Application of amide derivative compound in treatment of anxiety

Abstract

The invention relates to application of amide compounds in treatment of anxiety. The amide derivative disclosed by the invention has nNOS-CAPN protein-protein interaction inhibition activity, and can be used for preparing an anti-anxiety drug.

Description

Technical Field

[0001] The present invention belongs to the field of pharmaceuticals and provides pharmaceutical uses of a class of amide derivatives. These compounds can inhibit the activity of nNOS-CAPON protein coupling and have application prospects in the preparation of antianxiety drugs. The anxiety disorders described include generalized anxiety disorder, panic attacks and panic disorder, and social phobia. Background Art

[0002] Anxiety disorders are the most common mental disorders worldwide. According to the World Health Organization (WHO), anxiety disorders can be categorized as generalized anxiety disorder, panic disorder, social anxiety disorder, agoraphobia, separation anxiety disorder, specific anxiety disorders, and selective mutism. Research indicates that people may suffer from more than one anxiety disorder simultaneously, with symptoms typically beginning in childhood or adolescence and persisting into adulthood. Due to its high prevalence, chronicity, and comorbidities, the WHO has listed anxiety disorders as the ninth leading cause of health-related disability. Therefore, the prevention and treatment of anxiety disorders have become a key focus in psychiatry.

[0003] At present, the main drug treatments for anxiety disorders include selective serotonin reuptake inhibitors (SSRIs), serotonin-norepinephrine reuptake inhibitors (SNRIs), monoamine oxidase inhibitors (MAOIs) and benzodiazepines (BZDs). Among them, BZDs have been used to treat anxiety disorders, but they are usually not used as first-line drugs because of their limited long-term efficacy and easy dependence [5]. In the treatment of major anxiety disorders, SSRIs and SNRIs are the first-line drugs. They increase the concentration of neurotransmitters in the synapse by inhibiting the reuptake of serotonin and norepinephrine by transporters in the presynaptic membrane. However, there are still a considerable number of patients who do not respond to these first-line treatment drugs, and there is a lack of guidance on the effectiveness of treating anxiety disorders. Therefore, the development of new anti-anxiety drugs has become an urgent issue. In the past few decades, more and more studies have shown that the main molecules that regulate anxiety behavior include γ-aminobutyric acid (GABA), N-methyl-D-aspartate (NMDA) and serotonin (5-HT). Studies on the upstream and downstream pathways of these molecules have revealed that NMDAR-mediated activation of neuronal nitric oxide synthase (nNOS) plays an important role in the regulation of anxiety. Enhanced interaction between hippocampal nNOS and its carboxyl-terminal PDZ ligand (CAPON) can induce anxiety-like behaviors.

[0004] Therefore, it is expected that new antianxiety drugs will be developed by regulating nNOS-CAPON coupling. Summary of the Invention

[0005] The object of the present invention is to provide a drug for treating anxiety through a new mechanism, wherein the drug is an amide derivative or a pharmaceutically acceptable salt thereof.

[0006] The above-mentioned drug is characterized in that by inhibiting the coupling of nNOS-CAPON proteins, it alleviates anxiety-like symptoms, exerts the effect of treating anxiety, and ultimately achieves the purpose of treating anxiety.

[0007] Therefore, the present invention provides a new use of an amide derivative or a pharmaceutically acceptable salt thereof and a pharmaceutical composition thereof for preparing an anti-anxiety drug.

[0008] Technical problem to be solved: The present invention provides an application of an amide compound having nNOS-CAPON protein coupling inhibitory activity in the preparation of a drug for treating anxiety.

