Antifungal activity and application of Paris polyphylla steroidal saponins

Abstract

This invention provides a compound called *Paris polyphylla* saponin A, its pharmaceutical composition, preparation method, and application, belonging to the fields of pharmaceutical technology and cosmetic technology. It provides *Paris polyphylla* saponin A, as well as *Paris polyphylla* saponins A-D and *Paris polyphylla* saponin A in the preparation of drugs for the prevention and treatment of superficial fungal infections of the skin. The above compounds are effective against *Trichophyton rubrum* (…). Trichophyton rubrum Epidermophyton floccosum ( Epidermophyton floccosum ) and Microsporum gypseum ( Microsporum gypseum It has a significant inhibitory effect on dermatophytes such as dermatophytes and can be used to prepare drugs for the prevention and treatment of superficial fungal infections of the skin.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of medicine, and in particular relates to a preparation method of Paris fargesii saponin A, a pharmaceutical composition thereof, and applications of Paris fargesii saponin A and Paris polyphylla saponins A-D in the preparation of medicines for preventing and treating dermatophytosis and in the preparation of cosmetics. BACKGROUND

[0002] Dermatophytosis is a kind of superficial mycosis caused by dermatophytes such as Trichophyton rubrum, Epidermophyton floccosum and Microsporum gypseum [Xie Z M, Huang J M, Liu H F, Zeng W Y, Xu Q Z, He Z Q, Fu B, Xue Y H. Analysis of 4018 cases of superficial mycosis and pathogenic bacteria. Journal of Dermatology and Venereology, 2022, 29(4): 337-341]. Dermatophytosis often has a long treatment course, and there are limited types of existing antifungal drugs, which have large side effects. Therefore, it is of great significance to develop new therapeutic drugs for the clinical treatment of dermatophytosis [Huang R Q, Wang S. Research progress of traditional Chinese medicine in resisting dermatophytes. Chinese Journal of Endemic Diseases Control, 2020, 35(4): 432-434].

[0003] Steroidal saponins are the main chemical components of Paris L. plants, and Paris saponins usually have antibacterial activity [Duan X Y, Yue M C, Yang J, Bai X, Luo J F, Li H, Wang Y T. Chemical constituents of the rhizomes of Paris fargesii and their antibacterial activity. Chinese Journal of Chinese Materia Medica, 2023, 48(11): 2981-2988; Yang Y L, Bai X, Yang J, Luo J F, Duan X Y, Wang Y T. Chemical constituents of the rhizomes of Paris polyphylla var. tibetica and their antibacterial activity. Chinese Herbal Drugs, 2024, 55(5): 1466-1476]. However, at present, there is no report on the antibacterial activity of Paris polyphylla saponins A-D and Paris fargesii saponin A in the prior art. SUMMARY

[0004] The purpose of the present application is to provide a preparation method of a new compound Paris fargesii saponin A (PFV92), and applications of Paris polyphylla saponins A-D and Paris fargesii saponin A in the preparation of medicines for preventing and treating dermatophytosis.

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

[0006] A compound Paris fargesii saponin A as shown in the following structural formula,

[0007]

[0008] The preparation method of the compound PFE92, which comprises the following steps: crushing the dried roots of P. multifida, extracting the crushed roots with 70% ethanol at 78-80°C for 4h, 3h and 3h respectively, filtering to obtain a filtrate, concentrating the filtrate to obtain a crude extract, suspending the crude extract in distilled water, extracting the suspension with an equal volume of ethyl acetate for 3 times, recovering the solvent from the organic phase to obtain an ethyl acetate extraction part, and continuously extracting the water phase with an equal volume of n-butanol for 4 times, recovering the solvent from the organic phase to obtain an n-butanol extraction part;

