Preparation and application of twin drug with synergistic anti-depression and anti-anxiety effects

Abstract

The invention discloses preparation and application of a twin drug with synergistic anti-depression and anti-anxiety effects, the twin drug is formed by connecting a drug A and a drug B through a covalent bond, the drug A is desmethylvenlafaxine, and the drug B is a non-steroidal anti-inflammatory drug. On one hand, the twin drug increases the content of neurotransmitters (5-hydroxytryptamine and noradrenaline) in hippocampus, on the other hand, the twin drug plays a synergistic anti-depression and anti-anxiety role through playing anti-inflammatory, anti-oxidation, neuronal cell protection and other roles, and the anti-depression and anti-anxiety effects of the twin drug are superior to those of any single drug or a composition of drugs A and B. The twin drug has the potential of treating depression, anxiety, neuroinflammation and nervous system related diseases; meanwhile, the medicine taking compliance of a patient can be improved.

Description

TECHNICAL FIELD

[0001] The present application relates to a compound having synergistic antidepressant and anxiolytic effects, a preparation method and uses. BACKGROUND

[0002] Depressive disorder (also known as depression) is a common mental disorder that involves a long time of low mood or loss of pleasure or interest in activities. Anxiety disorder is a common psychological disorder, which is characterized by excessive worry and fear in daily situations, interfering with daily activities, and difficult to control. Although anxiety and depression are different, but the two diseases can coexist. According to the survey, about 33%-95% of patients with depression in clinical also have anxiety. Moreover, the symptoms of the two diseases overlap, such as decreased appetite, sleep disorders, heart and gastrointestinal discomfort, irritability, fatigue, etc. Depression and anxiety, the main causes of the disease mainly involve the interaction of biological, psychological and social factors. The complexity of the disease determines its difficulty to cure, but with the in-depth scientific research and the promotion of multi-disciplinary cooperation, it is expected to develop more precise and effective treatment drugs in the future to improve the quality of life of patients.

[0003] The role of neuroinflammation in the pathogenesis of depression and anxiety has attracted extensive attention. Neuroinflammation, as an abnormal immune response in the central nervous system (CNS), has been confirmed to be closely related to the pathogenesis of depression in recent years. Clinical studies have shown that the levels of pro-inflammatory cytokines in the serum and cerebrospinal fluid of patients with major depressive disorder are significantly elevated, among which the abnormal expression of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) is the most typical. These inflammatory mediators can affect the central nervous function through three main pathways: directly penetrating the weak areas of the blood-brain barrier; transmitting afferent signals through the peripheral vagus nerve; inducing secondary inflammatory factors from brain vascular endothelial cells. Pro-inflammatory factors entering the CNS disrupt neural homeostasis through multiple dimensions: first, they interfere with the synthesis and reuptake of monoamine neurotransmitters (5-HT, NE, DA), leading to an imbalance in synaptic transmitter concentration; second, they inhibit the expression of brain-derived neurotrophic factor (BDNF), hindering neural plasticity in brain regions such as the hippocampus; meanwhile, they continuously activate the hypothalamic-pituitary-adrenal axis (HPA axis), exacerbating glucocorticoid resistance. Notably, the abnormal activation of microglia forms a vicious cycle, as the reactive oxygen species (ROS) released by microglia not only exacerbate oxidative stress but also trigger gut microbiota disorders through bidirectional regulation of the gut-brain axis. Animal experiments have also provided strong evidence for the above mechanisms: when depressive model animals exhibit characteristics such as behavioral despair, anhedonia, and anxiety, the levels of IL-1β and TNF-α are abnormally elevated in emotion-regulating brain regions such as the prefrontal cortex and amygdala, accompanied by glutamatergic system disorders and reduced dendritic spine density. These cross-species evidence reveals a cascade amplification of pathological networks between neuroinflammation and depressive and anxious phenotypes, providing a key theoretical basis for the development of new antidepressant and anxiolytic therapies.

[0004] Currently, the main antidepressants used in clinical practice are based on the monoamine neurotransmitter hypothesis, including selective serotonin reuptake inhibitors (SSRIs, such as fluoxetine and sertraline), serotonin and norepinephrine reuptake inhibitors (SNRIs, such as venlafaxine), tricyclic antidepressants (TCAs, such as amitriptyline), and monoamine oxidase inhibitors (MAOIs). These drugs improve depressive symptoms by regulating synaptic neurotransmitter concentrations or modulating synaptic plasticity, but have the following significant limitations: traditional drugs usually require continuous administration for 2-4 weeks to take effect, and the risk of suicide in the acute phase may increase during this period. About 30-50% of patients have an inadequate response to first-line drugs, and even with combination therapy, 20% of patients still cannot achieve clinical remission. Existing drugs mainly target the monoamine neurotransmitter system, but depression involves multiple pathological mechanisms such as neuroinflammation, HPA axis dysfunction, and mitochondrial dysfunction. For example, elevated pro-inflammatory cytokines (IL-6, TNF-α) may weaken the efficacy of SSRIs by reducing tryptophan hydroxylase activity. In addition, drugs are difficult to reverse structural damage such as reduced hippocampal neurogenesis or prefrontal cortex synapse atrophy.

[0005] O-Desvenlafaxine is a serotonin and norepinephrine reuptake inhibitor (SNRIs) antidepressant, which is the active metabolite of Venlafaxine. It is mainly used in the treatment of various depressive disorders and generalized anxiety disorder, especially for depression with anxiety. It can increase the concentration of neurotransmitters in the synaptic cleft by inhibiting the reuptake of serotonin (5-HT) and norepinephrine (NE) at the presynaptic membrane, thereby exerting antidepressant and anxiolytic effects and improving mood and cognitive function. Compared with traditional SNRIs, O-Desvenlafaxine has more stable pharmacokinetics, does not require liver metabolism (is excreted directly through the kidneys), has a longer half-life (about 11 hours), and can maintain blood drug concentrations with once-daily administration. O-Desvenlafaxine has high drug safety and good body tolerance. When combined with other drugs, the risk of drug interactions is low. However, this drug has the disadvantages of long onset period and limited relief of depressive and anxiety symptoms.

[0006] Studies have shown that elevated COX-2 expression levels in the central nervous system are associated with depression and anxiety. COX-2's involvement in depression includes multiple aspects, such as neuroinflammation, changes in gut microbiota, neurotransmitter imbalance, hypothalamic-pituitary-adrenal (HPA) axis dysfunction, mitochondrial dysfunction, and hippocampal neuronal damage. Currently, methods for preventing and treating depression focus on inhibiting the in vivo expression of COX-2 using COX-2 inhibitors. This approach aims to reduce the production of pro-inflammatory mediators and related factors, thereby reducing the damage caused by inflammation, neurotransmitter imbalance, gut microbiota disorder, and neuronal damage.

[0007] Non-steroidal anti-inflammatory drugs (NSAIDs) are a class of drugs that reduce prostaglandin synthesis by inhibiting cyclooxygenase (COX), and have anti-inflammatory, analgesic, and antipyretic effects. Ibuprofen, naproxen, diclofenac, ketoprofen, and indomethacin are common non-steroidal drugs (the following formula), which mainly inhibit COX-1 and COX-2, reducing prostaglandin synthesis, thereby reducing inflammatory response and pain perception. When the drug enters the body, it reduces the levels of inflammatory factors (such as IL-6, IL-1β, TNF-α) and improves the levels of oxidative stress factors (MDA, SOD) by inhibiting COX-2 and the NF-κB inflammatory signaling pathway, which are related to the improvement of depressive behavior. A large number of relevant literature supports the role of non-steroidal anti-inflammatory drugs in the field of antidepressants, and there have been cases of non-steroidal anti-inflammatory drugs combined with antidepressants for treatment. Studies have found that the increased COX-2 content in the brain of patients with depression and the overactivation of the inflammatory signaling pathway, as well as animal experiments have shown that inhibiting the activity of COX-2 and NF-κB to reduce inflammation and oxidative stress levels and increase neurotransmitter levels have antidepressant and anxiolytic effects in depression models.

[0008]

[0009] Drug conjugate is a kind of drug molecule formed by connecting two same or different drug efficacy groups through covalent bond, which belongs to the classical drug design method. The synthesized new molecule is metabolized in vivo to generate the above two drugs to enhance the activity by synergistic effect, or produce new pharmacological activity, or improve the selectivity of action. SUMMARY

[0010] The first object of the present application is to provide a drug conjugate having synergistic antidepressant effect or a pharmaceutically acceptable salt thereof or a tautomer thereof, the second object is to provide a preparation method of the compound, and the third object is to provide a use of the compound.

[0011] TECHNICAL SCHEME

[0012] A drug conjugate having synergistic antidepressant effect or a pharmaceutically acceptable salt thereof or a tautomer thereof, characterized in that the drug conjugate is connected by a chemical bond between drug A and drug B, wherein the drug A is nortriptyline, and the drug B is a non-steroidal anti-inflammatory drug.

[0013] The non-steroidal anti-inflammatory drug is selected from ibuprofen, naproxen, aspirin, diclofenac, ketoprofen, indomethacin and flurbiprofen.

[0014] Preferably, the chemical structure of the drug conjugate or a pharmaceutically acceptable salt thereof or a tautomer thereof according to the present application is as follows:

[0015]

[0016] The present application prepares a drug conjugate by connecting nortriptyline and a non-steroidal anti-inflammatory drug (ibuprofen, naproxen, diclofenac, ketoprofen or indomethacin) through a covalent bond. The drug conjugate is released into the body by hydrolysis to generate nortriptyline and ibuprofen, naproxen, diclofenac, ketoprofen or indomethacin single drug, which produces an antidepressant effect through multiple mechanisms, so as to obtain better antidepressant and anxiolytic effect, reduce dose-related side effects, and better meet the clinical drug demand. Therefore, the drug conjugate of nortriptyline and a non-steroidal anti-inflammatory drug has a broad application prospect for the treatment of depression and anxiety.

