High adamantane-type phloroglucinol compounds, their preparation methods and applications

Abstract

The present invention belongs to the field of pharmaceutical technology, and specifically discloses a high adamantane type phloroglucinol compound, a preparation method thereof and an application thereof; the high adamantane type phloroglucinol compound Hyperichoisin C provided by the present invention has remarkable antidepressant activity, which is superior to the positive drugs fluoxetine and St. John's wort extract, and can be used for developing drugs for treating depression.

Description

Technical Field

[0001] The present invention belongs to the technical field of medicine, and more specifically, relates to a preparation method of a homoadamantane-type phloroglucinol compound, a pharmaceutical composition, and an application in antidepressant drugs. Background Art

[0002] The phloroglucinol components of the homoadamantane type are phloroglucinol components containing a homoadamantane carbon framework in the parent nucleus skeleton. These components have a "diamond-like" cage core and a common tricyclo[4.3.1.1]undecane core skeleton. This kind of skeleton is formed by an intramolecular cyclization reaction in which the isoprenyl substituent side chain C-7a at the C-7 position attacks C-3 on the parent nucleus based on the bicyclic polyisoprenyl-substituted acylphloroglucinol skeleton to form a cage-like adamantane type.

[0003] The inventor isolated the homoadamantane-type phloroglucinol compound Hyperichoisin C from the dried aerial parts of Hypericum perforatum, and found that it has significant antidepressant activity, superior to the positive drugs Fluoxetine and Neurostan, and can be used for the treatment of depression. Summary of the Invention

[0004] Aiming at the deficiencies of the existing technology, the present invention provides a homoadamantane-type phloroglucinol compound, its preparation method, a pharmaceutical composition, and an application.

[0005] To achieve the above object, the technical solution of the present invention is as follows:

[0006] A homoadamantane-type phloroglucinol compound (abbreviated as compound Hyperichoisin C) has the structure shown in the following formula (1):

[0007]

[0008] Among them, the tautomer of the phloroglucinol compound or a pharmaceutically acceptable salt also belongs to the protection scope of the present invention.

[0009] Another aspect of the present invention also provides a preparation method of the above phloroglucinol compound Hyperichoisin C, and the phloroglucinol compound is obtained by extraction and separation from Hypericum perforatum.

[0010] In the above technical solution, the preparation method of the phloroglucinol compound includes the following steps:

[0011] S1. Take the dried aerial parts of Hypericum perforatum, add a solvent for reflux extraction, combine the extraction solutions and concentrate to obtain an extract;

[0012] S2. After adding the extract into water and suspending it, extract with petroleum ether to obtain an extract.

[0013] S3. Use a petroleum ether - ethyl acetate solution for gradient elution of the extract through silica gel column chromatography to obtain fraction C; when using silica gel thin - layer chromatography for detection, the Rf value corresponding to fraction C is 0.75 - 0.77.

[0014] S4. Use a petroleum ether - ethyl acetate mixed solution for gradient elution of fraction C through silica gel column chromatography to obtain fraction C4; when using silica gel thin - layer chromatography for detection, the Rf value corresponding to fraction C4 is 0.72 - 0.74.

[0015] S5. Use a methanol - aqueous solution for gradient elution of fraction C4 through MCI column chromatography to obtain fraction C4C; when using silica gel thin - layer chromatography for detection, the Rf value corresponding to fraction C4C is 0.58 - 0.62.

[0016] S6. Use HPLC to separate from fraction C4C a homoadamantane - type phloroglucinol compound having the structure shown in formula (1); the mobile phase used in the HPLC method is a methanol - water mixed solution.

[0017] In this application, the Rf value is the Rf value of the fluorescent spots observed at scanning wavelengths of 254 nm and 365 nm.

[0018] Further, in step S6, further separate fraction C4C according to the characteristic ultraviolet absorption of phloroglucinol (λ max = 220, 280 nm) observed by HPLC analysis; the retention time of the obtained phloroglucinol compound is 28 - 33 min.

[0019] Further, in step S6, the volume ratio of methanol to water in the methanol - water mixed solution is (83:17) - (87:13); preferably 85:15.

[0020] Further, in step S3, based on a total volume of 100 as a reference, the volume ratio of petroleum ether to ethyl acetate in the petroleum ether - ethyl acetate solution when eluting fraction C (i.e., corresponding to fraction C) is (75:25) - (85:15); preferably, the volume ratio of petroleum ether to ethyl acetate is 80:20.

