A genipin derivative, preparation method and application thereof

Abstract

The present invention discloses a genipin derivative or a pharmaceutically acceptable salt thereof. The genipin derivative is of formula (I a ) or (I b ) shown in the compound: #imgabs0#. The present invention has better antidepressant and neuroprotective effects than the existing clinical depression drugs fluoxetine and genipin, and has a fast onset of action, few side effects, and broad clinical application prospects.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a genipin derivative, a preparation method and application thereof. Background Art

[0002] Depression is a common mental disorder with obvious and persistent low mood as its main clinical feature. Studies have found that the incidence of depression has been on the rise worldwide in recent years.

[0003] At present, the commonly used antidepressants in clinical practice include monoamine oxidase inhibitors, tricyclic antidepressants, selective serotonin and norepinephrine reuptake inhibitors, etc. Among them, monoamine oxidase inhibitors were the first antidepressants discovered in the 1950s. They improve the patient's mood and interest in things by increasing the levels of norepinephrine, dopamine and 5-HT in the brain, achieving the antidepressant effect. They were once widely used, but because they are potentially dangerous when used in combination with a variety of drugs, they have gradually been replaced by tricyclic antidepressants. Tricyclic antidepressants have been used to treat depression since the 1960s. Common varieties include amitriptyline, imipramine, chlorpromazine, doxepin and clomipramine. Currently, clomipramine is the most widely used. This type of drug has a significant antidepressant effect, but the dosage is relatively large and there are many side effects, including dry mouth, constipation, difficulty urinating, blurred vision, dizziness, drowsiness, etc., so it is used less and less. Selective serotonin (5-HT) reuptake inhibitors are new antidepressants that are currently widely used in clinical practice. They can selectively inhibit 5-HT transporters, block the reuptake of 5-HT by the presynaptic membrane, prolong and increase the effect of 5-HT, and thus produce antidepressant effects. They have low affinity for adrenergic, histaminergic, and cholinergic receptors, and have weak effects, so they produce fewer adverse reactions. Representative drugs include fluoxetine, etc. The above existing clinical treatment drugs all have obvious defects such as delayed onset of action (2-6 weeks), low or even no efficacy for some refractory depression, and unsatisfactory clinical application effects. Summary of the invention

[0004] In view of the above problems, the present invention provides a rapid antidepressant drug genipin derivative with improved antidepressant efficacy, fewer side effects and rapid onset of action, as well as a preparation method and application thereof.

[0005] In a first aspect of the present invention, there is provided a genipin derivative or a pharmaceutically acceptable salt thereof, wherein the genipin derivative is of formula (I a ) or (I b ) shown in:

[0006]

[0007] Wherein: X and Y independently represent CH or N;

[0008] Z represents O or N-R2;

[0009] R1 and R2 each independently represent a C1-C3 straight or branched chain alkyl group;

[0010] R3 represents any one of H, C1-C6 alkyl, halogen, C1-C3 alkoxy, haloalkyl or nitro;

[0011] n represents 0, 1, 2 or 3.

[0012] As a specific embodiment of the present invention, the Z represents O.

[0013] As a specific embodiment of the present invention, the R3 represents any one of H, C1-C3 alkyl, and methoxy.

[0014] As a specific embodiment of the present invention, the pharmaceutically acceptable salt is selected from any one of hydrochloride, sulfate, phosphate, hydrobromide, maleate, fumarate, citrate, methanesulfonate, p-toluenesulfonate, tartrate or acetate, preferably hydrochloride.

[0015] As a specific embodiment of the present invention, it is selected from any one of the compounds shown in the following W-1 to W-8:

[0016]

[0017] In a second aspect of the present invention, a method for preparing the aforementioned genipin derivative or a pharmaceutically acceptable salt thereof is provided, the method comprising: mixing compound 1, compound 2a or compound 2b, a reaction aid and an organic solvent, and allowing the reaction to complete at room temperature, wherein the structural formulas of compound 1, compound 2a and compound 2b are as follows:

[0018]

[0019] Wherein: X and Y independently represent CH or N;

[0020] Z represents O or N-R2;

[0021] R1 and R2 each independently represent a C1-C3 straight or branched chain alkyl group;

[0022] R3 represents any one of H, C1-C6 alkyl, halogen, C1-C3 alkoxy, haloalkyl or nitro;

[0023] n represents 0, 1, 2 or 3;

[0024] Preferably, the molar ratio of the compound 1, compound 2a, and reaction aid is 1:1:2; the molar ratio of the compound 1, compound 2b, and reaction aid is 1:1:2;

[0025] Preferably, the reaction aid comprises potassium carbonate;

[0026] Preferably, the organic solvent comprises N,N-dimethylformamide;

[0027] Preferably, the reaction time is 4h;

[0028] Preferably, the method further comprises extracting the reaction product with an organic solvent, washing the obtained organic phase and drying, concentrating and purifying the organic phase.

[0029] The third aspect of the present invention provides a pharmaceutical composition, which is composed of a therapeutically effective amount of the aforementioned genipin derivative or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable carriers.

[0030] In a fourth aspect of the present invention, there is provided a use of the aforementioned genipin derivative or a pharmaceutically acceptable salt thereof in the preparation of a medicament for preventing or treating depression. The depression includes any one of mild, moderate or severe unipolar depression, bipolar depression, psychotic depression, reactive depression, secondary depression, seasonal depression, postpartum depression and menopausal depression.

[0031] The fifth aspect of the present invention provides use of the aforementioned genipin derivative or a pharmaceutically acceptable salt thereof in the preparation of a drug having a neuroprotective effect.

[0032] The beneficial effects of the present invention are:

[0033] The genipin derivative or the pharmaceutically acceptable salt thereof provided by the present invention has better antidepressant and neuroprotective effects than the existing clinical depression drugs fluoxetine and genipin, and has a rapid onset of action, few side effects, and broad prospects for clinical application. BRIEF DESCRIPTION OF THE DRAWINGS

[0034] Figure 1 Comparison of the results of forced swimming immobility time of behaviorally desperate mice in each group;

[0035] Figure 2 Comparison of the tail suspension immobility time of behaviorally desperate mice in each group;

[0036] Figure 3 Schedule for CUMS modeling, drug administration, and behavioral testing;

[0037] Figure 4A The open field experiment was used to observe the open field routes of the mice in each group for autonomous activity.

[0038] Figure 4B The open field experiment was used to observe the total distance and average speed of the autonomous activities of mice in each group.