[0017] The pharmaceutical composition according to the present application contains the compound according to the present application and a pharmaceutically acceptable carrier.

[0018] Preferably, the pharmaceutically acceptable carrier can be an excipient that is widely used in the pharmaceutical production field. The excipient is mainly used to provide a safe, stable and functional pharmaceutical composition, and can also provide a method for allowing the active ingredient to be dissolved at a desired rate after the subject receives the administration, or to facilitate the active ingredient to be effectively absorbed after the subject receives the administration of the composition. The pharmaceutical excipient can be an inert filler, or provide a certain function, such as stabilizing the overall pH value of the composition or preventing the degradation of the active ingredient of the composition. The pharmaceutical excipient can include one or more of the following excipients: binders, suspending agents, emulsifying agents, diluents, fillers, granulating agents, adhesives, disintegrants, lubricants, anti-adhesion agents, flow aids, wetting agents, gelling agents, absorption retardants, dissolution inhibitors, enhancers, adsorbents, buffers, chelating agents, preservatives, colorants, flavorings and sweeteners.

[0019] The pharmaceutical composition of the present application can be prepared according to the disclosure using any method known to those skilled in the art. For example, conventional mixing, dissolving, granulating, emulsifying, dragee-making, encapsulating, entrapping or lyophilizing processes.

[0020] The pharmaceutical composition of the present application can be administered in any form, including injection (intravenous), mucosal, oral (solid and liquid preparations), inhalation, ocular, rectal, topical or parenteral (infusion, injection, implantation, subcutaneous, intravenous, intra-arterial, intramuscular) administration. The pharmaceutical composition of the present application can also be a controlled or sustained release dosage form (such as liposomes or microspheres). Examples of solid oral preparations include, but are not limited to, powders, capsules, caplets, soft capsules and tablets. Examples of liquid preparations for oral or mucosal administration include, but are not limited to, suspensions, emulsions, elixirs and solutions. Examples of topical preparations include, but are not limited to, emulsions, gels, ointments, creams, patches, pastes, foams, lotions, drops or serums. Examples of preparations for parenteral administration include, but are not limited to, injection solutions, dry powder preparations that can be dissolved or suspended in a pharmaceutically acceptable carrier, injection suspensions and injection emulsions. Examples of other suitable preparations of the pharmaceutical composition include, but are not limited to, eye drops and other ophthalmic preparations; aerosols, such as nasal sprays or inhalants; liquid dosage forms suitable for parenteral administration; suppositories; and lozenges.

[0021] The method for preparing the compound of the present application comprises the following steps:

[0022]

[0023] (1) condensing nortriptyline and the non-steroidal anti-inflammatory drug in the presence of a catalyst to obtain the final product.

[0024] wherein the drug B is as defined above.

[0025] Preferably, the catalyst in step 1 is selected from 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 4-dimethylaminopyridine, N,N'-dicyclohexylcarbodiimide, N,N'-diisopropylcarbodiimide. The molar ratio between norepinephrine, non-steroidal anti-inflammatory drug, catalyst is 1:1:(0.01-10), and the reaction solvent is selected from dichloromethane, acetonitrile, acetone or tetrahydrofuran.

[0026] The use of the twin drug and its pharmaceutically acceptable salt or tautomer thereof according to the present application is to prepare a drug for preventing or treating depression, anxiety, nervous system diseases.

[0027] Preferably, the drug is a drug for preventing or treating major depression, bipolar depression, psychotic depression, reactive depression, menopausal depression, child depression, secondary depression, postpartum depression, seasonal depression, persistent depression, postmenopausal and perimenopausal depression, and anxiety depression.

[0028] Preferably, the drug is a drug for preventing or treating generalized anxiety disorder, panic disorder, social anxiety disorder and separation anxiety disorder.

[0029] Beneficial effects: Compared with the prior art, the present application has the following obvious advantages:

[0030] The compound according to the present application can increase the content of neurotransmitters (5-hydroxytryptamine and norepinephrine) in the hippocampus on the one hand, and play a synergistic role in anti-depression and anti-anxiety by exerting anti-inflammatory, anti-oxidative and neuron cell protective effects on the other hand. The anti-depression effect is superior to that of any single drug or drug composition. The twin drug has the potential to treat depression, anxiety, neuroinflammation and nervous system related diseases, and can also increase patient medication compliance. BRIEF DESCRIPTION OF DRAWINGS

[0031] Figure 1 Figure 1 is a schematic diagram of the behavioral effects of compounds 1-5 in mice in the M-CPP-induced anxiety model in the embodiments of the present application. Figure 1 A is the movement distance of the mouse in the open field within the first 5 minutes, Figure 1 B is the movement time of the mouse in the open field within the first 5 minutes, Figure 1 C is the number of times the mouse enters the center area in the open field within the first 5 minutes;

[0032] Figure 2 Figure 2 is a schematic diagram of the behavioral effects of compounds 1-5 on acute LPS-induced depression in mice in the embodiments of the present application. Figure 2 A is the immobility time of the mouse in the tail suspension test, Figure 2 B is the immobility time of the mouse in the forced swimming test, Figure 2 C is the total movement distance of the mouse in the open field within the first 5 minutes,Figure 2 D is the total movement distance of mice in the open field within the first 5 min, Figure 2 E is the number of times mice cross the center within the first 5 min in the open field;

[0033] Figure 3 is a schematic diagram of the effect of O-desmethylvenlafaxine- ibuprofen (ODV-IBU, compound 1) on the behavior of mice in a LPS-induced depression model Figure 3 A is the percentage of sucrose water intake of mice, Figure 3 B is the immobile time of mice in the tail suspension test, Figure 3 C is the immobile time of mice in forced swimming, Figure 3 D is the total movement distance of mice in the open field within the first 5 min, Figure 3 E is the total movement time of mice in the open field within the first 5 min, Figure 3 F is the number of times mice cross the center within the first 5 min in the open field;

[0034] Figure 4 is a schematic diagram of the effect of O-desmethylvenlafaxine- ibuprofen (ODV-IBU, compound 1) on inflammatory factors in the brain of LPS-induced depression mice Figure 4 A is the content of TNF-α in the hippocampus of mice, Figure 4 B is the content of IL-1β in the hippocampus of mice, Figure 4 C is the content of IL-6 in the hippocampus of mice;

[0035] Figure 5 is a schematic diagram of the effect of O-desmethylvenlafaxine- ibuprofen (ODV-IBU, compound 1) on oxidative stress in the brain of LPS-induced depression mice Figure 5 A is the content of SOD in the hippocampus of mice, Figure 5 B is the content of MDA in the hippocampus of mice;

[0036] Figure 6 is a schematic diagram of the effect of O-desmethylvenlafaxine- ibuprofen (ODV-IBU, compound 1) on neurotransmitters in the brain of LPS-induced depression mice Figure 6 A is the content of 5-HT in the hippocampus of mice, Figure 6 B is the content of NE;

[0037] Figure 7 is a schematic diagram of the effect of O-desmethylvenlafaxine- ibuprofen (ODV-IBU, compound 1) on brain neuron cell damage in LPS-induced depression mice Figure 7 A is the Nissl staining of the DG region of the hippocampus of mice, Figure 7 B is the Nissl staining of the CA1 region of the hippocampus, Figure 7 C is the Nissl staining section of the cerebral cortex, Figure 7 D is the neuron cell count in the CA1 region, Figure 7E is the brain cortex neuron cell count;

[0038] Figure 8 Figure is the effect of ODV-NAP (compound 2) on the behavior of LPS-induced depression model mice Figure 8 A is the percentage of sucrose intake of mice, Figure 8 B is the immobility time of mice in the tail suspension test, Figure 8 C is the immobility time of mice in forced swimming, Figure 8 D is the total distance of movement of mice in the open field within the first 5 minutes, Figure 8 E is the total movement time of mice in the open field within the first 5 minutes, Figure 8 F is the number of times mice cross the central area in the open field within the first 5 minutes;

[0039] Figure 9 Figure is the effect of ODV-NAP (compound 2) on inflammatory factors in the brain of LPS-induced depression mice Figure 9 A is the content of TNF-α in the hippocampus of mice, Figure 9 B is the content of IL-1β in the hippocampus of mice, Figure 9 C is the content of IL-6 in the hippocampus of mice;

[0040] Figure 10 Figure is the effect of ODV-NAP (compound 2) on oxidative stress in the brain of LPS-induced depression mice Figure 10 A is the content of SOD in the hippocampus of mice, Figure 10 B is the content of MDA in the hippocampus of mice;

[0041] Figure 11 Figure is the effect of ODV-NAP (compound 2) on neurotransmitters in the brain of LPS-induced depression mice Figure 11 A is the content of 5-HT in the hippocampus of mice, Figure 11 B is the content of NE in the hippocampus of mice;

[0042] Figure 12 Figure is the effect of ODV-NAP (compound 2) on the damage of cerebral neurons in LPS-induced depression mice Figure 12 A is the Nissl staining of the DG region of the hippocampus of mice, Figure 12 B is the Nissl staining of the brain cortex, Figure 12 C is the neuron cell count in the DG region, Figure 12 D is the cell count in the cortex; DETAILED DESCRIPTION

[0043] The technical solutions of the present application are further described below in combination with examples.

[0044] The structure of the example compounds is determined by nuclear magnetic resonance (NMR) or / and mass spectrometry (MS). NMR shifts (δ) are given in ppm. NMR is measured by a Bruker AVANCE III HD 400 NMR spectrometer, with deuterated dimethyl sulfoxide (DMSO-d6) as the solvent and tetramethylsilane (TMS) as the internal standard.

[0045] MS is measured by an Agilent 1260 HPLC-6520 Accurate-Mass Q-Tof mass spectrometer, with the following test conditions: electrospray ion source (ESI), positive ion mode.

