Pharmaceutical composition for treating and / or preventing depression as well as preparation method and application thereof

Abstract

The invention discloses a pharmaceutical composition for treating and / or preventing depression, and belongs to the technical field of biological medicines. The technical problem to be solved is to significantly improve depression-like behaviors and protect nerve cells, and has an obvious effect on treatment and / or prevention of depression. According to the key point of the technical scheme, the pharmaceutical composition for treating and / or preventing depression comprises a secoiridoid glycoside compound, a xanthone glycoside compound and a beta-amyrin type pentacyclic triterpenoid compound in a mass ratio of 1: (0.25-6): (0.1-5), and one or more medically acceptable auxiliary materials.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and mainly to a pharmaceutical composition for treating and / or preventing depression, its preparation method, and its application. Background Technology

[0002] Depression, the fourth leading cause of disability worldwide, continues to grow rapidly. Currently, the most common treatments for depression are medication and psychotherapy. Medication mainly involves chemical drugs, which have significant side effects. Traditional Chinese medicine, guided by the principles of syndrome differentiation and holistic treatment, is gaining acceptance from doctors and patients due to its characteristics of targeting multiple points, acting through multiple pathways, regulating the whole body, and having fewer adverse reactions.

[0003] Sesocytic ether glycosides are a class of natural organic compounds with specific cyclic structures and glycosyl modifications. They belong to an important branch of iridoids and are named for the cleavage of cyclic ether bonds in their core nucleus. Their core is based on the "cleavage" characteristic of the iridoid skeleton. Gentiopiroside is an iridoid compound mainly found in plants of the Gentianaceae family (such as Gentiana and Swertia), possessing broad pharmacological activities and rich application value. Recent studies have shown that gentiopicroside can exert antidepressant activity through mechanisms such as regulating neurotransmitter levels (such as serotonin and norepinephrine), improving neuroplasticity, and inhibiting neuroinflammation, making it a potential candidate natural antidepressant. It also has sedative and analgesic effects, acting on the central nervous system to provide sedation, anti-anxiety, and analgesia, and can help relieve symptoms such as neuralgia and insomnia.

[0004] Xanthones are a class of natural organic compounds possessing both specific aromatic nuclei and glycosyl modifications. They belong to an important branch of the xanthones and are named for the simultaneous presence of an xanthone nucleus and a glycosyl group in their molecules. Widely found in higher plants, they are core active ingredients in many medicinal plants. Mangiferin, in particular, is mainly found in plants of the *Mangifera* genus (such as the leaves, fruit, and bark of mangoes) and other plants (such as *Anemarrhena asphodeloides* and *Curculigo orchioides*). Mangiferin, a type of compound (flavonoid derivative), possesses rich pharmacological activity and broad application potential. Studies have shown that mangiferin can exert potential antidepressant and anti-anxiety effects by regulating neurotransmitter levels (such as serotonin and dopamine) in the brain, improving neuroplasticity, and inhibiting neuroinflammation and oxidative stress.

[0005] β-amyrin-type pentacyclic triterpenoids are an important subclass of triterpenoids with well-defined structural characteristics and wide distribution. They belong to one of the core types of pentacyclic triterpenoids and are named for their β-amyrin-based carbon skeleton. They hold significant research value in phytochemistry and medicinal chemistry. Oleanolic acid, a triterpenoid widely found in plants, possesses diverse pharmacological activities and applications. Its antidepressant activity may be related to its multi-target regulatory effects, enhancing the function of monoamine neurotransmitters: deficiency or impaired function of monoamine neurotransmitters (such as serotonin, norepinephrine, and dopamine) is closely related to depression. Studies have found that oleanolic acid may exert its antidepressant effect by inhibiting the activity of monoamine oxidase (MAO, the enzyme responsible for degrading monoamine neurotransmitters), reducing the breakdown of serotonin and norepinephrine, thereby increasing the level of monoamine neurotransmitters in the brain.

[0006] Publication No. CN1562054A, published on January 12, 2005, discloses a novel use of oleanolic acid in anti-anxiety and antidepressant drugs. It is characterized by low toxicity and avoids the sedative, ataxia, and epileptiform induction effects common in anti-anxiety and antidepressant drugs. The novel use of oleanolic acid in anti-anxiety and antidepressant drugs allows for its application in the preparation of these drugs. The use of oleanolic acid and its pharmaceutically acceptable salts, esters, or glycosides in the preparation of drugs for the prevention or treatment of anxiety or depression is also discussed. Pharmaceutical compositions containing oleanolic acid and its pharmaceutically acceptable salts, esters, or glycosides in the preparation of drugs for the prevention or treatment of anxiety or depression are also included, including combinations with pharmaceutical carriers.

