Lycoctine-type diterpene alkaloid compounds, preparation method and application thereof

Abstract

The application discloses a lycoctine type diterpenoid alkaloid compound and a preparation method and application thereof. 20 The diterpenoid alkaloid compound Carmaloidline E shows significant antidepressant activity in a mouse behavioral despair model experiment, is obviously superior to fluoxetine as a positive medicine, and can be used for developing antidepressant drugs.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, and more specifically, relates to a lycoctine-type diterpenoid alkaloid compound, its preparation method, and its application. Background Technology

[0002] In today's increasingly competitive society, the high-intensity, high-pressure lifestyle is seriously impacting people's physical and mental health, leading to a rise in depression and severely affecting people's normal work and life. Currently, medications for treating depression generally have significant side effects, limiting their clinical application.

[0003] The inventors prepared a novel lycoctine-type C from the traditional Chinese medicine Aconitum carmichaelii. 20 The diterpenoid alkaloid compound Carmaloidline E was found to exhibit significant antidepressant activity in a mouse behavioral despair model, outperforming the positive control drug fluoxetine, and could be used to develop antidepressant drugs. Summary of the Invention

[0004] The purpose of this invention is to overcome the shortcomings of the existing technology and provide a novel lycoctine-type C 20 Carmaloidline E, a diterpenoid alkaloid compound, and its preparation methods, pharmaceutical compositions, and applications.

[0005] To achieve the above objectives, the present invention provides a lycoctine-type diterpenoid alkaloid compound (referred to as compound Carmaloidline E), which has the structure shown in formula (1):

[0006]

[0007] In another aspect, the present invention provides a method for preparing the above-mentioned lycoctine-type diterpenoid alkaloid compound, wherein the lycoctine-type diterpenoid alkaloid compound is extracted and isolated from Aconitum carmichaelii.

[0008] Furthermore, the extraction and separation steps include the following:

[0009] S1. Extract Aconitum carmichaelii by reflux with solvent, combine the extracts and concentrate to obtain extract;

[0010] S2. Dissolve the extract obtained in step S1 in an acidic solution, suspend and filter to remove impurities to obtain a filtered acidic aqueous solution; extract the filtered acidic aqueous solution with dichloromethane to obtain acidic aqueous solution A and extract A;

[0011] The pH of the acidic aqueous solution A was adjusted to be alkaline, and then extracted with dichloromethane to obtain extract B;

[0012] S3. The extract B is subjected to column chromatography and high performance liquid chromatography in sequence to obtain the lycoctine-type diterpenoid alkaloid compound.

[0013] The column chromatography separation includes sequential silica gel column chromatography, ODS column chromatography, and gel column chromatography.

[0014] Among them, the preferred gel column chromatography is Sephadex LH-20 gel column chromatography.

[0015] Further, in step S1, the solvent is one or more of methanol, ethanol, acetone, chloroform and petroleum ether.

[0016] Further, in step S1, the solvent is an 88-98% aqueous ethanol solution;

[0017] The solvent added should be 8-10 times the mass of Aconitum carmichaelii; the reflux extraction should be performed 2-4 times, with each extraction lasting 1-3 hours.

[0018] Further, in step S2, the acidic solution is 0.1-1.0% dilute hydrochloric acid; the amount of acidic solution added is 8-15 times the mass of the extract.

[0019] Furthermore, in step S2, the pH of the acidic aqueous solution A is adjusted to 8-11 by adding alkali; preferably, pH = 9.

[0020] Furthermore, based on a total volume of 100, in step S3, the silica gel column chromatography employs gradient elution, and the mobile phase is a mixed solvent composed of solvent A and solvent B in a volume ratio of (0:100)-(100:0).

[0021] Solvent A is any one of dichloromethane, chloroform, and petroleum ether;

[0022] Solvent B is any one of chloroform, acetone, ethyl acetate, and methanol.

[0023] Preferably, the mobile phase is dichloromethane-methanol.