[0046] Thin layer chromatography silica gel plates are used Yantai Huanghai HSGF254 or Qingdao GF254 silica gel plates.

[0047] Silica gel column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier.

[0048] Unless otherwise specified in the examples, the reaction temperature is room temperature, which is 20-30°C.

[0049] The reaction progress in the examples is monitored by thin layer chromatography (TLC).

[0050] Example 1: Synthesis of O-desmethylvenlafaxine-ibuprofen (ODV-IBU, compound 1)

[0051] Reaction formula:

[0052]

[0053] First, add O-desmethylvenlafaxine (0.45 g, 1.71 mmol, 1.0 eq), ibuprofen (0.345 g, 1.67 mmol, 1.0 eq), 15 mL of dichloromethane, DMAP (20 mg, 0.16 mmol, 0.2 eq) into a 100 mL volumetric flask, after stirring, add EDCI (0.39 g, 2.04 mmol, 1.2 eq), stir at room temperature for 3 h, and then monitor the reaction by TCL. Add 10 mL of dichloromethane and transfer the solution to a separatory funnel, add 10 mL of water, separate by extraction for 3 times, and collect the organic phase. Dry the organic phase with anhydrous sodium sulfate, filter, and rotary evaporate to obtain the crude product; the crude product is purified by column chromatography (eluted with DCM:MeOH=30:1) to obtain the oil product 0.48 g in a yield of 62.59%. 1H NMR (400 MHz, CDC13) δ 7.32 - 7.29 (m, 2H), 7.20 - 7.14 (m, 4H), 7.01 - 6.97 (m, 2H), 3.94 (q, J=7.1 Hz, 2H), 3.36 (t, J=5.4 Hz, 1H), 3.01 (dd, J=13.0, 5.1 Hz, 1H), 2.66 (s, 6H), 2.48 (d, J=7.2 Hz, 2H), 1.88 (dt, J=13.5, 6.8 Hz, 1H), 1.69 (ddd, J=12.4, 8.0, 3.4 Hz, 2H), 1.61 (d, J=7.2 Hz, 3H), 1.59 - 1.42 (m, 5H), 1.29 - 1.09 (m, 2H), 0.92 (d, J=6.7 Hz, 6H), 0.91 - 0.83 (m, 1H). 13 C NMR (126 MHz, CDC13) δ 173.20, 150.12, 140.93, 137.17, 137.05, 130.05, 129.54, 127.18, 121.57, 73.59, 60.57, 52.71, 45.26, 45.04, 36.90, 30.17, 25.37, 22.38, 21.48, 21.20, 18.42. (+)-ESI-MS: m / z 452.3160 (calcd. 452.3159 for C 29 H 42 NO3 + [M+H] + )

[0054] Example 2: Synthesis of norvenlafaxine-naproxen (ODV-NAP, compound 2)

[0055] Reaction Scheme:

[0056]

[0057] Firstly, in a 100ml volumetric flask, norvenlafaxine (0.265g, 1.01mmol, 1.0 eq), naproxen (0.233g, 1.01mmol, 1.0 eq), 15mL of dichloromethane, DMAP (18mg, 0.15mmol, 0.2 eq) were stirred and then EDCI (0.234g, 1.22mmol, 1.2 eq) was added. After stirring at room temperature for 3h, the reaction was monitored by TCL, norvenlafaxine had reacted completely. The reaction solution was added with 10mL of dichloromethane and transferred to a separatory funnel, 10mL of water was added, and 3 times of separation extraction was carried out, and the organic phase was reserved. The organic phase was dried with anhydrous sodium sulfate, filtered, and rotary evaporated to obtain a white crude product; column chromatography (DCM:MeOH=30:1) purification to obtain a white solid product 0.30g, yield 71.84%. 1 H NMR (400 MHz, CDC13) δ 7.78 - 7.70 (m, 3H), 7.50 (dd, J = 8.6, 1.8 Hz, 1H), 7.19 - 7.13 (m, 4H), 7.00 - 6.96 (m, 2H), 4.11 (q, J = 7.1 Hz, 1H), 3.99 (dd, J = 12.8, 4.8 Hz, 1H), 3.93 (s, 3H), 3.38 (t, J = 5.3 Hz, 1H), 3.08 (dd, J = 12.8, 5.6 Hz, 1H), 2.68 (s, 6H), 1.70 (d, J = 7.1 Hz, 3H), 1.66 - 1.41 (m, 7H), 1.27 - 1.19 (m, 1H), 1.10 (tt, J = 12.0, 3.5 Hz, 1H), 0.92 - 0.81 (m, 1H). 13 C NMR (126 MHz, CDC13) δ 173.13, 157.84, 150.16, 137.06, 134.93, 133.88, 130.08, 129.30, 128.99, 127.44, 126.16, 126.03, 121.62, 119.19, 105.68, 105.64, 73.51, 60.43, 55.35, 52.77, 45.58, 36.72, 25.29, 21.50, 21.14, 18.39. (+)-ESI-MS: m / z 476.2797 (calcd. 476.2795 for C 30 H 38 NO4 + [M+H] + )

[0058] Example 3: Synthesis of norvenlafaxine-diclofenac (ODV-DIC, compound 3)

[0059] Reaction scheme:

[0060]

[0061] In a 100 ml volumetric flask, norvenlafaxine (90 mg, 0.34 mmol, 1.0 eq), diclofenac (101 mg, 0.34 mmol, 1.0 eq), DMAP (10 mg, 0.08 mmol, 0.2 eq), 10 mL dichloromethane, after stirring, EDCI (78 mg, 0.41 mmol, 1.5 eq) was added, after stirring at room temperature for 3 h, TLC monitoring, norvenlafaxine has been reacted. The reaction solution was transferred to a separatory funnel, 10 mL water was added, extracted 3 times, the organic phase was collected and anhydrous sodium sulfate was added, after filtration, the solvent was removed by rotary evaporation to obtain an oily crude product, which was further purified by silica gel column chromatography (DCM:MeOH = 50:1) to obtain red solid product 78 mg, yield 40.53%. 1 H NMR (400 MHz, CDC13) δ 7.26 (s, 1H), 7.19 (s, 1H), 7.12-7.04 (m, 3H), 6.97-6.90 (m, 4H), 6.66 (s, 1H), 6.52 (d, J = 8.0 Hz, 1H), 3.97 (s, 2H), 3.25 (t, J = 12.3 Hz, 1H), 2.94 (dd, J = 11.9, 3.4 Hz, 1H), 2.29 (d, J = 7.0 Hz, 1H), 2.26 (s, 6H), 1.62 (ddt, J = 25.6, 12.8, 3.7 Hz, 3H), 1.45 (s, 1H), 1.41-1.24 (m, 2H), 1.24-1.14 (m, 2H), 0.92-0.77 (m, 2H).1 3 C NMR (101 MHz, CDC13) δ 170.61, 149.35, 142.72, 138.35, 137.77, 131.03, 130.16, 129.53, 128.88, 128.29, 124.19, 123.90, 122.32, 120.98, 118.55, 74.12, 60.89, 52.07, 45.33, 38.65, 37.98, 31.21, 25.87, 21.55, 21.32. (+)-ESI-MS: m / z 541.2021 (calcd. 541.2019 for C 30 H 35 Cl2N2O3 + [M+H] + )。

[0062] Example 4: Synthesis of norvenlafaxine-ketoprofen (ODV-KET, compound 4)

[0063] Reaction Scheme:

[0064]

[0065] In a 100ml volumetric flask, norvenlafaxine (100mg, 0.38mmol, 1.0 eq), ketoprofen (98mg, 0.38mmol, 1.0 eq), DMAP (10mg, 0.08mmol, 0.2 eq), 10mL dichloromethane, after stirring, EDCI (82mg, 0.43mmol, 1.5 eq) was added, after 3h reaction at room temperature, TLC monitoring, norvenlafaxine has been reacted completely. The reaction solution was transferred to a separatory funnel, 10mL dichloromethane was added, 10mL water was added, extracted 3 times, the organic phase was collected and anhydrous sodium sulfate was added, after filtration, the solvent was removed by rotary evaporation to obtain an oily crude product, which was further purified by silica gel column chromatography (DCM:MeOH = 50:1) to obtain a yellow oily product 93mg, yield 48.06%. 1 H NMR (400 MHz, CDC13) δ 7.86 (d, J = 1.9 Hz, 1H), 7.83 - 7.79 (m, 2H), 7.72 (dd, J = 7.8, 1.5 Hz, 1H), 7.65 - 7.57 (m, 2H), 7.48 (td, J = 7.6, 5.7 Hz, 3H), 7.12 (s, 1H), 7.10 (s, 1H), 6.93 (d, J = 8.3 Hz, 2H), 4.04 (q, J = 7.1 Hz, 1H), 3.28 (t, J = 12.5 Hz, 1H), 2.99 (dd, J = 12.2, 3.4 Hz, 1H), 2.32 (s, 6H), 2.29 (d, J = 3.6 Hz, 1H), 1.77 - 1.68 (m, 2H), 1.66 (d, J = 7.3 Hz, 3H), 1.59 - 1.45 (m, 4H), 1.35 (d, J = 13.0 Hz, 1H), 1.27 (dd, J = 13.0, 4.3 Hz, 1H), 0.96 - 0.82 (m, 2H). 13 C NMR (101 MHz, CDC13) δ 196.43, 172.57, 149.40, 140.36, 138.34, 138.14, 137.46, 132.58, 131.52, 130.10, 129.27, 128.76, 128.36, 120.74, 77.36, 77.05, 76.73, 74.12, 60.95, 52.00, 45.55, 45.43, 38.08, 31.20, 25.91, 21.56, 21.30, 18.48. (+)-ESI-MS: m / z 500.2797 (calcd. 500.2795 for C32 H 38 NO4 + [M+H] + )。

[0066] Example 5: Synthesis of norvenlafaxine-indomethacin (ODV-IND, compound 5)