[0007] Publication No. CN101099770A, published on February 9, 2008, discloses a pharmaceutical composition with imidazoline receptor agonist activity, as well as its preparation method and application. The pharmaceutical composition refers to extracts of Prunella vulgaris, Gentiana scabra, Leonurus japonicus, Paeonia lactiflora, and Glycyrrhiza uralensis, wherein the active ingredients include (by weight) 5-100 parts oleanolic acid, 2-50 parts ursolic acid, 0.01-80 parts Prunella vulgaris saponin A, and Prunella vulgaris saponin A. The composition contains 0.01-200 parts of calciferine B, 0.01-150 parts of gentiopicrin, 0.01-300 parts of gentianine, 0.01-230 parts of stachydrine, 0.01-170 parts of quercetin, 0.01-290 parts of paeoniflorin, 0.01-450 parts of glycyrrhizic acid, and 0.01-400 parts of glycyrrhizin flavonoids. This composition can be used to prepare drugs for treating congestive heart failure, antiarrhythmic drugs, antidementia drugs, antidepressants, and antihypertensive drugs.

[0008] Publication No. CN103191141A, published on October 22, 2014, discloses a novel use of 3-methoxymangiferin in the pharmaceutical field, as well as pharmaceutical compositions containing 3-methoxymangiferin. 3-methoxymangiferin can promote the gene and protein expression of serotonin 1A receptors in hippocampal dentate gyrus neurons, mediating the neurotrophic effects of serotonin, such as hippocampal remodeling, thereby significantly improving post-stroke hippocampal neuroregeneration impairment. It can also selectively inhibit the reuptake of 5-HT by the presynaptic membrane, reduce the expression of 5-HT-positive cells in the hypothalamus, and alleviate 5-HT depletion and the resulting anxiety and depressive symptoms. Therefore, it can be used to prepare drugs for the prevention or treatment of post-stroke depression.

[0009] Publication No. CN112494595A, published on March 16, 2021, discloses a traditional Chinese medicine composition, its preparation method, and its application. The optimal conditions for preparing the composition using a water extraction and alcohol precipitation method are: 10 times the amount of water added, two extractions, an extraction time of 2 hours, and an alcohol precipitation concentration of 60%. The oral liquid prepared using this method, with an optimal mass ratio of Anemarrhena asphodeloides to Lilium brownii of 1:2, contains a high content of the active ingredient mangiferin and exhibits antidepressant effects, making it suitable for the preparation of antidepressant drugs.

[0010] The journal *Drug Evaluation Research*, with the article titled "Research Progress on the Neuropsychopharmacological Effects of Oleanolic Acid and Ursolic Acid," Volume 38, published in October 2025, reports that oleanolic acid and ursolic acid belong to the pentacyclic triterpenoid acid class of compounds and are isomers with almost identical pharmacological effects. Both possess antioxidant, anti-inflammatory, immunomodulatory, antitumor, antimicrobial, hepatoprotective, antidiabetic, lipid-regulating, weight-loss, and anti-atherosclerotic effects. This review summarizes the neuropsychopharmacological effects of oleanolic acid and ursolic acid, including protecting nerve cells, analgesia, anti-anxiety, anti-schizophrenia, antidepressant, and improving learning and memory, providing a basis for the research and development of oleanolic acid and ursolic acid in the prevention and treatment of Alzheimer's disease, Parkinson's disease, and depression.

[0011] The prior art represented by the aforementioned literature has at least the following unresolved technical problems or defects: (1) The composition is complex, the controllability is poor, and the cost is high. The relevant evidence is the publication number CN101099770A. The composition described in this document contains active ingredients from 11 kinds of plant extracts. Therefore, it is necessary to extract from multiple plants and separate and purify multiple components. The quality control during the production process is difficult and the cost is relatively high.

[0012] (2) The antidepressant is not targeted enough. The relevant evidence is the publication number CN101099770A. The literature mentions that the prepared drug composition can be used for antidepressant, but it is based on the imidazoline receptor agonist activity. Its antidepressant mechanism may be relatively broad. Moreover, the literature does not test the antidepressant effect of its drug composition. The antidepressant mechanism may be less targeted and not very convincing.

[0013] (3) There may be side effects between the drugs. The relevant evidence is CN101099770A. The drug composition in this literature is complex. Some of the components may produce unnecessary side effects, or the interaction between different components may increase the risk of adverse reactions. The synergistic effect between multiple components is more complex. Some components may interfere with each other, or they may not form a good synergistic effect in antidepressant, making it difficult to achieve the best antidepressant effect. Summary of the Invention

[0014] The purpose of this invention is to address the deficiencies in the prior art by providing a pharmaceutical composition with synergistic effects for the treatment and / or prevention of depression, which has excellent efficacy in both the prevention and / or treatment of depression.