[0039] Figure 5Forced swimming test to observe the effects of each group on the balance ability of CUMS mice ( n=10);

[0040] Figure 6 The tail suspension experiment was used to observe the effects of each group on the balance ability of CUMS mice. n=10);

[0041] Figure 7 Effects of W-3 on the PKA-CREB-BDNF signaling pathway in brain tissue ( n=3)( # P<0.05, compared with the normal group; *p<0.05, **p<0.01, compared with the CUMS group);

[0042] Fig. 8A This is the color comparison of each group of ribbons in the mouse hippocampus;

[0043] Figure 8B Comparison of 5HT1A content in each group of mouse hippocampus;

[0044] Figure 8C Comparison of GluN2B content in each group of mouse hippocampus;

[0045] Fig.8D Comparison of GluN2A content in each group of mouse hippocampus;

[0046] Fig.9A This is the color comparison of each group of ribbons in the mouse hippocampus;

[0047] Fig. 9B Comparison of PKA content in each group of mouse hippocampus;

[0048] Fig. 9C Comparison of BDNF content in each group of mouse hippocampus;

[0049] Fig.9D Comparison of CREB content in each group of mouse hippocampus;

[0050] Fig. 10A Comparison of Glu content in each group of mouse brain tissue;

[0051] Fig. 10B Comparison of BDNF content in each group of mouse brain tissue;

[0052] Fig. 10C Comparison of 5-HT content in each group of mouse brain tissue;

[0053] Fig. 10D Comparison of DA content in each group of mouse brain tissue;

[0054] Fig.10E Comparison of NE content in each group of mouse brain tissue;

[0055] Fig.11 is the MS spectrum of compound W-1;

[0056] Fig.12 is compound W-1 1 H-NMR spectrum;

[0057] Fig.13 is compound W-1 13 C-NMR spectrum;

[0058] Fig.14 is the MS spectrum of compound W-2;

[0059] Fig.15 For compound W-2 1 H-NMR spectrum;

[0060] Fig.16 is compound W-2 13 C-NMR spectrum;

[0061] Fig.17 is the MS spectrum of compound W-3;

[0062] Fig.18 is compound W-3 1 H-NMR spectrum;

[0063] Fig.19 is compound W-3 13 C-NMR spectrum;

[0064] Fig. 20 is the MS spectrum of compound W-4;

[0065] Fig.21 For compound W-4 1 H-NMR spectrum;

[0066] Fig. 22 For compound W-4 13 C-NMR spectrum;

[0067] Fig.23 is the MS spectrum of compound W-5;

[0068] Fig.24 is compound W-5 1 H-NMR spectrum;

[0069] Fig.25 is compound W-5 13 C-NMR spectrum;

[0070] Fig.26 is the MS spectrum of compound W-6;

[0071] Fig. 27 For compound W-6 1 H-NMR spectrum;

[0072] Fig.28 For compound W-6 13 C-NMR spectrum;

[0073] Fig.29 is the MS spectrum of compound W-7;

[0074] Fig.30 For compound W-7 1 H-NMR spectrum;

[0075] Fig.31 For compound W-7 13 C-NMR spectrum;

[0076] Fig.32 is the MS spectrum of compound W-8;

[0077] Fig.33 For compound W-8 1 H-NMR spectrum;

[0078] Fig.34 For compound W-8 13 C-NMR spectrum. DETAILED DESCRIPTION

[0079] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments in the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. The experimental methods described in the following test examples are all conventional methods unless otherwise specified; the reagents and materials, unless otherwise specified, can be obtained from commercial channels.

[0080] Example 1

[0081] Compound W-3: Methyl (1S,4aR)-1-ethoxy-7-((4-((3,5,6-trimethylpyrazin-2-yl)methyl)piperazin-1-yl)methyl)-1,4a,5,7a-tetrahydrocyclopenta[c]pyran-4-carboxylate hydrochloride

[0082]

[0083] The synthesis method is as follows:

[0084] Step 1: Synthesis of methyl 1-ethoxy-7-(hydroxymethyl)-1,4a,5,7a-tetrahydrocyclopenta[c]pyran-4-carboxylate

[0085]

[0086] In a 50 mL round-bottom flask, genipin (500.0 mg, 2.21 mmol), p-toluenesulfonic acid monohydrate (504.5 mg, 2.65 mmol) and 3.87 mL of ethanol were added in sequence. After reacting at 80 °C for 1 h, TLC [V (petroleum ether): V (ethyl acetate) = 1:1 as the developing solvent] showed that the reaction was basically complete. 20 mL of saturated NaHCO3 aqueous solution was added to quench the reaction, and the mixture was extracted with dichloromethane (3×50 mL). The organic phases were combined, washed with saturated brine (3×50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a yellow oil. The mixture was purified by silica gel column chromatography [V (petroleum ether): V (ethyl acetate) = 4:1 as the eluent] to obtain 497.8 mg of a light yellow oil with a yield of 88.6%.

[0087] Step 2: Synthesis of methyl 1-ethoxy-7-((methylsulfonyl)oxy)-1,4a,5,7a-tetrahydrocyclopentyl[c]pyran-4-carboxylate

[0088]

[0089] In a 50 mL round-bottom flask, methyl 7-(hydroxymethyl)-1-ethoxy-1,4a,5,7a-tetrahydrocyclopenta[c]pyran-4-carboxylate (200.0 mg, 786.5 μmol), 5 mL of dichloromethane, and Et3N (0.2 mL, 1.4 mmol) were added in sequence. Methanesulfonyl chloride (0.060 mL, 786.5 μmol) was slowly added dropwise under ice bath conditions. After the addition was complete, the reaction was continued under ice bath conditions for 2 h. TLC [V (petroleum ether): V (ethyl acetate) = 1:1 as the developing solvent] showed that the reaction was basically complete. The reaction was quenched with 10% dilute hydrochloric acid, and extracted with dichloromethane (3×50 mL). The organic phases were combined, washed with saturated brine (3×50 mL), dried over anhydrous sodium sulfate, and concentrated to obtain 250 mg of a light yellow oil with a yield of 95.6%.

[0090] Step 3: Synthesis of 2,3,5-trimethyl-6-(piperazin-1-ylmethyl)-p-pyrazine

[0091]

[0092] In a 100mL round-bottom flask, ligustrazine (4.00g, 29.37mmol) was added, dissolved with 30mL of glacial acetic acid, and then 30% H2O2 (5.93mL, 58.74mmol) was added. The reaction was carried out at 70°C for 4h. TLC [V(petroleum ether):V(ethyl acetate)=1:1 as the developing solvent] showed that the reaction was basically complete. 20mL of saturated NaHCO3 aqueous solution was added to quench the reaction, and the mixture was extracted with dichloromethane (3×50mL). The organic phases were combined, washed with saturated brine (3×50mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 2.479g of a white solid with a yield of 75%.

[0093]

[0094] 2,3,5,6-ligustrazine 1-oxide (2.50 g, 16.41 mmol) was added to a 100 mL round-bottom flask, and 16 mL of acetic acid was added to dissolve it. Then 10 ml of acetic anhydride was added and the reaction was carried out at 110 ° C for 6 h. TLC [V (petroleum ether): V (ethyl acetate) = 1:1 as the developing solvent] showed that the reaction was basically complete. 20 mL of saturated NaHCO3 aqueous solution was added to quench the reaction, and it was extracted with dichloromethane (3×50 mL). The organic phases were combined, washed with saturated brine (3×50 mL), dried over anhydrous sodium sulfate, and the filtrate was concentrated under reduced pressure to obtain 2.92 g of brown oily liquid. The yield was 91.5%.