[0067] Reaction scheme:

[0068]

[0069] First, add norvenlafaxine (97 mg, 0.38 mmol, 1.0 eq), indomethacin (131 mg, 0.37 mmol, 1.0 eq), DMAP (10 mg, 0.08 mmol, 0.2 eq), 10 mL of dichloromethane into a 100 mL volumetric flask, after stirring, add EDCI (84 mg, 0.44 mmol, 1.2 eq), stir at room temperature for 3 h, monitor by TCL, norvenlafaxine has reacted completely, transfer the reaction solution to a separatory funnel, add 10 mL of dichloromethane, 10 mL of water, extract 3 times, collect the organic phase. Add anhydrous sodium sulfate to the organic phase, filter and rotary evaporate to remove the solvent to obtain an oily crude product, then purify by silica gel chromatography column

[0070] (DCM:MeOH=65:1) to obtain 98 mg of green oily product with a yield of 44.12%. 1H NMR (400 MHz, CDC13) δ 7.71 - 7.65 (m, 2H), 7.50 - 7.45 (m, 2H), 7.20 - 7.12 (m, 2H), 7.07 - 6.98 (m, 3H), 6.88 (d, J = 9.0 Hz, 1H), 6.70 (dd, J = 9.0, 2.6 Hz, 1H), 3.90 (s, 2H), 3.84 (s, 3H), 3.62 (dd, J = 12.7, 8.2 Hz, 1H), 3.14 (dd, J = 8.2, 4.9 Hz, 1H), 2.67 (dd, J = 12.8, 4.9 Hz, 1H), 2.46 (s, 6H), 2.04 (s, 3H), 1.72 - 1.60 (m, 2H), 1.55 - 1.34 (m, 3H), 1.33 - 1.11 (m, 3H), 1.10 - 0.81 (m, 2H).13C NMR (101 MHz, CDC13) δ 176.09, 169.25, 156.13, 149.77, 139.43, 137.73, 136.30, 133.78, 131.22, 130.88, 130.49, 130.17, 129.19, 121.32, 115.04, 111.89, 111.70, 101.34, 73.73, 60.30, 55.78, 52.64, 44.50, 37.06, 31.44, 30.59, 25.51, 21.58, 13.42. (+)-ESI-MS: m / z 603.2621 (calcd. 603.2620 for C 35 H 40 ClN2O5 + [M+H] + ).

[0071] Example 6: Metabolic stability experiment of compounds 1-5 in artificial gastric juice and artificial intestinal juice

[0072] The artificial gastric juice was prepared: 10 g of pepsin was contained in 1 L of phosphate buffer (pH = 1.2).

[0073] The artificial intestinal juice was prepared: 10 g of pancreatin was contained in 1 L of phosphate buffer (pH = 6.8). 2 mg of each of compounds 1-5 was dissolved in 100 μL of acetonitrile solution, and after complete dissolution, it was transferred to 10 ml of artificial intestinal juice and artificial gastric juice, and incubated under shaking at 37°C. 400 μL was sampled at different time intervals and 400 μL of acetonitrile was added to inactivate the enzyme; after centrifugation at 4°C, 10000 rpm for 10 min, the upper clear liquid was taken, filtered, and the remaining percentage of compounds 1-5 was analyzed by high performance liquid chromatography to calculate the half-life of the compounds in artificial gastric juice and intestinal juice.

[0074] Table 1 Stability of compounds 1-5 in artificial gastric juice and artificial intestinal juice (n=3)

[0075]

[0076] According to the remaining percentage of the drug in each time period in artificial gastric juice and intestinal juice, the half-life of compounds 1-5 in artificial intestinal juice was calculated, see Table 2.

[0077] Table 2 Half-life of compounds 1-5 in artificial intestinal juice

[0078]

[0079] The data in Table 1 shows that compounds 1-5 can be metabolized in artificial gastric juice and intestinal juice, among which compounds 1 and 4 are metabolized faster in artificial gastric juice, and compounds 2, 3 and 5 are metabolized slower in artificial gastric juice. In addition, compounds 2, 3 and 4 are metabolized faster in artificial intestinal juice, and compound 5 is metabolized slowest.

[0080] Example 6: Evaluation of the anti-anxiety activity of compounds 1-5 in the m-chlorophenylpiperazine (M-CPP) induced anxiety model

[0081] M-chlorophenylpiperazine (M-CPP) is a 5-HT 2C agonist, which non-selectively stimulates 5-HT receptors and is prone to produce anxiety-like behavior after administration. M-CPP 4 mg / kg was injected into mice together with the compounds, and 30 min later the mice were placed in the center of the open field, and the movement time, movement distance, and the number of times entering the center area of the mice in the open field were recorded to determine whether the drug had anti-anxiety effect by analyzing the indicators.

[0082] ICR mice, male, 25-30 g. The mice were divided into 8 groups, 6 in each group. The animals were randomly divided into a solvent group (Control), a model group (M-CPP) 4 mg / kg, nortriptyline (ODV) 16 mg / kg, nortriptyline-ibuprofen (compound 1, ODV-IBU) 27.44 mg / kg, nortriptyline-naproxen (compound 2, ODV-NAP) 28.89 mg / kg, nortriptyline-diclofenac (compound 3, ODV-DIC) 32.82 mg / kg, nortriptyline-ketoprofen (compound 4, ODV-KET) 30.33 mg / kg, and nortriptyline-indomethacin (compound 5, ODV-IND) 36.59 mg / kg; the doses of the drugs were converted according to the equivalent molar ratio. The drug administration volume was 10 ml / kg, and the drugs were dissolved in physiological saline containing 10% DMSO.

[0083] Figure 1A [F(7, 40) = 11.64, p < 0.0001], 18.94 ± 0.70 m in the vehicle group, 10.14 ± 0.72 m in the model group, 13.30 ± 0.25 m in the norvenlafaxine group, 15.54 ± 0.86 m in the compound 1 group, 16.53 ± 1.18 m in the compound 2 group, 14.78 ± 0.78 m in the compound 3 group, 13.58 ± 0.63 m in the compound 4 group, and 15.00 ± 0.99 m in the compound 5 group. m-CPP can cause anxiety in mice in an unfamiliar environment, thereby causing a decrease in movement. After treatment with compounds 1-5, the movement distance of mice in the open field within the first 5 min was significantly increased. Figure 1 B [F(7, 40) = 6.663, p = 0.0001], 197.8 ± 5.83 s in the vehicle group, 167.4 ± 3.10 s in the model group, 180.0 ± 2.09 s in the norvenlafaxine group, 193.3 ± 4.52 s in the compound 1 group, 191.4 ± 4.73 s in the compound 2 group, 189.4 ± 3.98 s in the compound 3 group, 183.4 ± 2.82 s in the compound 4 group, and 185.7 ± 3.78 s in the compound 5 group. m-CPP-induced anxiety can cause a decrease in the motor ability of mice, and a decrease in the movement time. After treatment with compounds 1-5, the movement time of mice in the open field within the first 5 min was increased, and there were significant differences between the compound groups and the m-CPP group, except for the compound 4 group. Figure 1 C [F(7, 40) = 8.386, p < 0.0001] is the number of times that mice cross the center within the first 5 min in the open field, 30.8 ± 2.46 in the vehicle group, 12.33 ± 1.14 in the model group, 21.00 ± 1.55 in the norvenlafaxine group, 26.67 ± 2.26 in the compound 1 group, 30.17 ± 1.25 in the compound 2 group, 22.83 ± 2.22 in the compound 3 group, 22.23 ± 1.78 in the compound 4 group, and 22.00 ± 2.92 in the compound 5 group. It can be observed that the number of times that mice cross the center is significantly increased after treatment.

[0084] Based on the above data, compounds 1-5 have certain anti-anxiety effects in the m-CPP-induced anxiety model, and among them, compounds 1 and 2 have the best anti-anxiety effects in the treatment of m-CPP-induced anxiety, and the mice treated with them show the most obvious behavioral changes.

[0085] Example 7: Evaluation of the antidepressant activity of compounds 1-5 in the acute LPS-induced depression model

[0086] Lipopolysaccharide is commonly used to rapidly induce depression in depression models and provides experimental basis for the screening and efficacy evaluation of antidepressants; this model is considered to be effective in inducing depression-like symptoms, and the behavioral characteristics of animals after stress change, and this model has high reference value as a depression model.

[0087] This experiment is a preliminary screening of the antidepressant activity of the drug, to determine whether it has antidepressant activity and drug toxicity, the experimental period of 4 days, 3 consecutive days given treatment drug, the third day after injection of drug injection of LPS 0.83mg / kg, the fourth day of behavioral testing. Test experiment mainly for, forced swimming, tail suspension test, open field test.