[0021] Further, in step S4, based on a total volume of 100 as a reference, the volume ratio of petroleum ether to ethyl acetate in the petroleum ether - ethyl acetate mixed solution when eluting fraction C4 (i.e., corresponding to fraction C4) is (65:35) - (75:25); preferably, the volume ratio of petroleum ether to ethyl acetate is 70:30.

[0022] Further, in step S5, the volume ratio of methanol to water in the methanol-aqueous solution when washing out fraction C4C (i.e., corresponding to fraction C4C) is (77:23)-(85:15); preferably, the volume ratio of methanol to water is 80:20.

[0023] Further, in step S1, the solvent is an ethanol aqueous solution of 88-98V%, and its addition mass is 8-10 times that of Hypericum perforatum, the number of reflux extractions is 1-3, and each extraction is for 1-3 h.

[0024] Further, in step S2, the extract is added to water with a mass 8-15 times that of the extract and suspended, and then extracted with petroleum ether.

[0025] Another aspect of the present invention also provides a pharmaceutical composition containing the above phloroglucinol compound Hyperichoisin C.

[0026] Further, the pharmaceutical composition includes a synergist and a pharmaceutically acceptable carrier or excipient.

[0027] That is to say, a pharmaceutical composition containing the high adamantane-type phloroglucinol compound of the present invention as an active ingredient and a conventional pharmaceutical excipient or adjuvant or carrier is also included in the present invention.

[0028] Further, the synergist is one or several of the following substances:

[0029] Fluoxetine, paroxetine, fluvoxamine, sertraline, citalopram, escitalopram, venlafaxine, duloxetine, mirtazapine, bupropion, agomelatine, trazodone, reboxetine, imipramine, amitriptyline, clomipramine, doxepin, maprotiline, moclobemide, Shugan Jieyu Capsule, Hypericum perforatum extract, Flupentixol Melitracen.

[0030] Further, the dosage form of the pharmaceutical composition is tablets, capsules, granules, oral liquids, extracts, dripping pills or pellets.

[0031] Another aspect of the present invention also provides the use of the above phloroglucinol compound Hyperichoisin C in anti-depressant drugs.

[0032] Compared with the prior art, the present invention has the following advantages:

[0033] (1) The present invention provides a method for extracting the high adamantane-type phloroglucinol compound Hyperichoisin C from Hypericum perforatum with simple operation, good reproducibility and high extraction purity;

[0034] (2) Experimental results show that the high adamantane type phloroglucinol compound Hyperichoisin C provided by the present invention has a neuroprotective effect on corticosterone-induced SH-SY5Y cells, which is superior to the positive drugs fluoxetine and St. John's wort, and can be used for developing drugs for treating depression. Description of the Drawings

[0035] Figure 1 1H NMR spectrum (400 MHz, CDCl3) of Hyperichoisin C prepared in Example 1 of the present invention; 1

[0036] Figure 2 13C NMR spectrum (100 MHz, CDCl3) of Hyperichoisin C prepared in Example 1 of the present invention. 13 Detailed Embodiments

[0037] In order to make the objectives, technical solutions and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention. In the embodiments, unless otherwise specified, the means used are all conventional means in the art. The term "comprising", "including" or any other variation thereof used herein is intended to cover a non-exclusive inclusion.

[0038] For those technical or conditions not specified in the embodiments, they shall be in accordance with the technologies or conditions described in the literature in this field or in accordance with the product specifications. For the reagents or instruments whose manufacturers are not specified, they are all conventional products that can be obtained through regular channels.

[0039] In the ranges disclosed herein, the endpoints and any values are not limited to the exact ranges or values. These ranges or values should be understood to include values close to these ranges or values. For numerical ranges, between the endpoint values of each range, between the endpoint values and individual point values of each range, and between individual point values, they can be combined with each other to obtain one or more new numerical ranges, and these numerical ranges should be regarded as specifically disclosed herein.

[0040] In addition, the technical features involved in the various embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0041] Example 1

[0042] This example discloses a preparation method of a high adamantane type phloroglucinol compound, which includes the following steps:

[0043] ​​S1. Take the dried aerial parts of Hypericum perforatum, add a solvent for reflux extraction, combine the extraction solutions and concentrate to obtain an extract. Among them, the solvent is an aqueous ethanol solution with 88 - 98V% ethanol, and its added mass is 8 - 10 times that of Hypericum perforatum. The number of times of reflux extraction is 1 - 3, and each extraction lasts for 1 - 3 h.