[0095]

[0096] In a 100 mL round-bottom flask, (3,5,6-trimethylpyrazin-2-yl)acetic acid methyl ester (2.92 g, 15.03 mmol) was added and dissolved with 15 mL of anhydrous ethanol. Then, an aqueous solution of NaOH (3.61 g, 90.20 mmol) was added and reacted at 75 °C for 2 h. TLC [V (petroleum ether): V (ethyl acetate) = 1:1 as the developing solvent] showed that the reaction was basically complete. 20 mL of distilled water was added to quench the reaction, and the mixture was extracted with dichloromethane (3 × 50 mL). The organic phases were combined, washed with saturated brine (3 × 50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain 1.587 g of a yellow solid with a yield of 69.4%.

[0097]

[0098] In a 50 mL round-bottom flask, (3,5,6-trimethylpyrazine-2-yl)methanol (1.59 g, 10.43 mmol) was added and dissolved in 10 mL of anhydrous dichloromethane. Then, SOCl2 (1.86 g, 15.64 mmol) was added under an ice bath and reacted at room temperature for 3 h. TLC [V (petroleum ether): V (ethyl acetate) = 1:1 as the developing solvent] showed that the reaction was basically complete. 20 mL of ice water was added to quench the reaction, and the mixture was extracted with dichloromethane (3×50 mL). The organic phases were combined, washed with saturated brine (3×50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a light yellow oily liquid. The mixture was purified by silica gel column chromatography [V (petroleum ether): V (ethyl acetate) = 6:1 as the eluent] to obtain 900.0 mg of a colorless oily liquid with a yield of 50.6%.

[0099]

[0100] Anhydrous piperazine (2.73 g, 31.65 mmol) was added to a 50 mL round-bottom flask and dissolved in 15 mL of dichloromethane. A dichloromethane solution of 2-(bromomethyl)-3,5,6-trimethylpyrazine (900.0 mg, 5.27 mmol) was slowly added dropwise at 0°C. After completion of the addition, the reaction was continued for 6 h in an ice bath. TLC [V (petroleum ether): V (ethyl acetate) = 1:1 as the developing solvent] showed that the reaction was basically complete. 20 mL of distilled water was added to quench the reaction, and the mixture was extracted with dichloromethane (3×50 mL). The organic phases were combined, washed with saturated brine (3×50 mL), dried over anhydrous sodium sulfate, and concentrated under reduced pressure to obtain a white solid crude product, which was purified by silica gel column chromatography [V (dichloromethane): V (methanol) = 10:1 as the eluent] to obtain 432.0 mg of a white solid with a yield of 37.2%.

[0101] Step 4: Synthesis of methyl 1-ethoxy-7-((methylsulfonyl)oxy)-1,4a,5,7a-tetrahydrocyclopenta[c]pyran-4-carboxylate

[0102]

[0103] Take a 250mL dry single-necked flask, add 1-ethoxy-7-((methylsulfonyl)oxy)methyl)-1,4a,5,7a-tetrahydrocyclopenta[c]pyran-4-carboxylic acid methyl ester (361.22mg, 1.13mmol), 2-(piperazinemethyl)-3,5,6-trimethylpyrazine (500mg, 2.27mmol), K2CO3 (313.65mg, 2.27mmol) in sequence, dissolve in 15mL DMF, and heat at room temperature. After 4 hours of reaction, TLC [V (petroleum ether): V (ethyl acetate) = 1:1 as the developing solvent] showed that the reaction was basically complete, 20 mL of distilled water was added to quench the reaction, and dichloromethane (3×50 mL) was used for extraction. The organic phases were combined, washed with saturated brine (3×50 mL), dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by silica gel column chromatography [V (dichloromethane): V (methanol) = 20:1 as the eluent] to obtain 87.2 mg of a yellow solid with a yield of 58%. mp 273.8~274.9℃. 1 H NMR(300MHz,Chloroform-d)δ7.52(s,1H,3-H),5.74(s,1H,7-H),4.71(d,J=7.3Hz, 1H,1-H),3.75(s,4H,CH2),3.63(d,J=7.1Hz,3H,COOCH3),3.19(q,J=8.5,8.1Hz,2H ,CH2),3.04(d,J=14.2Hz,1H,9-H),2.85(dd,J=15.6,9.1Hz,1H,5-H),2.71–2.63(m ,2H,CH2),2.60(s,8H,piperazine-H),2.52(s,9H,CH3),1.26(d,J=7.1Hz,3H,CH3). 13 C NMR(101MHz,Chloroform-d)δ168.01,152.31,149.87,149.47,148.03,129.52,110.95,101.29,65.21 ,61.76,57.93,53.22,51.14,46.60,38.78,35.11,21.55,21.45,20.97,15.13.ESI-HRMS(m / z):[M+H] + Calculate for C 25 H 36 N4O4:457.2809found457.2804.

[0104] Example 2

[0105] Compound W-1: 1-methoxy-7-((4-(4-methoxybenzyl)piperazin-1-yl)methyl)-1,4a,5,7a-tetrahydrocyclopentane[c]pyran-4-carboxylic acid methyl ester hydrochloride

[0106]

[0107] Methyl 1-methoxy-7-((methylsulfonyl)oxy)-1,4a,5,7a-tetrahydrocyclopentyl[c]pyran-4-carboxylate was synthesized by referring to the method of steps 1 to 2 of Example 1, and W-1 was obtained as a white solid with mp 226.8-227.9°C by combining it with 1-(4-methoxybenzyl)piperazine as raw materials and referring to the method of step 4 of Example 1. 1 H NMR(300MHz,D2O)δ7.39(s,1H,3-H),7.32(d,J=8.1Hz,2H,ArH),6.94(d,J=8.5Hz,2H,ArH),6.18(b rs,1H,7-H),5.11(d,J=2.7Hz,1H,1-H),4.31(brs,1H,ArCH2-Ha),4.29(s,3H,ArOCH3),4.02~3.82( m,1H,ArCH2-Hb),3.71(s,6H,COOCH3,OCH3),3.65–3.47(m,8H,piperazine-H),3.46(s,2H,10-H),3 .41(brs,1H,9-H),3.30(brs,1H,5-H),3.18~2.98(m,1H,6-Ha),2.71(dd,J=17.6,7.9Hz,1H,6-Hb). 13 C NMR(75MHz,D2O)δ:169.9,160.5,151.8,143.1,132.9,128.6,119.7,114.7, 111.3,99.1,59.9,56.5,55.4,51.8,47.8,47.7,45.8,38.7,33.0; IR(KBr,cm -1 )υ:2975.1,2904.6,2834.2,1708.9,1613.3,1517.1,1465.2,1439.6,1381.9,1253.2,1182.5,1074.4,1032.0,848.9; ESI-Mass for C 24 H 32 N2O5:m / z 429.25(M + +H).