[0024] Further, step S3 includes:

[0025] S31. Extract B was subjected to gradient elution using dichloromethane-methanol silica gel column chromatography to obtain fraction C; when identified by silica gel thin-layer chromatography, the Rf value corresponding to fraction C was 0.72-0.75.

[0026] S32. Using a mobile phase of methanol-water, fraction C was eluted by ODS column chromatography to obtain fraction C3; when identified by silica gel thin-layer chromatography, the Rf value corresponding to fraction C3 was 0.50-0.60.

[0027] S33. Using a mobile phase of methanol-water, fraction C3 was subjected to gel column chromatography to obtain fraction M1; the Rf value of fraction M1 was 0.30-0.39.

[0028] S34. The fraction M1 was subjected to RP-HPLC reversed-phase high-performance liquid chromatography with acetonitrile-water as the mobile phase to obtain lycoctine-type diterpenoid alkaloids with the structural formula shown in formula (1). The retention time of the lycoctine-type diterpenoid alkaloids was 10-15 min.

[0029] Further, in step S31, the mobile phase for silica gel column chromatography is dichloromethane-methanol with a volume ratio of (93:7)-(91:9).

[0030] Preferably, the volume ratio of the mobile phase dichloromethane to methanol is 92:8.

[0031] Furthermore, in step S32, the volume ratio of methanol to water in the ODS column chromatography mobile phase is (35:65)-(45:55); the mobile phase also contains formic acid at a volume percentage of 0.01-0.1%.

[0032] Preferably, the volume ratio of methanol to water in the mobile phase in step S32 is 40:60;

[0033] And / or, the formic acid volume percentage in the mobile phase of the ODS column chromatography is 0.05%.

[0034] Further, in step S33, the volume ratio of methanol to water in the mobile phase of gel column chromatography is (0:100)-(10:90).

[0035] Preferably, in step S33, the volume ratio of the mobile phase methanol to water is 0:100.

[0036] Further, in step S34, the mobile phase of RP-HPLC reversed-phase high-performance liquid chromatography is acetonitrile-water with a volume ratio of (4:96)-(10:90), and the mobile phase also contains 0.01-0.5% trifluoroacetic acid by volume.

[0037] Further, in step S34, the RP-HPLC reversed-phase high-performance liquid chromatography uses an acetonitrile-water mobile phase with a volume ratio of 5:95; the volume percentage of trifluoroacetic acid in the mobile phase is 0.1%.

[0038] Preferably, the retention time of lycoctine-type diterpenoid alkaloids with the structural formula shown in formula (1) is 12 min.

[0039] Tautomers of the lycoctine-type diterpenoid alkaloids or pharmaceutically acceptable salts are also within the scope of protection of this invention.

[0040] A third aspect of this application discloses a pharmaceutical composition comprising the aforementioned lycoctine-type diterpenoid alkaloid compound.

[0041] Furthermore, it also includes pharmaceutically acceptable carriers or excipients.

[0042] In other words, it contains the lycoctine-type C of the present invention as an active ingredient. 20 Pharmaceutical compositions comprising diterpenoid alkaloids and conventional pharmaceutical excipients, adjuvants, or carriers are also included in this invention.

[0043] Furthermore, it also includes synergists;

[0044] The synergist is one or more of the following substances:

[0045] Fluoxetine, paroxetine, fluvoxamine, sertraline, citalopram, escitalopram, venlafaxine, duloxetine, mirtazapine, bupropion, agomelatine, trazodone, reboxetine, imipramine, amitriptyline, clomipramine, doxepin, maprotiline, moclobemide, liver-soothing and mood-regulating capsules, St. John's wort extract, flupentixol melitracen.

[0046] Furthermore, the dosage form of the pharmaceutical composition is tablets, capsules, granules, oral liquids, powders, drops, microcapsules, intravenous injections, or intramuscular injections.

[0047] The present invention also provides the use of the above-mentioned lycoctine-type diterpenoid alkaloids or the above-mentioned pharmaceutical compositions in the preparation of antidepressant drugs.