[0009] Technical solution: The application of a compound represented by Formula I, Formula II and Formula III or a pharmaceutically acceptable salt thereof in the preparation of a drug for treating anxiety,

[0010]

[0011] wherein,

[0012] R1 is hydrogen, C1-C6 alkyl, C3-C4 cycloalkyl, trifluoroethyl, nitrogen-benzylpiperidinyl, methanesulfonyl or p-fluorobenzenesulfonyl;

[0013] R2 is hydrogen or methyl;

[0014] R3 and R4 are independently selected from hydrogen, C1-C5 alkyl, methoxy, fluorine, chlorine, trifluoromethyl, trifluoromethoxy, phenyl or nitrogen-methylpiperazinyl;

[0015] X is selected from carbon or nitrogen;

[0016] n is selected from the number 0 or 1;

[0017] m is selected from the number 1 or 3.

[0018] Preferably, the compound is selected from:

[0019]

[0020] Compound 1, as shown in N1:

[0021]

[0022] Compound 2, as shown in N2:

[0023]

[0024] Compound 3, as shown in N3:

[0025]

[0026] Compound 4, as shown in N4:

[0027]

[0028] Compound 5, as shown in N5:

[0029]

[0030] Compound 6, as shown in N6:

[0031]

[0032] Compound 7, as shown in N7:

[0033]

[0034] Compound 8, as shown in N8:

[0035]

[0036] Compound 9, as shown in N9:

[0037]

[0038] Compound 10, as shown in N10:

[0039]

[0040] Compound 11, as shown in N11:

[0041]

[0042] Compound 12, as shown in N12:

[0043]

[0044] Compound 13, as shown in N13:

[0045]

[0046] Compound 14, as shown in N14:

[0047]

[0048] Compound 15, as shown in N15:

[0049]

[0050] Compound 16, as shown in N16:

[0051]

[0052] Compound 17, as shown in N17:

[0053]

[0054] Compound 18, as shown in N18:

[0055]

[0056]

[0057] Compound 19, as shown in N19:

[0058]

[0059] Compound 20, as shown in N20:

[0060]

[0061] Compound 21, as shown in N21:

[0062]

[0063] Compound 22, as shown in N22:

[0064]

[0065] Compound 23, as shown in N23:

[0066]

[0067] Compound 24, as shown in N24:

[0068]

[0069]

[0070] Compound 25, as shown in N25:

[0071]

[0072] Compound 26, as shown in N26:

[0073]

[0074] Compound 27, as shown in N27:

[0075]

[0076] Compound 28, as shown in N28.

[0077] The compounds provided by the present invention also include pharmaceutically acceptable equivalents of the compound or a mixture of two or more thereof.

[0078] Preferably, the compounds provided by the present invention may include one or a mixture of two or more of pharmaceutically acceptable salts, hydrates, solvates, metabolites, and prodrugs.

[0079] Preferably, the compounds provided by the present invention include acid salts or base salts of the compounds provided by the present invention. The pharmaceutically acceptable salts have the pharmaceutical activity of the compounds and meet the requirements both biologically and in practical applications.

[0080] The present invention provides a class of amide compounds or pharmaceutically acceptable salts thereof for treating diseases such as anxiety.

[0081] Beneficial effects:

[0082] A class of amide compounds described in the present application is characterized by having nNOS-CAPON protein coupling inhibitory activity and can be used to prepare drugs for treating anxiety. Detailed implementation manners

[0083] The present invention discloses a class of amino acid ester derivatives and their uses. Those skilled in the art can draw on the content of this article and appropriately improve process parameters to achieve them. The methods and applications of the present invention have been described through preferred embodiments. Relevant personnel can obviously make changes or appropriate modifications and combinations to the methods and applications described herein without departing from the content, spirit, and scope of the present invention to implement and apply the technology of the present invention.

[0084] In order to enable those skilled in the art to better understand the technical solutions of the present invention, the present invention will be further described in detail below with reference to specific embodiments.