[0088] ICR male rats, 25-30g, the experiment is divided into 7 groups, 6 mice in each group. Except for the model group, the compound groups are the same as in Example 6. Among them, the model group was injected with normal saline for the first 3 days, and LPS 0.83mg / kg was injected on the fourth day. The drug dose, preparation and administration volume are the same as in Example 6. Figure 2 A[F(6, 35) = 6.222, p = 0.0002] is the immobility time of mice in the tail suspension test after treatment with compounds 1-5, the control group (Control) 97.28 ± 3.584s, the model (LPS) group 162.57 ± 9.265s, the nortriptyline (ODV) group 129.1 ± 4.61s, the nortriptyline-ibuprofen (compound 1, ODV-IBU) group 108.23 ± 9.506s, the nortriptyline-naproxen (compound 2, ODV-NAP) group 106.03 ± 6.804s, the nortriptyline-diclofenac (compound 3, ODV-DIC) group 123.73 ± 9.144s, the nortriptyline-ketoprofen (compound 4, ODV-KET) group 118.57 ± 12.81s, the nortriptyline-indomethacin (compound 5, ODV-IND) group 122.0 ± 4.42. After receiving LPS injection, the immobility time of mice in the tail suspension test is longer, and the survival desire is significantly decreased; Figure 2B[F(5, 34) = 17.85, p < 0.0001] is the immobility time of mice in forced swimming after receiving compound 1-5 treatment, the solvent group (Control) is 98.02 ± 4.66s, the model group (LPS) is 163.5s, the norepinephrine (ODV) group is 134.6 ± 7.44s, the norepinephrine-ibuprofen (compound 1, ODV-IBU) group is 119.9 ± 4.75s, the norepinephrine-naproxen (compound 2, ODV-NAP) group is 114.03 ± 4.61s, the norepinephrine-diclofenac (compound 3, ODV-DIC) group is 133.93 ± 5.61s, the norepinephrine-ketoprofen (compound 4, ODV-KET) group is 131.63 ± 9.72s, and the norepinephrine-indomethacin (compound 5, ODV-IND) group is 130.2 ± 3.79s. LPS also causes an increase in the immobility time of mice in forced swimming, and after receiving treatment, the immobility time of mice is significantly reduced, increasing the survival desire of mice in the behavioral experiment; from the experimental data, compounds 1-5 can reduce the immobility time of mice in the tail suspension and swimming, increase the survival desire of mice, and improve the despair behavior of mice. The above data results show that compounds 1-5 can improve the LPS-induced depressive behavior, and the antidepressant effect of compound 1 and compound 2 is the best.

[0089] Figure 2 C[F(5, 31) = 20.58, p < 0.0001] is the first 5 minutes of movement distance of mice, the solvent group (Control) is 19.88 ± 0.409m, the model group (LPS) is 6.00 ± 1.03m, the norepinephrine group is 13.14 ± 0.67s, the norepinephrine-ibuprofen (compound 1, ODV-IBU) group is 13.78 ± 0.82m, the norepinephrine-naproxen (compound 2, ODV-NAP) group is 14.30 ± 1.05m, the norepinephrine-diclofenac (compound 3, ODV-DIC) group is 12.14 ± 0.99m, the norepinephrine-ketoprofen (compound 4, ODV-KET) group is 13.20 ± 1.38m, and the norepinephrine-indomethacin (compound 5, ODV-IND) group is 12.33 ± 0.74m; Figure 2D[F(5, 31) = 15.67, p < 0.0001] is the first 5 minutes of the mouse movement time, the control group (Control) 220.72 ± 5.46s, the model group 130.92 ± 6.48s, the norepinephrine group 180.0 ± 4.79s, the norepinephrine-ibuprofen (compound 1, ODV-IBU) group 186.65 ± 3.97s, the norepinephrine-naproxen (compound 2, ODV-NAP) group 195.19 ± 5.14s, the norepinephrine-diclofenac (compound 3, ODV-DIC) group 174.72 ± 10.31s, the norepinephrine-ketoprofen (compound 4, ODV-KET) group 175.90 ± 10.82s, the norepinephrine-indomethacin (compound 5, ODV-IND) 180 ± 5.26s; Figure 2 E[F(5, 32) = 8.402, p < 0.0001] is the number of times each group of mice crosses the center area in the open field, the control group is 29.7 ± 1.56, the model group is 12.3 ± 2.60, the norepinephrine group is 26.2 ± 2.93, the norepinephrine-ibuprofen (compound 1, ODV-IBU) group is 27.4 ± 1.97, the norepinephrine-naproxen (compound 2, ODV-NAP) group is 30.3 ± 2.19, the norepinephrine-diclofenac (compound 3, ODV-DIC) group is 20.2 ± 2.93, the norepinephrine-ketoprofen (ODV-KET) group is 24.2 ± 2.64, and the norepinephrine-indomethacin (compound 5, ODV-IND) group is 21.5 ± 1.26. From the results in the open field experiment, it can be seen that compounds 1-5 can increase the exploration desire and movement ability of LPS model mice, and can alleviate the anxiety and depression behavior of mice, and among them, compounds 1 and 2 have better effects on alleviating anxiety.

[0090] From the behavior data, compounds 1-5 can improve the depression behavior of mice, increase the survival desire, enhance the exploration desire and movement ability, but the anti-depression effects of each compound are different, and among them, compounds 1 and 2 have the best effect, so the above two drugs are selected for further experiment evaluation.

[0091] Example 9: Evaluation of the antidepressant activity of norepinephrine-ibuprofen (ODV-IBU, compound 1) in the LPS-induced depression model for 8 days

[0092] ICR male mice, 25-30 g, were randomly divided into vehicle group (Control), model group (LPS), nortriptyline group (ODV) 16 mg / kg, ibuprofen group (IBU) 12.53 mg / kg, nortriptyline + ibuprofen group (ODV+IBU), nortriptyline-ibuprofen group (compound 1, ODV-IBU) 27.44 mg / kg, 8-10 mice in each group; the vehicle group and the model group were injected intraperitoneally with the corresponding dose of normal saline, and the drug treatment groups were given the corresponding dose of drugs intraperitoneally, and the drug volume, drug dose and preparation were the same as in Example 6.

[0093] In this experiment, the mice were modeled with lipopolysaccharide and administered for 8 days; except for the control group, each group of mice was administered continuously for 8 days, and 0.83 mg / kg of lipopolysaccharide solution was injected 45 minutes after treatment on the 6th day, and injected continuously for 3 days. The analysis of the depression model is mainly carried out by behavioral evaluation, and the commonly used evaluation methods include: sucrose preference test, tail suspension test, forced swimming test, open field test, etc., and secondly by evaluating inflammatory factors, oxidative stress, neurotransmitter content and brain neuron cell damage.

[0094] Experimental results

[0095] (1) The effect of nortriptyline-ibuprofen (compound 1, ODV-IBU) on the behavior of LPS-induced depression model mice

[0096] As Figure 3 As shown in A [F(5, 48) = 31.20, p < 0.0001], the sucrose preference of the vehicle group (Control), the model group (LPS), the nortriptyline group (ODV), the ibuprofen group (IBU), the nortriptyline + ibuprofen group (ODV+IBU), and the nortriptyline-ibuprofen group (compound 1, ODV-IBU) was 81.87 ± 0.97%, 53.49 ± 2.49%, 65.32 ± 2.67%, 63.66 ± 2.08%, 74.51 ± 0.79%, and 76.41 ± 1.12%, respectively, wherein the sucrose preference of the mice after drug treatment increased significantly, and the sucrose preference of the nortriptyline-ibuprofen group (compound 1, ODV-IBU) was significantly higher than that of the two single-drug groups and the nortriptyline + ibuprofen group (ODV+IBU); in the tail suspension and forced swimming tests, as shown in B [F(5, 54) = 16.11, p < 0.0001], Figure 3 B[F(5, 54) = 16.11, p < 0.0001], Figure 3C[F(5, 54) = 18.52, p < 0.0001] The immobility time of the last 4 min in the Control group was 105.7 ± 6.77 s / 98.32 ± 5.53 s, in the LPS group was 165.7 ± 3.74 s / 151.3 ± 4.82 s, in the ODV group was 135.3 ± 5.30 s / 133.9 s, in the IBU group was 122.8 ± 5.42 s / 130.1 s, in the ODV+IBU group was 113.5 ± 5.44 s / 123.8 ± 1.73 s, and in the ODV-IBU group was 112.0 ± 5.87 s / 121.3 ± 2.91 s. From the data, the immobility time of the LPS group was higher than that of the Control group, indicating that the LPS mice had reduced survival desire and exhibited behavioral despair, while the treatment groups could significantly reduce the immobility time, alleviate the despair behavior of the mice, and enhance the survival desire. Moreover, the immobility time of the ODV-IBU group and the ODV+IBU group was the least, indicating that ODV-IBU and ODV+IBU were superior to ODV and IBU in improving the depressive behavior of mice.

[0097] The open field test is usually to place mice in a strange open field and observe their movement ability and desire to explore the strange environment; for example Figure 3 D[F(5, 54) = 19.68, p < 0.0001] is the movement distance of mice in the area within the first 5 min, the Control group was 17.63 ± 0.72 m, the LPS group was 9.38 ± 0.61 m, the ODV group was 12.11 ± 0.51 m, the IBU group was 13.33 ± 0.88 m, the ODV+IBU group was 14.55 ± 0.58 m, and the ODV-IBU group was 16.05 ± 0.58 m. The movement distance of the mice receiving treatment increased significantly, and the movement distance of the mice receiving ODV-IBU treatment was the longest; Figure 3 E[F(5, 54) = 23.46, p < 0.0001] is the movement time of mice in the open field within the first 5 min, the Control group was 206.5 ± 6.55 s, the LPS group was 138.7 ± 3.53 s, the ODV group was 163.3 ± 5.21 s, the IBU group was 178.4 ± 3.35 s, the ODV+IBU group was 186.1 ± 5.16 s, and the ODV-IBU group was 190.9 ± 4.78 s.Figure 3 F[F(5, 54) = 19.27, p < 0.0001] is the number of times the mouse crosses the central area within the first 5 min, the mice in the control group (Control) group crosses the area 31.3 ± 2.13, the model group (LPS) 10.7 ± 2.14, the nortriptyline (ODV) group 20.1 ± 1.73, the ibuprofen (IBU) group 24 ± 1.56, the nortriptyline + ibuprofen (ODV + IBU) group 26.3 ± 1.17, the nortriptyline-ibuprofen (compound 1, ODV-IBU) group 29.4 ± 1.23, from the behavioral data, LPS can significantly reduce the mouse's ability to move and explore in the open field, mainly manifested as a decrease in movement distance, a decrease in movement time, and a decrease in the number of times entering the central area. After receiving four kinds of treatment drugs, the movement ability and exploration ability of mice can be restored, among which the effect of the nortriptyline-ibuprofen (ODV-IBU) group is the best, which can significantly increase the movement distance, movement time and central area crossing times;

[0098] In the LPS model, LPS causes the mouse's sucrose preference to decrease, the resting time to increase in the tail suspension and forced swimming tests, the total moving distance to decrease, the total moving time to decrease, and the number of times crossing the central area to decrease, but after being given the nortriptyline and ibuprofen composition or the nortriptyline-ibuprofen (ODV-IBU) treatment, the mouse's sucrose preference is improved, the resting time is reduced in the tail suspension test and the forced swimming test, the total moving distance is improved, the total moving time is increased, and the number of times crossing the central area is increased, indicating that the mouse's pleasure, survival desire, curiosity and exploration desire are improved, and the despair behavior and anxiety situation are alleviated; Among them, the composition and the nortriptyline-ibuprofen (ODV-IBU) have obvious advantages in the above behavioral evaluation compared with the single drug group. Therefore, it also proves that the nortriptyline-ibuprofen (ODV-IBU) has good antidepressant and anxiolytic effects in the LPS model.