[0009] The n-butanol extraction part is subjected to gradient elution with a 200-300 mesh normal phase silica gel column, and the eluent is 10:1-0:1 CH2Cl2-MeOH. After the eluent is detected by thin layer chromatography, 7 parts are combined: Fr.I, Fr.II, Fr.III, Fr.IV, Fr.V, Fr.VI and Fr.VII. Fr.VII is subjected to gradient elution with a 200-300 mesh normal phase silica gel column, and the eluent is 10:1-0:1 EtOAc-MeOH. After the eluent is detected by thin layer chromatography, 4 parts are combined: Fr.VII-1, Fr.VII-2, Fr.VII-3 and Fr.VII-4. 10.0g of Fr.VII-3 is subjected to RP C 18 silica gel column chromatography, MeOH-H2O 10:90-100:0, and 5 parts are obtained: Fr.VII-3-1, Fr.VII-3-2, Fr.VII-3-3, Fr.VII-3-4 and Fr.VII-3-5. Fr.VII-3-3 is subjected to Sephadex LH-20 gel column chromatography with MeOH, and then subjected to semi-preparative HPLC with Agilent Zorbax SB-C 18 , φ9.4x250mm; MeCN-H2O, 43:57; v=2mL / min, to purify PFE92.

[0010] A pharmaceutical composition taking PFE92 as an effective ingredient.

[0011] A cosmetic taking PFE92 as an effective ingredient.

[0012] Use of PFE92 in the preparation of a medicine for treating superficial mycosis of the skin.

[0013] Use of PFE92 in the preparation of a cosmetic.

[0014] Use of PFE52, PFE53, PFE56 and PFE59A in the preparation of a medicine for treating superficial mycosis of the skin.

[0015] Use of the compounds PFE52, PFE53, PFE56, PFE59A in the preparation of cosmetics.

[0016] The structural formula is as follows:

[0017]

[0018] In the present application, when the pharmaceutical composition is used for the preparation of a medicine, the content of the compound or its composition in the medicine is preferably 0.1-99%; in the pharmaceutical composition, the content of any one of the active ingredients or any combination thereof in the pharmaceutical composition is preferably 0.5-90%. The pharmaceutical composition of the present application is preferably used in the form of unit dose. In the present application, the prepared medicine can be administered in two forms of injection (intravenous injection, intramuscular injection) and oral administration. The pharmaceutical preparation includes tablets, capsules, nasal sprays, pills, dripping pills, etc., including but not limited to the above preparations.

[0019] In the present application, when the compounds PFE52, PFE53, PFE56, PFE59A and PFV92 are used for the preparation of cosmetics, the preparation of the cosmetics is not limited, and can be prepared by the conventional preparation method of cosmetics.

[0020] Compared with the prior art, the present application has the following advantages:

[0021] 1. The compound PFV92 is a new compound.

[0022] 2. The antibacterial activity of the compounds PFE52, PFE53, PFE56, PFE59A and PFV92 is reported for the first time.

[0023] 3. The compounds PFE59A and PFV92 selectively inhibit the superficial fungi Trichophyton rubrum, Epidermophyton floccosum and Microsporum gypseum.

[0024] 4. The inhibitory effect of the compounds PFE52, PFE53, PFE56, PFE59A and PFV92 on 5 strains of fungi is shown in Table 2. The compounds PFE59A and PFV92 have significant inhibitory activity on the superficially infected fungi Epidermophyton floccosum, Trichophyton rubrum and Microsporum gypseum. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 The chemical structural formula of PFE59A (PFV92) is shown in the following schematic diagram.

[0026] Figure 2Chemical structural formula of PFE52.

[0027] Figure 3 Key 2D NMR correlation diagram of PFV92. DETAILED DESCRIPTION

[0028] The essential content of the present application is further illustrated by the embodiments of the present application in conjunction with the accompanying drawings, but the present application is not limited by the embodiments.

[0029] Example 1

[0030] Sources and preparation of compounds.

[0031] Five PFE52, PFE53, PFE56, PFE59A and PFV92 were tested, respectively, and their sources or preparation methods are as follows. Figure 1 and Figure 2 ), respectively, and their sources or preparation methods are as follows.

[0032] 1. Preparation of PFV92.

[0033] Paris rugosa H. Li & Kurita was purchased from a Paris planting base in Shiyueliang Township, Fugong County, Yunnan Province in October 2020. After the dried rhizomes of Paris rugosa H. Li & Kurita (4.1 kg) were crushed, 70% ethanol (50 L x 3) was used to extract the crushed rhizomes at 78-80°C for 3 times, and the extraction time was 4 h, 3 h and 3 h, respectively. After filtration, the filtrate was obtained, and the filtrate was concentrated to obtain a crude extract (1.4 kg). The crude extract was suspended in 16 L distilled water, and an equal volume of ethyl acetate was used to extract the suspension 3 times. After recovering the solvent from the organic phase, the ethyl acetate extraction part (123.4 g) was obtained. The water phase was continuously extracted with an equal volume of n-butanol 4 times, and the organic phase was recovered to obtain the n-butanol extraction part (219.3 g).

