6-Shogaol analogues, their compositions and preparation methods

Abstract

This invention discloses a 6-shogaol analogue and its derivatives, compositions comprising the same and methods of preparation thereof, as well as methods of using said compounds / compositions to treat diseases or conditions, such as depression.

Description

Technical Field

[0001] This disclosure belongs to the field of medicinal chemistry, and particularly relates to a 6-shogaol analogue and its derivatives, compositions comprising the same and methods of preparation thereof, and methods of using said compounds / compositions to treat diseases or conditions such as depression. Background Technology

[0002] According to statistics from the World Health Organization in 2015, depression is one of the major global health problems, with a prevalence of 2.9%-6.31% in different countries. Patients with depression suffer both physical and psychological pain, which also increases the economic burden on the healthcare system (1). The causes of depression involve a variety of genetic, epigenetic, and environmental risk factors (2). Current treatments include psychotherapy and drug therapy, which are only effective for some patients with depression and are often accompanied by adverse side effects (3, 4). Among existing antidepressants, selective serotonin reuptake inhibitors (SSRIs), serotonin-norepinephrine reuptake inhibitors (SNRIs), and monoamine oxidase inhibitors (MAOIs) mainly work by regulating the signal transduction of monoamine neurotransmitters at the synapse (2). On the other hand, neuroinflammation is closely related to the occurrence and development of depression (5, 6). Patients with major depressive disorder (MDD) have significantly elevated levels of pro-inflammatory cytokines—tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) (7, 8). Similarly, neuroinflammation is associated with depressive-like symptoms in various mouse models of depression, including those induced by chronic restraint stress (CRS), chronic mild stress (CMS), chronic unpredictable mild stress (CUMS), or corticosterone (CORT) (9-12). In fact, nonsteroidal anti-inflammatory drugs (NSAIDs) are effective in some patients with major depressive disorder (MDD), and some existing antidepressants also have anti-inflammatory effects (13). Therefore, developing novel multi-target antidepressants is an important direction for current research.

[0003] On the other hand, in recent years, a number of anti-inflammatory and antioxidant natural compounds have been evaluated as potential therapeutic agents for antidepressants (14, 15). For example, 6-shogaol (6Sh), derived from ginger root (Rhizoma Zingiberis Recens), can inhibit neuroinflammation in a variety of neurodegenerative diseases (16). However, the medical application of 6Sh is greatly limited due to its poor water solubility, low bioavailability, and high cost (17).

[0004] In addition, 6-shogaol exhibited strong cytotoxicity in various cell lines, including KB cells (IC50). 50 =7.4±2.2μM) and HL60 cells (IC 50=7.9±2.0 μM). 10 μM of 6-shogaol can induce significant cell death in mouse microglia BV2.

[0005] Therefore, it is of great significance to develop a novel antidepressant 6-shogaol analogue (6SA) with lower toxicity, better water solubility, higher bioavailability and better cost-effectiveness. Summary of the Invention

[0006] This invention provides a novel 6-shogaol analog and its derivatives, exhibiting lower toxicity, higher water solubility, and a more cost-effective preparation method compared to 6-shogaol. In terms of chemical structure, the 6-shogaol analog comprises a nitrogen linkage between an aromatic moiety and an unsaturated alkyl carbonyl side chain. This structural modification reduces synthesis costs by 99% and significantly increases water solubility and bioavailability.

[0007] According to a preferred embodiment of the present invention, a 10 µM 6-shogaol analogue exhibits no significant cytotoxicity to BV2 cells compared to 6-shogaol. The 6-shogaol analogue provided by the present invention is a safer and more effective 6-shogaol-like substance.

[0008] According to a preferred embodiment of the present invention, the 6-shogaol analog provided by the present invention has better water solubility and oral bioavailability.

[0009] According to a preferred embodiment of the present invention, the 6-shogaol analogue is non-toxic at a concentration of 10 µM.

[0010] According to a preferred embodiment of the present invention, the 6-shogaol analogue exhibits better antidepressant activity in a mouse model of depression.

[0011] According to a preferred embodiment of the present invention, the 6-shogaol analogue and its derivatives can be used to treat obesity and mental illnesses (e.g., anxiety and depressive-like behaviors).

[0012] According to a preferred embodiment of the present invention, the 6-shogaol analogue and its derivatives are used to treat pain, anxiety, mood disorders or depression.

[0013] Specifically, the first aspect of the present invention provides compounds of formula I or derivatives thereof, pharmaceutically acceptable salts, enantiomers or solvates:

[0014] Among them, R is selected from H and C. 1-7 H 3-15 C6H5, C6H5O, C6H5O2 or C7H8O2.

[0015] According to a preferred embodiment of the present invention, the C 1-7 H 3-15 Preferred C 3-7 Alkenyl, more preferably 1-heptenyl.

[0016] According to a preferred embodiment of the present invention, the C6H5O2 is selected from 3,4-dihydroxyphenyl.

[0017] According to a preferred embodiment of the present invention, the C7H8O2 is selected from 3-hydroxy-4-methoxyphenyl.

[0018] According to this invention, the term "derivative" refers to a compound having one or more modifications or substitutions in its parent compound. In this invention, the parent benzene ring structure is retained, and its carbon chain portion is substituted. The "derivative" as used in this invention has properties or activities similar to the parent compound.

[0019] According to a preferred embodiment of the present invention, the compound of formula I has the structure shown in formula II:

[0020] A second aspect of the invention provides a pharmaceutical composition comprising the compounds described above according to the invention and a pharmaceutically acceptable carrier.

[0021] A third aspect of the invention provides a pharmaceutical composition comprising: (a) the above-described compound according to the invention and a pharmaceutically acceptable carrier; and (b) an additional medicament for treating mental or psychological illnesses or conditions.

[0022] According to a preferred embodiment of the invention, the pharmaceutically acceptable carrier is selected from the group consisting of: liquid carriers; isotonic agents; wetting agents, solubilizers or emulsifiers; preservatives; buffers; acidifiers; antioxidants; alkalizing agents; carriers; chelating agents; colorants; complexing agents; solvents; suspensions and / or thickeners; oils; penetration enhancers; polymers; hardeners; thickeners; proteins; carbohydrates; fillers; and lubricants.

[0023] According to a preferred embodiment of the present invention, the pharmaceutical composition is formulated to be suitable for administration via a route selected from the group consisting of oral administration, intravenous administration, intraperitoneal administration, and intramuscular administration.

[0024] A fourth aspect of the invention provides a method for treating a subject's mental or psychological illness or condition, comprising administering an effective amount of the compound of the invention to the subject in need, wherein the mental or psychological illness or condition is selected from mood disorders, depression, anxiety, pain, post-traumatic stress disorder, bipolar disorder, substance abuse disorder, eating disorder, obsessive-compulsive disorder, and irritability. According to a preferred embodiment of the invention, the pain is acute pain, chronic pain, inflammatory pain, or neuropathic pain.

[0025] According to a preferred embodiment of the present invention, the subject is a human.

[0026] According to a preferred embodiment of the invention, the method further includes administering a therapeutically effective amount of an additional agent for treating depression, said agent being selected from selective serotonin reuptake inhibitors, serotonin-norepinephrine reuptake inhibitors, serotonin modulators, serotonin antagonists and reuptake inhibitors, norepinephrine reuptake inhibitors, tricyclic antidepressants, tetracyclic antidepressants, monoamine oxidase inhibitors, atypical antidepressants, and antidepressants acting through one or more other mechanisms.

[0027] A fifth aspect of the invention provides a commercial package containing the disclosed compound, along with instructions for use in treating pain, anxiety, mood disorders, or depression.

[0028] A sixth aspect of the present invention provides a method for preparing a compound of formula II, comprising the following steps: . Attached Figure Description

[0029] Figure 1 (A) The chemical structure of 6Sh and the structural design of 6SA; (B) The chemical synthesis of 6SA.

