Novel phloroglucinol compound Hyperforidiol F and its preparation method and application

Abstract

The invention belongs to the field of medical technology, and specifically discloses a novel phloroglucinol compound Hyperforidiol F and a preparation method and application thereof; the novel phloroglucinol compound Hyperforidiol F provided by the invention has significant antidepressant activity, is superior to positive drugs fluoxetine and rulotane, and can be used for the research and development of drugs for treating depression.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and more specifically, relates to a novel phloroglucinol compound Hyperforidiol F and a preparation method thereof, a pharmaceutical composition and application thereof in treating antidepressants. Background Art

[0002] Polycyclic polyisopentenyl-substituted acylphloroglucinol natural products are a class of distinctive natural products derived from polyketide and isopentenyl complex pathways and specifically distributed in plants of the broad family Guttiferae.

[0003] Hypericum perforatum is a perennial herb of the genus Hypericum of the family Garcinia, and is widely distributed in Hebei, Shanxi, Shaanxi, Gansu, Xinjiang, etc. In order to find a new drug for treating depression, the inventor isolated a new phloroglucinol compound Hyperforidiol F from the dried aerial parts of Hypericum perforatum, and found that it has significant antidepressant activity. Summary of the invention

[0004] In view of the deficiencies in the prior art, the present invention provides a novel phloroglucinol compound Hyperforidiol F and a preparation method and application thereof.

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

[0006] A novel phloroglucinol compound Hyperforidiol F has a structure as shown in the following formula (1):

[0007]

[0008] Among them, the tautomers or pharmaceutically acceptable salts of the novel phloroglucinol compound Hyperforidiol F also fall within the protection scope of the present invention.

[0009] Another aspect of the present invention provides a method for preparing the novel phloroglucinol compound Hyperforidiol F. The novel phloroglucinol compound Hyperforidiol F is obtained by extraction and separation from Hypericum perforatum.

[0010] Furthermore, the preparation method of the novel phloroglucinol compound Hyperforidiol F comprises the following steps:

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

[0012] S2, adding the extract to water for suspension, and then extracting with petroleum ether to obtain an extract;

[0013] S3, gradient eluting the extract by silica gel column chromatography using petroleum ether-ethyl acetate solution to obtain fraction E; when detected by silica gel thin layer chromatography, the Rf value corresponding to fraction E is 0.65-0.68;

[0014] S4, using a petroleum ether-ethyl acetate mixed solution to gradiently elute fraction E through silica gel column chromatography to obtain fraction E4; when detected by silica gel thin layer chromatography, the Rf value corresponding to fraction E4 is 0.63-0.69;

[0015] S5. Fraction E4 is subjected to gradient elution by MCI column chromatography using methanol-water solution to obtain fraction E4B. When fraction E4B is identified by silica gel thin layer chromatography, the Rf value corresponding to fraction E4B is 0.47-0.49;

[0016] S6. Separating the phloroglucinol compound Hyperforidiol F having a structure as shown in formula (1) from fraction E4B by HPLC;

[0017] The mobile phase used in HPLC was a mixed solution of acetonitrile and water.

[0018] This application uses acetonitrile-water as the mobile phase and adopts HPLC method using C 18 The chromatographic column prepared a novel phloroglucinol compound having a structural formula as shown in formula (1) in fraction E4B.

[0019] Further, in step S6, the volume ratio of acetonitrile to water in the acetonitrile-water mixed solution is (55:45)-(65:35). Preferably, the volume ratio of acetonitrile to water in the acetonitrile-water mixed solution is 62:38.

[0020] Further, in step S6, according to the characteristic ultraviolet absorption (λ max =220, 280nm) fraction E4B was further separated to obtain a novel phloroglucinol compound Hyperforidiol F having a structural formula as shown in formula (1) and a retention time of 40-50min.

[0021] Furthermore, in step S2, the extract is added to 8-15 times the mass of water for suspension, and then extracted with petroleum ether.

[0022] The Rf values ​​in this application refer to the Rf values ​​of the fluorescent spots observed at scanning wavelengths of 254 nm and 365 nm.

[0023] Further, 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 when fraction E is washed out (i.e. corresponding to fraction E) is (45:55)-(55:45); preferably, the volume ratio of petroleum ether to ethyl acetate is 50:50.

