a new research trend.

[0003] Paris polyphylla (wrinkled leaves) Paris rugosaH. Li & Kurita) belongs to the genus Paris. According to different plant classification systems, it can be classified into Liliaceae, Trilliumaceae, or Melanthiaceae [1. Li Heng. Plants of the genus Paris. Beijing: Science Press. 1998; 2. Chen X, Chen SC, Liang S, Liang SJ, Xu J, Tamura MN. Liliaceae, in Flora of China, Wu ZY, Hong DY, Raven PH, eds. Science Press, Beijing; Missouri Botanical Garden Press, Saint Louis). 2000, 73–263; 3. Ji Y. A Monograph of Paris (Melanthiaceae). SciencePress, Beijing; Springer, Singapore. 2021]. The Nu people use the whole plant to treat unnamed boils and mumps [Dao Zhiling, Long Chunlin, Liu Yitao. A preliminary study on the traditional use of plants by the Nu people in Gaoligong Mountain, Yunnan. Biodiversity, 2003, 11 (3), 231–239]. Previously, compounds in the rhizome of Paris polyphylla were found to have certain antibacterial activity [Duan Xiaoyan, Yue Meicen, Yang Jun, Bai Xue, Luo Jifeng, Li Heng, Wang Yuehu. Study on chemical composition and antibacterial activity of the rhizome of Paris polyphylla. Chinese Journal of Traditional Chinese Medicine, 2023, 48 (11), 2981–2988]. However, there are currently no reports on the collagen-promoting activity of Paris polyphylla extract. Summary of the Invention

[0004] The purpose of this invention is to provide extracts of Paris polyphylla var. ...

[0005] To achieve the above-mentioned objectives of the present invention, the present invention provides the following technical solution:

[0006] Application of Paris polyphylla extract PFVB6 and / or PFEA6 in the preparation of collagen secretion activators.

[0007] Application of Paris polyphylla extract PFVB6 and / or PFEA6 in the preparation of anti-aging drugs.

[0008] Application of Paris polyphylla extract PFVB6 and / or PFEA6 in the preparation of skin care products.

[0009] According to the described application, the concentration of PFVB6 in the Paris polyphylla extract is 0.1%. μ g / mL, the concentration of PFEA6 is 10 μ g / mL.

[0010] According to the application described, the Paris polyphylla extracts PFVB6 and PFEA6 are prepared by the following method:

[0011] After the dried rhizomes of Paris polyphylla were pulverized, they were extracted with 70% ethanol at 60 °C using ultrasound-assisted extraction for 2 h. The filtrate was then filtered. The residue was extracted again under the same conditions for a total of 3 extractions. The filtrates were combined, and the solvent was recovered under reduced pressure using a rotary evaporator to obtain a crude extract. The crude extract was suspended in distilled water and extracted three times each with equal volumes of ethyl acetate and n-butanol. After the solvent was recovered under reduced pressure, the ethyl acetate extract and the n-butanol extract PFVB6 were obtained, respectively.

[0012] After grinding the dried rhizomes of Paris polyphylla, the extract was obtained by ultrasonic-assisted extraction with 70% ethanol at 60 °C for 30 min. The filtrate was then filtered. The residue was extracted again under the same conditions for a total of 3 extractions. The filtrates were combined, and the solvent was recovered under reduced pressure using a rotary evaporator to obtain the crude extract. The crude extract was then suspended in distilled water and extracted 3 times with an equal volume of n-butanol. After the solvent was recovered under reduced pressure, the n-butanol extract was obtained. The aqueous phase after the n-butanol extraction was then subjected to reduced pressure and solvent recovery to obtain the water-soluble fraction PFEA6.

[0013] An anti-aging pharmaceutical composition comprising, as an active ingredient, PFVB6 and / or PFEA6 extracted from Paris polyphylla, wherein the concentration of PFVB6 is 0.1%. μ g / mL, the concentration of PFEA6 is 10 g / mL. μ g / mL.

[0014] The cosmetic contains Paris polyphylla extract PFVB6 and / or PFEA6 as its active ingredient, with a PFVB6 concentration of 0.1%. μ g / mL, the concentration of PFEA6 is 10 g / mL. μ g / mL.