[0015] Terminology Explanation: Unless otherwise defined, all technical terms in this document have the same meanings as commonly understood by one of ordinary skill in the art to which the subject matter of the claims pertains. Unless otherwise stated, all patents, patent inventions, and publications cited in this document are incorporated herein by reference in their entirety. If multiple definitions exist for terms in this document, the definitions in this chapter shall prevail.

[0016] It should be understood that the above brief description and the following detailed description are exemplary and for illustrative purposes only, and do not limit the subject matter of the invention in any way. In this invention, the singular is used in conjunction with the plural unless otherwise specifically stated. It should also be noted that, unless otherwise stated, the use of “or” or “or” means “and / or”. Furthermore, the use of the term “comprising” and other forms such as “including,” “containing,” and “contains” are not limiting.

[0017] Definitions of standard chemical terms can be found in the references Chinese Pharmacopoeia 2025 edition and Complete Explanation of Pharmacological Terminology.

[0018] Unless otherwise stated, conventional methods within the scope of the art, such as forced swimming test, tail suspension test, conditioned position preference test, neurotransmitter detection method, inflammatory factor detection method, etc., shall be used.

[0019] Unless specifically defined herein, the use of all commercially available products herein employs standard techniques. For example, it may be carried out using the manufacturer's instructions for use with the kit, or in accordance with methods known in the art or the description of this invention. The techniques and methods described herein can generally be implemented according to conventional methods well known in the art, based on the descriptions in the various summary and more specific documents cited and discussed in this specification.

[0020] The term "secoiridoid glycosides" as used in this article refers to a class of natural organic compounds with specific cyclic structures and glycosyl modifications. They belong to an important branch of iridoid glycosides. The structure of secoiridoid glycosides can be broken down into two parts: a "secoiridoid nucleus" and a "glycosyl group". The two parts are connected by glycosidic bonds. The nucleus is based on the "secoiridoid" feature of the iridoid skeleton.

[0021] The term "oxanthonone glycosides" used in this article refers to a class of natural organic compounds with specific aromatic cores and glycosyl modifications. They belong to an important branch of the xanthonone class and are named for the simultaneous presence of an "oxanthonone core" and a "glycosyl group" in their molecules. They are widely found in higher plants and are the core active ingredients of many medicinal plants. The structure of xanthonone glycosides can be clearly separated into two parts: an "oxanthonone aglycone (core)" and a "glycosyl group," which are linked by glycosidic bonds. They play an important role in the research of natural product chemistry and pharmacology.

[0022] The term "β-amyrane-type pentacyclic triterpenoids" used in this article refers to an important subclass of triterpenoids with well-defined structural characteristics and wide distribution. It belongs to one of the core types of pentacyclic triterpenoids and is named for its basic carbon skeleton, β-amyrane (C30H). 52 Tetracyclic triterpenoids are a class of secondary metabolites widely found in plants. They have a parent nucleus structure containing five six-membered rings (A / B / C / D / E rings, where A / B / C / D rings are fused in a chair conformation, and D / E rings are fused in a trans conformation). These rings often undergo substitution of functional groups such as hydroxyl, carboxyl, ester, and ketone groups at specific positions (e.g., C-3, C-12, C-28). Some structures may also contain double bonds (e.g., a double bond at C-12). They are important research products in phytochemistry and medicinal chemistry.

[0023] The term "gentiopicroside" as used in this article refers to a naturally occurring iridoid glycoside compound, primarily derived from the dried roots and rhizomes of plants in the Gentianaceae family (such as Gentiana, Gentiana macrophylla, and Angelica dahurica), and is one of the hallmark active components of the Gentiana genus. Its chemical structure is based on an iridoid nucleus, bound to glucose via a glycosidic bond, exhibiting clear pharmacological activity and broad research value. Its molecular formula is C1. 16 H 20O9, with a molecular weight of 356.32, has a chemical structure containing an iridoid skeleton, an ester group, and a glucose residue. It is characterized by its high polarity and easy solubility in polar solvents such as water and methanol.

[0024] The term "mangiferin" as used in this article refers to a naturally occurring tetrahydroxypyranocoumarin compound, belonging to the phenolic acid derivative class. It is primarily derived from the leaves, fruits, bark, and roots of plants in the genus *Mangifera* (family Anacardiaceae), such as mango, and is also found in various other plants in the families Liliaceae and Gentianaceae. It has a unique chemical structure with the molecular formula C2. 19 H 18 O 11 With a molecular weight of 422.34, its chemical structure uses pyranocoumarin as the parent nucleus, linking multiple hydroxyl and glucuronic acid residues. It is characterized by strong polarity and easy solubility in polar solvents such as hot water, methanol, and ethanol, but poor solubility in non-polar solvents (such as ether and chloroform). It has a wide range of pharmacological activities and development value, and is one of the research hotspots in the field of natural product chemistry.