[0024] Furthermore, in step S4, the volume ratio of petroleum ether to ethyl acetate in the petroleum ether-ethyl acetate mixed solution when fraction E4 is washed out (i.e., corresponding to fraction E4) is (45:55)-(55:45); preferably, the volume ratio of petroleum ether to ethyl acetate is 50:50.

[0025] Furthermore, in step S5, the volume ratio of methanol to water in the methanol-water solution when fraction E4B is washed out (i.e., corresponding to fraction E4B) is (65:35)-(72:28); preferably, the volume ratio of methanol to water is 70:30.

[0026] Furthermore, in step S1, the solvent is 88-98V% ethanol aqueous solution; the mass of the added solvent is 8-10 times that of Hypericum perforatum, the number of reflux extractions is 1-3, and each extraction lasts 1-3 hours.

[0027] In another aspect, the present invention provides a pharmaceutical composition containing the novel phloroglucinol compound.

[0028] Furthermore, the pharmaceutical composition comprises a synergist and a pharmacodynamically acceptable carrier or excipient.

[0029] That is to say, the pharmaceutical composition containing the novel phloroglucinol compound of the present invention as an active ingredient and conventional pharmaceutical excipients or adjuvants or carriers is also included in the present invention.

[0030] Furthermore, the synergist is one or more of the following substances:

[0031] Fluoxetine, paroxetine, fluvoxamine, sertraline, citalopram, escitalopram, venlafaxine, duloxetine, mirtazapine, bupropion, agomelatine, trazodone, reboxetine, imipramine, amitriptyline, clomipramine, doxepin, maprotiline, moclobemide, Shugan Jieyu capsule, St. John's wort extract, flupentixol-melitracen.

[0032] Furthermore, the dosage form of the pharmaceutical composition is tablets, capsules, granules, oral liquid, granules, pills or micropills.

[0033] In another aspect, the present invention also provides the use of the novel phloroglucinol compound Hyperforidiol F and / or the above composition in antidepressant drugs.

[0034] Compared with the prior art, the present invention has the following advantages:

[0035] (1) The present invention provides a novel phloroglucinol compound Hyperforidiol F, which has not been reported so far. The experimental results show that the compound Hyperforidiol F has a significant protective effect on SH-SY5Y cells induced by corticosterone, which is significantly better than the positive drugs fluoxetine and ruxolitinib, and can be used to develop drugs for the treatment of depression.

[0036] (2) The preparation method of the new phloroglucinol compound Hyperforidiol F is simple to operate, has good reproducibility and high extraction purity. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] Figure 1 Hyperforidiol F prepared in Example 1 of the present invention 1 H NMR spectrum (400 MHz, CDCl3);

[0038] Figure 2 Hyperforidiol F prepared in Example 1 of the present invention 13 C NMR spectrum (100 MHz, CDCl3);

[0039] Figure 3 This is the HMBC spectrum of Hyperforidiol F prepared in Example 1 of the present invention. DETAILED DESCRIPTION

[0040] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with embodiments.

[0041] 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.

[0042] In the examples, unless otherwise specified, all the means used are conventional means in the art.

[0043] As used herein, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion. For example, a composition, step, method, article, or apparatus that comprises the listed elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, step, method, article, or apparatus.

[0044] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0045] The endpoints and any values ​​of the ranges disclosed in this article are not limited to the precise ranges or values, and these ranges or values ​​should be understood to include values ​​close to these ranges or values. For numerical ranges, the endpoint values ​​of each range, the endpoint values ​​of each range and the individual point values, and the individual point values ​​can be combined with each other to obtain one or more new numerical ranges, which should be considered as specifically disclosed in this article.

[0046] 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.

[0047] Example 1

[0048] This embodiment discloses a method for preparing a novel phloroglucinol compound Hyperforidiol F, comprising the following steps:

[0049] S1. Take the dried aerial part of Hypericum perforatum, add a solvent for reflux extraction, combine the extracts and concentrate to obtain an extract; specifically, the solvent is 88-98V% ethanol aqueous solution; the mass of the added solvent is 8-10 times that of Hypericum perforatum, the number of reflux extractions is 1-3, and each extraction is 1-3h.