[0048] The advantages and beneficial effects of this invention are as follows:

[0049] (1) This invention provides a novel lycoctine-type C that has not been reported before. 20 The study also provided a simple, reproducible, and highly pure method for extracting Carmaloidline E from Aconitum carmichaelii.

[0050] (2) Experimental results show that the novel lycoctine-type C provided by this invention... 20The diterpenoid alkaloid compound Carmaloidline E demonstrated significant antidepressant activity in a mouse behavioral despair model, outperforming the positive control drug fluoxetine (a first-line clinical treatment for depression). These results suggest that Carmaloidline E could be used to develop antidepressant drugs. Attached Figure Description

[0051] Figure 1 Compound 1 obtained in Example 1 of this invention 1 1H NMR spectrum (400MHz, DMSO-d6);

[0052] Figure 2 Compound 1 obtained in Example 1 of this invention 13 C NMR spectrum (100MHz, DMSO-d6);

[0053] Figure 3 The HMBC spectrum of compound 1 obtained in Example 1 of this invention;

[0054] Figure 4 This is the NOESY spectrum of compound 1 obtained in Example 1 of the present invention. Detailed Implementation

[0055] The specific embodiments of the present invention will be further described below with reference to the accompanying drawings and examples. The following examples are only used to more clearly illustrate the technical solutions of the present invention and should not be construed as limiting the scope of protection of the present invention.

[0056] (I) Preparation of lycoctine-type diterpenoid alkaloids

[0057] Example 1

[0058] A method for preparing a lycoctine-type diterpenoid alkaloid compound includes the following steps:

[0059] S1. Take dried Aconitum carmichaelii (about 200.5 kg), use 95% ethanol aqueous solution (10 times the weight of Aconitum carmichaelii) as solvent for reflux extraction three times, each extraction for 2 hours, combine the extracts and concentrate to obtain extract (about 6 kg).

[0060] S2. Add the extract to 10 times its weight of 0.5% dilute hydrochloric acid (60L) to fully dissolve and suspend it. Filter to remove impurities. Extract the filtered acidic aqueous solution three times with 1.5 times its volume of dichloromethane to obtain acidified dichloromethane extract A (about 2.5kg, discarded) and acidic aqueous solution A (about 1.2kg).

[0061] The pH of the acidic aqueous solution A was adjusted to 9 with sodium hydroxide, and then extracted with dichloromethane to obtain extract B, i.e., dichloromethane extract (about 500g).

[0062] S3 and extract B were sequentially separated by column chromatography and high performance liquid chromatography to obtain the lycoctine-type diterpenoid alkaloid compound.

[0063] Specifically, the steps include the following:

[0064] S31. Extract B is subjected to gradient elution using dichloromethane-methanol silica gel column chromatography to obtain fraction C; when identified by silica gel thin-layer chromatography, the Rf value corresponding to fraction C is 0.72-0.75. The mobile phase for silica gel column chromatography is dichloromethane-methanol with a volume ratio of (93:7)-(91:9), preferably 92:8.

[0065] It should be noted that during gradient elution, dichloromethane-methanol at volume ratios of (93:7) to (91:9) can elute fraction C. Fraction C is identified by silica gel thin-layer chromatography with an Rf value of 0.72-0.75. The dichloromethane-methanol volume ratio of 92:8 exhibits the highest elution efficiency. The principle behind the "preferred volume ratio" described in the following gradient elution processes is the same.