[0025] The term "oleanolic acid" as used in this article refers to a pentacyclic triterpenoid compound widely found in plants, named after its initial isolation from the leaves of the olive tree (Olea europaea). It is one of the main active ingredients in many traditional Chinese medicines, with the molecular formula C2. 30 H 48 O3, with a molecular weight of 456.71, has a chemical structure with a pentacyclic triterpenoid as the parent nucleus (oleanane-type skeleton). The molecule contains 3 hydroxyl groups (one of which is a carboxyl group). It is characterized by strong lipid solubility and is easily soluble in organic solvents such as methanol, ethanol, chloroform, and ether, but poorly soluble in water. It has diverse pharmacological activities and important medicinal value, and has attracted much attention in natural product chemistry, pharmacology, and clinical research.

[0026] The term "Tibetan Artemisia capillaris" used in this article refers to a general term for medicinal plants characteristic of the Qinghai-Tibet Plateau, not a single species. According to records such as *Chinese Tibetan Medicine* and *Illustrated Handbook of Tibetan Plateau Medicinal Herbs*, it mainly comes from various plants in the genera *Gentiana* and *Swertia* of the Gentianaceae family. Common species include: *Swertia szechuanensis*, *Swertia claspingii*, *Swertia rubra*, *Swertia serratifolia*, and *Gentiana linearis*. These plants mostly grow in alpine meadows, riverbanks, and gravelly slopes at altitudes of 2000-5000 meters, exhibiting strong adaptability and serving as an important resource for traditional Tibetan medicine. It has a long history of application in Tibetan medicine and is one of the core medicinal materials for treating liver and gallbladder diseases. Due to its growth in the cold, oxygen-deficient environment of high altitudes, its unique medicinal effects have earned it the reputation of being the "guardian of the liver and gallbladder" among the Tibetan people. Modern research has also confirmed its rich medicinal value.

[0027] The term "depressive-like behavior" used in this article refers to a common term in psychology, neuroscience, and pharmacology research, referring to behavioral patterns or physiological responses exhibited by animals in experimental settings that are similar to the core symptoms of human depression. It is not a "disease diagnosis" in the strict sense, but rather a behavioral indicator observed through standardized experimental methods, used to simulate the pathological mechanisms of depression, screen potential therapeutic drugs, or explore the causes of the disease.

[0028] The term "depression" as used in this article refers to a common mental disorder characterized by persistent low mood and loss of interest, accompanied by significant impairment in cognitive, physical, and social functions, and in severe cases, suicidal ideation or behavior. It is not simply a matter of "bad mood," but a complex illness caused by the combined effects of biological, psychological, and social factors. It is characterized by high prevalence, high relapse rate, and high disabling potential, and is listed by the World Health Organization (WHO) as one of the leading causes of disability globally.

[0029] The term "chemical drug" used in this article refers to drugs with a defined chemical structure that are prepared through chemical synthesis or semi-synthesis, or extracted and purified from natural products. It is one of the core components of the modern drug system and, together with "biological drugs" and "traditional Chinese medicine / natural drugs," constitutes the three major categories of clinical drugs.

[0030] The term "target" as used in this article refers to the biomolecule or structure that a drug or other bioactive molecule specifically binds to or regulates when exerting its effects in the body; it is the core bridge connecting drugs and disease treatment. Essentially, a target is a key biomolecule or physiological structure in the body related to the occurrence and development of a disease. Abnormal function of a target (such as overactivation, abnormal expression, or structural mutation) is often the core mechanism of the disease. Drugs correct the abnormal function of targets through specific interactions (such as binding, inhibition, and activation), thereby achieving the goal of treating the disease. In modern drug development, the discovery and validation of targets is the starting point for drug design, directly determining the efficacy, selectivity, and safety of the drug.

[0031] The term "HT22 cells (mouse hippocampal neurons)" used in this article refers to an immortalized neuronal cell line derived from mouse hippocampal tissue. These cells originate from the mouse hippocampus (a key brain region responsible for learning, memory, and emotion regulation) and, after in vitro culture and immortalization, can be passaged in the laboratory for extended periods. Due to their neuron-like characteristics, culture stability, and functional suitability, HT22 cells have become an important bridge connecting in vitro experiments with in vivo physiological mechanisms in neuroscience research. Their widespread application in neurodegenerative diseases, neuronal damage protection, and drug screening provides crucial support for elucidating the mechanisms of neurological diseases and developing novel therapeutic strategies.

[0032] The term "DMEM medium" used in this article refers to Dulbecco's Modified Eagle Medium. DMEM medium is a synthetic medium widely used in cell biology research and biopharmaceutical fields. It was improved by American scientist Renato Dulbecco based on the classic Eagle Medium (MEM). By optimizing the concentration of nutrients (especially glucose, amino acids, and vitamins), it provides an ideal growth environment for the in vitro culture of various mammalian cells and is one of the most commonly used mediums for cell culture.