[0050] Specific implementation plan is as follows: weigh the dry aerial part of Hypericum perforatum (208 kg), add 95% ethanol aqueous solution as solvent which is 10 times the mass of Hypericum perforatum, reflux and extract three times, each extraction is 2 hours, combine the extracts and concentrate to obtain an extract (about 9.0 kg).

[0051] S2, adding the extract to water for suspension, and then extracting with petroleum ether to obtain an extract;

[0052] Furthermore, in step S2, the extract is added to 8-15 times the mass of water for suspension, and then extracted with petroleum ether.

[0053] The specific steps are as follows: the extract (9.0 kg) is added to 10 times the mass of water (90 L) for suspension, and then extracted with 1.5 times the volume of petroleum ether of the suspension, and extracted 3 times to obtain a petroleum ether extract (about 3100 g).

[0054] S3. Use petroleum ether-ethyl acetate solution to gradient elute the extract through silica gel column chromatography to obtain fraction E; when detected by silica gel thin layer chromatography, the Rf value corresponding to fraction E is 0.65-0.68; based on the total volume of 100, the volume ratio of petroleum ether to ethyl acetate in the petroleum ether-ethyl acetate solution when fraction E is eluted (i.e., corresponding to fraction E) is (45:55)-(55:45); preferably, the volume ratio of petroleum ether to ethyl acetate is 50:50.

[0055] It should be noted that during gradient elution, petroleum ether-ethyl acetate solutions with a volume ratio of (45:55)-(55:45) can elute fraction E. Using silica gel thin layer chromatography, the fraction with an Rf value of 0.65-0.68 is fraction E. Among them, the elution efficiency of petroleum ether-ethyl acetate solution with a volume ratio of 50:50 is the highest. The principle of "preferably" described in the following gradient elution process is the same.

[0056] The Rf values ​​in this application refer to the Rf values ​​of the fluorescent spots observed at scanning wavelengths of 254 nm and 365 nm.

[0057] The specific steps are as follows: the petroleum ether extract is subjected to silica gel column chromatography, using a petroleum ether-ethyl acetate gradient elution with a volume ratio of 100:0, 90:10, 80:20, 60:40, 50:50, 40:60, 30:70, 20:80, 10:90 and 0:100, each 500 mL volume is one fraction, and 56 fractions 1-5, 6-10, 11-25, 26-29, 30-40, 41-44, 45-47, 48-51, 52-53, 54-56 are obtained in sequence. The Rf values ​​observed at 254 nm and 365 nm are 0.81-0.83 (fractions 1-5), 0. The fluorescent spot results of 78-0.80 (fraction 6-10), 0.75-0.77 (fraction 11-25), 0.72-0.74 (fraction 26-29), 0.65-0.68 (fraction 30-40), 0.64-0.63 (fraction 41-44), 0.57-0.62 (fraction 45-47), 0.52-0.54 (fraction 48-51), 0.46-0.47 (fraction 52-53), and 0.35-0.39 (fraction 54-56) were merged with similar fractions to obtain 10 fractions, which were named A, B, C, D, E, F, G, H, I and J respectively; among them, fraction E was obtained by merging fractions 30-40.

[0058] According to the characteristic ultraviolet absorption (λ max =220, 280 nm) and select fraction E for the next separation.

[0059] S4. Use a petroleum ether-ethyl acetate mixed solution to gradient elute fraction E through silica gel column chromatography to obtain fraction E4; when detected by silica gel thin layer chromatography, the Rf value corresponding to fraction E4 is 0.63-0.69; when eluting fraction E4 (i.e., the volume ratio of petroleum ether to ethyl acetate in the petroleum ether-ethyl acetate mixed solution (i.e., corresponding to fraction E4) is (45:55)-(55:45); preferably, the volume ratio of petroleum ether to ethyl acetate is 50:50.