[0030] Figure 2 : Structural verification of 6SA. (A) UPLC-UV spectrum of 6SA (280nm); (B) LC-MS spectrum of 6SA.

[0031] Figure 3: 1H NMR spectrum of 6SA (600MHz, CDCl3), δ 0.88 (t, 3H, 8-CH3), 1.25 (m, 4H,6,7-CH2), 1.39 (q, 2H, 5-CH2), 2.11 (qd, 2H, 4-CH2), 3.80 (s, 3H, 8'-CH3), 4.35 (d, 2H, 7'-CH2), 6.23 (t, 1H, 2-CH), 6.71 (dd, 1H, 3-CH), 6.77 (d, 1H,6'-CH), 6.81 (m, 2H, 2',5'-CH).

[0032] Figure 4 13C NMR spectrum of 6SA (151MHz, CDCl3): δ 13.99 (8-CH3), 22.47 (7-CH2), 27.96 (5-CH2), 31.36 (6-CH2), 32.04 (4-CH2), 43.54 (7'-CH2), 55.96 (8'-CH3), 111.00 (2'-CH), 114.53 (5'-CH), 120.85 (2'-CH), 123.50 (6'-CH), 130.26 (1'-C), 145.11 (s-CH), 145.27 (4'-C), 146.91 (3'-C), 166.05 (1-C).

[0033] Figure 5 Cytotoxicity of 6Sh and 6SA on BV2 cells. (A) Morphology of BV2 cells: BV2 cells were examined under a microscope. (B) Effects of 6Sh and 6SA on BV2 cell viability: BV2 cells were treated with 0, 5, 10, 20, 40, and 80 μM 6Sh or 6SA for 24 hours, and cell viability of BV2 cells (n=5) was examined by MTT assay. Data are expressed as mean ± SD. (n = 3) p<0.05 p<0.01, p<0.001, , p<0.0001.

[0034] Figure 6Flow cytometry analysis of cell cycle and apoptosis. (AB) Cell cycle distribution of BV2 cells: BV2 cells were treated with solvent, 10 μM 6Sh or 6SA for 24 hours, stained with PI, and analyzed by flow cytometry. (CD) Apoptosis assay of BV2 cells: BV2 cells were treated with solvent, 10 μM 6Sh or 6SA for 24 hours, stained with Annexin V-FITC and PI, and detected by flow cytometry. Data are expressed as mean ± SD (n=3). p < 0.05 p < 0.01.

[0035] Figure 7 In vivo safety of 6SA in C57BL / 6N mice. Mice were randomly divided into five groups: control group, CORT (20 mg / kg / day), 6Sh (20 mg / kg / day) + CORT (20 mg / kg / day), 6SA (20 mg / kg / day) + CORT (20 mg / kg / day), and Cita (10 mg / kg / day) + CORT (20 mg / kg / day); (A) H&E staining: major organs were stained with H&E 21 days after drug treatment, scale bar: 200 μm; (B) Measurement of liver and kidney injury markers in mouse serum.

[0036] Figure 8 Pharmacokinetics and brain distribution of 6Sh and 6SA: (A) Plasma concentration-time curves of 6Sh and 6SA; (B) Brain concentration-time curves of 6Sh and 6SA; (C) Pharmacokinetic parameters of 6Sh and 6SA.

[0037] Figure 9 The antidepressant effect of 6SA: (A) Animal experimental timeline, mice were randomly divided into five groups (n=6): control group, CORT (20 mg / kg / day), 6Sh (20 mg / kg / day) + CORT (20 mg / kg / day), 6SA (20 mg / kg / day) + CORT (20 mg / kg / day), and Cita (10 mg / kg / day) + CORT (20 mg / kg / day); (B) Tail suspension test (TST); (C) Forced swimming test (FST); (D) Open field test (OFT); (E) Weight change, data are expressed as mean ± SEM (n=6). p < 0.05; p < 0.01; p < 0.001; , p<0.0001.

[0038] Figure 10Effects of 6SA on CORT-induced neuronal damage, pro-inflammatory cytokines, and serum 5-HT levels. (A) Nissl staining, arrow: black shrunken neurons; (BC) RT-qPCR analysis of IL-1β and TNF-α mRNA levels in mouse PFC; (D) Serum 5-HT levels in mice. Data are expressed as mean ± SD (n=3). p < 0.05; p < 0.01; p < 0.001.

[0039] Figure 11 The effect of 6SA on TRPV1: A) Tail suspension test (TST), mice were randomly divided into four groups (n=5): control group, CORT (20 mg / kg / day), 6SA (20 mg / kg / day) + CORT (20 mg / kg / day), and 6SA (20 mg / kg / day) + CORT (20 mg / kg / day) + CPZ (1 mg / kg / day); (B) Forced swimming test (FST); (CD) Open field test (OFT). Data are expressed as mean ± SEM. p < 0.05; p < 0.01; (E) Detection of intracellular calcium ion fluorescence imaging using fluorescence microscopy; (F) Quantitative detection of fluorescence intensity read by an ELISA reader. Data are expressed as mean ± SD (n=3). p < 0.05; p < 0.01; p < 0.001. Detailed Implementation

[0040] The present invention will be further described in detail below through specific embodiments.

[0041] 6-Shogaol is a potent anti-inflammatory and antioxidant compound derived from ginger root (dried ginger). Its medical application in treating neuroinflammation associated with various neurological disorders is limited by its poor bioavailability and high cost.

[0042] This invention describes a novel 6-shogaol analogue, its preparation method, and a pharmacological evaluation of its antidepressant and anti-neuroinflammatory effects. The chemical structure of the compound is characterized by the incorporation of nitrogen bonds between the aromatic moiety and the unsaturated alkyl carbonyl side chain.

[0043] Specifically, according to the present invention, compounds of formula I or derivatives thereof, pharmaceutically acceptable salts, enantiomers or solvates are provided:

[0044] Among them, R is selected from H and C. 1-7 H 3-15 C6H5, C6H5O, C6H5O2 or C7H8O2.

[0045] According to this invention, the term "derivative" refers to a compound having one or more modifications or substitutions in its parent compound. In this invention, the parent benzene ring structure is retained, and its carbon chain portion is substituted. The "derivative" as used in this invention has properties or activities similar to the parent compound.

[0046] In some embodiments, the compounds of the present invention may comprise mixtures of stereoisomers, single stereoisomers, mixtures of enantiomers, mixtures of multiple stereoisomers, double bond isomers (i.e., geometric E / Z isomers), or diastereomers (e.g., enantiomers (i.e., (+) or (-)) or cis / trans isomers). In some embodiments, the compounds of the present invention, and therefore compounds according to the content of the present invention, comprise the corresponding stereoisomers, i.e., stereoisomerically pure forms (e.g., geometrically pure, enantiomerically pure, or diastereomerically pure) and mixtures of enantiomers and stereoisomers, such as racemates. Typically, the compounds can be provided with any desired chiral purity. In some embodiments, the compounds of the present invention may comprise stereoisomers or mixtures of stereoisomers, including D- and L-configurations of amino acids.

[0047] According to the present invention, mixtures of enantiomers and stereoisomers of a compound can generally be resolved into their constituent enantiomers or stereoisomers by well-known methods, such as chiral gas chromatography, chiral high-performance liquid chromatography, crystallizing the compound into a chiral salt complex, or crystallizing the compound in a chiral solvent. Enantiomers and stereoisomers can also be obtained by well-known asymmetric synthetic methods from stereoisomer or enantiomer-pure intermediates, reagents, and catalysts.

[0048] According to a preferred embodiment of the present invention, the C 1-7 H 3-15 Preferred C 3-7 Alkenyl, more preferably 1-heptenyl.