[0044] Preferably, use 10 times the mass of Hypericum perforatum of 95% aqueous ethanol solution as the solvent for reflux extraction three times, each extraction for 2 h, combine the extraction solutions and concentrate to obtain an extract.

[0045] S2. Suspend the extract in 8 - 15 times the mass of water and then extract with petroleum ether to obtain an extract. Further preferably, suspend the extract in 10 times the mass of water and then extract with 1.5 times the volume of petroleum ether of the suspension for 3 times to obtain a petroleum ether extract.

[0046] S3. Use a petroleum ether - ethyl acetate solution for gradient elution of the extract through a silica gel column chromatography to obtain fraction C. When detected by silica gel thin - layer chromatography, the Rf value corresponding to fraction C is 0.75 - 0.77. Based on a total volume of 100, the volume ratio of petroleum ether to ethyl acetate in the petroleum ether - ethyl acetate solution when eluting fraction C (i.e., corresponding to fraction C) is (75:25) - (85:15); preferably, the volume ratio of petroleum ether to ethyl acetate is 80:20.

[0047] It should be noted that during gradient elution, petroleum ether - ethyl acetate solutions with a volume ratio of (75:25) - (85:15) can all elute fraction C. When detected by silica gel thin - layer chromatography, the fraction with an Rf value of 0.75 - 0.77 is fraction C. Among them, the petroleum ether - ethyl acetate solution with a volume ratio of 80:20 has the highest elution efficiency. The principle of "preferably" in the following gradient elution process is the same.

[0048] The specific operations include: subjecting the petroleum ether extract to silica gel column chromatography and eluting it with a petroleum ether-ethyl acetate gradient with volume ratios of 100:0, 90:10, 80:20, 60:40, 50:50, 40:60, 30:70, 20:80, 10:90, and 0:100 to successively obtain 56 fractions numbered 1-8, 9-18, 19-25, 26-29, 30-40, 41-44, 45-47, 48-51, 52-53, and 54-56. Then, silica gel thin-layer chromatography was used for identification. According to the Rf values observed at 254 nm and 365 nm, which were 0.81-0.83 (fractions 1-8), 0.78-0.80 (fractions 9-18), 0.75-0.77 (fractions 19-25), 0.72-0.74 (fractions 26-29), 0.65-0.68 (fractions 30-40), 0.64-0.63 (fractions 41-44), 0.57-0.62 (fractions 45-47), 0.52-0.54 (fractions 48-51), 0.46-0.47 (fractions 52-53), and 0.35-0.39 (fractions 54-56), similar fractions were combined to successively obtain 10 fractions, named A, B, C, D, E, F, G, H, I, and J respectively; among them, fractions 19-25 were combined to obtain fraction C.

[0049] S4. Use a petroleum ether-ethyl acetate mixed solution to perform gradient elution on fraction C through silica gel column chromatography to obtain fraction C4; when using silica gel thin-layer chromatography for identification, the Rf value corresponding to fraction C4 is 0.72-0.74; the volume ratio of petroleum ether to ethyl acetate in the petroleum ether-ethyl acetate mixed solution when eluting fraction C4 (i.e., corresponding to fraction C4) is (65:35)-(75:25); preferably, the volume ratio of petroleum ether to ethyl acetate is 70:30.

[0050] The specific steps include: based on the characteristic ultraviolet absorption of phloroglucinol observed by HPLC analysis (λ max= 220, 280 nm), fraction C was selected for the next separation. Fraction C was subjected to silica gel column chromatography and eluted with a gradient of petroleum ether - ethyl acetate with volume ratios of 100:0, 90:10, 80:20, 70:30, 60:40, 40:60, 50:50, 30:70, 20:80, 10:90, 0:100, successively obtaining 111 fractions numbered 1 - 10, 11 - 27, 28 - 35, 36 - 56, 57 - 61, 62 - 65, 66 - 70, 71 - 75, 76 - 80, 81 - 90, 91 - 111. Then, silica gel thin - layer chromatography was used for identification. Based on the Rf values of the fluorescent spots observed at wavelengths of 254 nm and 365 nm, which were 0.85 - 0.82 (fraction 1 - 10), 0.79 - 0.81 (fraction 11 - 27), 0.75 - 0.78 (fraction 28 - 35), 0.72 - 0.74 (fraction 36 - 56), 0.68 - 0.70 (fraction 57 - 61), 0.65 - 0.67 (fraction 62 - 65), 0.63 - 0.64 (fraction 66 - 70), 0.60 - 0.62 (fraction 71 - 75), 0.50 - 0.52 (fraction 76 - 80), 0.45 - 0.49 (fraction 81 - 90), 0.38 - 0.39 (fraction 91 - 111), the fractions were combined to obtain 11 fractions C1 - C11; among them, fractions 36 - 56 were combined to obtain fraction C4.