[0085] Example 1: Screening of the inhibitory activity of amide derivatives on the protein-protein interaction of nNOS-CAPON

[0086] 1 Materials and methods

[0087] 1.1 Recombinant proteins and antibodies

[0088] Name Manufacturer Article No. Lot No. his-nNOS(1-299) Nanjing Bioroid Biotechnology Co., Ltd. N / A N / A FITC-CAPON-C20 Nanjing Genscript Biotechnology Co., Ltd. N / A A01632 Tb-anti-his Antibody Thermo Fisher PV5863 1370618D

[0089] 1.2 PPI detection buffer (Assay buffer)

[0090] Formulation: 20 mM PB, pH 7.4, 1 mM EDTA, 50 mM NaCl, and 0.05% pluronic F-68

[0091] 1.3 Main instruments and consumables

[0092] Name Manufacturer Model Specification Multi-channel Pipette (8 channels) Rainin 0.5 - 10μL Microplate Reader MD SpectraMax i3x 96 well plate Greiner 784075

[0093] 1.4 Amide derivatives numbering and structure

[0094]

[0095]

[0096]

[0097] 1.5 Experimental system

[0098] In this experiment, a 20 μL reaction system was used. Before the experiment, each group was diluted to 4 times the working solution concentration with Assay Buffer to prepare 4×His-nNOS(1-299) reaction solution, 4×FITC-CAPON-C20 reaction solution, 4×Tb-anti-hisantibody reaction solution, and 4×test sample reaction solution. Then, 5 μL was added to a 384-well plate in sequence and reacted at 25 °C for 6 h. Fluorescence was detected using a microplate reader: 337 nm - 488 nm - 520 nm.

[0099] 1.6 Data analysis

[0100] The emission light ratio of the FRET system (337 - 488 - 520 nm) is Ratio = F520 nm / F490 nm.

[0101] The inhibition rate of the compound inhibiting PPI is calculated according to the formula Inhibition(%) = 100%×(Ratio sample - Ratio 0％inhibition ) / (Ratio 100％inhibition - Ratio 0％inhibition ); where Ratio sample is the emission light ratio of the compound well, Ratio 0％inhibition is the emission light ratio of the DMSO control well, and Ratio 100％inhibition is the emission light ratio of the well without his-NOS1-299.

[0102] Use Prism 7 (GraphPad Software) log(inhibitor) vs. response--Normalized response-Variable slope to fit the S curve of compound concentration - inhibition rate, and calculate the IC 50 value of the compound inhibiting PPI.

[0103] 2 Experimental results

[0104] All the measured amide derivatives showed certain Ppi inhibitory activities (Table 1), among which, N2 had the strongest PPI inhibitory activity.

[0105] Table 1 Inhibition of PPI by incubated compounds in the PPI reaction system, IC 50 Summary

[0106]

[0107] Example 2: Elevated Plus Maze Test of Compound N2

[0108] 1.1 Experimental animals

[0109] C57BL / 6J mice, male, SPF grade, body weight 20 - 30 g, from Beijing Vital River Laboratory Animal Technology Co., Ltd.

[0110] 1.2 Experimental equipment

[0111]

[0112]

[0113] 1.3 Experimental methods

[0114] 1.3.1 Model establishment method

[0115] Normal C57BL / 6J mice have anxiety-like symptoms and do not require model establishment.

[0116] 1.3.2 Animal grouping and administration

[0117] This experiment was divided into 5 groups, namely the solvent control group, N2 administration group (10 mg / kg), N2 administration group (20 mg / kg), FLX administration group (5 mg / kg), and FLX administration group (5 mg / kg). C57BL / 6J mice were randomly and blindly divided into each group with equal probability. Mice in each group were accurately drawn with the corresponding amount of the control group administration preparation or N2 administration preparation or FLX solution by a 1 mL syringe and administered by oral gavage. N2 was administered 2 times a day (interval about 8 hours), and FLX and the solvent control group were administered 1 time a day for 8 consecutive days.

[0118] 1.3.3 Elevated Plus Maze (Elevate Plus Maze test, EPM) experiment

[0119] After continuous administration for 8 days according to body weight, the elevated plus maze experiment was carried out 1.0 h after administration on the 8th day.