[0034] The n-butanol extraction part (209.2 g) was subjected to gradient elution using normal phase silica gel (200-300 mesh) column chromatography, and the eluent was CH2Cl2-MeOH (10:1-0:1). After the eluent was detected by thin layer chromatography, 7 parts were combined: Fr. I, Fr. II, Fr. III, Fr. IV, Fr. V, Fr. VI and Fr. VII. Fr. VII (162.0 g) was subjected to gradient elution using normal phase silica gel (200-300 mesh) column chromatography, and the eluent was EtOAc-MeOH (10:1-0:1). After the eluent was detected by thin layer chromatography, 4 parts were combined: Fr. VII-1 (0.3 g), Fr. VII-2 (6.1 g), Fr. VII-3 (22.2 g) and Fr. VII-4 (112.0 g). 10.0 g of Fr. VII-3 was subjected to RP C18 Silica gel column chromatography (MeOH-H2O, 10:90~100:0) was divided into five parts: Fr.VII-3-1 (63.4 mg), Fr.VII-3-2 (312.3 mg), Fr.VII-3-3 (334.0 mg), Fr.VII-3-4 (56.0 mg) and Fr.VII-3-5 (7.6 g). Fr.VII-3-3 (334.0 mg) was purified by Sephadex LH-20 gel column chromatography (MeOH) and then by semi-preparative HPLC (Agilent Zorbax SB-C 18 , φ 9.4 x 250 mm; MeCN-H2O, 43:57; v = 2 mL / min) to give Pachysandersonin A (PFV92; 4.5 mg, t R = 19.4 min).

[0035] 2. Sources of PFE52, PFE53, PFE56 and PFE59A.

[0036] Pachysandersonin B (PFE52), Pachysandersonin A (PFE53), Pachysandersonin D (PFE56) and Pachysandersonin C (PFE59A) were extracted and isolated from the dried rhizomes of Paris dulongensis H. Li & Kurita, which was purchased from the Paris cultivation base in Longjiang Township, Gongshan County, Yunnan Province in September 2021. The preparation method of the compounds is described in the literature [Jia J.-K., Yang J., Yang X.-Z., Luo J.-F., Duan X.-Y., Yang Y.-L., Wan J.-F., Wang Y.-H. Polyhydroxylated spirostanol saponins from the rhizomes of Paris dulongensis. Chem. Biodivers., 2024, e202400980. https: / / doi.org / 10.1002 / cbdv.202400980].

[0037] Example 2:

[0038] Structure elucidation of Pachysandersonin A (PFV92).

[0039] Pachysandersonin A (PFV92): white amorphous powder, (c 0.2, MeOH); UV (MeOH) λ max(log ε) 323 (2.03), 274 (2.51), 216 (3.54) nm; ECD (c, 0.08, MeOH) λ max (Δε) 217 (+10.04) nm; ESI-MS m / z 935 [M + Na] + ; HR-ESI-MS m / z 935.4245 [M + Na] + (C 45 H 68 NaO 19 , calculated 935.4247); 1 H and 13 C NMR data are listed in Table 1.

[0040] In the high resolution electrospray ionization mass spectrum (HR-ESI-MS) of compound PFV92, the quasi-molecular ion peak m / z 935.4245 [M + Na] + (calcd for C 45 H 68 NaO 19 , 935.4247), combined with 13 C NMR data (Table 1), suggested the molecular formula of C 45 H 68 O 19 with 12 degrees of unsaturation. According to 1 H and 13 C NMR data (Table 1), there were 5 methyl groups [δ H 1.73 (3H, d, J = 6.2 Hz), 1.21 (3H, s), 1.05 (d, J = 6.8 Hz), 0.89 (3H, s), 0.67 (d, J = 5.4 Hz)], 1 disubstituted double bond [δ H 6.67 (1H, d, J = 8.5 Hz) and 6.33 (1H, d, J = 8.5 Hz); δ C 136.6 (CH) and 130.9 (CH)], 1 trisubstituted double bond [δ H 5.48 (1H, dd, J = 5.3, 1.7 Hz); δ C 144.0 (C) and 119.4 (CH)], and 3 sugars [δ H 6.39 (1H, br s), 5.09 (1H, d, J = 7.6 Hz), 4.80 (1H, d, J = 7.5 Hz); δ C104.5 (CH), 102.1 (CH), 100.6 (CH)]. After acid hydrolysis of compound PFV92, D-glucopyranose and L-rhamnopyranose were detected. Combined with NMR data, it was determined that compound PFV92 contained 2 β-D-glucopyranose [δ H 5.09 (1H, d, J = 7.6 Hz) and 4.80 (1H, d, J = 7.5 Hz)] and 1 a-L-rhamnopyranose [δ H 6.39 (1H, br s)]. By comparing with the NMR data of Paris vanioside A [Yan H., Ni W., Yu L.-L., Xiao L.-G., Ji Y.-H., Liu H.-Y. Paris vaniosides A-E, five new steroidal saponins from Paris vaniotii. Steroids, 2022, 177: 108949], it was speculated that compound PFV92 was a steroidal saponin with the same aglycone.