[0033] The term "fetal bovine serum" as used in this article refers to a natural biological product isolated and purified from the blood of fetal bovine cells (unborn bovine fetuses). Rich in various nutrients and bioactive substances, it is one of the most commonly used additives in in vitro cell culture. It provides the critical nutrients and signaling support required for the growth, proliferation, and survival of the vast majority of mammalian cells, making it an indispensable core reagent in cell biology, biopharmaceutical, and medical research.

[0034] The term "dual antibiotic" used in this article refers to penicillin-streptomycin solution, one of the most commonly used antibiotic additives in cell culture. By combining the antibacterial spectra of two antibiotics, it effectively prevents bacterial contamination and maintains a sterile environment in the cell culture system, making it a key reagent for ensuring cell survival and experimental stability in cell experiments.

[0035] The term "glutamate" used in this article refers to a non-essential amino acid widely found in nature, and also an important neurotransmitter and metabolic intermediate in living organisms. Glutamate's chemical name is 2-aminoglutaric acid, with the molecular formula C5H9NO4 and a molecular weight of 147.13 g / mol. It is a white crystalline powder, readily soluble in water, slightly soluble in ethanol, and insoluble in ether. Glutamate plays a dual role in living organisms: nutritional metabolism and neurotransmission, both of which work together to maintain normal physiological activities. In-depth research on glutamate not only helps in understanding the mechanisms of life activities but also provides important evidence for the treatment of related diseases and the development of health products.

[0036] The term "oxidative stress injury" used in this article refers to the pathological process in which the body or cells experience an imbalance between the oxidative and antioxidant systems, leading to an excessive accumulation of oxidative substances such as reactive oxygen species (ROS), resulting in damage to the structure and function of biomolecules (such as nucleic acids, proteins, and lipids), and consequently causing cellular metabolic disorders, tissue damage, and even disease. It is widely present in various physiological and pathological processes, including aging, inflammation, neurodegenerative diseases, and cardiovascular diseases.

[0037] The term "CCK8 method" used in this article refers to a cell viability assay based on water-soluble tetrazolium salt. The core component of the CCK-8 kit is water-soluble tetrazolium salt (WST-8), chemically named 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfonic acid benzene)-2H-tetrazole monosodium salt. Its core principle is that WST-8 is reduced to an orange-yellow formazan product by intracellular dehydrogenases (such as succinate dehydrogenase). The amount of formazan produced is positively correlated with cell viability: the more viable cells and the stronger their metabolic activity, the more formazan is produced; dead cells or metabolically inhibited cells cannot participate in the reaction, resulting in reduced formazan production. By measuring the absorbance (OD value) of formazan at a wavelength of approximately 450 nm using a microplate reader, the number or activity of viable cells can be indirectly reflected.

[0038] The term "OD value" used in this article refers to optical density, a quantitative indicator that measures the ability of a material to absorb light of a specific wavelength. The OD value represents the degree to which light is absorbed when it passes through a sample, and the calculation formula is: OD = log 10 (I0 / I), where I0 is the incident light intensity and I is the transmitted light intensity. A higher OD value indicates stronger absorption of light at that wavelength by the substance, and more significant attenuation of transmitted light. It measures the degree of attenuation of light after passing through the sample using a spectrophotometer, indirectly reflecting the concentration of the target substance in the sample or the physiological state of cells, and is one of the fundamental parameters for experimental data analysis.

[0039] The term "cell viability" as used in this article refers to a quantitative indicator that measures the proportion of surviving cells to the total cell population under certain conditions. It is the percentage of surviving cells to the total number of cells tested, and the calculation formula is: Cell viability = (Number of surviving cells / Total number of cells) × 100%. In this invention, the calculation formula is: Cell viability = [(OD drug-containing wells - OD blank wells) / (OD control wells - OD blank wells)] × 100%.

[0040] The term "efficiency" as used in this article refers to the proportion of an intervention (such as a drug, treatment plan, policy, technology, etc.) that achieves the expected effect in practical application. Efficiency specifically refers to the percentage of subjects who reach the preset "effective standard" after receiving the intervention in a particular research or application scenario, out of the total number of intervention subjects. The calculation formula is: Efficiency = (Number of subjects reaching the effective standard / Total number of subjects receiving the intervention) × 100%. In this invention, the formula for calculating efficiency is: Efficiency = (Survival rate of drug-containing well cells - Survival rate of model well cells) / (Survival rate of control well cells - Survival rate of model well cells) × 100%.

[0041] The term "CI Joint Index" used in this paper refers to the quantification of the strength and nature of a combined effect by comparing the "actual effect of the combined intervention" with the "theoretical additive effect of each intervention acting alone." A smaller value indicates a stronger synergistic effect; a value of 1 indicates an additive effect; and a value greater than 1 indicates an antagonistic effect.