[0051] S5. Fraction C4 was subjected to gradient elution with a methanol - aqueous solution on an MCI column chromatography to obtain fraction C4C; when using silica gel thin - layer chromatography for identification, the Rf value corresponding to fraction C4C was 0.58 - 0.62; the volume ratio of methanol to water in the methanol - aqueous solution when eluting fraction C4C (i.e., corresponding to fraction C4C) was (77:23) - (85:15); preferably, the volume ratio of methanol to water was 80:20.

[0052] The specific operation is as follows:

[0053] Based on the characteristic ultraviolet absorption of phloroglucinol observed by HPLC analysis (λ max = 220, 280 nm), C4 was selected for further separation.

[0054] The fraction C4 was subjected to MCI column chromatography and eluted with a methanol-water gradient of volume ratios 70:30, 75:25, 80:20, 85:15, 90:10, 95:5, and 100:0. A total of 35 fractions were successively collected, namely fractions 1-3, 4-10, 11-17, 18-23, 24-25, 26-30, and 31-35. They were identified by silica gel thin-layer chromatography. Based on the Rf values of the fluorescent spots observed at wavelengths 254 nm and 365 nm, which were 0.31 - 0.34 (fraction 1-3), 0.47 - 0.49 (fraction 4-10), 0.58 - 0.62 (fraction 11-17), 0.65 - 0.69 (fraction 18-23), 0.70 - 0.73 (fraction 24-25), 0.76 - 0.78 (fraction 26-30), and 0.80 - 0.82 (fraction 31-35), they were combined to obtain 7 fractions, namely C4A - C4G. Among them, fraction 11-17 was combined to obtain fraction C4C.

[0055] S6. The high adamantane-type phloroglucinol compound with the structure shown in formula (1) was isolated from fraction C4C by HPLC. The mobile phase used in the HPLC method was a methanol-water mixed solution. The volume ratio of methanol to water in the methanol-water mixed solution was (83:17) - (87:13).

[0056] Fraction C4C was selected for the next separation based on the characteristic ultraviolet absorption of phloroglucinol (λ max = 220, 280 nm) observed by HPLC analysis; the retention time of the obtained adamantane-type phloroglucinol compound was 28 - 33 min;

[0057] Further preferably, the volume ratio of methanol to water in the methanol-water mixed solution was 85︰15. The retention time of compound Hyperichoisin C prepared from fraction C4C was 31 min (purity 99%).

[0058] In this application, the Rf value is the Rf value of the fluorescent spots observed at scanning wavelengths of 254 nm and 365 nm.

[0059] For the compound Hyperichoisin C prepared in Example 1, its structure, physical properties, and detection data are as follows:

[0060] Colorless oily liquid. HR-ESI-MS m / z: 587.3692 [M+H] + , indicating that the molecular formula of this compound is C 32 H 46 O5.

[0061] As Figure 1 shown, at 1In the ¹H NMR (CDCl₃, 400 MHz) spectrum, δ H 7.40 (1H, t, J = 7.4 Hz, H-10d), 7.29 (2H, m, H-10c / e), 7.07 (2H, d, J = 7.3 Hz, H-10b / f) are the proton signals on the benzene ring; 5.25 (1H, t, J = 7.4 Hz, H-5b), 5.06 (1H, t, J = 6.5 Hz, H-5g) are the proton signals on the two double bonds; δ H 2.75 (1H, dd, J = 12.6, 7.5 Hz, H-3a), 2.23 (1H, d, J = 13.3 Hz, H-3a'), 2.18 (1H, m, H-3b) are the proton signals of the methylene and methine on the five-membered ring; δ H 2.61 (1H, m, H-5a), 2.60 (1H, m, H-5a'), 2.49 (1H, dd, J = 14.7, 6.8 Hz, H-6), 2.19 (1H, m, H-6'), 2.05 (5H, m, H-5d / f, 7a), 1.75 (1H, m, H-7a') are the proton signals of the methylene on the ring and on the side chain; δ H 2.29 (1H, J = 12.0, 7.5 Hz, H-7b), 1.75 (1H, m, H-7) are the proton signals of the methine; δ H 1.66 (6H, s, H-5i / e), 1.59 (3H, s, H-5j), 1.41 (1H, s, H-11), 1.39 (1H, s, H-12), 1.36 (6H, s, H-3d / e), 1.21 (1H, s, H-7e), 1.07 (1H, s, H-7d) are the proton signals of the methyl groups.