[0015] A method for preparing PFVB6 and PFEA6 from Paris polyphylla extract, comprising the following steps:

[0016] After the dried rhizomes of Paris polyphylla were pulverized, they were extracted with 70% ethanol at 60 °C using ultrasound-assisted extraction for 2 h. The filtrate was then filtered. The residue was extracted again under the same conditions for a total of 3 extractions. The filtrates were combined, and the solvent was recovered under reduced pressure using a rotary evaporator to obtain a crude extract. The crude extract was suspended in distilled water and extracted three times each with equal volumes of ethyl acetate and n-butanol. After the solvent was recovered under reduced pressure, the ethyl acetate extract and the n-butanol extract PFVB6 were obtained, respectively.

[0017] After grinding the dried rhizomes of Paris polyphylla, the extract was obtained by ultrasonic-assisted extraction with 70% ethanol at 60 °C for 30 min. The filtrate was then filtered. The residue was extracted again under the same conditions for a total of 3 extractions. The filtrates were combined, and the solvent was recovered under reduced pressure using a rotary evaporator to obtain the crude extract. The crude extract was then suspended in distilled water and extracted 3 times with an equal volume of n-butanol. After the solvent was recovered under reduced pressure, the n-butanol extract was obtained. The aqueous phase after the n-butanol extraction was then subjected to reduced pressure and solvent recovery to obtain the water-soluble fraction PFEA6.

[0018] In this invention, when the pharmaceutical composition is used to prepare a drug, the content of the compound or its combination in the drug is preferably 0.1% to 99%; in the pharmaceutical composition, the content of any one or any combination of the active ingredients is preferably 0.5% to 90%. The pharmaceutical composition of this invention is preferably used in the form of a dose per unit body weight. In this invention, the prepared drug is preferably administered by both injection (intravenous injection, intramuscular injection) and oral administration. The pharmaceutical preparations include tablets, capsules, nasal sprays, pills, drops, etc., including but not limited to the above-mentioned preparations.

[0019] In this invention, when the extracts PFVB6 and PFEA6 of Paris polyphylla are used to prepare cosmetics, the formulation of the cosmetics is not limited and can be prepared using conventional cosmetic preparation methods.

[0020] Compared with the prior art, the present invention has the following advantages.

[0021] 1. Even at low concentrations, the extracts PFVB6 and PFEA6 from Paris polyphylla can promote collagen secretion.

[0022] 2. At the concentrations at which the extracts of Paris polyphylla var. rubrum are effective, PFVB6 and PFEA6 are not toxic to adult dermal fibroblasts. Detailed Implementation

[0023] The following embodiments of the present invention will further illustrate the substantive content of the present invention, but are not intended to limit the present invention.

[0024] Example 1:

[0025] Preparation of PFVB6 and PFEA6 from Paris polyphylla extract.

[0026] Dried Paris polyphylla (wrinkled leaves) Paris rugosa 4.1 kg of rhizomes were crushed and extracted with 70% ethanol (15 L) at 60 °C using ultrasound-assisted extraction for 2 h. The filtrate was obtained after filtration, and the residue was extracted again under the same conditions. The extraction was repeated three times. The filtrates were combined, and the solvent was recovered under reduced pressure using a rotary evaporator to obtain a crude extract (350 g). The crude extract was suspended in 1.5 L of distilled water and extracted three times each with equal volumes of ethyl acetate and n-butanol. After solvent recovery under reduced pressure, ethyl acetate extract (123.0 g) and n-butanol extract (209.0 g) were obtained, respectively. The n-butanol extract was designated PFVB6, with a yield of 5.1%.

[0027] Dried Paris polyphylla (wrinkled leaves) Paris. rugosa 7.2 kg of rhizomes were ground and extracted with 70% ethanol (70 mL) at 60℃ using ultrasound-assisted extraction for 30 min. The filtrate was then filtered, and the residue was extracted again under the same conditions. This process was repeated three times. The filtrates were combined, and the solvent was recovered under reduced pressure using a rotary evaporator to obtain a crude extract (1.2 g). The crude extract was suspended in 10 mL of distilled water and extracted three times with an equal volume of n-butanol. After solvent recovery under reduced pressure, 427.4 mg of the n-butanol extract was obtained. The aqueous phase after n-butanol extraction was further extracted under reduced pressure to obtain a water-soluble fraction of 421.3 mg, designated PFEA6, with a yield of 5.9%.