[0066] The specific procedure included: Extract B was subjected to silica gel column chromatography, eluted with a dichloromethane-methanol gradient at volume ratios of 100:0, 95:5, 92:8, 90:10, 85:15, 75:25, 65:35, 50:50, and 0:100, respectively, to obtain fractions 1-10, 11-20, 21-30, 31-40, 41-50, 51-60, 61-70, 71-80, and 81-90, for a total of 90 fractions. These fractions were then identified using silica gel thin-layer chromatography, and the Rf values ​​were observed based on the brick-red spots revealed by potassium bismuth iodide reagent, ranging from 0.82 to 0.86 (fractions). Fractions 1-10, 0.76-0.80 (fractions 11-20), 0.72-0.75 (fractions 21-30), 0.65-0.70 (fractions 31-40), 0.56-0.62 (fractions 41-50), 0.50-0.54 (fractions 51-60), 0.42-0.48 (fractions 61-70), 0.30-0.38 (fractions 71-80), and 0.25-0.28 (fractions 81-90) were combined to obtain fractions A, B, C, D, E, F, G, H, and I; fractions 21-30 were combined to obtain fraction C.

[0067] Based on the brick-red spots identified by thin-layer chromatography and the characteristic ultraviolet absorption (λ) of diterpenoid alkaloids observed by HPLC analysis, bismuth iodide was detected. max =260nm) Select fraction C for the next separation step.

[0068] S32. Elute fraction C using ODS column chromatography with a mobile phase of methanol-water to obtain fraction C3; when identified by silica gel thin-layer chromatography, the Rf value corresponding to fraction C3 is 0.50-0.60; the volume ratio of methanol-water in the ODS column chromatography mobile phase is (35:65)-(45:55); the mobile phase also contains 0.01-0.1% formic acid by volume. Preferably, in step S32, the volume ratio of methanol-water in the mobile phase is 40:60; the volume percentage of formic acid in the mobile phase is 0.05%.

[0069] Specifically, fraction C was subjected to ODS column chromatography using a methanol-water gradient elution (containing 0.05% formic acid, v / v) at ratios of 20:80, 30:70, 40:60, 50:50, 60:40, 70:30, 80:20, 90:10, and 100:0, yielding fractions 1-5, 6-15, 16-25, 26-35, 36-45, 46-50, 51-55, 56-60, and 61-68, for a total of 68 fractions. These fractions were then analyzed using silica gel thin-layer chromatography. Identification was performed using potassium bismuth iodide reagent. Based on the brick-red spots observed, the Rf values ​​were 0.35-0.44 (fractions 1-5), 0.45-0.49 (fractions 6-15), 0.50-0.60 (fractions 16-25), 0.61-0.69 (fractions 26-35), 0.70-0.74 (fractions 36-55), 0.75-0.79 (fractions 56-60), and 0.80-0.89 (fractions 61-68). Similar fractions were combined into seven fractions, C1 to C7. Fractions 16-25 were combined to obtain fraction C3.

[0070] Based on the brick-red spots observed by potassium bismuth iodide reagent in thin-layer chromatography and the characteristic ultraviolet absorption (λ) of diterpenoid alkaloids observed by HPLC analysis, [further details are needed]. max =260nm) Select fraction C3 for the next separation step.

[0071] S33. Using a mobile phase of methanol-water, fraction C3 is subjected to gel column chromatography to obtain fraction M1; the Rf value of fraction M1 is 0.30-0.39; the volume ratio of the mobile phase of methanol-water for gel column chromatography is (0:100)-(10:90). Preferably, it is 0:100.

[0072] The specific procedure included: fraction C3 was subjected to dextran gel LH-20 column chromatography, using a methanol-water gradient elution with volume ratios of 0:100, 30:70, 70:30, and 100:0, collecting fractions 1-6, 7-14, 15-20, and 21-29 sequentially, for a total of 29 fractions. These fractions were then identified using silica gel thin-layer chromatography. Based on the brick-red spots observed with potassium bismuth iodide reagent, the Rf values ​​were 0.30-0.39 (fractions 1-6), 0.40-0.54 (fractions 7-14), 0.55-0.61 (fractions 15-20), and 0.62-0.85 (fractions 21-29), respectively. Similar fractions were combined to obtain four fractions, M1 to M4. Fractions 1-6 were combined to obtain fraction M1.

[0073] Based on the brick-red spots observed by potassium bismuth iodide reagent and the characteristic ultraviolet absorption (λ) of diterpenoid alkaloids observed by HPLC analysis. max =260nm) Select fraction M1 for the next separation step.