[0062] As Figure 2 shown, in the 13 ¹³C NMR (CDCl₃, 100 MHz) spectrum, δ C 205.8 (C-9), 204.6 (C-4), 204.4 (C-2), 193.0 (C-10) are the four carbonyl carbon signals; δ C 135.1 (C-10a), 132.4 (C-10d), 128.6 (C-10b / f), 128.4 (C-10c / e) are the carbon signals on the benzene ring; δ C 139.2 (C-5c), 131.6 (C-5h), 124.3 (C-5g), 118.9 (C-5b) are the double bond carbon signals on the side chain; δ C81.7 (C-1), 73.4 (C-3c), 73.3 (C-3), 67.9 (C-5), 47.8 (C-8), 46.9 (C-7c) are quaternary carbon signals; δ C 59.2 (C-3b), 54.9 (C-7b), 42.5 (C-7) are quaternary carbon signals on the ring; δ C 40.2 (C-5d), 35.3 (C-6), 31.3 (C-3a), 29.2 (C-5a), 26.7 (C-5f), 25.0 (C-7a) are secondary carbon signals; Another δ C 31.8 (C-3e), 30.5 (C-3d), 27.7 (C-7d), 27.1 (C-7e), 25.9 (C-5i), 25.3 (C-11), 22.1 (C-12), 17.8 (C-5j), 16.5 (C-5e) are methyl carbon signals. The compound was identified as Hyperichoisin C based on the above data.

[0063] 1 H NMR, 13 The signal assignments of 13C NMR are shown in Table 1.

[0064] Table 1 NMR data of Compound 1 δ (ppm)

[0065]

[0066]

[0067] In summary, the structure of the new compound 1 (Compound Hyperichoisin C) is determined as follows:

[0068]

[0069] Comparative Example 1

[0070] Comparative Example 1 of the present invention provides a method for preparing a high adamantane type phloroglucinol compound. The steps are similar to those in Example 1, except that in step S5, when fraction C4 is passed through an MCI column chromatography, methanol-water is used for elution, and the volume ratio of methanol to water is less than (77:23) or greater than (85:15).

[0071] The results showed that Compound 1 could not be prepared.

[0072] Comparative Example 2

[0073] Comparative Example 2 of the present invention provides a method for preparing a high adamantane-type phloroglucinol compound, and its steps are similar to those of Example 1, except that in step S6, in the mobile phase used for separating fraction C4C by HPLC method, the volume ratio of methanol to water is less than (83:17) or greater than (87:13).

[0074] The results showed that compound 1 could not be prepared.

[0075] To better understand the essence of the present invention, the following describes the new applications of the above phloroglucinol compounds in the pharmaceutical field in combination with pharmacological tests and results.

[0076] Test Example

[0077] This test example discloses an antidepressant activity experiment of the above compound Hyperichoisin C in corticosterone-induced SH-SY5Y cell (a common cell model for antidepressants) injury.

[0078] (1) Experimental materials and instruments

[0079] Human neuroblastoma SH-SY5Y was purchased from the Institute of Basic Medicine, Chinese Academy of Medical Sciences;

[0080] DMEM medium, PBS buffer, fetal bovine serum (FBS) and trypsin were all purchased from Gibco, USA;

[0081] Fluoxetine (Shanghai Yuanye Bio-Technology Co., Ltd.);

[0082] Neurostan (Dr. Willmar Schwabe Pharmaceuticals GmbH, Germany);

[0083] CCK-8 cell viability detection kit (Elabscience);

[0084] Corticosterone, DMSO (Sigma, USA);

[0085] 96-well plates (Corning, USA);

[0086] Carbon dioxide cell incubator (Thermo Fisher Scientific, USA);

[0087] Full wavelength microplate reader (Thermo Fisher Scientific, USA);

[0088] BIOFUGE STRATOS centrifuge (Thermo Fisher Scientific, USA);

[0089] IX73 inverted electron microscope (Olympus, Japan);

[0090] Pipette (Eppendorf, Germany);

[0091] Electronic balance (Sartorius, Germany);

[0092] Ultra-clean workbench, centrifuge tubes, pipettes and other related consumables.