[0028] Example 2:

[0029] Experiments on the collagen secretion-promoting effects of PFVB6 and PFEA6.

[0030] 1. Cell

[0031] Human dermal fibroblasts (HDFa) were purchased from BeiNa Bio.

[0032] 2. Reagents

[0033] 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 Thermo Fisher Scientific; and transfer growth factor was also provided. β (transforming growth factor beta, TGF- βThe collagen ELISA kit was purchased from Peprotech; the collagen ELISA kit was purchased from TaKaRa; and the MTS reagent was purchased from Promega.

[0034] 3. Experimental Methods

[0035] In a 96-well cell culture plate, HDFa cells were mixed with the test sample, and a drug-free blank control and TGF-β- were set up. β Positive control; cultured at 37 °C, 5% CO2 for 3 days, collected cell culture supernatant and stored at −80 °C; added MTS, and detected the OD value at 490 nm using the MTS colorimetric method; collagen secretion was detected according to the method provided in the collagen ELISA kit, and the OD value was measured using an ELISA reader at a wavelength of 450 nm. The collagen secretion increase rate was calculated.

[0036] Collagen secretion increase rate (%) = (OD of experimental wells) 450 nm / Cell viability / Octopic pore OD 450 nm -1)×100%

[0037] 4. Experimental Results

[0038] Table 1. The promoting effects and cytotoxic effects of PFVB6 and PFEA6 on HDFa collagen secretion.

[0039] sample Concentration (g / mL) Cell viability (%) Increased collagen secretion (%) TGF- (positive control) 0.01 123.1 118.2 ± 0.1 PFVB6 100 17.2 Untested PFVB6 0.1 103.0 12.1 ± 2.0 PFEA6 10 91.0 37.8 ± 0.9

[0040] Example 3:

[0041] The excipients were added to the Paris polyphylla extract at a weight ratio of 1:1 (PFVB6 and / or PFEA6) and granulated and compressed into tablets.

[0042] Example 4:

[0043] The excipients were added to the Paris polyphylla extract at a weight ratio of 1:2 (PFVB6 and / or PFEA6) and granulated and compressed into tablets.

[0044] Example 5:

[0045] Extracts of Paris polyphylla var. ...

[0046] Example 6:

[0047] Prepare tablets using the following method.

[0048] Tablets: Paris polyphylla extract PFVB6 and / or PFEA6 100mg

[0049] Starch as needed

[0050] Corn syrup as needed

[0051] Magnesium stearate (appropriate amount)

[0052] Example 7:

[0053] Capsules: Paris polyphylla extract PFVB6 and / or PFEA6 100mg

[0054] Starch as needed

[0055] Magnesium stearate (appropriate amount)

[0056] Preparation method: Mix the Paris polyphylla extract PFVB6 and / or PFEA6 with the excipients, sieve, mix evenly in a suitable container, and fill the resulting mixture into hard gelatin capsules.

[0057] Example 8:

[0058] Nasal spray: Paris polyphylla extract PFVB6 and / or PFEA6 80 mg

[0059] Sodium chloride 8 mg

[0060] EDTA 1 mg

[0061] Sodium phosphate buffer (pH 6.5) 10 mg

[0062] Polyethoxy ether 10 mg

[0063] 2ml of redistilled water

[0064] Preparation method: Add one component at a time to an appropriate volume of double-distilled water while stirring until completely dissolved, then add the next component. After adding water to 2 ml, filter the solution through a sterile filter, bottle it, and divide it according to appropriate dosages.

[0065] Example 9:

[0066] Droplets: 1g of Paris polyphylla extract PFVB6 and / or PFEA6

[0067] Polyethylene glycol 6000 9g

[0068] Preparation: Preparation of the melt of Paris polyphylla extract PFVB6 or / and PFEA66000: Weigh the Paris polyphylla extract PFVB6 or / and PFEA6 according to the above prescription, add an appropriate amount of anhydrous ethanol, dissolve by gentle heating, and then add to the prescribed amount of polyethylene glycol melt (keep warm in a 60℃ water bath). Stir and mix evenly until the ethanol evaporates completely. Let it stand in a 60℃ water bath for 30 minutes until the air bubbles are removed. Then transfer the above-mentioned well-mixed melt with the air bubbles removed into a storage cylinder. Under the condition of keeping warm at 80-85℃, control the dripping rate and drip it drop by drop into the condensate. After complete condensation, pour off the condensate, collect the pellets, drain them, remove the condensate from the pellets with filter paper, and place them in a silica gel desiccator or air dry.