[0049] According to a preferred embodiment of the present invention, the C6H5O2 is selected from 3,4-dihydroxyphenyl.

[0050] According to a preferred embodiment of the present invention, the C7H8O2 is selected from 3-hydroxy-4-methoxyphenyl.

[0051] According to a preferred embodiment of the present invention, the compound of formula I has the structure shown in formula II:

[0052] In some embodiments, one or more compounds of the present invention, such as those represented by formula (I), may be provided in combination with a carrier (e.g., a pharmaceutically acceptable carrier).

[0053] Preparation of the compound – Compounds of formula (I) may be prepared by any suitable method known to those skilled in the art. Representative methods can be found, for example, in Francis A. Carey et al., Advanced Organic Chemistry: Part B: Reactions and Synthesis (5th edition, 2005).

[0054] In a preferred embodiment, the present invention synthesizes 6SA (formula II compound) by coupling vanillin with trans-2-octenic acid and characterizes it using UPLC-UV / Vis, LC-MS / MS and NMR.

[0055] In some embodiments, the present invention provides a pharmaceutical composition comprising the compounds described herein, such as those represented by formula (I), or pharmaceutically acceptable salts thereof, and pharmaceutically acceptable excipients. In some embodiments, the present invention provides a pharmaceutical composition comprising an effective amount of the compounds disclosed herein, such as those represented by formula (I), preferably a pharmaceutical composition comprising the compound with the structure shown in formula II (6SA).

[0056] This invention uses the shake-flask method to determine the n-octanol / water partition coefficient. The safety of 6SA was evaluated through cytotoxicity assessment in BV2 cells and toxicological assessment in C57BL / 6J mice. Furthermore, the antidepressant and anti-inflammatory effects of 6SA were investigated in a corticosterone (CORT)-induced depression mouse model. Results showed that compared to 6-shogaol, 6SA exhibited better water solubility, improved oral bioavailability, reduced toxicity, and cost-effectiveness. Moreover, 6SA demonstrated more potent antidepressant and anti-inflammatory effects without toxicity in mice. Therefore, 6SA is a safer and more effective antidepressant candidate.

[0057] In some embodiments, the pharmaceutical composition of the present invention can be used to treat pain. "Pain" refers to, but is not limited to, acute pain, chronic pain, inflammatory pain, neuropathic pain, etc. In some embodiments, the pharmaceutical composition can be used to treat anxiety, mood disorders, or depression.

[0058] The compounds described in this invention, such as those of formula (I), and their pharmaceutically acceptable salts, enantiomers, solvates, or derivatives are of practical use because they may have pharmacological activity in animals, including humans. In some embodiments, one or more of the compounds disclosed herein may remain stable in plasma when administered to a subject, such as a human.

[0059] Typically, the compounds described in this invention can be given to a subject in need, or cells or samples can be contacted with a pharmaceutical composition containing a therapeutically effective amount of the compounds described in this invention (such as the compound shown in formula (I)).

[0060] In some embodiments, the compounds of the present invention, such as those shown in formula (I), may be used in combination with any other active agent or pharmaceutical composition, and such combination therapy may help treat pain, anxiety, mood disorders or depression.

[0061] Combinations of compounds described in this invention, or combinations of compounds for treatment as described in this invention, as well as other medications that can be used to treat pain, anxiety, mood disorders, or depression, may be administered alone or in combination. The administration of one medication may precede, coincide with, or follow other medications.

[0062] Combinations of compounds described in this invention, or combinations of compounds for treatment as described herein, may be provided alone or in combination with other compounds in the presence of liposomes, nanoparticles, adjuvants, or any pharmaceutically acceptable carrier, diluent, or excipient, in a form suitable for administration to a subject. If desired, treatment with compounds provided according to this invention may be combined with more conventional and existing therapies for the therapeutic indications described herein. Compounds provided according to this invention may be administered long-term or intermittently. "Long-term" administration means administration in a continuous rather than acute manner to maintain the initial therapeutic effect (activity) over a longer period. "Intermittent" administration means that treatment is not continuous and uninterrupted but periodic. The terms "administered," "administerable," or "administerable" as used herein should be understood as providing the compounds of this invention to a subject requiring treatment.

[0063] "Pharmaceutically acceptable carriers, diluents, or excipients" may include, but is not limited to, any adjuvant, carrier, excipient, gliding agent, sweetener, diluent, preservative, dye / coloring agent, flavor enhancer, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier that has been approved by the U.S. Food and Drug Administration or other government agencies for use in humans or animals.

[0064] The compounds described in this invention can be administered in the form of pharmaceutically acceptable salts. In this case, pharmaceutical compositions according to the invention may comprise salts of such compounds, preferably physiologically acceptable salts, which are known in the art. In some embodiments, the term "pharmaceutically acceptable salt" as used herein refers to the active ingredient comprising a compound of formula I, used in salt form, particularly where the salt form imparts improved pharmacokinetic properties to the active ingredient compared to its free form or other previously disclosed salt forms.

[0065] "Pharmaceutically acceptable salts" can include acid addition salts and base addition salts. "Pharmaceutically acceptable acid addition salts" refer to those salts that retain the bioavailability and properties of the free base, have no adverse effects on biology or otherwise, and can form with inorganic acids (such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, etc.) and organic acids (such as acetic acid, trifluoroacetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc.).

[0066] "Pharmaceutically acceptable base addition salts" refer to salts that retain the bioavailability and properties of the free acid without adverse effects on biology or otherwise. These salts can be prepared by adding an inorganic or organic base to the free acid. Salts derived from inorganic bases may include, but are not limited to, sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, and aluminum salts.

[0067] Preferred inorganic salts may be ammonium, sodium, potassium, calcium, and magnesium salts. Salts derived from organic bases may include, but are not limited to, salts of primary, secondary, and tertiary amines, as well as salts of substituted amines (including naturally occurring substituted amines), cyclic amines, and basic ion exchange resins, such as isopropylamine, trimethylamine, diethylamine, triethylamine, tripropylamine, ethanolamine, 2-dimethylaminoethanol, 2-diethylaminoethanol, dicyclohexylamine, lysine, arginine, histidine, caffeine, procaine, benzyltetramethylammonium, choline, betaine, ethylenediamine, glucosamine, methylglucosamine, theobromine, purine, piperazine, piperidine, N-ethylpiperidine, polyamine resins, etc. Particularly preferred organic bases may be isopropylamine, diethylamine, ethanolamine, trimethylamine, dicyclohexylamine, choline, and caffeine.

[0068] Therefore, the term "pharmaceutically acceptable salt" encompasses all acceptable salts, including but not limited to acetates, lactobionates, benzenesulfonates, laurates, benzoates, malates, bicarbonates, maleates, bisulfates, mandelates, tartrates, methanesulfonates, borates, methyl bromide, bromides, methyl nitrite, calcium EDTA, methyl sulfate, camphorsulfonates, mucilages, carbonates, naphthalenesulfonates, chlorides, nitrates, clavulanates, N-methylglucosamine, citrates, ammonium salts, dihydrochlorides, oleates, and EDTA. Oxalate, diethanesulfonate, bis(hydroxynaphthyl)ate (ambrotate), ester salts, palmitate, ethanesulfonate, pantothenate, fumarate, phosphate / bisphosphate, gluconate, polygalacturonate, gluconate, salicylate, glutamate, stearate, ethylene glycol benzylhydrazine, sulfate, hexylresorcinate, acetite, hydrated ammonium salts, succinate, hydrobromide, tannate, hydrochloride, tartrate, hydroxynaphthyl, theanate, iodide, toluenesulfonate, isothiocyanate, triiodide, lactate, pantothenate, valerate, etc. Pharmaceutically acceptable salts of the compounds described in this invention can be used as dosage forms to modify solubility or hydrolysis properties, or in sustained-release or prodrug formulations. Furthermore, pharmaceutically acceptable salts of the compounds described in this invention may include salts formed from cations such as sodium, potassium, aluminum, calcium, lithium, magnesium, and zinc, as well as salts formed from bases such as ammonia, ethylenediamine, N-methyl-glutamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphenylethylamine, diethylamine, piperazine, tris(hydroxymethyl)aminomethane, and tetramethylammonium hydroxide.