[0041] According to 1 H- 1 H COSY correlations( Figure 3 ), fragments a (H-1 / H2-2 / H-3 / H2-4), b (H-6 / H-7), c (H-11 / H2-12), d (H-14 / H2-15 / H-16 / H-17 / H-20 / H3-21), e [H2-23 / H2-24 / H-25 / (H2-26) / H3-27], f (H-1’ / H-2’ / H-3’ / H-4’ / H-5’ / H2-6’), g (H-1” / H-2” / H-3” / H-4” / H-5” / H3-6”) and h (H-1”’ / H-2”’ / H-3”’ / H-4”’ / H-5”’ / H2-6”’) can be deduced. In the HMBC correlation spectrum( Figure 3 ), H3-19 was correlated with C-1, C-5, C-9 and C-10, H2-4 was correlated with C-10, H-6 was correlated with C-4 and C-10, H-7 was correlated with C-9, H3-18 was correlated with C-12, C-13, C-14 and C-17, and H3-21 and H2-26 were correlated with C-22, indicating that the double bond was located at C-6; H2-15 was correlated with C-8 and H3-19 was correlated with C-5, indicating that C-5 and C-8 were oxidized. In the ROESY correlation spectrum( Figure 3), it can be seen that H-1 β / H3-19, H3-19 / H-6, H-7 / H3-18, H3-18 / H-20, H-20 / H2-23 and H3-27 / H2-26 have correlations, indicating that these hydrogen atoms are in the same plane, and assuming β orientation, the orientation of the peroxide bridge and H-17 is α orientation; H-17 / H-14, H-14 / H-16 and H-1 α / H-3 have correlations, indicating that these hydrogen atoms are in α orientation, H-26 ax / H3-27 and H-26 eq / H3-27, indicating that 25-Me is in the equatorial position of the chair conformation of the six-membered ring, thus confirming that the aglycone of compound PFV92 is (25R)-Spirost-6,11-diene-5α,8α-epidioxy-3β-ol. According to the HMBC correlations of H-3 with C-1', H-1' with C-3, H-2' with C-1", H-1" with C-2', H-3' with C-1'", and H-1'" with C-3', it is shown that β-D-glucopyranose [δ H 5.09 (1H, d, J = 7.6 Hz) is located at C-2', α-L-rhamnopyranose [δ H 6.39 (1H, br s)] is located at C-3', thus, it is determined that the sugar chain is α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→3)]-β-D-glucopyranosyl, located at the 3-OH of the aglycone. Therefore, the chemical structure of PFV92 is determined as (25R)-Spirost-6,11-diene-5α,8α-epidioxy-3β-ol 3-α-L-rhamnopyranosyl-(1→2)-O-[β-D-glucopyranosyl-(1→3)]-β-D-glucopyranoside, named as rugosaroside A.

[0042] Table 1 PFV92 in deuterated pyridine 1 H (500 MHz) and 13 C (126 MHz) NMR data (δ in ppm, J in Hz)

[0043]

[0044] Example 3:

[0045] The test method for anti-dermatophyte activity is as follows:

[0046] Epidermophyton floccosum CBS 566.94, Trichophyton rubrum ATCC4438, Microsporum gypseum CBS118893 were purchased from China Medical University Mycological Culture Collection Center; Candida albicans ATCC10231 was purchased from Microbiologics, USA; Candida albicans fluconazole-resistant strain was a gift from the Department of Dermatology, the First Affiliated Hospital of Kunming Medical College.