[0108] Example 3

[0109] Compound W-2: 1-methoxy-7-((4-(pyridin-3-ylmethyl)piperazin-1-yl)methyl)-1,4a,5,7a-tetrahydrocyclopenta[c]pyran-4-carboxylic acid methyl ester hydrochloride

[0110]

[0111] Methyl 1-methoxy-7-((methylsulfonyl)oxy)-1,4a,5,7a-tetrahydrocyclopentyl[c]pyran-4-carboxylate was synthesized by referring to the method of steps 1 to 2 of Example 1, and W-2 was obtained as a white solid with mp 197.1-198.3°C by using it and 1-(pyridin-3-ylmethyl)piperazine as raw materials. 1 H NMR(300MHz,D2O)δ8.98(s,1H,Pyridine-H),8.84(d,J=5.7Hz,1H,Pyridine-H),8.74(d,J=8.2Hz,1H,Pyridine-H),8 .11(t,J=7.8Hz,1H,Pyridine-H),7.45(s,1H,3-H),6.23(d,J=18.1Hz,1H,7-H),4.55(d,J=8.3Hz,1H,1-H),3.97(brs ,2H,CH2),3.83~3.61(m,8H,piperazine-H),3.59(s,3H,COOCH3),3.46(s,2H,10-CH2),3.30(s,3H,OCH3),3.09(t,J= 8.1Hz, 1H, 9-H), 2.79 (ddd, J=11.1, 8.7, 2.7Hz, 1H, 5-H), 2.70~2.56 (m, 1H, 6-Ha), 2.04 (dd, J=17.1, 9.4Hz, 1H, 6-Hb). 13 C NMR(75MHz,D2O)δ:169.8,153.1,151.7,149.7,143.4,129.8,129.1,128.2,110.4,102 .5,99.1,57.3,56.48,56.0,55.6,55.1,51.8,48.6,46.0,38.7,35.0,32.9.IR(KBr,cm -1 )υ:2978.7,2956.3,2824.1,1701.8,1633.2,1532.7,1436.2,1460.2,1379.3,1289.1,1179.3,1083.5,946.7,891.0,686.1; ESI-Mass for C 22 H 29N3O4: m / z 400.22 (M + +H).

[0112] Example 4

[0113] Compound W-4: 1-isopropoxy-7-((4-(4-methoxybenzyl)piperazin-1-yl)methyl)-1,4a,5,7a-tetrahydrocyclopentane[c]pyran-4-carboxylic acid methyl ester hydrochloride

[0114]

[0115] Methyl 1-isopropoxy-7-((methylsulfonyl)oxy)-1,4a,5,7a-tetrahydrocyclopentyl[c]pyran-4-carboxylate was synthesized by referring to the method of steps 1 to 2 of Example 1, and W-4 was obtained as a white solid with mp 225.3-226.5°C by using it and 1-(4-methoxybenzyl)piperazine as raw materials and referring to the method of step 4 of Example 1. 1 H NMR(300MHz,D2O)δ7.45(s,1H,3-H),7.38(d,J=8.7Hz,2H,ArH),6.97(d,J=8.7Hz,2H,ArH),6.32(s,1H,7-H),4.68(d,J =8.4Hz,1H,1-H),4.35(s,2H,CH2),4.02(dd,J=12.7,6.3Hz,2H,10-CH2),3.94~3.85(m,1H,CH),3.75(s,3H,COOCH3),3 .62(s,3H,OCH3),3.61~3.45(m,8H,piperazine-H),3.12(q,J=8.4Hz,1H,6-Ha),2.82(ddd,J=11.4,8.7,2.7Hz,1H,5-H ), 2.55 (t, J = 8.0Hz, 1H, 9-H), 2.07 (dd, J = 17.4, 9.4Hz, 1H, 6-Hb), 1.17 (d, J = 6.0Hz, 3H, CH3), 1.07 (d, J = 6.0Hz, 3H, CH3). 13 C NMR(75MHz,D2O)δ170.0,160.5,153.4,143.1,132.9,130.0,119.7,114.8,110.3,100.1,73 .8,72.9,60.0,55.6,55.4,51.8,48.3,47.8,46.2,38.8,35.4,33.8,22.4,20.9; IR(KBr,cm -1)υ:2972.1,2830.7,1712.0,1631.0,1518.0,1437.8,1382.9,1251.4,1157.9,1104.3,945.2,857.9; ESI-Mass for C 26 H 36 N2O5: m / z 457.21 (M + +H).

[0116] Example 5

[0117] Compound W-5: 1-methoxy-7-((4-(4-methylbenzyl)piperazin-1-yl)methyl)-1,4a,5,7a-tetrahydrocyclopentane[c]pyran-4-carboxylic acid methyl ester hydrochloride

[0118]

[0119] Methyl 1-methoxy-7-((methylsulfonyl)oxy)-1,4a,5,7a-tetrahydrocyclopentyl[c]pyran-4-carboxylate was synthesized by referring to the method of steps 1 to 2 of Example 1. It was reacted with 1-(4-methylbenzyl)piperazine as raw materials and W-5 was obtained as a white solid by referring to the method of step 4 of Example 1, mp 231.3-232.4°C. 1 HNMR(300MHz,D2O)δ:7.40(s,1H,3-H),7.29(d,J=8.1Hz,2H,ArH),7.23(d,J=8.1Hz,2H,ArH),6. 18(brs,1H,7-H),5.12(d,J=3.0Hz,1H,1-H),4.33(s,2H,CH2),3.97(d,J=13.8Hz,1H,10-Ha),3.8 8(d,J=13.8Hz,1H,10-Hb),3.60(s,3H,COOCH3),3.58~3.35(m,8H,piperazine-H),3.30(s,3H,OC H3),3.09(t,J=8.1Hz,2H,6-H),2.76~2.68(m,1H,9-H),2.24(s,3H,CH3),2.12~2.05(m,1H,5-H). 13 CNMR(75MHz,D2O)δ:169.8,151.7,143.2,141.3,131.1,130.0,128.5,124.3,11 1.3,99.1,60.2,56.4,55.1,51.8,48.3,47.8,45.8,38.7,33.0,20.3;IR(KBr,cm -1)υ:2981.3,2950.6,1712.5,1637.4,1433.6,1368.0,1315.7,1288.9,1271.8,1197.4,1167.2,1074.0,1024.6,954.1,890.0; ESI-Mass for C 24 H 32 N2O4:m / z 413.30(M + +H).