[0074] S34. Fraction M1 is subjected to RP-HPLC reversed-phase high-performance liquid chromatography with acetonitrile-water as the mobile phase to obtain lycoctine-type diterpenoid alkaloids with the structural formula shown in formula (1). The retention time of the lycoctine-type diterpenoid alkaloids is 10-15 min. The mobile phase is acetonitrile-water with a volume ratio of (4:96)-(10:90); and the mobile phase also contains 0.01-0.5% trifluoroacetic acid by volume; more preferably, the volume percentage of trifluoroacetic acid is 0.1%.

[0075] Preferably, HPLC is used with a mobile phase of acetonitrile-water (volume ratio 5:95, containing 0.1% trifluoroacetic acid by volume) as the mobile phase, utilizing C 18 The chromatographic column was prepared in fraction M1 to obtain lycoctine-type C 20 Diterpenoid alkaloid compound 1 (compound Carmaloidline E)(t) R =12min, purity is 99%).

[0076] The physical properties and detection data of compound 1 obtained in Example 1 are as follows:

[0077] Physical properties: White amorphous powder, readily soluble in methanol.

[0078] High-resolution mass spectrometry (HR-ESI-MS) m / z 378.2632 ([M+H] + The calculated value is 378.2639), combined with 1 HNMR ( Figure 1 ), 13 CNMR spectrum ( Figure 2Its molecular formula was determined to be C. 22 H 35 O4N has an unsaturation degree of 6.

[0079] 1 HNMR (400MHz, DMSO-d6) spectrum Figure 1 The data shows that there are hydrogen signals [δ] on two oxygen / nitromethyl groups. H 3.67 (1H,s), 2.71 (1H,s)], hydrogen signals on the four oxygen / nitromethylene groups [δ H 3.30 (1H, adqd, J = 11.3, 5.6 Hz), 3.23 (1H, adqd, J = 11.3, 5.6 Hz), 2.31 (1H, overlapped), 1.91 (1H, d, J = 8.4 Hz), 2.36 (1H, m), 2.24 (1H, s)] and two methyl signals [δ H 0.97 (3H, d, J = 7.2 Hz), 0.70 (3H, s)] and in δ H There are 5 methine and 6 methylene signals between 2.08 and 1.14.

[0080] 13 CNMR (100MHz, DMSO-d6) spectrum Figure 2 The display shows a total of 22 carbon signals, including 7 oxygen / nitrogen sp signals. 3 Hybrid carbon [δ] C 66.6(C-1),77.0(C-8),82.0(C-14),66.7(C-17),56.9(C-19),62.6(C-20),48.2(N C H2CH3)] and 15 aliphatic sps 3 Hybridized carbon.

[0081] Combination 1 H NMR and 13 Based on the C1NMR spectrum (Table 1), compound 1 is presumed to contain 7 carbon signals bonded to oxygen / nitrogen and 2 methyl groups. Combining this information with comparisons to the references, it is speculated that compound 1 may be a lycoctine-type C1NMR compound. 20 Type diterpenoid alkaloids.