[0093] (2) Cell resuscitation and culture

[0094] Take out the cryopreservation tube of SH-SY5Y cells marked in liquid nitrogen, immediately put it into a 37°C water bath, and try to complete rapid thawing and melting within 1 min; after disinfecting the cryopreservation tube with alcohol, transfer the cryopreservation solution in the tube to a 15 mL sterile centrifuge tube, add the corresponding culture medium, mix well, and then centrifuge to remove the supernatant; repeat the above steps once for washing, then add 10 mL of culture medium to mix and resuspend the cells, and then transfer them to a 10 mL culture dish and place them in a constant temperature incubator at 37°C and 5% CO2 for culture.

[0095] When the cell density reaches 70%-80%, passage the cells; first aspirate the old culture medium, add PBS and wash 2 times, add trypsin containing EDTA to the culture dish and place it in a 37°C incubator for 3 min, then add culture medium to terminate digestion, centrifuge at 1000 rpm for 5 min, aspirate the digestion solution, add culture medium containing serum again and pipette the cells repeatedly to form a cell suspension, and finally transfer it to a new culture dish according to the number of cells to complete cell passage.

[0096] (3) Drug administration and evaluation of cell viability by CCK-8 method

[0097] Seed SH-SY5Y cells at 1×10 5 cells / well in a 96-well plate, with 6 replicates per group, 80 μL / well. After plating, culture for 24 hours. Dissolve and dilute the positive drugs fluoxetine, hypericum perforatum, hypericin, and tetrahydrohypericin to 10 μM, and dissolve and dilute compound Hyperichoisin C to 5 μM and 10 μM respectively. Add 10 μL / well to treat the cells for 1 h, then add 10 μL of corticosterone at a concentration of 25 μM, culture at 37°C for 48 h, then add 10 μL of CCK-8, and after 1 h, use an enzyme-linked immunosorbent assay (ELISA) reader to detect the absorbance at 450 nm and calculate the cell survival rate.

[0098] Cell survival rate (%) = [(Acontrol - Asample) / (Acontrol - Ablank)] × 100%.

[0099] (4) Experimental results

[0100] The experimental results (Table 2) showed that, compared with the model group, low and high doses (5 μM, 10 μM) of Hyperichoisin C had significant protective effects on corticosterone-induced SH-SY5Y neuronal cell damage. The neuroprotective activity of low-dose Hyperichoisin C was significantly better than that of the positive drugs St. John's wort extract (the first natural antidepressant plant drug in the world, 10 μM), hypericin (10 μM), and tetrahydrohypericin (10 μM); the neuroprotective activity of high-dose Hyperichoisin C was better than that of the positive drug fluoxetine (a first-line chemical drug for clinical treatment of depression, 10 μM).

[0101] Table 2 Neuroprotective effects of compounds on corticosterone-induced SH-SY5Y cell damage( n = 5)

[0102] Group Concentration (μM) Cell survival rate (%) Blank 0 100.00±7.89 Model 0 <![CDATA[52.49±2.10 #### > Fluoxetine 10 <![CDATA[65.79±2.33 ** > St. John's Wort 10 <![CDATA[60.03±3.33 * > Hypericin 10 55.83±2.71 Hypohypericin 10 <![CDATA[58.03±3.33 * > Hyperichoisin C 5 <![CDATA[62.52±3.59 ** > Hyperichoisin C 10 <![CDATA[67.52±6.59 *** >

[0103] Note: Compared with the blank group: #### P < 0.0001; compared with the model group: * P < 0.05, ** P < 0.01, *** P < 0.001

[0104] In summary, Hyperichoisin C described in the present invention exhibits significant antidepressant activity and is superior to the first-line antidepressant drugs fluoxetine and St. John's wort extract, and can be used as a drug for the treatment of depression.

[0105] Application Example 1

[0106] The application example of the present invention discloses a capsule using Hyperichoisin C as the raw material drug, and its components are as follows:

[0107] Hyperichoisin C 18.0 mg

[0108] Starch 6.0 g

[0109] Sodium metabisulfite 0.2 g

[0110] Magnesium stearate 0.2 g

[0111] Absolute ethanol appropriate amount

[0112] Manufactured into 100 capsules.