[0120] Example 6

[0121] Compound W-6: 1-isopropoxy-7-((4-(3-methoxybenzyl)piperazin-1-yl)methyl)-1,4a,5,7a-tetrahydrocyclopentane[c]pyran-4-carboxylic acid methyl ester hydrochloride

[0122]

[0123] Methyl 1-isopropoxy-7-((methylsulfonyl)oxy)-1,4a,5,7a-tetrahydrocyclopentyl[c]pyran-4-carboxylate was synthesized by referring to the method of steps 1 to 2 of Example 1, and W-6 was obtained as a white solid with mp 208.0-209.2°C by using it and 1-(3-methoxybenzyl)piperazine as raw materials and referring to the method of step 4 of Example 1. 1 H NMR(300MHz,D2O)δ7.44(s,1H,3-H),7.36(t,J=8.3Hz,1H,ArH),7.07~6.97(m,3H,ArH),6.26(s,1H,7-H),4.67(d,J=8 .4Hz,1H,1-H),4.38(s,2H,CH2),4.03(dd,J=13.5,7.8Hz,2H,10-CH2),3.96~3.85(m,1H,CH),3.74(s,3H,COOCH3),3.6 6(s,3H,OCH3),3.61~3.47(m,8H,piperazine-H),3.11(q,J=8.4Hz,1H,6-Ha),2.81(ddd,J=11.4,8.4,2.7Hz,1H,5-H) ,2.55(t,J=7.5Hz,1H,9-H),2.06(dd,J=17.6,9.5Hz,1H,6-Hb),1.16(d,J=6.0Hz,3H,CH3),1.06(d,J=6.0Hz,3H,CH3). 13C NMR(75MHz,D2O)δ170.0,159.4,153.4,143.3,130.8,129.9,128.8,123.8,116.6,116.2,110.3 ,100.1,73.8,60.3,55.6,55.5,51.8,48.3,48.1,46.2,38.8,35.4,23.7,22.4,20.9; IR(KBr,cm -1 )υ:2973.2,2947.8,2883.6,2831.9,1706.6,1629.8,1371.3,1437.1,1270.1,1160.3,1101.6,940.3,888.5,787.1,697.6; ESI-Mass for C 26 H 36 N2O5:m / z457.16(M + +H).

[0124] Example 7

[0125] Compound W-7: 1-methoxy-7-((4-(3-methylbenzyl)piperazin-1-yl)methyl)-1,4a,5,7a-tetrahydrocyclopentane[c]pyran-4-carboxylic acid methyl ester hydrochloride

[0126]

[0127] Methyl 1-methoxy-7-((methylsulfonyl)oxy)-1,4a,5,7a-tetrahydrocyclopentyl[c]pyran-4-carboxylate was synthesized by referring to the method of steps 1 to 2 of Example 1. It was reacted with 1-(3-methylbenzyl)piperazine as raw materials and W-7 was obtained as a white solid by referring to the method of step 4 of Example 1, mp 230.5-231.6°C. 1 H NMR(300MHz,D2O)δ:7.48(s,1H,3-H),7.36~7.18(m,4H,ArH),6.28(brs,1H,7-H),4.56(d,J=8 .2Hz,1H,1-H),4.36(s,2H,CH2),4.08~3.90(m,2H,10-CH2),3.63(s,3H,COOCH3),3.61~3.50( m,8H,piperazine-H),3.48(s,3H,OCH3),3.12(dd,J=15.9,7.5Hz,1H,9-H),2.81(dd,J=17.0, 8.5Hz, 1H, 6-Ha), 2.64~2.49 (m, 1H, 6-Hb), 2.27 (s, 3H, CH3), 2.06 (dd, J=16.5, 8.7Hz, 1H, 5-H).13 C NMR(75MHz,D2O)δ:170.00,153.10,142.64,139.77,131.72,131.23,129.86,129.33,128.11,127.3 1,110.44,102.57,60.43,57.30,55.65,51.82,48.33,47.94,45.98,38.75,35.06,20.33.IR(KBr,cm -1 )υ:2977.1,2947.6,2629.5,1708.2,1627.6,1438.2,1369.9,1287.6,1178.7,1107.6,957.8,939.5,890.1,766.6,700.3.ESI-Mass for C 24 H 32 N2O4: m / z 413.26 (M + +H).

[0128] Example 8

[0129] Compound W-8: 1-isopropoxy-7-((4-(4-methylbenzyl)piperazin-1-yl)methyl)-1,4a,5,7a-tetrahydrocyclopentane[c]pyran-4-carboxylic acid methyl ester hydrochloride

[0130]

[0131] Methyl 1-isopropoxy-7-((methylsulfonyl)oxy)-1,4a,5,7a-tetrahydrocyclopentyl[c]pyran-4-carboxylate was synthesized by referring to the method of steps 1 to 2 of Example 1. It was reacted with 1-(4-methylbenzyl)piperazine as raw materials and W-8 was obtained as a white solid by referring to the method of step 4 of Example 1, mp 223.1-224.4°C. 1H NMR (300MHz, D2O) δ7.42(s,1H,3-H),7.28(d,J=8.0Hz,2H,ArH),7.21(d,J=8.0Hz,2H,ArH),6.28(s,1H,7-H),4.67(d,J=8. 4Hz,1H,1-H),4.33(s,2H,CH2),4.00(d,J=14.1Hz,1H,10-Ha),3.93(d,J=14.1Hz,1H,10-Hb),3.79(s,3H,COOCH3),3.63~3 .39(m,8H,piperazine-H),3.14(m,1H,CH),2.77(ddd,J=11.1,8.1,2.7Hz,1H,5-H),2.52(t,J=8.1Hz,1H,9-H),2.22(s,3H , CH3), 2.20~2.16 (m, 1H, 6-Ha), 2.04 (dd, J=17.7, 9.4Hz, 1H, 6-Hb), 1.13 (d, J=6.3Hz, 3H, CH3), 1.02 (d, J=6.3Hz, 3H, CH3). 13 C NMR(75MHz,D2O)δ170.0,153.4,143.3,141.3,131.2,130.0,124.2,110.3,100.1,95.6, 73.8,60.2,55.6,51.8,47.9,47.0,46.2,38.8,35.4,23.7,22.4,20.9,20.4; IR(KBr,cm -1 )υ:3031.5,2972.9,2945.6,2873.2,1712.3,1630.8,1436.1,1384.2,1371.4,1284.7,1242.9,1157.0,1103.4,941.9,849.0; ESI-Mass for C 26 H 36 N2O4: m / z 441.26 (M + +H).

[0132] Test example

[0133] 1. Detection of the protective activity of the genipin derivatives of the present invention against glutamate-induced HT-22 cell damage

[0134] Mouse hippocampal neuron HT-22 cells were selected for digestion, collection and centrifugation. After the cells were resuspended and counted, 2×10 4 / well HT-22 cells were inoculated in a 96-well cell culture plate, the volume of the cell suspension in each well was 100 μL, 6 replicate wells were set up in each group, and a normal control group, a model group and a drug intervention group were set up. The cell culture plate was placed in an incubator at 37°C and 5% CO2 for 24 hours. After 24 hours, the mouse hippocampal neuron HT-22 cells were Glu-induced to establish an injury model. After the supernatant in each well of the 96-well cell culture plate was sucked out with a pipette, the cells were grouped and processed: ① The normal control group was cultured with normal medium replacement for 24 hours; ② The model group was added with 150 μL of 5mM Glu solution and the cells were placed in a 37°C, 5% CO2 incubator for 24 hours. 4 hours before the end of incubation, 10 μL of CCK-8 solution was added to each well of the well plate; ③ The drug-treated group was added with 150 μL of compound solution containing 5mM final concentration of Glu, and the final concentrations of the compounds were 0.1 μM, 1 μM and 10 μM. The well plate was placed in a 37°C, 5% CO2 incubator for 24 hours. 4 hours before the end of incubation, 10 μL of CCK-8 solution; ④ In the fluoxetine group, 150 μL of 0.1 μM, 1 μM and 10 μM fluoxetine solutions of different concentrations were added and placed in a 37°C, 5% CO2 incubator for incubation for 24 h. 4 h before the end of incubation, 10 μL of CCK-8 solution was added to each well; after adding CCK-8 reagent, the 96-well plate was wrapped with tin foil and placed in an incubator for incubation for 4 h, and then placed in a multi-function microplate reader to detect the OD value of each well at a wavelength of 570 nm. The experiment was repeated 3 times.