[0082] like Figure 3 As shown, in the HMBC spectrum, H3-18 (δ H 0.70) and C-4(δ C 33.9), C-5 (δ) C 45.9), C-19 (δ) C The related peaks of H-3 (δ 56.9) H1.20) and C-1(δ C 66.6), C-5 (δ) C The related peaks of H-6 (δ) at 45.9) H 1.30) and C-4(δ) C 33.9), C-8 (δ) C 77.0), C-11(δ) C The related peaks of H-10 (δ) are 49.4). H 2.31) and C-8(δ) C 77.0), C-9 (δ) C 45.7), C-11(δ) C The related peaks of H-13 (δ) are 49.4). H 1.67) and C-10(δ C 38.9), C-12 (δ) C 31.5), C-14 (δ) C 82.0), C-15 (δ) C The related peaks of 32.9) and H-20 (δ) H 2.71) and C-5(δ) C 45.9), C-6 (δ) C 24.7), C-8(δ) C 77.0), C-19 (δ) C The correlation peak at 56.9 indicates that compound 1 may have a conventional Aconitine-type C ... 19 The skeleton of diterpenoid alkaloids, but H2-17 (δ H 3.30, 3.23) and C-9 (δ C 45.7), C-13 (δ) C 35.6), C-14 (δ) C The relevant peaks at 82.0 indicate that compound 1 is not an Aconitine-type C12 compound. 19 Diterpenoid alkaloids, but more commonly lycoctine-type C 20 Diterpenoid alkaloids, NCH2 CH3 (δ H 0.97) and N C H2CH3(δ C 48.2) related peaks and N CH2 CH3(δ H 2.36, 2.24) and C-19 (δ C 56.9), C-20 (δ) C The relevant peak at 62.6 indicates that an ethylamine group is attached to C-19 and C-20. Combined with the HMBC spectrum and chemical shift, the planar configuration of compound 1 was confirmed.

[0083] The relative stereoconfiguration of compound 1 was determined by NOESY spectroscopy (see [link]). Figure 4 H-1(δ) has been confirmed. H 3.67) and H-20 (δ) H 2.71) related peaks and H-1(δ H 3.67) and H-12α(δ H 1.14) related peaks, H-12β (δ H 1.78) and H-10β(δ H The correlation peak at 2.31 indicates that the proton at the C-1 position is α-oriented. H-17 (δ) H 3.30) and H-13β(δ) H The relevant peak at 1.67 indicates that the hydroxyl group at C-14 is α-oriented.

[0084] In summary, the structure of the new compound 1 (compound Carmaloidline E) was determined as follows:

[0085]

[0086] 1 H NMR, 13 The signal assignments for C NMR are shown in Table 1.

[0087] Table 1 1 H NMR, 13 Signal assignment δ(ppm) for C NMR

[0088]

[0089] Comparative Example 1

[0090] The preparation method is the same as in Example 1, except that in step S34, the aqueous solution of the mobile phase used to separate fraction M1 by HPLC does not contain trifluoroacetic acid.

[0091] The compound prepared in Comparative Example 1 had severe peak tailing during the preparation process, and liquid chromatography analysis after preparation revealed a large number of impurities. The purity of the prepared compound 1 was less than 55%.

[0092] Comparative Example 2

[0093] The preparation method is the same as that in Example 1, except that in step S32, when fraction C is subjected to ODS column chromatography, methanol-water (without formic acid) with an initial volume ratio of 40:60 is used for elution.

[0094] Comparative Example 2 was found to contain a large number of impurities during subsequent fraction preparation by liquid chromatography analysis, and the purity of Compound 1 obtained was less than 20%.

[0095] (2) Verification experiments

[0096] Experimental Example 1: Mouse behavioral despair model experiment of Carmaloidline E (the gold standard for antidepressant drug screening)

[0097] 1. Experimental materials and instruments

[0098] Fluoxetine hydrochloride was purchased from Shanghai Yuanye Bio-Technology Co., Ltd.; Higenamine and Methyllycaconitine were purchased from Shanghai Standard Technology Co., Ltd.; Total alkaloids of Aconitum carmichaeli Debx.; Carmaloidline E.

[0099] ZIL-2 type mouse locomotor activity box (Shanghai Xinman Science and Education Equipment Co., Ltd.); YLS-18A type mouse tail suspension apparatus (Anhui ZhengHua Bio-Instrument Equipment Co., Ltd.); Electronic balance (Sartorius Scientific Instruments Co., Ltd., Beijing).

[0100] 2. Experimental animals

[0101] ICR mice, male, weighing 18 - 22 g, SPF grade, provided by Vital River Laboratory Animal Technology Co., Ltd., animal license number: SYXK(Beijing)2023 - 0001. The animals were housed in an environment with a temperature of 23 ± 2°C and a humidity of 50% ± 10%, and the lighting time was 12 h per day (7:00 - 19:00 lighting). After the animals were housed for 3 days, various experiments were carried out. 6 h before the start of the behavioral experiments, the animals were fasted and allowed free access to water.