[0042] The term "μM" as used in this article refers to a unit for measuring solution concentration, used to express the molar concentration of solute contained in a given volume of solution. It is an abbreviation for micromoles per liter, indicating that each liter of solution contains 10 μM of solute. 6 Moles of solute. Conversion to other concentration units: 1 mol / L = 10 3 mmol / L=10 6 μmol / L=10 9 nmol / L.

[0043] In a first aspect, the present invention provides: a composition for treating and / or preventing depression, wherein the active ingredients of the composition include secoiridoid glycosides, xanthonesin glycosides, and β-amyrin-type pentacyclic triterpenoids.

[0044] Preferably, the secoiridoid glycosides are selected from at least one of gentiopicroside and strychnoside; the xanthone glycosides are selected from at least one of mangiferin, 7-O-methylmangiferin, polygalactone XI, and gambogeylic acid; and the β-amyrin-type pentacyclic triterpenoids are selected from at least one of oleanolic acid, glycyrrhizic acid, and crataegolic acid.

[0045] More preferably, the secoiridoid glycoside is gentiopicroside, the xanthone glycoside is mangiferin, and the β-amyrin-type pentacyclic triterpenoid is oleanolic acid.

[0046] Preferably, the active ingredients in the composition include secoiridoid glycosides, xanthones glycosides, and β-amyrin-type pentacyclic triterpenoids in a mass ratio of 1:0.25-6:0.1-5. Specifically, the mass ratio of sepsis-enol glycosides, xanthonesin glycosides, and β-amyrin-type pentacyclic triterpenoids can be any value within the aforementioned range. For example, the mass ratio of sepsis-enol glycosides to xanthonesin glycosides to β-amyrin-type pentacyclic triterpenoids can be any value from 1 to 0.25 (e.g., 0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6) to any value from 0.1 to 5 (e.g., 0.1, 0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6) of sepsis-enol glycosides to β-amyrin-type pentacyclic triterpenoids. The ratio formed by any point values ​​of 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5) can be, specifically, the mass ratio of separacyl ether glycosides, xanthones glycosides, and β-amyrin-type pentacyclic triterpenoids can be 1:0.25:0.1, 1:0.3:0.2, 1:0.4:0.3, 1:0.5:0.4, 1:0.6:0.5, 1:0.7:0.6, 1:0.8:0.7, 1:0.9:0.8, 1:1:0.9, 1:1:1, 1:2:2, 1:3:3, 1:4:4, 1:5:5, 1:6:5, etc. Furthermore, the preferred mass ratio of the secoiridoid glycosides, xanthonesin glycosides, and β-amyrin-type pentacyclic triterpenoids in the composition is 1:1-6:1-5; Furthermore, the mass ratio of the secoiridoid glycosides, xanthonesone glycosides, and β-amyrin-type pentacyclic triterpenoids in the composition is 1:1:1, 1:6:5, and 1:3:2.

[0047] Furthermore, the active ingredients in the composition include gentiopicroside, mangiferin, and oleanolic acid in a mass ratio of 1:0.25-6:0.1-5; Specifically, the mass ratio of gentiopicroside, mangiferin, and oleanolic acid can be any value within the aforementioned range. For example, the mass ratio of gentiopicroside, mangiferin, and oleanolic acid can be any value between 1% and 0.25-6% of mangiferin (e.g., 0.25, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6) and any value between 0.1-5% of oleanolic acid (e.g., 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7). The ratio formed by any point values ​​of gentiopicroside, mangiferin, and oleanolic acid (0.8, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5) can be, specifically, the mass ratio of gentiopicroside, mangiferin, and oleanolic acid can be 1:0.25:0.1, 1:0.3:0.2, 1:0.4:0.3, 1:0.5:0.4, 1:0.6:0.5, 1:0.7:0.6, 1:0.8:0.7, 1:0.9:0.8, 1:1:0.9, 1:1:1, 1:2:2, 1:3:3, 1:4:4, 1:5:5, 1:6:5, etc. Furthermore, the preferred mass ratio of gentiopicroside, mangiferin and oleanolic acid in the composition is 1:1-6:1-5; Most preferably, the mass ratio of gentiopicroside, mangiferin and oleanolic acid in the composition is 1:1:1, 1:6:5, or 1:3:2.

[0048] Among them, the "secoiridoid glycosides, xanthone glycosides and β-amyrin-type pentacyclic triterpenoids", "gentiopicrin, mangiferin and oleanolic acid" can be purchased through conventional channels as commercial products or extracted from Tibetan Artemisia capillaris plants. Other active substances can also be obtained by purchasing commercial products or extracting from Tibetan Artemisia capillaris plants.

[0049] Secondly, the present invention provides a method for obtaining the active ingredient in the composition by extraction from Artemisia capillaris plants.