[0060] The specific operation steps include: subjecting fraction E to silica gel column chromatography, using petroleum ether-ethyl acetate gradient elution with a volume ratio of 100:0, 80:20, 60:40, 50:50, 30:70, 20:80, 10:90, 0:100, to obtain fractions 1-14, 15-30, 31-55, 56-75, 76-77, 78-80, 81-100, 101-115, a total of 115 fractions, and then using silica gel thin layer chromatography to identify, according to the fluorescence spot Rf value observed at wavelengths of 254nm and 365nm The fractions E1 to E8 were respectively 0.79-0.85 (fraction 1-14), 0.75-0.78 (fraction 15-30), 0.71-0.74 (fraction 31-55), 0.63-0.69 (fraction 56-75), 0.50-0.62 (fraction 76-77), 0.45-0.49 (fraction 78-80), 0.35-0.44 (fraction 81-100), and 0.30-0.34 (fraction 101-115). Fractions 56-75 were combined to form fraction E4.

[0061] S5. Fraction E4 is gradient eluted using a methanol-water solution through MCI column chromatography to obtain fraction E4B; when detected by silica gel thin layer chromatography, the Rf value corresponding to fraction E4B is 0.47-0.49; the volume ratio of methanol to water in the methanol-water solution when fraction E4B is eluted (i.e., corresponding to fraction E4B) is (65:35)-(72:28); preferably, the volume ratio of methanol to water is 70:30.

[0062] The specific operation includes: according to the characteristic ultraviolet absorption (λ max =220, 280nm) E4 was selected for further separation. Fraction E4 was subjected to MCI column chromatography and gradient elution with methanol-water in volume ratios of 60:40, 70:30, 75:25, 80:20 and 90:10 was used. 35 fractions, namely 1-6, 7-16, 17-25, 26-30 and 31-35, were collected in sequence and identified by silica gel thin layer chromatography. The Rf values ​​of the fluorescent spots observed at wavelengths of 254nm and 365nm were 0.31-0.34 (fraction 1-6), 0.47-0.49 (fraction 7-16), 0.65-0.62 (fraction 17-25), 0.74-0.70 (fraction 26-30) and 0.82-0.85 (fraction 31-35), respectively. The fractions were combined to obtain 5 fractions, namely E4A to E4E. Among them, fractions 7-16 were combined to form fraction E4B.

[0063] S6. Separate the phloroglucinol compound Hyperforidiol F (referred to as Compound 1) having a structure as shown in Formula (1) from fraction E4B using HPLC; the mobile phase used in the HPLC method is an acetonitrile-water mixed solution.

[0064] The volume ratio of acetonitrile to water in the acetonitrile-water mixed solution is (55:45)-(65:35).

[0065] According to the characteristic ultraviolet absorption (λ max =220, 280nm) fraction E4B was further separated to obtain the phloroglucinol compound Hyperforidiol F having a structural formula as shown in formula (1) with a retention time of 40-50min.

[0066] Further preferably, the volume ratio of acetonitrile to water in the acetonitrile-water mixed solution is 62:38; the retention time of the compound Hyperforidiol F prepared in fraction E4B is 45 min (purity 98%).

[0067] The physical properties and test data of the compound 1 prepared in Example 1 are as follows:

[0068] White oil, easily soluble in chloroform and methanol. According to high-resolution mass spectrometry, its molecular weight is estimated to be 586, and its molecular formula is determined to be C 35 H 54 O7, its unsaturation is calculated to be 9.

[0069] like Figure 1 shown 1 H-NMR (CDCl3, 400MHz) spectrum H 5.02 (2H, m) indicates that there are two double bond proton signals connected to the methylene group in the structure. In addition, δ H 3.81 (1H, d, J = 9.4 Hz) indicates a signal of an oxygen-linked methine; δ H 3.21 (1H, dd, J = 9.6, 2.0 Hz) indicates an oxygen-linked methine signal connected to a methylene group; δ H 3.28 (1H, dd, J = 15.1, 6.2 Hz), 3.16 (1H, dd, J = 15.1, 6.0 Hz) indicate a group of methylene signals connected to the olefin bond, and 10 groups of methyl hydrogen signals [δ H1.70(3H,s), 1.68(3H,s), 1.67(3H,s), 1.65(3H,s), 1.24(3H,s), 1.21(3H,s), 1.1 7(3H,s), 1.17(3H,d,J=6.2Hz), 1.12(3H,s), 1.10(3H,d,J=6.6Hz), 0.98(3H,s)].