[0102] 3. Experimental methods

[0103] 70 male ICR mice were arranged in a snake-like pattern and divided into 7 groups in descending order of the number of spontaneous activities during animal screening, namely the blank group (normal saline of the same volume), the positive drug fluoxetine hydrochloride group (10 mg / kg), the higenamine group (10 mg / kg), the methyllycaconitine group (10 mg / kg), the total alkaloids of Aconitum carmichaeli Debx. group (10 mg / kg), the low-dose Carmaloidline E group (0.03 mg / kg), and the high-dose Carmaloidline E group (0.3 mg / kg). Each group had 10 mice, which were numbered separately. The drugs were all added to normal saline to form a suspension. Each group was administered by gavage once a day, and the administration volume was 0.1 ml / 10 g body weight. After continuous administration for 3 days, the locomotor activity experiment was carried out on the 4th day, and the tail suspension experiment was carried out on the 5th day.

[0104] 4. Experimental results

[0105] As shown in Table 2, compared with the blank group, both the low- and high-dose Carmaloidline E groups (0.03, 0.3 mg / kg) significantly shortened the immobility time of mice in the tail suspension test, demonstrating a significant antidepressant effect, which was significantly better than the positive control drugs fluoxetine (10 mg / kg, a first-line chemotherapeutic drug for the clinical treatment of depression), norcodonol (10 mg / kg), methylcodonol (10 mg / kg), and total alkaloids of Aconitum carmichaelii (10 mg / kg).

[0106] Table 2. Effects of each group on immobility time in the tail suspension test.

[0107]

[0108] (mean ± standard deviation, compared with the blank control group) *** P<0.001)

[0109] As shown in Table 3, in the open field experiment, none of the groups had a significant effect on the spontaneous activity of mice compared with the blank group, indicating that the effect of each group on the immobility time of mice in the tail suspension experiment was not related to nerve excitability.

[0110] Table 3. Effects of each group on the number of horizontal grids traversed by mice in the open field experiment.

[0111]

[0112]

[0113] In summary, the novel lycoctine-type C described in this invention... 20 Carmaloidline E, a diterpenoid alkaloid, exhibits significant antidepressant activity, which is markedly superior to fluoxetine, a first-line clinical drug, and can be used as a prodrug for the treatment of depression.

[0114] Application Example 1

[0115] This invention discloses a capsule formulation using Carmaloidline E as the active pharmaceutical ingredient, the components of which are as follows:

[0116] Carmaloidline E 3.5mg

[0117] 6.0g of starch

[0118] Sodium metabisulfite 0.2g

[0119] Magnesium stearate 0.2g

[0120] Anhydrous ethanol appropriate amount

[0121] Make 100 pills.

[0122] The specific preparation process is as follows:

[0123] Carmaloidline E, starch, and sodium metabisulfite are mixed evenly, and anhydrous ethanol is added to make a soft material. The mixture is then passed through a 24-mesh sieve to form granules, dried, and magnesium stearate is added. The mixture is then mixed and filled into capsules.

[0124] Application Example 2

[0125] This invention discloses a granule formulation using Carmaloidline E as the active pharmaceutical ingredient, the components of which are as follows:

[0126] Carmaloidline E 4.0mg

[0127] 6.0g of starch

[0128] Sodium bisulfite 0.2g

[0129] Magnesium stearate 0.2g

[0130] Anhydrous ethanol appropriate amount

[0131] Make 100 bags.

[0132] The specific preparation process is as follows:

[0133] Carmaloidline E was mixed with starch and sodium bisulfite, and then anhydrous ethanol was added to make a soft material. The material was passed through a 24-mesh sieve to form granules, dried, and then magnesium stearate was added. The mixture was then mixed and packaged.