[0113] The specific preparation process is as follows:

[0114] Take Hyperichoisin C, starch, and sodium metabisulfite and mix them evenly, add absolute ethanol to make soft materials, pass through a 24-mesh sieve, make granules, dry, add magnesium stearate, mix evenly, and fill into capsules.

[0115] Application Example 2

[0116] This application example of the present invention discloses a granule with compound Hyperichoisin C as the raw material medicine, and its components are as follows:

[0117] Hyperichoisin C 35.0mg

[0118] Starch 6.0g

[0119] Sodium bisulfite 0.2g

[0120] Magnesium stearate 0.2g

[0121] Absolute ethanol appropriate amount

[0122] Manufactured into 100 bags.

[0123] The specific preparation process is as follows:

[0124] Take Hyperichoisin C, mix it with starch and sodium bisulfite, add absolute ethanol to make soft material, pass through a 24-mesh sieve, make granules, dry, add magnesium stearate, mix well, and bag.

[0125] Application Example 3

[0126] This application example of the present invention discloses an oral liquid with compound Hyperichoisin C as the raw material medicine, and its components are as follows:

[0127] Hyperichoisin C 25.0 mg

[0128]

[0129]

[0130] The specific preparation process is as follows:

[0131] After mixing the above components, adopt the conventional preparation method for oral liquid and subpackage it.

[0132] Application Example 4

[0133] This application example of the present invention discloses an injection with compound Hyperichoisin C as the raw material medicine, and its components are as follows:

[0134] Hyperichoisin C 45.0 mg

[0135] Vitamin C 0.2g

[0136] Sodium chloride 6.0g

[0137] Sodium bicarbonate 0.1 g

[0138] Water for injection 1000 mL

[0139] Manufacture 100 ampoules

[0140] The specific preparation process is as follows:

[0141] After mixing the above components, using the conventional preparation method for injections, 100 ampoules can be obtained.

[0142] Application Example 5

[0143] The application example of the present invention discloses a tablet using compound Hyperichoisin C and fluoxetine as raw materials, and its components are as follows:

[0144] Hyperichoisin C 20.0 mg

[0145] Fluoxetine 5 g

[0146] Hydroxypropyl methylcellulose 18 g

[0147] Talcum powder 0.4 g

[0148] Lactose 0.2 g

[0149] Magnesium stearate 0.2 g

[0150] Absolute ethanol appropriate amount

[0151] Manufacture 100 tablets

[0152] The specific preparation process is as follows:

[0153] Take Hyperichoisin C, fluoxetine and hydroxypropyl methylcellulose, talcum powder, lactose, magnesium stearate, mix evenly, add absolute ethanol to make soft material, pass through a 24-mesh sieve, make granules, dry, add magnesium stearate, mix evenly, and press tablets.

[0154] Application Example 6

[0155] The application example of the present invention discloses a capsule using compound Hyperichoisin C and sertraline as raw materials, and its components are as follows:

[0156] Hyperichoisin C 18.0 mg

[0157] Sertraline 2.0 g

[0158] Starch 6.0 g

[0159] Sodium metabisulfite 0.2 g

[0160] Magnesium stearate 0.2 g

[0161] Anhydrous ethanol q.s.

[0162] Manufacture 100 capsules.

[0163] The specific preparation process is as follows:

[0164] Take Hyperichoisin C, sertraline, starch, and sodium metabisulfite, mix them evenly, add anhydrous ethanol to make soft material, pass through a 24-mesh sieve, make granules, dry, add magnesium stearate, mix evenly, and fill into capsules.

[0165] Application Example 7

[0166] The application example of the present invention discloses an injection using compound Hyperichoisin C and escitalopram as raw materials, and its components are as follows:

[0167] Hyperichoisin C 30.0 mg

[0168] Escitalopram 2.0 g

[0169] Vitamin C 0.2 g

[0170] Sodium chloride 6.0 g

[0171] Sodium bicarbonate 0.1 g

[0172] Water for injection 1000 mL

[0173] Manufacture 100 vials.

[0174] The specific preparation process is as follows:

[0175] After mixing the above components, using the conventional preparation method for injections, 100 vials can be obtained.

[0176] The above embodiments only represent several implementation manners of the present invention, and the description is relatively specific and detailed, but it should not be construed as a limitation to the scope of the invention patent.

[0177] It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several deformations and improvements can still be made, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the present invention patent should be subject to the appended claims.