[0069] Pharmaceutical formulations typically contain one or more carriers suitable for the route of administration of the formulation. Suitable carriers may be those known in the art for such routes of administration.

[0070] Suitable pharmaceutical compositions can be formulated using methods known in the art, and their route of administration and dosage are determined by a skilled practitioner. For parenteral administration, the compound can be dissolved in sterile water or saline, or in a pharmaceutically acceptable carrier for the administration of non-water-soluble compounds, such as those for vitamin K. For enteral administration, the compound can be formulated as tablets, capsules, or in liquid form. Tablets or capsules can be enteric-coated or formulated as sustained-release preparations. Many suitable formulations are known, including polymer or protein microparticles, ointments, gels, hydrogels, or solutions encapsulating the compound to be released, which can be used for topical or localized administration. Sustained-release patches or implants can be used to achieve prolonged release. Many techniques known to skilled practitioners are described in Alfonso Gennaro's *Remington: Pharmaceutical Theory and Practice* (20th edition, Williams & Wilkins, 2000). Parenteral formulations may contain excipients, polyalkylene glycols (such as polyethylene glycol), vegetable oils, or hydrogenated naphthalene. The release of the compound can be controlled using biocompatible, biodegradable lactide polymers, lactide / glycolic acid copolymers, or polyoxyethylene-polyoxypropylene copolymers. Other possible parenteral delivery systems for modulating the compound may include ethylene-vinyl acetate copolymer particles, osmotic pumps, implantable infusion systems, and liposomes. Inhaled formulations may contain excipients such as lactose, or may be aqueous solutions containing polyoxyethylene-9-lauryl ether, glycocholate, and deoxycholate, or may be oily solutions administered as nasal drops or gels.

[0071] The compounds or pharmaceutical compositions described in this invention can be administered via oral or non-oral routes, such as intramuscular injection, intraperitoneal injection, intravenous injection, intracerebrospinal injection or infusion, subcutaneous injection, percutaneous or mucosal routes. In some embodiments, the compounds or pharmaceutical compositions according to the present invention can be administered via medical devices or apparatuses, such as implants, grafts, prostheses, stents, etc. Implants designed to contain and release such compounds or compositions can be developed. One example is an implant made of a polymer material that releases the compound over a period of time. The compound can be administered alone or in combination with a pharmaceutically acceptable carrier, for example, in the form of solid dosage forms (such as tablets, capsules, granules, powders, etc.), liquid dosage forms (such as syrups, injections, etc.), injections, drops, suppositories, etc. In some embodiments, the compounds or pharmaceutical compositions according to the present invention can be administered via inhalation sprays, nasal, vaginal, rectal, sublingual, or local routes, and can be formulated alone or in combination into suitable dosage unit formulations containing conventional, non-toxic, pharmaceutically acceptable carriers, adjuvants, and excipients suitable for each route of administration.

[0072] The compounds described in this invention can be used to treat animals, including mice, rats, horses, cattle, sheep, dogs, cats, and monkeys. However, the compounds disclosed in this invention can also be used in other organisms, such as birds (e.g., chickens). One or more compounds disclosed in this invention can also be effectively used in humans. The term "subject," or as otherwise used herein, "patient," refers to an animal, such as a mammal (e.g., a human), as a subject of treatment, observation, or experimentation. However, one or more compounds, methods, and pharmaceutical compositions disclosed in this invention can also be used to treat animals. Therefore, as used herein, "subject" can refer to humans, non-human primates, rats, mice, cattle, horses, pigs, sheep, goats, dogs, cats, etc. Subjects may be suspected of having or at risk of having pain, anxiety, mood disorders, or depression.

[0073] The "effective amount" of the compounds described in this invention may include a therapeutically effective amount or a preventatively effective amount. A "therapeuticly effective amount" refers to an amount sufficient, within the necessary dose and time frame, to achieve the desired therapeutic effect (e.g., treatment of pain, anxiety, mood disorders, or depression, or any condition described herein). Therapeuticly effective amounts of a compound may vary depending on factors such as an individual's disease state, age, sex, weight, and the compound's ability to elicit the desired response in that individual. Dosing regimens may be adjusted to provide optimal therapeutic effect. A therapeutically effective amount also refers to an amount in which any toxic or harmful effects of the compound are offset by its beneficial therapeutic effects. A "preventatively effective amount" refers to an amount sufficient, within the necessary dose and time frame, to achieve the desired preventative effect (e.g., prevention of pain, anxiety, mood disorders, or depression, or any condition described herein). Typically, preventative doses are used in subjects before or in the early stages of disease, and therefore, preventatively effective amounts may be lower than therapeutically effective amounts. Suitable ranges for the therapeutic or preventative effective amount of the compound may be, for example, 0.1 nM-0.1 M, preferably 0.1 nM-0.05 M, more preferably 0.05 nM-15 M, and most preferably any value in the range of 0.01 nM-10 μM.

[0074] In alternative implementations, when treating or preventing conditions that may require treatment for pain, anxiety, mood disorders, or depression, appropriate dose levels are typically from about 0.01 to 500 mg per kilogram of body weight per day, administered once or multiple times. In some implementations, dose levels are from about 0.1 to about 250 mg per kilogram of body weight per day. It should be understood that the specific dose level and frequency of administration may vary for any particular patient and may depend on a variety of factors, including the activity of the particular compound used, the metabolic stability and duration of action of the compound, age, weight, general health condition, sex, diet, route and time of administration, excretion rate, drug combination, severity of the specific condition, and the patient being treated.

[0075] It should be noted that dosage values ​​may vary depending on the severity of the condition to be alleviated. For any given subject, the specific dosing regimen may be adjusted over time based on individual needs and the professional judgment of the person administering or supervising the dosing. The dosage ranges described herein are merely examples and do not limit the dosage ranges that a medical practitioner may choose. The amount of the active compound in the composition may vary depending on factors such as disease state, subject age, sex, and weight. Dosing regimens can be adjusted to provide the best therapeutic response. For example, the dosage may be administered in a single dose, in divided doses over time, or proportionally reduced or increased depending on the urgency of the treatment situation. For ease of administration and uniform dosage, it may be beneficial to formulate the injectable composition into dosage units. Generally, the compounds of the present invention should be used without causing significant toxicity, and as described herein, one or more compounds may exhibit safety characteristics suitable for therapeutic use. The toxicity of the compounds of the present invention can be determined using standard techniques, for example, by testing in cell culture or laboratory animals and determining the therapeutic index. However, in some cases, such as severe disease states, it may be necessary to administer a significantly excessive amount of the composition. In some embodiments, one or more compounds disclosed herein may not be observed to have toxicity at high doses (e.g., 1000 μmol / kg).

[0076] In some embodiments, one or more compounds described herein may exhibit stronger or higher potency than 6-shogaol (6Sh). In some embodiments, one or more compounds described herein may exhibit stronger or higher potency than 6Sh. In some embodiments, one or more compounds disclosed herein may exhibit greater stability compared to 6Sh, such as stability in plasma. In some embodiments, one or more compounds described herein may exhibit greater stability compared to 6Sh.