[0070] like Figure 2 shown 13 The C-NMR (CDCl3, 100 MHz) spectrum showed 35 carbon signals, including 3 carbonyl carbon signals (δ C 211.5, 206.4, 194.3), 1 carbon signal of olefin oxide δ C 165.7, 5 olefin carbon signals (δ C 134.3, 133.6, 129.9, 122.0, 119.5), 1 quarterly carbon signal δ C 45.7, 6 methylene carbon signals (δ C 42.3, 33.8, 30.7, 29.5, 24.1, 22.8), 2 methine carbon signals (δ C 41.1, 35.0), 10 methyl carbon signals (δ C 26.4, 26.1, 26.0, 25.6, 25.5, 23.2, 21.5, 20.7, 18.1, 18.1, 16.7). In addition, there are 2 tertiary carbon signals connected to oxygen (δ C 87.8, 79.0); 2 oxygen-linked quaternary carbon signals (δ C 73.3, 72.6); quaternary carbon signals of two carbonyl groups (δ C 74.5, 64.4).

[0071] 1 H NMR, 13 The signal assignments of C NMR are shown in Table 1.

[0072] Table 1 NMR data of compound 1 δ (ppm)

[0073]

[0074] like Figure 3 In the HMBC spectrum shown, H-11 (δ H 2.51) and C-12 (δ C 21.8), C-10(δ C 209.4), indicating the presence of an isobutyryl group in the compound; H-17 (δ H 5.00) and C-15 (δ C38.6), C-19(δ C 25.8), C-20(δ C 18.0) indicates that 4-methyl-3-pentyl is attached to the C-8 position; in addition, H-22 (δ H 4.90) and C-21(δ C 27.9), C-7(δ C 45.1), C-23(δ C 133.8), C-24(δ C 26.0), C-25(δ C 18.0), proving that 3-methyl-2-enebutyl is connected to carbon 7; H-31b (δ H 2.08) and C-6(δ C 41.4), C-5(δ C 63.5) and H-32 (δ H 3.40) and C-31(δ C 32.9), C-33(δ C 73.2), C-33(δ C The long-range correlation between 24.3) proves that 2,3-dihydroxy-3-methylbutyl is attached to the 5-carbon.

[0075] In summary, the structure of the new compound 1 (compound Hyperforidiol F) was determined as follows:

[0076]

[0077] Comparative Example 1

[0078] Comparative Example 1 of the present invention provides a method for preparing a novel phloroglucinol compound, and its steps are similar to those of Example 1, except that in step S5, when fraction E4 is subjected to MCI column chromatography, the volume ratio of methanol to water in the methanol-water solution used is lower than 65:35 or higher than 72:28.

[0079] The results showed that compound 1 could not be prepared.

[0080] Comparative Example 2

[0081] Comparative Example 2 of the present invention provides a method for preparing a novel 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 E4B by HPLC, the volume ratio of acetonitrile to water is lower than 55:45, or higher than 65:35.

[0082] The results showed that the purity of the prepared compound 1 was no higher than 70%.

[0083] In order to better understand the essence of the present invention, the new application of the novel phloroglucinol compound in the pharmaceutical field is explained below in combination with pharmacological tests and results.

[0084] Test example

[0085] This test example discloses an experiment on the antidepressant activity of the compound Hyperforidiol F in SH-SY5Y cells induced by corticosterone (a commonly used cell model for antidepressants).

[0086] (1) Experimental materials and instruments

[0087] Human neuroblastoma SH-SY5Y was purchased from the Institute of Basic Research, Chinese Academy of Medical Sciences;

[0088] DMEM medium, PBS buffer, fetal bovine serum (FBS), and trypsin were purchased from Gibco, USA;

[0089] Fluoxetine (Shanghai Yuanye Biotechnology Co., Ltd.);

[0090] Luyoutai (Dr. Weimar Shupe Pharmaceuticals, Germany);

[0091] CCK-8 cell activity detection kit (Elabscience);

[0092] 96-well plate (Corning, USA);

[0093] Carbon dioxide cell culture incubator (Thermo Fisher Scientific, USA);

[0094] Full wavelength microplate reader (Thermo Fisher Scientific, USA);

[0095] BIOFUGE STRATOS centrifuge (Thermo Fisher Scientific, USA);

[0096] IX73 inverted electron microscope (Olympus, Japan);

[0097] Pipette (Eppendorf, Germany);

[0098] Electronic balance (Sartorius, Germany);

[0099] Clean bench, centrifuge tubes, pipettes and other related consumables.