[0069] Example 10:

[0070] Tablets: Paris polyphylla extract PFVB6 and / or PFEA6 10 mg, lactose 180 mg, starch 55 mg, magnesium stearate 5 mg;

[0071] Preparation method: Mix Paris polyphylla extract PFVB6 and / or PFEA6, lactose and starch, moisten with water, sieve and dry the moistened mixture, sieve again, add magnesium stearate, and then compress the mixture into tablets, each weighing 250 mg and containing 10 mg of the compound.

[0072] Example 11:

[0073] Ampoule contents: Paris polyphylla extract PFVB6 and / or PFEA6 2 mg, sodium chloride 10 mg;

[0074] Preparation method: Dissolve the extract of Paris polyphylla PFVB6 and / or PFEA6 and sodium chloride in an appropriate amount of water for injection, filter the resulting solution, and fill it into ampoules under aseptic conditions.

[0075] Example 12:

[0076] Capsules: Paris polyphylla extract PFVB6 or / and PFEA6 10 mg, lactose 187 mg, magnesium stearate 3 mg;

[0077] Preparation method: Mix the extract of Paris polyphylla PFVB6 and / or PFEA6 with the adjuvants, sieve, mix evenly, and fill the resulting mixture into hard gelatin capsules, each capsule weighing 200 mg and containing 10 mg of active ingredient.

[0078] Example 13:

[0079] Cream formulations containing Paris polyphylla extract PFVB6 and / or PFEA6 (W%):

[0080] Paris polyphylla extract PFVB6 and / or PFEA6 0.0001

[0081] Stearic acid 8.0

[0082] C16 alcohol 2.0

[0083] Self-emulsifying monoglyceride 2.0

[0084] Hydrogenated lanolin 2.0

[0085] Liquid paraffin 12.0

[0086] Glycerin 7.0

[0087] Emulsifier 1.5

[0088] Preservative 0.2

[0089] Fragrance 0.2

[0090] Add deionized water to 100

[0091] The cosmetic product of the above-described formula of the present invention is prepared using conventional methods for manufacturing cosmetics.

[0092] Example 14:

[0093] Emulsion formulations containing Paris polyphylla extract PFVB6 and / or PFEA6 (W%):

[0094] Paris polyphylla extract PFVB6 and / or PFEA6 0.0001

[0095] Stearic acid 1.4

[0096] Cetyl alcohol 0.1

[0097] 2-Ethyl alcohol cetyl stearate 1.8

[0098] Isopropyl myristate 0.2

[0099] 2-Hexyl-1-decyl alcohol 1.0

[0100] Liquid paraffin 7.5

[0101] Glycerin 3.0

[0102] Propylene glycol 8.0

[0103] Triethanolamine 1.0

[0104] Carboxyvinyl polymer 0.35

[0105] Arlacel 165 2.0

[0106] Preservative 0.2

[0107] Fragrance 0.2

[0108] Add deionized water to 100

[0109] The cosmetic product of the above-described formula of the present invention is prepared using conventional methods for manufacturing cosmetics.

Claims

1. The use of PFEA6 extract from Paris fargesii Franch. as an active ingredient in the preparation of an active agent for promoting collagen secretion, which is prepared by the following method: The dried rhizome of Paris fargesii Franch. is ground, and then extracted with 70% ethanol at 60°C for 30 min under ultrasonic assistance, and the filtrate is obtained after filtration; the residue is continuously extracted under the same conditions for a total of 3 times, and the filtrate is combined; after the solvent is recovered by rotary evaporation under reduced pressure, a crude extract is obtained; the crude extract is suspended in distilled water, and then extracted with an equal volume of n-butanol for 3 times; after the solvent is recovered under reduced pressure, the n-butanol extraction part is obtained; and after the water phase after n-butanol extraction is recovered under reduced pressure, the water-soluble part PFEA6 is obtained.

2. Use of the Paris polyphylla var. chinensis extract PFEA6 as an active ingredient in the preparation of an active agent for promoting collagen secretion according to claim 1, characterized in that: wherein the concentration of the PFEA6 extract of Paris fargesii is 10 μ g / mL.