[0077] In some embodiments, one or more compounds of the present invention may exhibit increased membrane permeability compared to 6Sh. "Enhanced," "increased," or "elevated" means any increase between about 5% and about 90%, or any increase between about 30% and about 60%, or an increase of about 100%, or an increase of about 1, 2, 5, 10, 15, 25, 50, 100, or more times compared to one or more 6Sh compounds. In some embodiments, "enhanced," "increased," or "improved" stability means stability in plasma for at least 5 hours, for example, 5 hours, 10 hours, 15 hours, 24 hours, 37 hours, 48 ​​hours, or longer.

[0078] In some embodiments, one or more compounds of the present invention may exhibit reduced or decreased side effects compared to 6Sh. In some embodiments, one or more compounds of the present invention may exhibit reduced or decreased tolerability, physical dependence, respiratory depression potential, and / or constipation compared to 6Sh. Compared to 6Sh, “reduced” or “decreased” means a reduction of any value between about 5% and about 90%, or a reduction of any value between about 30% and about 60%, or a reduction of about 100%, or a reduction of about 1, 2, 5, 10, 15, 25, 50, 100, or more.

[0079] In some embodiments, one or more compounds described in this invention can be used to treat pain, anxiety, mood disorders, or depression.

[0080] In some embodiments, one or more compounds described in this invention can be used to manufacture medicaments for treating pain, anxiety, mood disorders, or depression.

[0081] In some embodiments, one or more compounds described herein may be used in methods for treating pain, anxiety, mood disorders, or depression, specifically (a) administering the compounds or pharmaceutical compositions described herein to a subject in need of such treatment.

[0082] The term "pain" as used in this article includes, but is not limited to, acute pain, chronic pain, inflammatory pain, neuropathic pain, nociceptive pain, psychogenic pain, visceral pain, thermal pain, musculoskeletal pain, and postoperative pain.

[0083] The term "anxiety" as used in this article refers to an emotion characterized by feelings of tension, worrying thoughts, and physical changes such as elevated blood pressure. Anxiety disorders or "anxiety" include, but are not limited to, panic attacks, generalized anxiety disorder, social anxiety disorder (social phobia), specific phobias, and separation anxiety disorder.

[0084] The term "mood disorder" as used in this article refers to a pervasive emotional state or mood that distorts or contradicts the patient's actual condition and may interfere with their normal functioning. Mood disorders include, but are not limited to, bipolar disorder, seasonal affective disorder (SAD), cyclical mood disorder, premenstrual anxiety disorder, postpartum depression, persistent depressive disorder (dysthymia disorder), disruptive mood regulation disorder, dysthymia disorder, medically related depression, depression caused by substance use or medication, and major depressive disorder.

[0085] In specific embodiments, the compositions of the present invention are used to treat conditions involving cognitive impairment, movement disorders, chronic pain, depression, loss of appetite, addiction, epileptic seizures and convulsions, as well as other conditions.

[0086] In specific embodiments, the compositions of the present invention are provided for treating diseases or conditions. In specific embodiments, the diseases or conditions are mental or psychological illnesses or conditions. In specific embodiments, the diseases or conditions are selected from depression, anxiety, post-traumatic stress disorder, bipolar disorder, substance abuse disorder, eating disorders, obsessive-compulsive disorder, and irritability.

[0087] In some embodiments, one or more compounds of the present invention may be provided in commercial packaging comprising (a) one or more compounds of the present invention; and (b) instructions for using said compounds to treat pain, anxiety, mood disorders, or depression. In some embodiments, the one or more compounds of the present invention provided in commercial packaging may be in the form of pharmaceutical compositions.

[0088] Example Compounds and reagents 6Sh (CAS 555-66-8) was purchased from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China); vanillin (CAS 1196-92-5) and benzotriazol-1-yloxytris(dimethylamino)phosphonium hexafluorophosphate (BOP, CAS 56602-33-6) were purchased from Shanghai Bid Pharmaceutical Technology Co., Ltd. (Shanghai, China); (E)-2-octenic acid (CAS 1871-67-6) was purchased from Shanghai Maclean Biochemical Technology Co., Ltd. (Shanghai, China); methyltrimethoxysilane (MTM, CAS 1185-55-3) was purchased from Thermo Fisher Scientific Co., Ltd. (Waltham, Massachusetts, USA). Capsaicin (CPZ, CAS 138977-28-3) was purchased from Shanghai Aladdin Biochemical Technology Co., Ltd. (Shanghai, China).

[0089] Example 1 Chemical synthesis of 6-shogaol analogue (6SA) Equal amounts of (E)-2-octenic acid and vanillin were mixed with 250 mol% MTM of the total reactants in 1M toluene, and the reaction mixture was refluxed and heated for 24 h. After the reaction was complete, the mixture was cooled to room temperature and concentrated by vacuum rotary evaporation. To remove non-volatile oligosiloxanes, methyl ester byproducts, and unreacted carboxylic acids, the residue was diluted with a 0.3 M THF / NaOH aqueous solution and stirred for 1 h. Then, diethyl ether was added, the aqueous phase was saturated with NaCl, and the layers were separated. To remove unreacted amines, the aqueous layer was extracted with diethyl ether, the organic layers were combined, and washed with HCl (1 M). The aqueous layer was extracted with diethyl ether, the organic extract was dried over anhydrous MgSO4, and evaporated using a rotary evaporator. The residue was dissolved in a small amount of DCM and purified by column chromatography (silica gel, 60A, 40-63 μm, DAVISIL). ®(Grace Company, Cambridge, Massachusetts, USA), with a yield of 61.09%. Ultra-high performance liquid chromatography (UPLC, UltiMate 3000 system with RS diode array detector, Thermo Fisher Scientific, Waltham, Massachusetts, USA), liquid chromatography-mass spectrometry (LC-MS, X500R QTOF system, AB Sciex Pte. Ltd., Singapore), and... 1 H, 13 6SA was characterized by nuclear magnetic resonance (NMR, Bruker Avance III 600 MHz NMR spectrometer, Bruker, Rheinstein, Germany). Figures 2-4 ).

[0090] UPLC-UV / Vis ultra-high performance liquid chromatography combined with ultraviolet-visible light chromatography, λmax: 200.72 nm, 280.49 nm. 1 H NMR (600HZ, CDCl3): δ 0.88 (t, 3H, 8-CH3), 1.25 (m, 4H, 6,7-CH2), 1.39(q, 2H, 5-CH2), 2.11 (qd, 2H, 4-CH2), 3.80 (s, 3H, 8'-CH3), 4.35 (d, 2H, 7'-CH2), 6.23 (t, 1H, 2-CH), 6.71 (dd, 1H, 3-CH), 6.77 (d, 1H, 6'-CH), 6.81 (m,2H, 2',5'-CH). 13 C NMR (600HZ, CDCl3): δ 13.99 (8-CH3), 22.47 (7-CH2), 27.96 (5-CH2), 31.36 (6-CH2), 32.04 (4-CH2), 43.54 (7'-CH2), 55.96 (8'-CH3), 111.00 (2'-CH), 114.53 (5'-CH), 120.85 (2'-CH), 123.50 (6'-CH), 130.26 (1'-C), 145.11(s-CH), 145.27 (4'-C), 146.91 (3'-C), 166.05 (1-C). LC-MS (m / z): 278.1554 [M+H] + 300.1350 [M+Na] + 577.2935[2M+Na] + .

[0091] The above results prove that 6SA is (E)-N-(4-hydroxy-3-methoxybenzyl)oct-2-enamide, with the molecular formula C. 16 H 23 NO3 has a molecular weight of 277.364.

[0092] Example 2 Determination of partition coefficients (n-octanol / water) of 6Sh and 6SA The partition coefficient was evaluated using the shake-flask method according to OECD guideline 107(18). In short, 5 mM 6Sh and 6SA were dissolved in presaturated n-octanol and mixed with presaturated ultrapure water at volume ratios of 1:2.5, 1:5, and 1:10, respectively. The mixture was shaken on a shaker at 25°C for 1 h and then centrifuged at 2000 rpm for 2 min at 25°C to separate the two phases. The partition coefficients were measured at 280 nm using a microplate reader (CLARIOstar). ® The absorbance of the samples was determined by a Plus, BMG LABTECH, Ottenberg, Germany, and quantification was performed using the external standard method.