[0050] Preferably, the Tibetan Artemisia species include, but are not limited to, Swertia thaliana, Swertia davidii, Swertia purpurea, Swertia sibiricum, or Swertia sibiricum.

[0051] More preferably, the Tibetan Artemisia species are Swertia macrophylla and Swertia spp.

[0052] The preparation method of the active ingredient is as follows: (1) Extraction: After crushing or cutting the Tibetan Artemisia capillaris, add 6-8 times the weight of the medicinal material in 80-90% ethanol solution, soak at 75-85℃ for 1-3 hours, and perform hot reflux extraction at 0.12-0.17MPa steam pressure for 2-3 hours. Filter the reflux liquid, add 5-7 times the weight of the unrefluxed residue in 80-90% ethanol solution, repeat the above soaking and extraction steps, combine the obtained reflux liquids, add 4-6 times the weight of the unrefluxed residue in 80-90% ethanol solution, repeat the above soaking and extraction steps, combine the obtained reflux liquids to obtain the extract; (2) Concentration: The extract was concentrated under reduced pressure twice. The first concentration was carried out for 17-20 hours, with a solid content of 10%-15%. The second concentration was carried out for 8-12 hours, with a solid content of 23%-40%. Furthermore, the solid content after the two vacuum concentrations can be any point within the above-mentioned numerical range. Specifically, the solid content after the first vacuum concentration can be 10%, 11%, 12%, 13%, 14%, or 15%, and the solid content after the second vacuum concentration can be 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, or 40%. Furthermore, the solid content after the first vacuum concentration is 11%-14%; the solid content after the second vacuum concentration is 25%-37%. This invention discovers that controlling the vacuum concentration time of Artemisia capillaris can affect the solid content of the extract; furthermore, different solid contents will affect the content of various active substances in the Artemisia capillaris extract, thereby affecting the efficacy. (3) Drying: Vacuum dry the concentrate to ensure that the moisture content is ≤5%, and pulverize the dried material through an 80-mesh sieve to obtain the Artemisia capillaris extract.

[0053] Thirdly, the present invention provides the use of the said therapeutic and / or preventive composition for the treatment of depression in the preparation of medicaments for the treatment and / or prevention of depression.

[0054] The manifestations of a depressive episode include, but are not limited to, three aspects: core symptom clusters, psychological symptom clusters, and somatic symptom clusters. Specifically, these include symptoms such as depressed mood, anxiety, slowed thinking, sleep disturbances, and loss of energy.

[0055] Fourthly, the present invention provides a pharmaceutical composition for treating and / or preventing depression, the active ingredient of which includes the above-described antidepressant composition and one or more medically acceptable excipients.

[0056] In pharmaceutical compositions, the term "composition" includes products comprising an active ingredient and an inert component (medically acceptable excipient) constituting a carrier, as well as any product obtained directly or indirectly from a combination, complexation, or aggregation of two or more components, or from the decomposition of one or more components, or from other types of reactions or interactions of one or more components. Therefore, the pharmaceutical compositions of the present invention include any composition prepared by mixing the said antidepressant composition, other active ingredients, and medically acceptable excipients.

[0057] In a pharmaceutical composition, the excipients include pharmaceutically acceptable carriers, diluents, excipients, or other additives; the dosage form of the pharmaceutical composition includes solid, liquid, or semi-solid dosage forms; the pharmaceutical composition includes compositions suitable for oral, rectal, topical, and parenteral (including subcutaneous, intramuscular, and intravenous) administration, although the most appropriate route in any particular case depends on the specific subject, the nature and severity of the disease to which the active ingredient is administered. Pharmaceutical compositions can be prepared by any method known in the field of pharmaceutics.

[0058] Solid dosage forms, such as the pharmaceutical composition, can be capsules, tablets, pills, lozenges, powders, and granules.

[0059] Liquid dosage forms, such as the pharmaceutical composition, can be solutions, emulsions, suspensions, syrups, and elixirs.

[0060] Other suitable dosage forms include suppositories, sprays, ointments, creams, gels, inhalers, skin patches, implants, etc.

[0061] The pharmaceutical compositions of the present invention may also include other active ingredients.

[0062] Other active ingredients include glutamate receptor modulators, neuropeptide modulators, inflammatory pathway inhibitors, natural products, and derived active components.

[0063] Preferably, the ratio of the composition of secoiridoid glycosides, xanthone glycosides and β-amyrin-type pentacyclic triterpenoids to the other active ingredients is 1:0.1-50; Preferably, the ratio of the composition of gentiopicroside, mangiferin and oleanolic acid to the other active ingredients mentioned above is 1:0.1-50; The aforementioned glutamate receptor modulators are a class of drugs or active substances that exert therapeutic effects by targeting and regulating the function of glutamate receptors, thereby affecting glutamate signal transmission in the central nervous system. Glutamate is the most important excitatory neurotransmitter in the central nervous system, and abnormal activation or dysfunction of its receptors is closely related to various neurological diseases, such as depression, Alzheimer's disease, epilepsy, and schizophrenia. Examples include esketamine (an NMDA receptor antagonist that rapidly repairs neural synapses and promotes BDNF expression to achieve rapid antidepressant effects, used for treatment-resistant depression) and lamotrigine (a glutamate release inhibitor used as adjunctive therapy for bipolar depression to reduce mood swings).