[0047] Terbinafine hydrochloride, DMSO were purchased from Sigma company; Agar powder, chloramphenicol were purchased from Scientific Research Special company.

[0048] Take 96-well culture plates, dilute the sample to be tested, add fungal liquid to each well, and the final concentration is 5x10 5 CFU / mL, 25℃ for 5 days, and detect the absorbance at 625nm by microplate reader. The culture medium blank control, fungal control and positive drug control were set up at the same time.

[0049] Example 4:

[0050] The inhibitory effect of compounds PFE52, PFE53, PFE56, PFE59A and PFV92 on 5 strains of fungi is shown in Table 2. Compounds PFE59A and PFV92 have significant inhibitory activity on superficially infected fungi Epidermophyton floccosum, Trichophyton rubrum and Microsporum gypseum, but have no inhibitory effect on deeply infected fungi Candida albicans and Candida albicans fluconazole-resistant strain.

[0051] Table 2 Inhibitory effect of compounds on 5 strains of fungi

[0052]

[0053] *Positive control

[0054] Example 5:

[0055] Take one or any combination of Pachysandra saponin A (PFV92), Dulong Pachysandra saponin B (PFE52), Dulong Pachysandra saponin A (PFE53), Dulong Pachysandra saponin D (PFE56), Dulong Pachysandra saponin C (PFE59A) and add excipient in a weight ratio of 1:1, granulate and tablet.

[0056] Example 6:

[0057] Take wrinkle leaf Paris ginseng saponin A (PFV92), Paris ginseng saponin B (PFE52), Paris ginseng saponin A (PFE53), Paris ginseng saponin D (PFE56), Paris ginseng saponin C (PFE59A) any one or any of them, with the weight ratio of 1:2 of the excipient, the excipient is added, granulation and tabletting.

[0058] Example 7:

[0059] Take wrinkle leaf Paris ginseng saponin A (PFV92), Paris ginseng saponin B (PFE52), Paris ginseng saponin A (PFE53), Paris ginseng saponin D (PFE56), Paris ginseng saponin C (PFE59A) any one or any of them, according to the conventional capsule preparation method to make capsules.

[0060] Example 8:

[0061] Tablets are prepared by the following method

[0062] Tablets: wrinkle leaf Paris ginseng saponin A (PFV92), Paris ginseng saponin B (PFE52), Paris ginseng saponin A (PFE53), Paris ginseng saponin D (PFE56), Paris ginseng saponin C (PFE59A)

[0063] Example 9:

[0064] Capsules: wrinkle leaf Paris ginseng saponin A (PFV92), Paris ginseng saponin B (PFE52), Paris ginseng saponin A (PFE53), Paris ginseng saponin D (PFE56), Paris ginseng saponin C (PFE59A) any one or any of them 100mg

[0065] Starch appropriate amount

[0066] Magnesium stearate appropriate amount

[0067] Preparation method: according to the wrinkle leaf Paris ginseng saponin A (PFV92), Paris ginseng saponin B (PFE52), Paris ginseng saponin A (PFE53), Paris ginseng saponin D (PFE56), Paris ginseng saponin C (PFE59A) any one or any of them, mixed with auxiliary agent, sieved, mixed uniformly in a suitable container, the obtained mixture is filled into hard gelatin capsules.

[0068] Example 10:

[0069] Nasal spray: Paridis saponin A (PFV92), Paridis saponin B (PFE52), Paridis saponin A (PFE53), Paridis saponin D (PFE56), Paridis saponin C (PFE59A)

[0070] Preparation method: under stirring, add one ingredient in proper volume of heavy distilled water at a time, until completely dissolved, then add another ingredient. After adding water to 2 ml, filter the solution on a sterile filter, fill into bottles and separate according to proper dosage.

[0071] Example 11

[0072] Spray: take any one or any combination of Paridis saponin A (PFV92), Paridis saponin B (PFE52), Paridis saponin A (PFE53), Paridis saponin D (PFE56), Paridis saponin C (PFE59A), soak in water for 4 hours, then perform water vapor distillation, collect the aromatic water in 4 times the amount of medicinal materials, then perform secondary distillation, collect the aromatic water in 1.5 times the amount of medicinal materials, then add peppermint oil and Tween-80, shake well to clarify, filter, and seal to obtain. External use.