[0120] A plus maze with a height of 50 cm from the ground and an arm length of 30 cm was used as the test tool. The maze consisted of two open arms (30×5 cm), two closed arms (30×5 cm). The periphery and the end of the closed arms were surrounded by a 15-cm-high fence. The open arms and the closed arms were connected by a central area (5×5 cm). At the beginning of the experiment, the mice were placed in the central area of the maze, facing the open arms and allowed to move freely. The activities of the animals were recorded within 5 min.

[0121] Observation indicators included: the number of entries into the open arms and the closed arms (counting as 1 time when all four limbs completely entered), the residence time in the open arms, and the residence time in the closed arms. The proportion of the number of entries into the open arms, the proportion of the residence time in the open arms, and the total number of entries into the maze were calculated.

[0122] 1.4 Data statistical analysis

[0123] Quantitative results were expressed as Mean±SEM. One-way analysis of variance (One-way ANOVA) was performed on each behavioral detection index using Prism6 software. After ANOVA test showed differences, Fisher's LSD test was performed between two groups. P<0.05 was defined as having significant differences.

[0124] 2 Experimental results

[0125] Through one-way analysis of variance, there were statistical differences in the residence time in the open arms among groups (F(4,65)=0.7125, **P=0.0072). Compared with the control group, the N2 (20 mg / kg) dose group could significantly increase the residence time of mice in the open arms (p<0.01). The results are shown in Table 2.

[0126] Table 2 Experimental results of compound N2 in the EPM

[0127] Group Sample Size (number) Open Arm Retention Time (s) Vehicle 14 21.62±4.10 N2(10mg / kg) 14 22.72±3.37 N2(20mg / kg) 14 36.75±3.32** FLX(5mg / kg) 14 17.78±3.19 FLX(10mg / kg) 14 20.09±4.82

[0128] Compared with the model group, *P<0.05, **P<0.01;

[0129] FLX: Fluoxetine hydrochloride.

Claims

1. A class of amide compounds represented by Formula I, Formula II and Formula III, or a pharmaceutically acceptable salt thereof, characterized in that, Use in the preparation of drugs for treating anxiety wherein R1 is hydrogen, C1-C6 alkyl, C3-C4 cycloalkyl, trifluoroethyl, nitrogen benzyl piperidyl, methanesulfonyl or p-fluorobenzenesulfonyl; R2 is hydrogen or methyl; R3 and R4 are independently selected from hydrogen, C1-C5 alkyl, methoxy, fluorine, chlorine, trifluoromethyl, trifluoromethoxy, phenyl or nitrogen methyl piperazinyl; X is selected from carbon or nitrogen; n is selected from the number 0 or 1; m is selected from the number 1 or 3.

2. The compound according to claims 1 to 2 or a pharmaceutically acceptable salt thereof, characterized in that, The compounds are selected from: Compound 1, as shown in N1: Compound 2, as shown in N2: Compound 3, as shown in N3: Compound 4, as shown in N4: Compound 5, as shown in N5: Compound 6, as shown in N6: Compound 7, as shown in N7: Compound 8, as shown in N8: Compound 9, as shown in N9: Compound 10, as shown in N10: Compound 11, as shown in N11: Compound 12, as shown in N12: Compound 13, as shown in N13: Compound 14, as shown in N14: Compound 15, as shown in N15: Compound 16, as shown in N16: Compound 17, as shown in N17: Compound 18, as shown in N18: Compound 19, as shown in N19: Compound 20, as shown in N20: Compound 21, as shown in N21: Compound 22, as shown in N22: Compound 23, as shown in N23: Compound 24, as shown in N24: Compound 25, as shown in N25: Compound 26, as shown in N26: Compound 27, as shown in N27: Compound 28, as shown in N28.

3. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt thereof, characterized in that Use of the compounds in the preparation of drugs for treating generalized anxiety disorder 4. The compound according to claim 1 or 2 or a pharmaceutically acceptable salt thereof, characterized in that, Use of the compounds in the preparation of drugs for treating panic attacks and panic disorder 5. The compound according to claims 1 to 2 or a pharmaceutically acceptable salt thereof, characterized in that, Use of the compounds in the preparation of drugs for treating social phobia

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