[0134] Application Example 3

[0135] This invention discloses an oral liquid using the compound Carmaloidline E as a raw material, the components of which are as follows:

[0136] Carmaloidline E 6.0mg

[0137] 3.0g sucrose

[0138] Sodium bisulfite 0.2g

[0139] Methylparaben 0.2g

[0140] Sodium bicarbonate 0.5g

[0141] 1000.0 mL of water for injection

[0142] Make 100 pieces.

[0143] The specific preparation process is as follows:

[0144] After the above components are mixed, they can be dispensed using conventional oral liquid preparation methods.

[0145] Application Example 4

[0146] This invention discloses an injectable formulation using Carmaloidline E as the active pharmaceutical ingredient, the components of which are as follows:

[0147] Carmaloidline E 4.0mg

[0148] Vitamin C 0.2g

[0149] Sodium chloride 6.0g

[0150] Sodium bicarbonate 0.1 mL

[0151] 1000.0 mL of water for injection

[0152] Make 100 pieces.

[0153] The specific preparation process is as follows:

[0154] After the above components are mixed, 100 vials can be obtained by using conventional injection preparation methods.

[0155] Application Example 5

[0156] This invention discloses a tablet using Carmaloidline E and fluoxetine as active pharmaceutical ingredients, the composition of which is as follows:

[0157] Carmaloidline E 3.5mg

[0158] Fluoxetine 20.0mg

[0159] Hydroxypropyl methylcellulose 18.0g

[0160] Talc powder 0.4g

[0161] Lactose 0.2g

[0162] Magnesium stearate 0.2g

[0163] Anhydrous ethanol appropriate amount

[0164] Make 100 pieces.

[0165] The specific preparation process is as follows:

[0166] Carmaloidline E, fluoxetine, hydroxypropyl methylcellulose, talc, lactose, and magnesium stearate are mixed evenly, and anhydrous ethanol is added to make a soft mass. The mass is then passed through a 24-mesh sieve to form granules, dried, and magnesium stearate is added. The mixture is then mixed evenly and compressed into tablets.

[0167] Application Example 6

[0168] This invention discloses a capsule formulation using Carmaloidline E and Luyoutai as active pharmaceutical ingredients, the components of which are as follows:

[0169] Carmaloidline E 3.5mg

[0170] Luyoutai 300.0mg

[0171] 6.0g of starch

[0172] Sodium metabisulfite 0.2g

[0173] Magnesium stearate 0.2g

[0174] Anhydrous ethanol appropriate amount

[0175] Make 100 pills.

[0176] The specific preparation process is as follows:

[0177] Carmaloidline E, Luyoutai, starch, and sodium metabisulfite were mixed and then anhydrous ethanol was added to form a soft material. The material was passed through a 24-mesh sieve to form granules, dried, and then magnesium stearate was added. The mixture was then filled into capsules.

[0178] Application Example 7

[0179] This invention discloses an injectable formulation using Carmaloidline E and paroxetine as active pharmaceutical ingredients, the components of which are as follows:

[0180] Carmaloidline E 5.0mg

[0181] Paroxetine 20.0mg

[0182] Vitamin C 0.2g

[0183] Sodium chloride 6.0g

[0184] Sodium bicarbonate 0.5g

[0185] 1000.0 mL of water for injection

[0186] Make 100 pieces.

[0187] The specific preparation process is as follows:

[0188] After the above components are mixed, 100 vials can be obtained by using conventional injection preparation methods.

[0189] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the technical principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Use of a lycoctine-type diterpene alkaloid compound or a pharmaceutical composition comprising a lycoctine-type diterpene alkaloid compound in the preparation of an antidepressant medicament, characterized in that, The lycoctine-type diterpene alkaloid compound has a structure as shown in formula (1): （1）。 2. Use according to claim 1, characterized in that, and further comprising a pharmaceutically acceptable carrier or excipient.

3. Use according to claim 1, characterized in that, The dosage form of the pharmaceutical composition is a tablet, a capsule, a granule, an oral solution, a powder, a dripping pill or a micro-pill.

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