[0073] Or, take any one or any combination of Paridis saponin A (PFV92), Paridis saponin B (PFE52), Paridis saponin A (PFE53), Paridis saponin D (PFE56), Paridis saponin C (PFE59A), prepare a spray according to the conventional preparation method of spray, for external use.

[0074] Example 12:

[0075] Dropping pills: 1 g of any one or any combination of Paridis saponin A (PFV92), Paridis saponin B (PFE52), Paridis saponin A (PFE53), Paridis saponin D (PFE56), Paridis saponin C (PFE59A)

[0076] Polyethylene glycol 6000 9 g

[0077] Preparation method: preparation of 6000 melting liquid: according to the above prescription, weigh one or any of parispory lily rhizome saponin A (PFV92), parispory lily rhizome saponin B (PFE52), parispory lily rhizome saponin A (PFE53), parispory lily rhizome saponin D (PFE56), parispory lily rhizome saponin C (PFE59A), add appropriate amount of anhydrous ethanol, dissolve under slight heat, then add the prescription amount of polyethylene glycol melting liquid (60℃ water bath), stir and mix evenly until the ethanol is evaporated, stand in the 60℃ water bath for 30 minutes, then transfer the above mixed and dissolved liquid without bubbles into the liquid storage cylinder, control the drop speed under the condition of 80-85℃, drop into the condensate drop by drop, wait for complete condensation, pour off the condensate, collect the dripping pills, drain and remove the condensate on the pills with filter paper, and place in a silica gel dryer or dry naturally.

[0078] Example 13:

[0079] Tablets: 10mg of one or any of parispory lily rhizome saponin A (PFV92), parispory lily rhizome saponin B (PFE52), parispory lily rhizome saponin A (PFE53), parispory lily rhizome saponin D (PFE56), parispory lily rhizome saponin C (PFE59A), 180mg of lactose, 55mg of starch, and 5mg of magnesium stearate;

[0080] Preparation method: mix one or any of parispory lily rhizome saponin A (PFV92), parispory lily rhizome saponin B (PFE52), parispory lily rhizome saponin A (PFE53), parispory lily rhizome saponin D (PFE56), parispory lily rhizome saponin C (PFE59A), lactose and starch, uniformly wet with water, sieve and dry the wet mixture, sieve again, add magnesium stearate, then press the mixture into tablets, each weighing 250mg, with a compound content of 10mg.

[0081] Example 14:

[0082] Ampoules: 2mg of one or any of parispory lily rhizome saponin A (PFV92), parispory lily rhizome saponin B (PFE52), parispory lily rhizome saponin A (PFE53), parispory lily rhizome saponin D (PFE56), parispory lily rhizome saponin C (PFE59A), and 10mg of sodium chloride;

[0083] Preparation method: dissolve one or any of parispory lily rhizome saponin A (PFV92), parispory lily rhizome saponin B (PFE52), parispory lily rhizome saponin A (PFE53), parispory lily rhizome saponin D (PFE56), parispory lily rhizome saponin C (PFE59A), and sodium chloride in an appropriate amount of water for injection, filter the resulting solution, and fill it into ampoules under sterile conditions.

[0084] Example 15:

[0085] Capsules: 10 mg of any one of or any combination of Parisporyae Rhizoma Saponin A (PFV92), Parisporyae Rhizoma Saponin B (PFE52), Parisporyae Rhizoma Saponin A (PFE53), Parisporyae Rhizoma Saponin D (PFE56), Parisporyae Rhizoma Saponin C (PFE59A), 187 mg of lactose, and 3 mg of magnesium stearate;

[0086] Preparation: any one of or any combination of Parisporyae Rhizoma Saponin A (PFV92), Parisporyae Rhizoma Saponin B (PFE52), Parisporyae Rhizoma Saponin A (PFE53), Parisporyae Rhizoma Saponin D (PFE56), Parisporyae Rhizoma Saponin C (PFE59A) is mixed with an auxiliary agent, sieved, uniformly mixed, and the obtained mixture is filled into hard gelatin capsules, each weighing 200 mg, with an active ingredient content of 10 mg.