[0093]

[0094] Example 3: Pharmacological Efficacy Cell culture Mouse microglia BV2 were purchased from the American Culture Collection (Manassas, Virginia, USA). Cells were cultured in DMEM supplemented with 10% FBS and 1% penicillin / streptomycin solution and incubated at 37°C and 5% CO2. For cell experiments, cells were sputtered at a rate of 1.5 × 10⁶ cells / mL. 5 Cells were seeded at a density of 12-well or 96-well plates and treated with different concentrations of the drug for 24 h.

[0095] Laboratory animals and drug administration All animal experiments were approved by the Committee on the Use of Live Animals in Teaching and Research (No. 5906-21) and conducted in accordance with the University of Hong Kong (HKU) Guidelines for the Use of Laboratory Animals. Male C57BL / 6J mice (6-8 weeks old, 18-25 g) were purchased from the Centre for Comparative Medicine, University of Hong Kong. Animals were housed in facilities with free access to food and water and under 12-hour light-dark cycles. To induce depressive-like behavior in mice using corticosterone (CORT), mice were subcutaneously injected with 20 mg / kg of corticosterone for 21 consecutive days, while control mice were subcutaneously injected with an equal volume of the solvent. Intervention and modeling were performed concurrently. Mice in the 6Sh group were orally administered 20 mg / kg of 6Sh daily, mice in the Cita group were orally administered 10 mg / kg of citalopram daily, mice in the 6SA group were orally administered 20 mg / kg of 6SA daily, and mice in the 6SA+CPZ group were orally administered 20 mg / kg of 6SA daily and intraperitoneally injected with 1 mg / kg of CPZ. The control and model mice were given 10 ml / kg of physiological saline containing 1% dimethyl sulfoxide (DMSO) orally for 21 days.

[0096] Behavioral testing At the end of drug treatment, as described above (19), depressive behavior in animals was assessed using the suspended tail test (TST), forced swimming test (FST), and open field test (OFT). In the TST, the mouse's tail was taped to a suspension bar, maintaining a distance of 20-25 cm between the mouse's nose and the ground. The immobility time of each mouse was recorded during the 6-minute test. In the FST, the mouse was placed in a cylindrical container filled with water (30 cm high, 20 cm high) at 22 ± 2°C, with the water level up to 15 cm. After acclimatization for 2 minutes, the immobility time of each mouse was recorded within 4 minutes. In the OFT, under the monitoring of a smart tracking system (SMART V3.0, Panlab, USA), the mouse was placed in an open box (40 × 40 × 40 cm), and the movement paths and times of the mouse in different areas were recorded.

[0097] MTT method The cytotoxicity of 6SA and 6Sh in BV2 cells was detected using the MTT assay. After treatment with 0, 5, 10, 20, 40, or 80 μM 6SA or 6Sh for 24 hours, BV2 cells were incubated with 0.5 mg / ml MTT for 4 hours. Cell viability was calculated by measuring the absorbance (OD) of DMSO crystals at 570 nm using a microplate reader.

[0098] Cell cycle assay Cell cycle stages were detected using a cell cycle analysis kit (Beyotime Biotechnology Co., Ltd., Shanghai, China). 1.5 × 10⁻⁶ cells were used. 5BV2 cells were seeded at 1000 cells / mL in 12-well plates and treated with solvent, 10 μM 6Sh or 10 μM 6SA for 24 hours. Cells were collected and fixed overnight at 4°C with 1 mL of pre-chilled 70% ethanol. After centrifugation at 1000 rpm for 5 min, the cell pellet was stained with propidium iodide (PI) staining solution at 37°C in the dark for 30 min and analyzed using an Agilent NovoCyte Advanteon BVYG analyzer (Agilent Technologies, California, USA).

[0099] Apoptosis assay Cells were analyzed using the Annexin V-FITC apoptosis detection kit (Beyotime Biotechnology Co., Ltd., Shanghai, China). 1.5 × 10⁶ cells were used. 5 BV2 cells were seeded at 100 cells / mL in 12-well plates and treated with solvent, 10 μM 6Sh or 10 μM 6SA for 24 hours. Cells were then collected and stained with Annexin V-FITC and PI staining solution at room temperature in the dark for 10 minutes. Analysis was performed using an Agilent NovoCyte Quanteon flow cytometer (Agilent Technologies, California, USA).

[0100] Hematoxylin-eosin (H&E) staining After drug treatment, mice were euthanized and major organs (heart, liver, spleen, lung, and kidney) were collected, fixed with 4% paraformaldehyde, dehydrated, and embedded in paraffin. Paraffin tissue sections with a thickness of 5 µm were prepared, dewaxed with xylene, and hydrated with ethanol of progressively decreasing concentrations. Cell nuclei were stained with hematoxylin, and the extracellular matrix and cytoplasmic compartments were stained with eosin. The sections were dehydrated and fixed by gradually increasing the concentrations of ethanol and xylene. Tissue micrographs were taken using an optical microscope (Olympus, Tokyo, Japan).

[0101] Serum biomarker assessment Blood was collected after the behavioral test and centrifuged at 4000 rpm for 15 minutes, and the supernatant was collected. Serum levels of serotonin (5-HT), alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and creatinine (CRE) were assessed using commercial kits according to the instructions.

[0102] Pharmacokinetics of 6Sh and 6SA Male C57BL / 6J mice (6–8 weeks old) were orally administered a single dose of 40 mg / kg 6Sh or 6SA. Blood samples were collected at 0, 5, 15, 30, 45, 60, 90, 120, 180, 240, 360, 480, and 720 minutes after administration. Plasma samples were collected after centrifugation at 4000 rpm for 15 minutes and stored at -80°C. The mice were perfused with saline at 0, 5, 15, 30, 45, 60, 90, 120, 180, 240, 360, 480, and 720 minutes after administration, and the brains were collected and stored at -80°C.

[0103] Add 100 μL of plasma sample to nonanoyl vanillin (internal standard, 50 ng / mL, CAS 2444-46-4) and extract with 1 mL of dichloromethane. After vortexing for 5 min, centrifuge the mixture at 12000 rpm / min for 15 min. Then collect the organic layer and dry it in a SpeedVac vacuum concentrator. Dissolve the residue in 100 μL of acetonitrile (ACN) and filter through a 0.22 μm filter. Homogenize the brain sample in saline (1 mL / g) containing 50 ng / mL nonanoyl vanillin. Extract the homogenate with 10 times its volume of dichloromethane. Collect the organic layer and dry it, then dissolve the residue in 100 μL of ACN and filter through a 0.22 μm filter.

[0104] UPLC-MS / MS analysis was performed using a Sciex LC-MS / MS QTOF X500R system and an Agilent Poroshell 120 EC-C18 column (2.1 mm × 100 mm, 2.7 µm). Samples were eluted at a flow rate of 0.4 ml / min with a gradient of water containing 0.1% formic acid (A) and ACN containing 0.1% formic acid (B), as follows: 10%–90% B for 0–6 min, 90% B for 6–8 min, 90%–10% B for 8–9 min, and 10% B for 9–10 min. MS detection parameters were as follows: scan range 50–500 m / z, spray voltage 5000 V (positive ion mode), declustering potential 60 V, collision energy 21 V, and collision energy spread 15 V. Multiple reaction monitoring (MRM) was used for scanning, and retention times (RT) were recorded. The monitored analytes and their specific MRM transitions were: Nonivamide, 294.2→137.1; 6Sh, 277.2→137.1; 6SA, 277.2→137.1. Pharmacokinetic parameters were calculated using Kinetica 3.0 software.