[0100] (2) Establishment and evaluation of the SH-SY5Y cell model induced by corticosterone

[0101] Take out the marked SH-SY5Y cell cryopreservation tube from liquid nitrogen and immediately put it into a 37°C water bath, and try to complete rapid thawing within 1 minute; after disinfecting the cryopreservation tube with alcohol, transfer the cryopreservation solution in the tube into a 15mL sterile centrifuge tube, add the corresponding culture medium, mix it, and then centrifuge to remove the supernatant; repeat the above steps once for washing, then add 10mL of culture medium to mix and resuspend the cells, then transfer them to a 10mL culture dish, and culture them in a constant temperature incubator at 37°C and 5% CO2.

[0102] When the cell density reaches 70%-80%, it is passaged; first, the old culture medium is aspirated, PBS is added to wash twice, trypsin containing EDTA is added to the culture dish and placed in a 37°C incubator for 3 minutes, then culture medium is added to terminate digestion, centrifuged at 1000rpm for 5 minutes, the digestion solution is aspirated, and the serum-containing culture medium is re-added and the cells are repeatedly blown to form a cell suspension. Finally, according to the number of cells, they are transferred to a new culture dish to complete cell passage.

[0103] SH-SY5Y cells were cultured at 1×10 5 The cells / well were plated in a 96-well plate, with 6 replicate wells in each group, 80 μL / well, and cultured for 24 hours after plating. The positive drugs fluoxetine, rulothione, hyperforin, tetrahydrohyperforin, and Hypericumoxide M were dissolved and diluted to 10 μM. The compound Hyperforidiol F was dissolved and diluted to 5 μM and 10 μM, respectively, and treated with 10 μL / well for 1 hour. Then, 10 μL of 25 μM corticosterone was added and cultured at 37°C for 48 hours. Then, 10 μL of CCK-8 was added. After 1 hour, the absorbance at 450 nm was detected using an enzyme reader, and the cell survival rate was calculated.

[0104] Cell viability (%) = [(A control - A sample) / (A control - A blank)] x 100%.

[0105] (3) Experimental results

[0106] The experimental results (Table 2) show that low and high doses (5μM, 10μM) of the compound Hyperforidiol F have a significant protective effect on corticosterone-induced SH-SY5Y neuronal cell damage, which is significantly better than the positive drugs fluoxetine (a first-line chemical drug for the clinical treatment of depression, 10μM), Luyoutai (the world's first natural antidepressant herbal medicine, 10μM), HypericumoxideM (10μM), hyperforin (10μM) and tetrahydrohyperforin (10μM).

[0107] Table 2 Neuroprotective effects of compounds on SH-SY5Y cell injury induced by corticosterone ( n=5)

[0108] 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 ** ]]> Lu Youtai 10 <![CDATA[60.03±3.33 * ]]> Hyperforin 10 55.83±2.71 Tetrahydrohyperforin 10 <![CDATA[58.03±3.33 * ]]> Hypericumoxide M 10 56.34±4.24 Hyperforidiol F 10 <![CDATA[74.22±7.14 *** ]]> Hyperforidiol F 5 <![CDATA[69.63±3.64 ** ]]>

[0109] Note: Compared with the blank group: #### P<0.0001; compared with the model group: * P<0.05, ** P<0.01, *** P<0.001

[0110] In summary, the Hyperforidiol F described in the present invention exhibits significant antidepressant activity and is superior to the first-line antidepressant drugs fluoxetine and Luyoutai, and can be used as a drug for treating depression.

[0111] Application Example 1

[0112] The application example of the present invention discloses a capsule preparation using Hyperforidiol F as a raw material drug, and the components thereof are as follows:

[0113]

[0114] The specific preparation process is as follows:

[0115] Hyperforidiol F, starch and sodium metabisulfite are mixed evenly, anhydrous ethanol is added to make a soft material, the mixture is passed through a 24-mesh sieve to form granules, the granules are dried, magnesium stearate is added, the mixture is mixed evenly and the granules are filled into capsules.