[0087] Example 16:

[0088] Ointment: An oily substance (e.g., vaseline, stearic acid) is heated to about 80 degrees to melt, filtered with a fine cloth; another water-soluble ingredient (e.g., borax, sodium hydroxide, triethanolamine, lauryl sulfate, a humectant, a preservative, etc.) is dissolved in water, and when the temperature is slightly higher than that of the oil phase, the water-soluble phase is slowly added to the oil phase while stirring, to prepare an emulsion base. The drug is added according to any one of or any combination of Parisporyae Rhizoma Saponin A (PFV92), Parisporyae Rhizoma Saponin B (PFE52), Parisporyae Rhizoma Saponin A (PFE53), Parisporyae Rhizoma Saponin D (PFE56), Parisporyae Rhizoma Saponin C (PFE59A), and stirred until condensation. Topical use.

[0089] Alternatively, any one of or any combination of Parisporyae Rhizoma Saponin A (PFV92), Parisporyae Rhizoma Saponin B (PFE52), Parisporyae Rhizoma Saponin A (PFE53), Parisporyae Rhizoma Saponin D (PFE56), Parisporyae Rhizoma Saponin C (PFE59A) is taken, and an ointment is obtained according to the conventional preparation method of an ointment, for topical use.

[0090] Example 17:

[0091] Cream formulation containing any one of or any combination of Parisporyae Rhizoma Saponin A (PFV92), Parisporyae Rhizoma Saponin B (PFE52), Parisporyae Rhizoma Saponin A (PFE53), Parisporyae Rhizoma Saponin D (PFE56), Parisporyae Rhizoma Saponin C (PFE59A) (W%):

[0092] Parisporyae Rhizoma Saponin A (PFV92), Parisporyae Rhizoma Saponin B (PFE52), Parisporyae Rhizoma Saponin A (PFE53), Parisporyae Rhizoma Saponin D (PFE56), Parisporyae Rhizoma Saponin C (PFE59A)

[0093] The cosmetic products of the above-mentioned formulations of the present application are prepared by the conventional method of preparing cosmetic products.

[0094] Example 18:

[0095] Emulsion formulation (W%) containing any one or any combination of Paridisin A (PFV92), Paridisin B (PFE52), Paridisin A (PFE53), Paridisin D (PFE56), Paridisin C (PFE59A) of Paris polyphylla var. chondosperma:

[0096] Paridisin A (PFV92), Paridisin B (PFE52), Paridisin A (PFE53), Paridisin D (PFE56), Paridisin C (PFE59A) of Paris polyphylla var. chondosperma

[0097]

[0098] The cosmetic products of the above-mentioned formulations of the present application are prepared by the conventional method of preparing cosmetic products.

Claims

1. A compound of the following structural formula: PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59A, PFE 52, PFE 53, PFE 56, PFE 59 。 2. The method for preparing the compound Paris polyphylla saponin A, i.e., PFV92, as described in claim 1, is characterized in that, ​ The n-butanol extracted part was subjected to gradient elution using 200-300 mesh normal phase silica gel column chromatography with CH2Cl2-MeOH (10:1-0:1) as eluent. The eluate was detected by thin layer chromatography and combined into 7 parts: Fr. I, Fr. II, Fr. III, Fr. IV, Fr. V, Fr. VI and Fr. VII. Fr. VII was subjected to gradient elution using 200-300 mesh normal phase silica gel column chromatography with EtOAc-MeOH (10:1-0:1) as eluent. The eluate was detected by thin layer chromatography and combined into 4 parts: Fr. VII-1, Fr. VII-2, Fr. VII-3 and Fr. VII-4. 10.0 g of Fr. VII-3 was subjected to RP C 18 18 column chromatography with MeOH-H2O (10:90-100:0) as eluent and divided into 5 parts: Fr. VII-3-1, Fr. VII-3-2, Fr. VII-3-3, Fr. VII-3-4 and Fr. VII-3-5. Fr. VII-3-3 was subjected to Sephadex LH-20 gel column chromatography with MeOH as eluent and then semi-preparative HPLC, Agilent Zorbax SB-C 18 18, ϕ 9.4 × 250 mm; MeCN-H2O, 43:57; v = 2 mL / min, to purify to obtain Chongye Chonglou Saponin A, i.e. PFV92.

3. A pharmaceutical composition, characterized by: ​ 4. Cosmetic product, characterized in that: ​ ​ ​ ​ 。 ​ 。

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