[0135] As shown in Table 1, the in vitro HT-22 cell protection activity experiment shows that the genipin derivatives prepared in each embodiment of the present invention have protective activity against HT-22 cells induced by Glu, and the results show a concentration-dependent increase, and the in vitro neuronal cell protection activity of the screened genipin derivatives is better than that of the control drug genipin. Among them, compound W-3 ​​exhibits good neuronal cell protection activity, and can significantly improve the cell survival rate in the Glu-induced mouse hippocampal neuron HT-22 cell injury model at 10 μM (cell survival rate 75.06 ± 1.12), and the results are better than the positive control fluoxetine (cell survival rate 74.84 ± 0.92) and the control drug genipin (57.03 ± 1.26).

[0136] Table 1 Effects of genipin derivatives on glutamate (Glu)-induced HT-22 cell injury ( n=6)

[0137]

[0138]

[0139] Note: The cell survival rate of the blank control group (100±2.88) and the drug-treated group was compared with the model control group.* p<0.05, ** p<0.01; drug treatment group compared with positive drug fluoxetine group, # p<0.05, ## p<0.01.

[0140] II. Effects of the Genipin Derivatives of the Present Invention on the Immobility Time of Mice in the Forced Swim Test and the Tail Suspension Test

[0141] The experimental animals were male ICR mice (purchased from Anhui Medical University, animal production license number scxk(Wan)-2017-001); genipin derivatives (W-3 and W-6) were dissolved in DMSO (purchased from Shanghai Biyuntian Biotechnology Co., Ltd.) and diluted to the required concentration with 5% DMSO in 9% Tween80 / normal saline solution before use for intraperitoneal injection; fluoxetine (purchased from Aladdin Biochemical Technology Co., Ltd.).

[0142] Experimental procedure: The experimental mice were raised in a 12h / 12h light-dark cycle and had free access to water. The drugs were administered after 5 days of adaptation. The mice were randomly divided into 8 groups, with 6 mice in each group. The drug solvent was a 5% DMSO solution prepared with 9% Tween80 in saline solution. The 8 groups of animals were (a) blank control group; (b) W-6 low, medium and high dose groups (1mg / kg, 10mg / kg and 30mg / kg); (c) W-3 low, medium and high dose groups (1mg / kg, 10mg / kg and 30mg / kg); (d) fluoxetine 30mg / kg group (dissolved in saline). Each group was given a single intraperitoneal injection, and a 5-min forced swimming test and a tail suspension test were performed 1 hour after administration, and the immobility time was recorded.

[0143] Forced swimming test: The forced swimming apparatus is made of transparent plexiglass (30 cm high, 15 cm in diameter, 15 ± 1 cm in water depth, 25 ± 1 °C), and the mice were allowed to adapt for 15 min the day before the test. During the test, the mice were placed in the apparatus and the floating time of the mice was recorded within 6 min and 4 min after the test. The floating time was defined as the time when the mice curled up slightly and were in a floating state (the forced swimming test is a classic antidepressant drug screening model, and effective antidepressant drugs can significantly reduce the immobility time of mice in forced swimming).

[0144] Tail suspension test: The experimental operation was carried out according to the method of Lucien Steru. A laboratory-made tail suspension box was used. A small clip was fixed on the top of each tail suspension box (25×25×25 cm). The mice were taped 2 cm away from the tip of the tail and then suspended for 6 minutes. Video recording was then used to analyze the immobility time of the mice for 4 minutes after the 6-minute tail suspension period using software or manual analysis.

[0145] Statistical analysis: GraphPadprism 9.0 was used to analyze the data. All quantitative data were expressed as mean ± standard deviation. The data were tested for normality and homogeneity of variance. The measurement data that met the conditions were tested with t-test (comparison between two groups) or analysis of variance (comparison between multiple groups). Dunnett's-T3 was used for pairwise comparison when the variance was unequal. If the data did not conform to the normal distribution, the Kruskal-Wills H test in the nonparametric test was used. p≤0.05 was considered statistically significant.

[0146] The results of the forced swimming test were as follows Figure 1 As shown, compared with the blank control group (160.0±12.98s), the forced swimming immobility time of mice in the W-6 10mg / kg single-dose group (115.3±25.14s) and fluoxetine 30mg / kg single-dose group (112.2±19.15s) was significantly reduced (p<0.05), and the forced swimming immobility time of mice in the W-31mg / kg single-dose group (84.5±24.93s), W-310mg / kg single-dose group (82±17.61s), and W-330mg / kg single-dose group (79.33±14.34s) was extremely significantly reduced (p<0.01).

[0147] The results of the tail suspension test are as follows Figure 2 As shown, compared with the vehicle control group (153.5±17.36s), the immobility time of the mice in the tail suspension test in the W-6 1mg / kg single administration group (93.83±20.85s), W-6 10mg / kg single administration group (63.67±14.61s), W-3 1mg / kg single administration group (107.2±17.00s), W-3 10mg / kg single administration group (60.83±16.87s), W-3 30mg / kg single administration group (56.33±10.27s) and fluoxetine 30mg / kg single administration group (76±20.14s) was significantly reduced (p<0.01). Compared with the model control group, W-6 and W-3 ​​and the positive drug fluoxetine can increase the immobility time of mice in each concentration group (p≤0.01), showing a certain antidepressant activity.

[0148] III. Effects of the genipin derivatives of the present invention on depressive-like behavior in mice induced by chronic unpredictable stress

[0149] The experimental process is as follows Figure 3As shown, 3-month-old male C57BL / 6J mice (purchased from Hangzhou Ziyuan Experimental Animal Technology Co., Ltd., animal use license number: SCXK (Su) 2018-0008) were used. Before the experiment, the mice were adapted to the animal room of Anhui University of Traditional Chinese Medicine for one week. The temperature and humidity of the breeding room were 25 ± 2 °C and 55 ± 5%, respectively, with a 12-h day and night cycle. All animal experiments were approved by the Experimental Animal Ethics Committee of Anhui University of Traditional Chinese Medicine (Hefei, Anhui, China). After 5 days of adaptation, chronic unpredictable stress (CUMS) was given; the mice were randomly divided into 6 groups, with 10 mice in each group; the drug solvent was normal saline; the 6 groups of animals were (a) normal group: normal saline (ip); (b) model group: normal saline (ip) + CUMS; (c) W-3 0.1 mg / kg + CUMS group; (d) W-3 1 mg / kg + CUMS group; (e) W-3 10 mg / kg + CUMS group; (f) fluoxetine 20 mg / kg group + CUMS. After the grouping, the group given CUMS was subjected to CUMS for 28 days, and the blank control group did not receive behavioral intervention. After the CUMS, each group was given a single intraperitoneal injection of the corresponding drug, and a sugar water preference test was performed on the first day after the CUMS, an open field test was performed on the second day, a forced swimming test was performed on the third day, and a sugar water preference test was performed again on the 7th day.