[0116] Application Example 2

[0117] The application example of the present invention discloses a granule preparation using the compound Hyperforidiol F as a raw material drug, and the components thereof are as follows:

[0118] Hyperforidiol F35.0 mg

[0119] Starch 6.0g

[0120] Sodium bisulfite 0.2g

[0121] Magnesium stearate 0.2g

[0122] Anhydrous ethanol

[0123] Makes 100 bags.

[0124] The specific preparation process is as follows:

[0125] Hyperforidiol F was mixed with starch and sodium bisulfite, and then anhydrous ethanol was added to make a soft material. The mixture was passed through a 24-mesh sieve to form granules, dried, magnesium stearate was added, mixed, and bagged.

[0126] Application Example 3

[0127] The application example of the present invention discloses an oral liquid using the compound Hyperforidiol F as a raw material drug, and its components are as follows:

[0128]

[0129] The specific preparation process is as follows:

[0130] After the above components are mixed evenly, they can be packaged using a conventional oral liquid preparation method.

[0131] Application Example 4

[0132] The application example of the present invention discloses an injection with the compound Hyperforidiol F as the raw material drug, and its components are as follows:

[0133] Hyperforidiol F45.0 mg

[0134] Vitamin C 0.2g

[0135] Sodium chloride 6.0g

[0136] Sodium bicarbonate 0.1g

[0137] Water for injection 1000mL

[0138] Made into 100 pieces.

[0139] The specific preparation process is as follows:

[0140] After the above components are mixed evenly, 100 vials can be obtained using a conventional injection preparation method.

[0141] Application Example 5

[0142] The application example of the present invention discloses a tablet with compound Hyperforidiol F and fluoxetine as raw materials, and its components are as follows:

[0143] Hyperforidiol F20.0 mg

[0144] Fluoxetine 0.5g

[0145] Hydroxypropyl methylcellulose 18g

[0146] Talc 0.4g

[0147] Lactose 0.2g

[0148] Magnesium stearate 0.2g

[0149] Anhydrous ethanol

[0150] Makes 100 pieces.

[0151] The specific preparation process is as follows:

[0152] Hyperforidiol F, fluoxetine, hydroxypropyl methylcellulose, talc, lactose and magnesium stearate were mixed evenly, anhydrous ethanol was added to make a soft material, the mixture was passed through a 24-mesh sieve to make granules, dried, magnesium stearate was added, mixed evenly and tableted.

[0153] Application Example 6

[0154] The application example of the present invention discloses a capsule preparation using the compound Hyperforidiol F and sertraline as raw materials, and the components thereof are as follows:

[0155] Hyperforidiol F18.0 mg

[0156] Sertraline 2.0g

[0157] Starch 6.0g

[0158] Sodium metabisulfite 0.2g

[0159] Magnesium stearate 0.2g

[0160] Anhydrous ethanol

[0161] Makes 100 tablets.

[0162] The specific preparation process is as follows:

[0163] Hyperforidiol F, sertraline, starch and sodium metabisulfite were mixed, anhydrous ethanol was added to make a soft material, the mixture was passed through a 24-mesh sieve to make granules, the mixture was dried, magnesium stearate was added, the mixture was mixed, and the mixture was put into capsules.

[0164] Application Example 7

[0165] The application example of the present invention discloses an injection with the compound Hyperforidiol F and escitalopram as raw materials, and its components are as follows:

[0166] Hyperforidiol F30.0 mg

[0167] Escitalopram 2.0g

[0168] Vitamin C 0.2g

[0169] Sodium chloride 6.0g

[0170] Sodium bicarbonate 0.1g

[0171] Water for injection 1000mL

[0172] Made into 100 pieces.

[0173] The specific preparation process is as follows:

[0174] After the above components are mixed evenly, 100 vials can be obtained using a conventional injection preparation method.

[0175] The above-mentioned embodiments only express several implementation modes of the present invention, and the description thereof is relatively specific and detailed, but it should not be understood as limiting the scope of the invention patent.