[0099] (2) Effect of nortriptyline-ibuprofen (compound 1, ODV-IBU) on the content of inflammatory factors in the hippocampus of LPS-induced depressed mice

[0100] Studies have shown that excessive expression of inflammatory factors is related to depression, and relevant literature has also reported that a large number of inflammatory factors occur in the body of patients with depression. In order to explore whether the nortriptyline-ibuprofen (compound 1, ODV-IBU) has a regulatory effect on inflammatory factors, we detected the inflammatory factors in the hippocampus of mice by ELISA; the results are as follows Figure 4TNF-α levels in each group were as follows: Control 129.7 ± 18.13 pg / ml, LPS 436.5 ± 27.22 pg / ml, ODV 285.1 ± 28.22 pg / ml, IBU 230.4 ± 23.74 pg / ml, ODV + IBU 204.9 ± 12.25 pg / ml, ODV-IBU (Compound 1) 200.8 ± 12.02 pg / ml; as shown by F(5, 30) = 24.37, p < 0.0001. Figure 4 IL-1β levels in each group were as follows: Control 37.84 ± 4.26 pg / ml, LPS 148.4 ± 10.27 pg / ml, ODV 101.9 ± 8.86 pg / ml, IBU 70.69 ± 8.65 pg / ml, ODV + IBU 60.82 ± 5.26 pg / ml, ODV-IBU (Compound 1) 43.93 ± 2.95 pg / ml; as shown by F(5, 30) = 33.11, p < 0.0001. Figure 4 IL-6 levels in each group were as follows: Control 36.95 ± 3.80 pg / ml, LPS 157.90 ± 6.39 pg / ml, ODV 104.50 ± 11.19 pg / ml, IBU 80.17 ± 13.46 pg / ml, ODV + IBU 71.12 ± 6.49 pg / ml, ODV-IBU (Compound 1) 63.48 ± 9.13 pg / ml; when the mice were given the therapeutic drugs, the inflammatory factors in the mice were significantly reduced, and the levels of inflammatory factors in the hippocampus of mice in the ODV + IBU group and the ODV-IBU (Compound 1) group were significantly lower than those in the two single-drug groups, indicating that norvenlafaxine and ibuprofen produced a synergistic anti-inflammatory effect in vivo.

[0101] Lipopolysaccharide administration activates the NF-kB inflammatory signaling pathway and COX-2, resulting in excessive expression of inflammatory factors. We can significantly reduce the levels of TNF-ɑ, IL-1β, and IL-6 in the hippocampus after administration of norvenlafaxine-ibuprofen (ODV + IBU) or norvenlafaxine-ibuprofen (Compound 1, ODV-IBU), thereby reducing the impact of inflammation on the behavior of mice.

[0102] (3) Effect of ODV-IBU on the content of oxidative stress in hippocampus of LPS-induced depression mice

[0103] The overexpression of inflammatory factors usually promotes oxidative stress, leading to the imbalance of oxidation and antioxidant mechanisms in vivo, and further leading to the excessive disorder of SOD and MDA and other substances. In order to explore whether ODV-IBU has the effect of anti-oxidative stress, we analyzed the content of SOD and MDA in the hippocampus of mice by ELISA. As shown in Figure 5 As shown in A [F(5, 30) = 9.199, p < 0.0001], the SOD content of each group was as follows: the solvent group (Control) 211.1 ± 6.30 U / mg, the model group (LPS) 168.6 ± 4.00 U / mg, the ODV group 186.5 ± 2.66 U / mg, the IBU group 190.4 ± 5.49 U / mg, the ODV + IBU group 196.1 ± 4.12 U / mg, and the ODV-IBU group 200.2 ± 4.92 U / mg; Figure 5 B [F(5, 29) = 8.181, p < 0.0001] was the MDA content of each group, the solvent group (Control) 2.39 ± 0.10 nmol / mg, the model group (LPS) 4.028 ± 0.24 nmol / mg, the ODV group 3.028 ± 0.24 nmol / mg, the IBU group 2.942 ± 0.26 nmol / mg, the ODV + IBU group 2.653 ± 0.16 nmol / mg, and the ODV-IBU group 2.650 ± 0.16 nmol / mg.

[0104] Oxidative stress and inflammation are important factors in the development of depression. In addition, oxidative stress can trigger inflammatory response. Compared with other organs, the central nervous system is more susceptible to oxidative stress. Therefore, neuroinflammation is closely related to excessive oxidative stress. The results showed that injection of LPS reduced the content of SOD and increased MDA, while O-desvenlafaxine (ODV) and ibuprofen (IBU) had the effect of regulating oxidative stress and had a regulatory effect on MDA and SOD. In addition, O-desvenlafaxine + ibuprofen (ODV + IBU) and O-desvenlafaxine-ibuprofen (compound 1, ODV-IBU) also have good antioxidant effects. From the experimental data, the content of SOD in the O-desvenlafaxine + ibuprofen (ODV + IBU) group and the O-desvenlafaxine-ibuprofen (compound 1, ODV-IBU) group was significantly higher than that in the two single drug groups, and the content of MDA was also significantly lower than that in the two single drug groups.

[0105] (4) Effect of O-desvenlafaxine-ibuprofen (compound 1, ODV-IBU) on the content of neurotransmitters in the brain of LPS-induced depression mice

[0106] The production of inflammation can lead to overexpression of NF-κB and COX-2. Studies have reported that inflammation can affect the level of neurotransmitters in the brain, leading to a decrease in the content of 5-HT, NE and GABA, and neurotransmitters are related to depression-like behavior. Therefore, we detected the content of neurotransmitters in the hippocampus of mice; Figure 6 A [F (5, 30) = 10.57, p < 0.0001] is the content of 5-HT in each group, respectively, the solvent group (Control) 8.572 ± 0.50 ng / ml, the model group (LPS) 5.598 ± 0.13 ng / ml, the O-desvenlafaxine group (ODV) 7.255 ± 0.42 ng / ml, the ibuprofen group (IBU) 6.962 ± 0.29 ng / ml, the O-desvenlafaxine + ibuprofen group (ODV + IBU) 7.588 ± 0.38 ng / ml, the O-desvenlafaxine-ibuprofen group (compound 1, ODV-IBU) 8.505 ± 0.15 ng / ml; Figure 6 B [F (5, 12) = 24.40, p < 0.0001] is the content of NE in the hippocampus of mice in each group, the solvent group (Control) 51.08 ± 3.00 pg / ml, the model group (LPS) 13.49 ± 1.71 pg / ml, the O-desvenlafaxine group (ODV) 42.45 ± 3.81 pg / g / ml, the ibuprofen group (IBU) 28.42 ± 3.23 pg / ml, the O-desvenlafaxine + ibuprofen group (ODV + IBU) 48.89 ± 2.42 pg / ml, the O-desvenlafaxine-ibuprofen group (compound 1, ODV-IBU) 47.26 ± 3.26 pg / ml.

[0107] In summary, LPS caused the decrease of 5-HT and NE in the hippocampus of mice, and the content of 5-HT and NE increased after drug treatment. The content of neurotransmitters in the hippocampus of mice treated with ODV+IBU and ODV-IBU was higher than that of single drug group, which indicated that ODV+IBU and ODV-IBU significantly reversed the decrease of neurotransmitters induced by LPS stress and increased the content of neurotransmitters in the brain.

[0108] (5) Effect of ODV-IBU (compound 1) on the damage of cerebral neurons of LPS stressed mice

[0109] In order to determine whether LPS-induced inflammation would induce damage to the synaptic structure of hippocampal neurons, Nissl staining was performed on brain sections of mice; as shown in Figure 7 D[F(5, 18) = 17.51, p < 0.001] showed that the number of neurons in the cerebral cortex and hippocampal CA1 region was counted and analyzed by ImageJ, and the cell count was counted under normal conditions, and the number of cells in the hippocampal CA1 region was 76.25 ± 1.49, 55.00 ± 1.87, 64.25 ± 2.46, 65.00 ± 1.96, 74.00 ± 1.87, and 75.75 ± 2.29, respectively, in the control group, the model group (LPS), the ODV group, the IBU group, the ODV+IBU group, and the ODV-IBU group (compound 1); Figure 7 E[F(5, 18) = 23.75, p < 0.0001] was the number of cells in the cerebral cortex of each group, and the number of cells was 149.7 ± 4.48, 100.0 ± 5.29, 122.7 ± 2.73, 129.3 ± 3.71, 141.3 ± 2.33, and 140.3 ± 2.19, respectively, in the control group, the model group (LPS), the ODV group, the IBU group, the ODV+IBU group, and the ODV-IBU group (compound 1).