[0150] CUMS protocol: Mice were randomly exposed to two different chronic unpredictable stresses every day for 28 days. The stress methods used are described in Table 2.

[0151] Table 2 Chronic Unpredictable Mild Stress (CUMS) Method

[0152]

[0153]

[0154] Open field test: Using the open field test video analysis system Supermaze (Shanghai Xinruan Information Technology), the mice were placed in the room waiting for the test and adapted for about 10 minutes, and then placed in the center of the open field stereo chamber (400mm (W) × 400mm (H) × 400mm (D)). The mice were allowed to move freely for 5 minutes. After the video recording, the system automatically analyzed the trajectory of the mice and obtained the total distance and average speed within 5 minutes. After each open field test, 10% ethanol solution was used to clean and deodorize before the next open field test.

[0155] Forced swimming test: The forced swimming apparatus is made of transparent plexiglass (30 cm high, 15 cm in diameter, 15 ± 1 cm in water depth, 25 ± 1 °C), and the mice were allowed to adapt for 15 min the day before the test. During the test, the mice were placed in the apparatus and the floating time of the mice was recorded within 6 min and 4 min after the test. The floating time was defined as the time when the mice curled up slightly and were in a floating state (the forced swimming test is a classic antidepressant drug screening model, and effective antidepressant drugs can significantly reduce the immobility time of mice in forced swimming).

[0156] Tail suspension test: The experimental operation was carried out according to the method of Lucien Steru. A laboratory-made tail suspension box was used. A small clip was fixed on the top of each tail suspension box (25×25×25 cm). The mice were taped 2 cm away from the tip of the tail and then suspended for 6 minutes. Video recording was then used to analyze the immobility time of the mice for 4 minutes after the 6-minute tail suspension period using software or manual analysis.

[0157] Statistical analysis: GraphPadprism 9.0 was used to analyze the data. All measured data were expressed as mean ± standard deviation. The data were tested for normality and homogeneity of variance. The measurement data that met the conditions were tested with t-test (comparison between two groups) or analysis of variance (comparison between multiple groups). Dunnett's-T3 was used for pairwise comparison when the variance was unequal. If the data did not conform to the normal distribution, the Kruskal-Wills H test in the nonparametric test was used. p≤0.05 was considered statistically significant.

[0158] The results of the open field test are as follows Figures 4A to 4B As shown in the figure, the open field test evaluates the autonomous activity ability of mice by measuring the total distance and average speed of mice. The results showed that compared with the normal group, the total distance and average speed of mice in the CUMS group were significantly reduced (p<0.01), and the fluoxetine hydrochloride and genipin derivative administration groups were able to prolong the total distance and average speed of mice (p<0.5).

[0159] The results of the forced swimming test were as follows Figure 5 As shown in the figure, the forced swimming test is to record and score the forced swimming time and movements of mice. Compared with the normal group, the immobility time of the CUMS group was prolonged (p<0.01). During the experiment, it can be observed that the immobility time of the mice in the CUMS group was prolonged. The immobility time of the mice in the genipin derivative group and the fluoxetine hydrochloride group in the water was reduced (p<0.01). The results are shown in the figure. Figure 6 .

[0160] The results of the tail suspension test are as follows Figure 6As shown in the figure, the tail suspension test is to record and score the hanging time and movements of mice. Compared with the normal group, the hanging score of the CUMS group was reduced (p<0.01). During the experiment, it can be observed that the inverted hanging immobility time of mice in the CUMS group was prolonged. The inverted hanging immobility time of mice in the genipin derivative administration group and the fluoxetine hydrochloride group was reduced (p<0.01).

[0161] The above experiments show that a single administration of W-3 has a therapeutic effect on depression, and the effect takes effect quickly and lasts for seven days or more.

[0162] IV. Effects of the Genipin Derivatives of the Present Invention on the BDNF / PKA / CREB Signaling Pathway

[0163] Place the samples of different groups on ice in a 6-well plate and wash them three times with pre-cooled PBS buffer; RNA extraction solution lysis: take 500 μL of pre-cooled RNA extraction solution, add the extraction solution to a small dish, place it on ice for 5 minutes to fully lyse it, and then transfer the sample to an EP tube, mix it, and let it stand at room temperature for 5 minutes; add chloroform: add chloroform (1 / 5 of the volume of the extraction solution) in proportion to another EP tube, shake and mix it, let it stand at room temperature for 10 minutes, and centrifuge it for 15 minutes (4°C, 12000rpm); isopropanol precipitation: on the upper layer Add an equal volume of isopropanol to the aqueous phase, mix well, let stand at room temperature for 20 minutes, and centrifuge for 10 minutes (4°C, 12000rpm / min); ethanol elution: discard the supernatant, add 1mL of pre-cooled 75% ethanol (anhydrous ethanol: DEPC water = 3:1), and centrifuge for 15 minutes (4°C 12000rpm / min); extract total RNA: discard the supernatant, dry for 10-30 minutes, add 20μL DEPC water to dissolve and precipitate, obtain total RNA, and store at -80°C; determine RNA purity (OD 260 / OD 280 ).

[0164] qRT-PCR results Figure 7 As shown: Compared with the normal group, the mRNA expression levels of PKA, CREB, and BDNF in the CUMS group mice were decreased (p<0.05). Compared with the CUMS group, the mRNA expression level of the compound W-3 ​​group was increased (p<0.05), and the increase was dose-dependent.

[0165] V. Effects of the genipin derivatives of the present invention on GluN2A, GluN2B, and 5-HT in mouse hippocampus 1A , BDNF, CREB, p-CREB and PKA protein expression

[0166] Among the numerous NMDA receptor subunits, GluN2A and GluN2B subunits have the greatest impact on receptor function and are closely associated with neurological diseases such as depression and cognitive dysfunction. Studies have shown that the protein expression of GluN2A and GluN2B in the hippocampus of mice in CUMS is significantly increased. NMDA receptors activate G protein-coupled receptors to increase cAMP concentration, activate intracellular calcium ion concentration, promote CREB phosphorylation, and then regulate the expression of genes associated with neuronal growth and survival, ultimately achieving the purpose of antidepressant. 5-HT 1A Heteromeric receptors are highly expressed in the axons of neurons in the hippocampus (CA1 and CA3 regions). Activates postsynaptic 5-HT 1A Heterologous receptors will cause hyperpolarization of the postsynaptic membrane and inhibit the excitability of neurons. Activation of the PKA-CREB signaling pathway can also upregulate the expression of BDNF. By increasing the expression of p-CREB protein in the hippocampus, the BDNF signaling pathway of CERB in depression model rats is regulated. CREB can activate the production of BDNF, and BDNF can also induce the phosphorylation of CREB. The interaction between the two forms an important positive feedback loop that affects neural plasticity and exerts a rapid antidepressant effect. Therefore, this experiment used immunoblotting to examine the expression of GluN2A, GluN2B, and 5-HT in the hippocampus of mice after W-3 administration. 1A , BDNF, CREB, p-CREB and PKA protein expression changes.