[0176] It should be noted that, for those skilled in the art, several modifications and improvements can be made without departing from the concept of the present invention, and these modifications and improvements all belong to the protection scope of the present invention. Therefore, the protection scope of the patent of the present invention shall be subject to the attached claims.

Claims

1. A novel phloroglucinol compound Hyperforidiol F, characterized in that: It has the structure shown in formula (1):

2. A method for preparing the novel phloroglucinol compound Hyperforidiol F as claimed in claim 1, characterized in that: The novel phloroglucinol compound Hyperforidiol F is obtained by extraction and separation from the aerial part of Hypericum perforatum. The following steps are involved: S1. Take the dried aerial part of Hypericum perforatum, add a solvent for reflux extraction, combine the extracts and concentrate to obtain an extract; the solvent is 88-98V% ethanol aqueous solution; S2, adding the extract to water and suspending it, and then extracting it with petroleum ether to obtain an extract; S3, gradient eluting the extract by silica gel column chromatography using petroleum ether-ethyl acetate solution to obtain fraction E; when detected by silica gel thin layer chromatography, the Rf value corresponding to fraction E is 0.65-0.68; S4, using a petroleum ether-ethyl acetate mixed solution to gradiently elute fraction E through silica gel column chromatography to obtain fraction E4; when detected by silica gel thin layer chromatography, the Rf value corresponding to fraction E4 is 0.63-0.69; S5. Fraction E4 is gradient eluted by MCI column chromatography using a methanol-water solution to obtain fraction E4B; when detected by silica gel thin layer chromatography, the Rf value corresponding to fraction E4B is 0.47-0.49; the volume ratio of methanol to water in the methanol-water solution when eluting fraction E4B is (65:35)-(72:28); S6, separating the phloroglucinol compound Hyperforidiol F having a structure as shown in formula (1) from fraction E4B by HPLC; The mobile phase used in the HPLC method is an acetonitrile-water mixed solution; the volume ratio of acetonitrile to water in the acetonitrile-water mixed solution is (55:45)-(65:35).

3. The preparation method according to claim 2, characterized in that: In step S6, fraction E4B is further separated according to the characteristic ultraviolet absorption of phloroglucinol observed by HPLC analysis to obtain a phloroglucinol compound Hyperforidiol F having a structural formula as shown in formula (1) and a retention time of 40-50 min.

4. The preparation method according to claim 2, characterized in that: In step S2, the extract is added into water with a mass of 8 to 15 times the mass, suspended, and then extracted with petroleum ether.

5. The preparation method according to claim 2, 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 when fraction E is eluted is (45:55)-(55:45).

6. The preparation method according to claim 2, 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 when fraction E is eluted is 50:

50.

7. The preparation method according to claim 2, characterized in that: In step S4, the volume ratio of petroleum ether to ethyl acetate in the petroleum ether-ethyl acetate mixed solution when fraction E4 is washed out is (45:55)-(55:45).

8. The preparation method according to claim 2, characterized in that: In step S4, the volume ratio of petroleum ether to ethyl acetate in the petroleum ether-ethyl acetate mixed solution when fraction E4 is washed out is 50:

50.

9. The preparation method according to claim 2, characterized in that: In step S5, the volume ratio of methanol to water in the methanol-water solution when fraction E4B is eluted is 70:

30.

10. The preparation method according to claim 2, characterized in that: In step S1, the mass of the added solvent is 8-10 times that of Hypericum perforatum, the number of reflux extractions is 1-3, and each extraction lasts 1-3 hours.

11. A pharmaceutical composition, characterized in that The invention comprises the novel phloroglucinol compound as claimed in claim 1.

12. The pharmaceutical composition according to claim 11, characterized in that The pharmaceutical composition comprises a synergist and a pharmacologically acceptable carrier or excipient.

13. The pharmaceutical composition according to claim 11, characterized in that The synergist is one or more 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, St. John's wort extract, flupentixol-melitracen.

14. The pharmaceutical composition according to claim 11, characterized in that The dosage form of the pharmaceutical composition is tablets, capsules, granules, oral liquid, granules, dropping pills or micropills.

15. Use of the novel phloroglucinol compound according to claim 1 and / or the pharmaceutical composition according to any one of claims 11 to 14 in the preparation of a drug for treating depression.

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