[0110] As Figure 7A-C, the neurons of the cerebral cortex and hippocampus (CA1, DG) of the mice in the control group were arranged in order, the cell outline was clear and complete, the number of cells was more, and the morphology was normal. The cells in the cortex and hippocampus of the mice in the model group (LPS) were severely damaged, the number of neurons was reduced, the cells were arranged loosely, the space between the cells was large, the cell morphology was different, and some cells showed nuclear pyknosis, cell rupture, and vacuolization. Compared with the model group (LPS), the degree of cell damage in the cortex and hippocampus of the mice in the nortriptyline (ODV) group and the ibuprofen (IBU) group was lighter, and the cell morphology was occasionally abnormal. The number of cortical cells and hippocampal cells was significantly increased. Compared with the model group (LPS), the cells in the cortex and hippocampus of the mice in the nortriptyline + ibuprofen group (ODV+IBU) and the nortriptyline-ibuprofen group (compound 1, ODV-IBU) were arranged in order, the number of neurons was significantly increased, and the cell morphology was less abnormal, the cytoplasmic staining of the cells was deeper, and the cell outline was clear. The above data can show that ODV-IBU effectively reversed the reduction of the number of neurons in the cortex and hippocampus of the mice caused by LPS, and the effect was better than that of nortriptyline and ibuprofen.

[0111] In summary, LPS can cause damage to the neurons of the cerebral cortex and hippocampus of mice, leading to cell death or cell death. After drug treatment, the damage to the neurons of the mice is alleviated, and nortriptyline + ibuprofen (ODV+IBU) and nortriptyline-ibuprofen (compound 1, ODV-IBU) have better protective effects on neurons than single-drug treatment, and the effect of ODV-IBU is more significant.

[0112] Example 10: Evaluation of the antidepressant effect of nortriptyline-naproxen (compound 2, ODV-NAP) in a LPS-induced depression model with a cycle of 8 days

[0113] ICR mice, male, 22-30g, the animals were randomly divided into control group (Control), model group (LPS), nortriptyline group (ODV) 16mg / kg, naproxen group (NAP) 13.99mg / kg, nortriptyline + naproxen group (ODV+NAP), nortriptyline-naproxen (compound 2, ODV-NAP) 28.89mg / kg, n=9-10 in each group; the control group and the LPS group were injected intraperitoneally with the corresponding dose of normal saline, and the drug dose and preparation were the same as in Example 9.

[0114] (1) The effect of nortriptyline-naproxen (compound 2, ODV-NAP) on the behavior of LPS-induced depression model mice

[0115] As Figure 8A [F(5, 42) = 11.54, p < 0.0001] showed that the sucrose preference of the control group (Control) was 71.08 ± 2.30%, the model group (LPS) was 50.23 ± 3.12%, the norvenlafaxine (ODV) group was 61.39 ± 2.02%, the naproxen (NAP) group was 59.99 ± 1.78%, the norvenlafaxine + naproxen group (ODV + NAP) was 68.59 ± 2.42%, and the norvenlafaxine-naproxen group (compound 2, ODV-NAP) was 69.09 ± 1.95%; wherein the sucrose intake of the mice after drug treatment was significantly increased, and the sucrose intake percentage of the ODV + NAP group and the ODV-NAP group was significantly better than that of the two single-drug groups; Figure 8 B [F(5, 54) = 10.56, p < 0.0001] showed that the immobility time of the control group (Control) in the last 4 min was 111.7 ± 6.45 s, the model group (LPS) was 173.3 ± 5.12 s, the norvenlafaxine (ODV) group was 146.0 ± 3.55 s, the naproxen (NAP) group was 143.8 ± 7.925 s, the norvenlafaxine + naproxen group (ODV + NAP) was 130.6 ± 6.94 s, and the norvenlafaxine-naproxen group (compound 2, ODV-NAP) was 128.9 ± 7.39 s, and the forced swimming test data showed that the immobility time of the model group (LPS) was higher than that of the normal group, indicating that the survival desire of the mice was reduced, and the behavior of the mice was desperate, while the treatment groups could significantly reduce the immobility time, reduce the desperate behavior of the mice, enhance the survival desire, and the immobility time of the combination group was the least among the four treatment groups, indicating that the norvenlafaxine-naproxen (compound 2, ODV-NAP) had a better treatment effect on improving the depressive behavior of the mice than the single-drug treatment groups. Figure 8 C [F(5, 54) = 12.51, p < 0.0001] showed that the immobility time of the control group (Control) in the last 4 min was 116.4 ± 5.95 s, the model group (LPS) was 166.2 ± 4.85 s, the norvenlafaxine (ODV) group was 144.6 ± 2.60 s, the naproxen (NAP) group was 141.8 ± 4.69 s, the norvenlafaxine + naproxen group (ODV + NAP) was 128.1 ± 5.77 s, and the norvenlafaxine-naproxen group (compound 2, ODV-NAP) was 123.9 ± 5.76 s, and the forced swimming test data showed that the immobility time of the model group (LPS) was higher than that of the normal group, indicating that the survival desire of the mice was reduced, and the behavior of the mice was desperate, while the treatment groups could significantly reduce the immobility time, reduce the desperate behavior of the mice, enhance the survival desire, and the immobility time of the combination group was the least among the four treatment groups, indicating that the norvenlafaxine-naproxen (compound 2, ODV-NAP) had a better treatment effect on improving the depressive behavior of the mice than the single-drug treatment groups.

[0116] As Figure 8D[F(5, 60) = 14.02, p < 0.0001] shows that the distance of mice in the open field in the first 5 min, the control group (Control) is 17.79 ± 0.76 m, the model group (LPS) is 10.51 ± 0.68 m, the norvenlafaxine (ODV) group is 13.49 ± 0.69 m, the naproxen (NAP) group is 14.03 ± 0.77 m, the norvenlafaxine + naproxen group (ODV + NAP) is 15.24 ± 0.42 m, and the norvenlafaxine-naproxen group (compound 2, ODV-NAP) is 15.22 ± 0.44 m; at the same time Figure 8 E[F(5, 54) = 9.769, p < 0.0001] is the total movement time of mice in the open field in the first 5 min, the control group (Control) is 200.2 ± 7.06 s, the model group (LPS) is 151.8 ± 5.08 s, the norvenlafaxine (ODV) group is 174.8 ± 4.19 s, the naproxen (NAP) group is 179.5 ± 5.58 s, the norvenlafaxine + naproxen group (ODV + NAP) is 190.7 ± 5.50 s, and the norvenlafaxine-naproxen group (compound 2, ODV-NAP) is 191.8 ± 5.11 s; Figure 8 F[F(5, 54) = 9.819, p < 0.0001] is the number of mice crossing the central area in the first 5 min, the control group (Control) is 30.5 ± 1.93, the model group (LPS) is 15.2 ± 1.61, the norvenlafaxine (ODV) group is 24.7 ± 1.86, the naproxen (NAP) group is 27.2 ± 1.86, the norvenlafaxine + naproxen group (ODV + NAP) is 29.0 ± 1.46, and the norvenlafaxine-naproxen group (compound 2, ODV-NAP) is 29.1 ± 1.99. In combination with other data, LPS can significantly reduce the movement ability and exploration ability of mice in the open field, mainly manifested as a decrease in movement distance, a decrease in movement time, and a decrease in the number of entering the central area. Among the three treatment groups, the movement ability and exploration ability of mice can be restored, and norvenlafaxine-naproxen (compound 2, ODV-NAP) treatment can significantly increase the movement distance, movement time, and central area number;

[0117] (2) Effect of norvenlafaxine-naproxen (compound 2, ODV-NAP) on inflammatory factors in the hippocampus of LPS-induced depressive mice

[0118] In order to explore whether norvenlafaxine-naproxen (compound 2, ODV-NAP) has a regulatory effect on inflammatory factors, we detected the inflammatory factors in the hippocampus of mice by ELISA; the results are as follows Figure 9TNF-α levels in each group, as shown by A [F(5, 30) = 18.28, p < 0.0001], were as follows: Control 187.7 ± 8.06 pg / ml, LPS 348.0 ± 18.42 pg / ml, ODV 262.6 ± 16.63 pg / ml, NAP 238.7 ± 16.17 pg / ml, ODV + NAP 224.1 ± 13.45 pg / ml, ODV-NAP (Compound 2) 191.2 ± 5.14 pg / ml; as Figure 9 IL-1β levels in each group, as shown by B [F(5, 30) = 34.56, p < 0.0001], were as follows: Control 49.05 ± 5.49 pg / ml, LPS 145.3 ± 7.46 pg / ml, ODV 82.12 ± 7.20 pg / ml, NAP 69.71 ± 5.78 pg / ml, ODV + NAP 58.32 ± 4.11 pg / ml, ODV-NAP (Compound 2) 54.40 ± 5.80 pg / ml; as Figure 9 IL-6 levels in each group, as shown by C [F(5, 30) = 36.60, p < 0.0001], were as follows: Control 56.12 ± 7.09 pg / ml, LPS 175.0 ± 10.03 pg / ml, ODV 134.8 ± 8.70 pg / ml, NAP 112.3 ± 10.37 pg / ml, ODV + NAP 69.66 ± 6.25 pg / ml, ODV-NAP (Compound 2) 43.44 ± 7.34 pg / ml; from the data, LPS significantly increased the levels of inflammatory factors in the hippocampus of mice, while ODV and NAP can reduce the levels of inflammatory factors, and the anti-inflammatory effect of NAP is better than that of ODV, and the anti-inflammatory effect of ODV-NAP (Compound 2) is significantly better than that of the two single drugs, which indicates that ODV-NAP (Compound 2) has good anti-inflammatory effect.