[0167] Expression levels of GluN2A and GluN2B proteins in mouse hippocampus

[0168] The results are as follows Fig. 8A , 8C As shown in Figure 8D, the expression of GluN2A and GluN2B proteins in the hippocampus of mice in the CUMS model group was upregulated. After intervention with compound W-3, the expression of the above two proteins was downregulated, and the effect of compound W-3 ​​in downregulating the expression of GluN2A and GluN2B proteins was significantly stronger than that of the positive control fluoxetine at a high dose (10 mg / kg).

[0169] Mouse hippocampal 5-HT 1A Protein expression level

[0170] The results are as follows Fig. 8A , 8B As shown, 5-HT in the hippocampus of mice in the CUMS model group 1A The expression of receptor protein decreased, and after the intervention of compound W-3, the 5-HT 1A The expression of receptor protein was enhanced (the results showed a dose-dependent increase), among which the intervention effect of compound W-3 ​​in the high-dose group (10 mg / kg) was better than that of the positive control fluoxetine.

[0171] BDNF protein expression level in mouse hippocampus

[0172] The results are as follows Fig.9A , 9C As shown in the figure, compared with the normal group, the color of the strips in the model group mice was lighter, indicating that the expression level of BDNF protein in the hippocampus of the model group mice was reduced. The results showed that depression would reduce the BDNF protein in the hippocampus of mice; compared with the model group, the color of the hippocampal strips in the drug-treated group mice was darker, and the BDNF level showed an obvious upward trend, indicating that the intervention treatment of the new genipin derivatives can reverse the decrease in BDNF protein expression caused by chronic stress, and may exert an anti-depressant effect by upregulating the BDNF protein level in the hippocampus.

[0173] CREB protein expression level in mouse hippocampus

[0174] The results are as follows Fig.9A , 9D As shown in the figure, compared with the normal group, the color of the strips in the model group mice was lighter, indicating that the expression level of CREB protein in the hippocampus of the model group mice was reduced. The results showed that depression would reduce the CREB protein in the hippocampus of mice; compared with the model group, the color of the hippocampal strips in the drug-treated group mice was darker, and the CREB level showed an obvious upward trend, indicating that the intervention treatment of the new genipin derivatives can reverse the decrease in CREB protein expression caused by chronic stress, and may exert an anti-depressant effect by upregulating the CREB protein level in the hippocampus.

[0175] PKA protein expression level in mouse hippocampus

[0176] The results are as follows Fig.9A , 9B As shown, compared with the normal group, the color of the strips in the model group mice was lighter, indicating that the expression level of PKA protein in the hippocampus of the model group mice was reduced. The results showed that depression would reduce the level of PKA protein in the hippocampus of mice; compared with the model group, the color of the hippocampal strips in the drug-treated group mice was darker, and the PKA level showed an obvious upward trend, indicating that the intervention treatment of the new genipin derivatives can reverse the decrease in PKA protein expression caused by chronic stress. In addition, the reversal of the depressive-like behavior of mice may be related to the upregulation of PKA protein levels in the hippocampus.

[0177] VI. Effects of the Genipin Derivatives of the Present Invention on Signal Factors in the Body

[0178] ELISA test results Figures 10A to 10EThe results showed that compared with the normal group, the Glu content in the brain tissue of mice in the model group was increased (p<0.05); compared with the model group, the Glu level in the brain tissue of mice in the compound W-3 ​​group was decreased (p<0.05); compared with the normal group, the 5-HT, DA, NE and BDNF contents in the brain tissue of mice in the model group were decreased (p<0.05); compared with the model group, the 5-HT, DA, NE and BDNF contents in the brain tissue of mice in the compound W-3 ​​group were increased (p<0.05).

[0179] The above test results show that the genipin derivatives of the present invention have good antidepressant activity, and some of the compounds have good neuroprotective activity. Some compounds can significantly upregulate the expression of PKA, pCREB / CREB, BDNF and 5-HT1A proteins, and downregulate the expression of GluN2A and GluN2B proteins. Among them, the effect of compound W-3 ​​on the expression of PKA, CREB and BDNF proteins is significantly better than the positive control fluoxetine at 10 mg / kg. Therefore, the genipin derivatives of the present invention can be used as therapeutic drugs for depression.

[0180] In summary, the genipin derivatives of the present invention have a good improvement effect on depression, and its mechanism may be related to the regulation of the PKA-CREB-BDNF signaling pathway. It can also exert a potential rapid antidepressant effect by affecting 5-HT1A, GluN2A and GluN2B receptor target proteins, making it have good application prospects.

[0181] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

Claims

1. A genipin derivative or a pharmaceutically acceptable salt thereof, characterized in that: The genipin derivative is a compound shown below: 。 2. The genipin derivative or a pharmaceutically acceptable salt thereof according to claim 1, characterized in that: The pharmaceutically acceptable salt is selected from any one of hydrochloride, sulfate, phosphate, hydrobromide, maleate, fumarate, citrate, methanesulfonate, p-toluenesulfonate, tartrate or acetate.

3. The genipin derivative or a pharmaceutically acceptable salt thereof according to claim 2, characterized in that: The pharmaceutically acceptable salt is hydrochloride.

4. The method for preparing the genipin derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3, characterized in that: The preparation method comprises: mixing compound 1, compound 2b, a reaction aid and an organic solvent, and allowing the reaction to complete at room temperature. The structural formulas of compound 1 and compound 2b are as follows: ； Compound 1 Compound 2b; The reaction aid is potassium carbonate.

5. The preparation method according to claim 4, characterized in that: The molar ratio of the compound 1, the compound 2b and the reaction auxiliary agent is 1:1:

2.

6. The preparation method according to claim 4, characterized in that: The organic solvent includes N,N-dimethylformamide.

7. The preparation method according to claim 4, characterized in that: The reaction time is 4 h.

8. The preparation method according to claim 4, characterized in that: The method also includes subjecting the reaction product to organic solvent extraction, washing the obtained organic phase and drying, concentrating and purifying the organic phase.

9. A pharmaceutical composition, characterized in that The pharmaceutical composition is composed of a therapeutically effective amount of the genipin derivative or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 3 and one or more pharmaceutically acceptable carriers.

10. Formula (I b ) Use of the genipin derivative or a pharmaceutically acceptable salt thereof as shown in ) in the preparation of a medicament for preventing or treating depression; ； （I b ） in: X and Y independently represent CH or N; Z stands for O; R1 represents a C1~C3 straight or branched chain alkyl group; R3 represents any one of H, C1~C6 alkyl, halogen, and C1~C3 alkoxy; n represents 0, 1, 2 or 3.

11. The use according to claim 10, characterized in that The depression includes any one of mild, moderate or severe unipolar depression, bipolar depression, psychotic depression, reactive depression, secondary depression, seasonal depression, postpartum depression and menopausal depression.

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