Claims

1. A method for preparing a high adamantane-type phloroglucinol compound, characterized in that, The high adamantane type phloroglucinol compound has the structure shown in formula (1): （1）； The phloroglucinol compound is obtained by extraction and separation from Hypericum perforatum; The preparation method comprises the following steps: S1. Take the dried aerial part of Hypericum perforatum, add a solvent for reflux extraction, combine the extraction solutions and concentrate to obtain an extract; the solvent is an ethanol aqueous solution of 88 - 98 V%; S2. Suspend the extract in water and extract with petroleum ether to obtain an extract; S3. Use a petroleum ether - ethyl acetate solution for gradient elution of the extract by silica gel column chromatography to obtain fraction C; when detected by silica gel thin - layer chromatography, the Rf value corresponding to fraction C is 0.75 - 0.77; S4. Use a petroleum ether - ethyl acetate mixed solution for gradient elution of fraction C by silica gel column chromatography to obtain fraction C4; when detected by silica gel thin - layer chromatography, the Rf value corresponding to fraction C4 is 0.72 - 0.74; S5. Use a methanol - aqueous solution for gradient elution of fraction C4 by MCI column chromatography to obtain fraction C4C; when detected by silica gel thin - layer chromatography, the Rf value corresponding to fraction C4C is 0.58 - 0.62; the volume ratio of methanol to water in the methanol - aqueous solution for eluting fraction C4C is (77:23) - (85:15); S6. Use HPLC method to separate from fraction C4C the high adamantane type phloroglucinol compound having the structure shown in formula (1); The mobile phase used in the HPLC method is a methanol - water mixed solution; the volume ratio of methanol to water in the methanol - water mixed solution is (83:17) - (87:13).

2. The preparation method according to claim 1, characterized in that, In step S6, fraction C4C is selected for further separation according to the characteristic ultraviolet absorption of phloroglucinol observed by HPLC analysis; the retention time of the obtained adamantane type phloroglucinol compound is 28 - 33 min.

3. The preparation method according to claim 1, characterized in that, In step S3, based on the total volume of 100, the volume ratio of petroleum ether to ethyl acetate in the petroleum ether - ethyl acetate solution for eluting fraction C is (75:25) - (85:15).

4. The preparation method according to claim 1, wherein In step S3, based on the total volume of 100, the volume ratio of petroleum ether to ethyl acetate in the petroleum ether - ethyl acetate solution for eluting fraction C is 80:

20.

5. The preparation method according to claim 1, characterized in that, In step S4, the volume ratio of petroleum ether to ethyl acetate in the petroleum ether - ethyl acetate mixed solution for eluting fraction C4 is (65:35) - (75:25).

6. The preparation method according to claim 1, characterized in that, In step S4, the volume ratio of petroleum ether to ethyl acetate in the petroleum ether - ethyl acetate mixed solution for eluting fraction C4 is 70:

30.

7. The preparation method according to claim 1, wherein In step S5, the volume ratio of methanol to water in the methanol - aqueous solution for eluting fraction C4C is 80:

20.

8. The preparation method according to claim 1, wherein In step S1, the mass of the solvent added is 8 - 10 times that of Hypericum perforatum, the number of times of reflux extraction is 1 - 3, and each extraction is for 1 - 3 h.

9. The preparation method according to claim 1, wherein, In step S2, the extract is suspended in 8 - 15 times the mass of water and then extracted with petroleum ether.

10. Use of adamantane-type phloroglucinol compounds or pharmaceutical compositions comprising adamantane-type phloroglucinol compounds in the preparation of antidepressant drugs, characterized in that, The high adamantane type phloroglucinol compound has the structure shown in formula (1): （1）。 11. The application according to claim 10, wherein The pharmaceutical composition comprises a synergist and a pharmaceutically acceptable carrier or excipient.

12. The application according to claim 11, wherein, The synergist is one or several of the following substances: Fluoxetine, paroxetine, fluvoxamine, sertraline, citalopram, escitalopram, venlafaxine, duloxetine, mirtazapine, bupropion, agomelatine, trazodone, reboxetine, imipramine, amitriptyline, clomipramine, doxepin, maprotiline, moclobemide, Shugan Jieyu Capsule, Flupentixol Melitracen.

13. The application according to claim 10, wherein The dosage form of the pharmaceutical composition is tablet, capsule, granule, oral liquid, infusion, dropping pill or pellet.

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