[0105] Nissl stain Following behavioral testing, mice were euthanized, and their brains were collected, fixed with 4% PFA, dehydrated, and embedded in paraffin. The tissue was cut into 5µm thick paraffin sections. Nissl bodies were stained with cresol violet. Photographs were taken using an Olympus microscope.

[0106] Quantitative reverse transcription polymerase chain reaction (qRT-PCR) Total RNA was extracted from brain tissue using TRIzol reagent (Thermo Fisher Scientific, USA) and converted to cDNA using the RevertAid first-strand cDNA synthesis kit (Thermo Fisher Scientific, USA). qRT-PCR was performed using ChamQ Universal SYBR qPCR Master Mix (Novizan, Nanjing, China). Specific primers were provided by QIAGEN (Valencia, California, USA): Il-1β (Mm_ll1b_2_SG; QT01048355), TNF-α (Mm_Tnf_1_SG; QT00104006), and GAPDH (Mm_Gapdh_3_SG; QT01658692).

[0107] Fluorescence assay of intracellular calcium ion levels To determine the effect of 6SA on TRPV1, intracellular calcium ion levels in BV2 cells were analyzed using a fluorescence assay. In short, 1×10⁻⁶ ions were used. 5 BV2 cells were seeded at 1.5 × 10⁶ cells / well in 6-well plates or 1.5 × 10⁶ cells / well. 4 Cells were seeded per well in 96-well plates and stained with 2.5 μM Fluo-4-AM for 30 min in Hanks balanced salt solution (HBSS) at 37 °C. Cells were washed three times with HBSS and incubated for 1 h with 10 μM 6SA / 6Sh / NV (with or without 10 μM CPZ). Fluorescence images were then captured using a Carl Zeiss fluorescence microscope (Germany). Fluorescence intensity was detected using a microplate reader (485 nm / 530 nm).

[0108] Experimental results BV2 cells are more tolerant to 6SA. To assess the cytotoxicity of 6Sh and 6SA, BV2 cells were treated with both drugs for 24 hours, and cell viability was assessed using the standard MTT assay as described previously (20). Figure 5As shown, when the concentration exceeded 10 μM, 6Sh significantly reduced cell viability in a concentration-dependent manner. In contrast, 6SA showed no significant cytotoxicity at concentrations up to 40 μM. Further mechanistic studies revealed that treatment with 10 μM 6Sh for 24 hours significantly increased the proportion of BV2 cells in the G2 / M phase, inducing G2 / M phase arrest, accompanied by an increase in the proportion of apoptotic cells. Figure 6 Conversely, the same concentration of 6SA did not induce cell cycle abnormalities or apoptosis. These results indicate that 6SA is far less toxic to BV2 cells than 6Sh, exhibiting better cell tolerance.

[0109] 6SA has higher safety in mice. To evaluate the in vivo toxicity of 6SA, C57BL / 6J mice were orally administered 20 mg / kg 6SA, 6Sh, or 10 mg / kg citalopram for 21 days, and major tissues (e.g., heart, liver, spleen, lung, kidney) were collected and subjected to H&E staining. Figure 7 As shown, compared with the control group, no significant histopathological changes were observed in the heart, liver, spleen, lungs and kidneys of mice at an oral dose of 20 mg / kg of 6SA, indicating that 6SA did not cause significant in vivo toxicity at this dose.

[0110] 6SA has higher bioavailability Quantitative analysis of drug concentrations in plasma and brain was performed using UPLC-MS / MS. Figure 8 AB shows the concentration-time curves of 6Sh and 6SA in plasma and brain. Over time, the drug concentration of 6SA is higher than that of 6Sh. Figure 8 C proves that 6SA has a longer half-life (T 1 / 2 ) and average length of stay (MRT) 0-t A larger area under the plasma concentration-time curve (AUC) 0-t ) and maximum plasma concentration (C max ).

[0111] 6SA improves behavioral disorders and weight loss in CORT-injured mice To evaluate the antidepressant effect of 6SA, mice were subcutaneously injected with CORT (20 mg / kg / day) and orally administered saline (10 ml / kg / day), 6Sh (20 mg / kg / day), 6SA (20 mg / kg / day), or Cita (10 mg / kg / day) for 21 days. Figure 9 As shown in Figure A. The severity of depression in mice was assessed using TST, FST, and OFT (21, 22). Results are as follows. Figure 9As shown in BD, 6Sh, 6SA, and Cita can reduce immobility time in FST and TST while increasing movement distance in OFT. Notably, 6SA more effectively improved CORT-induced depressive-like behavior in mice compared to 6Sh. Furthermore, 6SA and Cita significantly alleviated CORT-induced weight loss, while 6Sh had no such effect. Figure 9 E).

[0112] 6SA reduces CORT-induced neuronal damage in mice. Assess CORT-induced neuronal damage using Nissl staining. Figure 10 As shown in Figure A, compared with the control group, the number of black shrunken neurons was significantly increased in the CORT group, and interventions with 6Sh, 6SA, and Cita could alleviate CORT-induced damage. Notably, 6SA showed the best neuroprotective effect in reducing the number of damaged neurons caused by CORT.

[0113] 6SA reduces neuroinflammation in the prefrontal cortex (PFC) of mice. To verify the anti-inflammatory effect of 6SA, PFCs were collected from mice after drug treatment, and the mRNA levels of IL-1β and TNF-α were measured. Figure 10 As shown in BC, 6SA, 6Sh, and Cita effectively reversed the CORT-induced upregulation of pro-inflammatory cytokines in mouse PFC. These results confirm that 6SA and 6Sh have similar anti-inflammatory effects.

[0114] 6SA upregulates serotonin (5-HT) levels. To assess serum 5-HT levels, blood samples were collected from each group of mice, and the concentration of 5-HT in mouse serum was measured using a commercial 5-HT assay kit. Figure 10 As shown in Figure D, CORT significantly reduces serum 5-HT levels, while treatment with 6SA or 6Sh (20 mg / kg) can effectively reverse the CORT-induced decrease in 5-HT levels, restoring them to near-normal concentrations.

[0115] 6SA exhibits antidepressant effects through TRPV1 activation. To investigate whether the mechanism of action of 6SA is related to the TRPV1 channel, the TRPV1 antagonist CPZ was used in combination with 6SA in a CORT depression model. Figure 11 As shown in AD, the therapeutic effect of 6SA on a CORT-induced depressed mouse model could be reversed by CPZ. Further analysis using calcium ion imaging assays revealed TRPV1 activation, with NV serving as a positive control. Figure 11EF showed that 10 μM of 6SA, 6Sh, and NV significantly increased intracellular calcium ion levels. Notably, this effect was completely inhibited by the TRPV1 antagonist CPZ, suggesting that 6SA and 6Sh may exert their antidepressant effects by directly activating TRPV1.

[0116] in conclusion The increasing prevalence of depression and the side effects of existing antidepressants necessitate new, safe, and effective treatments for depression (23). While natural products have the potential to treat depression, they generally suffer from poor water solubility and low oral bioavailability (20, 24). 6Sh was selected as the lead compound for this study due to its significant potential in treating obesity and mental disorders (15, 25-27). We chemically modified the structure of 6Sh to improve its water solubility, oral bioavailability, and cost-effectiveness (28, 29). Therefore, a hydrophilic amide-containing 6SA was synthesized and evaluated as a safe and effective antidepressant candidate.