[0119] (3) Effect of ODV-NAP (Compound 2) on oxidative stress in the hippocampus of LPS-induced depressive mice

[0120] Overexpression of inflammatory factors can promote oxidative stress, leading to imbalance between oxidation and antioxidant mechanisms in the body, and thus leading to imbalance between SOD and MDA and other substances. In order to explore whether O-desmethylvenlafaxine-naproxen (compound 2, ODV-NAP) has an antioxidant stress effect, we performed ELISA analysis on the mouse hippocampus to detect the contents of SOD and MDA related to the contents of SOD and MDA. As shown in Figure 10 As shown in A [F(5, 30) = 15.90, p < 0.0001], the SOD content of each group was solvent group (Control) 234.3 ± 6.49 U / mg, model group (LPS) 170.2 ± 3.04 U / mg, O-desmethylvenlafaxine (ODV) group 192.1 ± 5.35 U / mg, naproxen (NAP) group 190.9 ± 3.37 U / mg, O-desmethylvenlafaxine + naproxen group (ODV + NAP) 204.4 ± 7.00 U / mg, O-desmethylvenlafaxine-naproxen group (compound 2, ODV-NAP) 208.5 ± 5.82 U / mg; as shown in Figure 10 As shown in B [F(5, 30) = 24.87, p < 0.0001], the MDA content of each group was solvent group (Control) 2.022 ± 0.20 nmol / mg, model group (LPS) 4.642 ± 0.22 nmol / mg, O-desmethylvenlafaxine group (ODV) 3.512 ± 0.19 nmol / mg, naproxen group (NAP) 3.265 ± 0.22 nmol / mg, O-desmethylvenlafaxine + naproxen group (ODV + NAP) 2.623 ± 0.15 nmol / mg, O-desmethylvenlafaxine-naproxen group (compound 2, ODV-NAP) 2.270 ± 0.14 nmol / mg.

[0121] Oxidative stress and inflammation are important factors in the development of depression. In addition, oxidative stress can trigger an inflammatory response. Compared with other organs, the central nervous system is more susceptible to oxidative stress. Therefore, neuroinflammation is closely related to excessive oxidative stress. The results of the study showed that injection of LPS reduced SOD and increased MDA, while O-desmethylvenlafaxine (ODV) and naproxen (NAP) had an effect of relieving oxidative stress, reducing the content of MDA in the hippocampus and increasing the content of SOD. In addition, the combination of O-desmethylvenlafaxine and naproxen and O-desmethylvenlafaxine-naproxen also had a good antioxidant stress effect. From the experimental data, the antioxidant stress effect of O-desmethylvenlafaxine-naproxen (compound 2, ODV-NAP) was significantly better than that of other treatment groups.

[0122] (4) Effect of O-desmethylvenlafaxine-naproxen (compound 2, ODV-NAP) on the content of neurotransmitters in the hippocampus of LPS-induced depressive mice

[0123] The production of inflammation can lead to overexpression of NF-κB and COX-2, and studies have reported that inflammation can affect the levels of neurotransmitters in the brain, leading to a decrease in the content of 5-HT and NE in the brain, and the decrease in neurotransmitters is associated with depressive-like behavior. Therefore, we detected the content of neurotransmitters in the hippocampus of mice; as Figure 11 A[F(5, 30) = 19.81, p < 0.0001], the content of 5-HT in each group was, solvent group (Control) 14.53 ± 0.35 ng / mg, model group (LPS) 8.167 ± 0.39 ng / mg, nortriptyline group (ODV) 11.37 ± 0.62 ng / mg, naproxen group (NAP) 10.31 ± 0.57 ng / mg, nortriptyline + naproxen group (ODV + NAP) 11.32 ± 0.62 ng / mg, nortriptyline-naproxen group (compound 2, ODV-NAP) 13.61 ± 0.47 ng / mg; as Figure 11 B[F(5, 12) = 71.91, p < 0.0001], the content of NE in each group was, solvent group (Control) 163.1 ± 10.28 pg / ml, model group (LPS) 50.80 ± 5.20 pg / ml, nortriptyline group (ODV) 124.0 ± 7.83 pg / ml, naproxen group (NAP) 78.79 ± 8.09 pg / ml, ODV + NAP group 128.7 ± 4.41 pg / ml, nortriptyline-naproxen group (compound 2, ODV-NAP) 153.4 ± 2.46 pg / ml.

[0124] From the experimental data, LPS can cause a decrease in the content of 5-HT and NE in the hippocampus of mice, and the content of 5-HT and NE is increased after drug treatment, among which nortriptyline significantly increases the content of 5-HT and NE in the hippocampus, in addition, the content of neurotransmitters after treatment with nortriptyline + naproxen (ODV + NAP) and nortriptyline-naproxen (compound 2, ODV-NAP) is better than that of single drug treatment, which indicates that ODV-NAP treatment can significantly reverse the neuron damage induced by LPS stress and increase the content of neurotransmitters in the brain.

[0125] (5) Effect of nortriptyline-naproxen (compound 2, ODV-NAP) on LPS stress-induced neuron cell damage in the brain of mice

[0126] In order to determine whether LPS-induced inflammation can induce neuron cell damage in the hippocampus, Nissl staining was performed on the brain sections of mice; as Figure 12 As shown, the neuron cells in the cerebral cortex and hippocampus DG region of the sections were counted and analyzed by Image J, wherein the cell count was under normal conditions, and Figure 12C [F(5, 12) = 34.63, p < 0.0001] is the cell number in the hippocampal DG region, the vehicle group (Control) is 814.7 ± 13.35, the model group (LPS) is 512.7 ± 16.60, the norepinephrine group (ODV) is 675.3 ± 23.85, the naproxen group (NAP) is 697.7 ± 31.35, the norepinephrine + naproxen group (ODV + NAP) is 851.7 ± 27.00, and the norepinephrine-naproxen group (compound 2, ODV-NAP) is 842.0 ± 15.39; Figure 12 D [F(5, 12) = 32.67, p < 0.0001] is the cell count in the cerebral cortex of each group, the vehicle group (Control) is 84.33 ± 2.33, the model group (LPS) is 50.67 ± 2.40, the norepinephrine group (ODV) is 63.0 ± 2.52, the naproxen group (NAP) is 65.33 ± 1.76, the norepinephrine + naproxen group (ODV + NAP) is 80.33 ± 2.67, and the norepinephrine-naproxen group (compound 2, ODV-NAP) is 81.0 ± 2.08.

[0127] As Figure 12 A, 12B shows that the cerebral cortex and hippocampal DG neurons of the vehicle group (Control) mice are arranged in order, the neuron cell profile is clear and complete, the cell number is more, and the morphology is mostly normal. The cortex and hippocampal region cells of the model group (LPS) mice are severely damaged, the cell number is reduced, the cell arrangement is loose and the space between cells is large, the cell morphology is different, and part of the cells have nuclear pyknosis, cell rupture and vacuolization and other pathological phenomena. Compared with the model group (LPS), the cortex and hippocampal cells of the norepinephrine group (ODV) and the naproxen group (NAP) mice are less damaged, and the cell morphology is occasionally abnormal, wherein the number of cortex cells and the number of hippocampal cells are significantly increased. Compared with the model group (LPS), the cerebral cortex and hippocampus of the norepinephrine + naproxen group (ODV + NAP) and the norepinephrine-naproxen group (compound 2, ODV-NAP) mice are arranged in order, the number of neurons is significantly increased, and the cell morphology is less abnormal, the cell cytoplasm is stained darker, and the cell profile is clear. The above data can show that norepinephrine-naproxen (compound 2, ODV-NAP) effectively reverses the reduction of neuron cell number in the cortex and hippocampal region of mice caused by LPS, and its effect is better than that of norepinephrine (ODV) and naproxen (NAP).

[0128] Through the above experiments, it is proved that the twin drugs and isomers thereof in the application have the effects of enhancing the anti-depression effect, anti-inflammation, anti-oxidation, increasing the content of neurotransmitters and protecting neuron cells. The composition can be used for treating depression, anxiety and nervous system related diseases, significantly enhancing the anti-depression effect and improving the patient's drug compliance.

[0129] The above examples only express several embodiments of the description, which are described in a more specific and detailed manner, but cannot be understood as limiting the scope of the description. It should be noted that for ordinary skilled persons in the art, several modifications and improvements can be made without departing from the concept of the description, and these all belong to the protection of the description.

Claims

1. A twin drug or a pharmaceutically acceptable salt thereof or a tautomer of a drug with synergistic antidepressant and anti-anxiety effects, characterized in that: The twin drug is obtained by connecting drug A and drug B through a chemical bond, wherein drug A is norvenlafaxine and drug B is a non-steroidal anti-inflammatory drug.

2. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a tautomer thereof, wherein The non-steroidal anti-inflammatory drug is selected from the group consisting of ibuprofen, naproxen, aspirin, diclofenac, ketoprofen, indomethacin, flurbiprofen.

3. The twin drug or its pharmaceutically acceptable salt or its tautomer according to claim 1-2, characterized as follows:

4. A process for preparing a compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a tautomer thereof, characterized in that, The preparation method comprises the following steps: Norvenlafaxine and the non-steroidal anti-inflammatory drug are condensed under the action of a catalyst to obtain the final product.

5. The preparation method according to claim 4, characterized in that, The catalyst in step 1 is selected from the group consisting of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 4-dimethylaminopyridine, N,N'-dicyclohexylcarbodiimide, N,N'-diisopropylcarbodiimide. The molar ratio of norvenlafaxine, non-steroidal anti-inflammatory drug and catalyst is 1:1:(0.01-10), and the reaction solvent is selected from the group consisting of dichloromethane, acetonitrile, acetone or tetrahydrofuran.

6. Use of a compound according to any one of claims 1-3 in the preparation of a medicament for preventing and / or treating depression, anxiety and nervous system diseases.

7. Use according to claim 6, characterized in that, The medicament is a medicament for preventing or treating major depression, bipolar depression, psychotic depression, reactive depression, menopausal depression, child depression, secondary depression, postpartum depression, seasonal depression, persistent depression, postmenopausal and perimenopausal depression, and anxiety depression.

8. Use according to claim 6, characterized in that, The medicament is a medicament for preventing or treating generalized anxiety disorder, panic disorder, social anxiety disorder and separation anxiety disorder. The medicament is a medicament for preventing or treating generalized anxiety disorder, panic disorder, social anxiety disorder and separation anxiety disorder.

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