[0117] The α,β-unsaturated carbonyl group in 6Sh endows it with unique physical, chemical, and biological properties (29). 6SA retains this α,β-unsaturated carbonyl group and introduces nitrogen bonds between the aromatic ring moiety and the α,β-unsaturated alkyl carbonyl side chain. The 6SA of this invention is synthesized by direct coupling of (E)-2-octenic acid with vanillin, and its structure was confirmed by UPLC-UV / Vis, NMR, and LC-MS / MS. The synthesis cost can be reduced by 99% using two low-cost raw materials. Compared to 6Sh, 6SA exhibits better water solubility, higher bioavailability, and better brain tissue distribution. As shown in Table 1, the partition coefficient of 6SA (log P, 1.63 ± 0.21) is lower than that of 6Sh (log P, 2.33 ± 0.03), indicating better hydrophilicity. According to Ribinski's five rules, the ideal log P value for central nervous system drugs is approximately 2, while the log P value for orally and intestinally absorbed drugs should be between 1.35 and 1.8 (31, 32). Therefore, the structural optimization of 6Sh in this invention meets the requirements for drug development. Studies have reported that 6Sh exhibits strong cytotoxicity in various cell lines, such as KB cells (ICV). 50 =7.4±2.2μM) and HL60 cells (IC 50 =7.9±2.0μM)(30). This invention evaluated the in vitro toxicity of 6Sh and 6SA in BV2 microglia and their systemic toxicity in mice. Results showed that 6SA at concentrations up to 40µM had no toxic effect in BV2 cells, while 6Sh exhibited cytotoxicity at 5µM. Similarly, continuous oral administration of 6SA at a dose of 20 mg / kg / day for 21 days did not cause significant toxicity in C57BL / 6J mice ( ). Figure 7The above results fully demonstrate that 6SA has good safety profile both in vitro and in vivo.

[0118] Table 1. Determination of partition coefficients (n-octanol / water) of 6Sh and 6SA

[0119] Furthermore, CORT, as a glucocorticoid, can induce neuroinflammation by mediating chronic stress-related cellular signaling, thereby inducing depressive-like behavior in mice (33,34). This invention demonstrates that 6SA can effectively alleviate CORT-induced depressive behavior in mice. Further mechanistic studies revealed that CORT upregulated the expression of pro-inflammatory cytokines IL-1β and TNF-α in mouse PFC, while both 6SA and 6Sh effectively inhibited the increase of these inflammatory factors ( ). Figure 10 The above results indicate that 6SA has the potential to treat depression through anti-inflammatory effects.

[0120] Previous reports have indicated that 6Sh can act as a TRPV1 agonist to induce Ca²⁺. + Influx (34). To investigate whether 6SA has the same effect on TRPV1 as 6Sh, and whether the activation of TRPV1 is involved in its antidepressant effect, behavioral tests and cellular calcium ion imaging experiments were conducted. Figure 11 As shown in AD, the TRPV1 antagonist CPZ reversed the antidepressant effect of 6SA. Meanwhile, Figure 11 Fluorescence microscopy images and fluorescence intensity measurements of EF cells stained with calcium ions indicate that both 6SA and 6Sh can increase intracellular calcium levels. 2+ The levels were elevated, and CPZ could block this effect. In summary, these findings suggest that TRPV1 activation may be a key mechanism by which 6SA exerts its antidepressant effect.

[0121] This invention develops a novel analogue of 6Sh, 6SA, which is significantly superior to the lead compound in terms of druggability and cost-effectiveness. Comprehensive evaluation indicates that the new product, 6SA, is a safer and more effective anti-inflammatory and antidepressant drug. Compared to 6Sh, 6SA exhibits lower toxicity in both in vitro and in vivo models, while maintaining comparable or even superior antidepressant and neuroprotective effects in a CORT-induced mouse depression model. Its therapeutic mechanism may be related to TRPV1 activation and anti-inflammation. In conclusion, 6SA is an antidepressant candidate drug with good safety, efficacy, and development potential.

[0122] The description of the specific embodiments above will fully reveal the general nature of the invention, enabling others to easily modify and / or adapt these specific embodiments for various applications without departing from the overall concept of the invention, by utilizing technical knowledge in related fields (including those cited herein and incorporated by reference) without extensive experimentation. Therefore, based on the teachings and guidance provided herein, these adaptations and modifications should be considered within the equivalent meaning and scope of the disclosed embodiments. It should be understood that the wording or terminology in this document is for descriptive purposes only and not for limitation; therefore, the terminology or terminology in this specification should be interpreted by those skilled in the art based on the teachings and guidance provided herein and in conjunction with the knowledge of those skilled in the art.

[0123] While various embodiments of the present invention have been described above, it should be understood that these embodiments are merely examples and not limitations. Those skilled in the art will understand that various modifications in form and detail can be made without departing from the spirit and scope of the invention. Therefore, the present invention should not be limited to any of the exemplary embodiments described above, but should be defined solely by the following claims and their equivalents. All references cited in this article are incorporated herein in their entirety, and their scope and effect are the same as if each individual publication, patent, or patent application were explicitly and separately indicated to be incorporated herein in its entirety for all purposes.

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Claims

1. A compound of Formula I or a derivative thereof, a pharmaceutically acceptable salt, an enantiomer, or a solvate: in, R is selected from H and C. 1-7 H 3-15 C6H5, C6H5O, C6H5O2 or C7H8O2.

2. The compound according to claim 1, characterized in that, The C 1-7 H 3-15 Selected from C 3-7 Alkenyl, preferably 1-heptenyl; And / or, the C6H5O2 is selected from 3,4-dihydroxyphenyl; And / or, the C7H8O2 is selected from 3-hydroxy-4-methoxyphenyl.

3. The compound according to claim 1, characterized in that, The compound shown in Formula I has the structure shown in Formula II: 。 4. A pharmaceutical composition comprising the compound of any one of claims 1-3 and a pharmaceutically acceptable carrier.

5. The pharmaceutical composition according to claim 4, characterized in that, The pharmaceutically acceptable carrier is selected from the group consisting of: liquid carriers; isotonic agents; wetting agents, solubilizers or emulsifiers; preservatives; buffers; acidifiers; antioxidants; alkalizing agents; carriers; chelating agents; colorants; complexing agents; solvents; suspending agents and / or thickeners; oils; penetration enhancers; polymers; hardeners; thickeners; a protein; carbohydrates; fillers; and lubricants.

6. The pharmaceutical composition according to claim 4, characterized in that, In addition to (a) the compound of claim 4 or 5 and a pharmaceutically acceptable carrier, it also includes (b) an adjunct medicament for treating a mental or psychological illness or condition.

7. The pharmaceutical composition according to any one of claims 4-6, formulated for administration via routes selected from: oral administration, intravenous administration, intraperitoneal administration, and intramuscular administration.

8. The use of the compound according to any one of claims 1-3 in the preparation of a medicament for treating a subject’s mental or psychological illness or condition; preferably, the mental or psychological illness or condition is selected from mood disorders, depression, anxiety, pain, post-traumatic stress disorder, bipolar disorder, substance abuse disorder, eating disorder, obsessive-compulsive disorder, and irritability.

9. The application according to claim 8, wherein, The pain is acute pain, chronic pain, inflammatory pain, or neuropathic pain.

10. The application according to claim 8, wherein, The subjects were humans.

11. The application according to claim 8, wherein, The drug comprises an effective amount of the compound according to any one of claims 1-3.

12. The application according to claim 11, wherein, The medication also includes a therapeutically effective amount of an additional agent for the treatment of depression, the agent being selected from selective serotonin reuptake inhibitors, serotonin-norepinephrine reuptake inhibitors, serotonin modulators, serotonin antagonists and reuptake inhibitors, norepinephrine reuptake inhibitors, tricyclic antidepressants, tetracyclic antidepressants, monoamine oxidase inhibitors, atypical antidepressants, and antidepressants acting through one or more other mechanisms.

13. A commercial package comprising the compound of any one of claims 1-3 and instructions for use in treating pain, anxiety, mood disorders, or depression.

14. A method for preparing a compound of formula II, comprising the following steps: 。

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