[0064] The aforementioned neuropeptide modulators are a class of drugs or active substances that exert therapeutic effects by targeting and regulating the function of neuropeptides and their receptors, thereby affecting signal transmission in the central nervous system. Neuropeptides are small molecule peptides secreted by neurons and are widely involved in various physiological processes such as mood regulation, stress response, pain perception, and cognitive function. Abnormal expression or function of neuropeptides is closely related to various diseases such as depression, anxiety, schizophrenia, and pain. Examples include brezanolone (a GABA-A receptor-positive allosteric modulator that targets and regulates the hypothalamus-pituitary-adrenal (HPA) axis, used for postpartum depression, with rapid onset of action via intravenous injection) and substance P antagonists (such as aprepitant: by blocking the binding of the neuropeptide substance P to the NK1 receptor, it relieves physical discomfort and anxiety associated with depression).

[0065] The aforementioned inflammatory pathway inhibitors are a class of drugs or active substances that exert neuroprotective and antidepressant effects by targeting and regulating signaling pathways related to central or peripheral inflammatory responses, inhibiting the excessive release of pro-inflammatory factors, and reducing neuroinflammation. Examples include tofacitinib (a JAK-STAT pathway inhibitor that inhibits the release of pro-inflammatory factors (such as IL-6) and is used for inflammation-related depression, such as depression associated with autoimmune diseases) and minocycline (a tetracycline antibiotic that has both inhibitory effects on microglia activation and anti-inflammatory effects and is being studied as an adjunct antidepressant component).

[0066] The above-mentioned natural products and derived active components are selected from swertiamarin, swertiamarin, swertiamarin, swertiamarin alcohol, neomangiferin, isomangiferin, luteolin, ursolic acid, isovitexin, swertiamarin, and 3-oxodesylbis(oxetine). 7-O-[α-L-rhamnopyranose-(1→2)-β-D-xylanose]-1,8-dihydroxy-3-methoxy 7-O-β-D-xylanose-1,8-dihydroxy-3-methoxy 3-O-β-D-glucopyranose-1,8-dihydroxy-5-methoxy Or one or more of isopropanols.

[0067] The active ingredient can be administered orally in solid or liquid dosage forms, such as capsules, tablets, lozenges, sugar lozenges, granules, and powders, and in liquid forms such as elixirs, syrups, emulsions, dispersions, and suspensions. The active ingredient can also be administered parenterally in sterile liquid dosage forms such as dispersions, suspensions, or solutions. Other dosage forms that can be used to administer the active ingredient include ointments, creams, drops, transdermal patches, or powders for topical administration; ophthalmic solutions or suspensions for ocular administration, i.e., eye drops; sprays or powder compositions for inhalation or intranasal administration; or creams, ointments, sprays, or suppositories for rectal or vaginal administration. Gelatin capsules contain the active ingredient and a powdered carrier, such as lactose, starch, cellulose derivatives, magnesium stearate, stearic acid, etc. Similar diluents can be used to prepare compressed tablets. Both tablets and capsules can be formulated as sustained-release products to provide sustained release of the drug over several hours. Compressed tablets can be sugar-coated or film-coated to mask any unpleasant taste and protect the tablet from air, or they can be enteric-coated for selective disintegration in the gastrointestinal tract. Liquid dosage forms for oral administration may contain colorants and flavoring agents to increase patient acceptability. Generally, water, suitable oils, saline solutions, aqueous solutions of dextran (glucose), and related sugar solutions, and glycols such as propylene glycol or polyethylene glycol are suitable carriers for parenteral solutions. Solutions for parenteral administration preferably contain water-soluble salts of the active ingredient, suitable stabilizers, and buffering agents as needed. Antioxidants such as sodium bisulfite, sodium sulfite, or ascorbic acid, alone or in combination, are suitable stabilizers. Citric acid and its salts, and sodium EDTA may also be used. Furthermore, parenteral solutions may contain preservatives such as benzalkonium chloride, methylparaben, or propylparaben, and chlorobutanol. For inhalation administration, the compounds of the present invention can be conveniently delivered in aerosol form from pressurized packaging or a nebulizer. The antidepressant composition can also be delivered in a formulated powder form, which can be inhaled with the aid of a powder inhaler device. A preferred delivery system for inhalation is a metered-dose inhaler (MDI) aerosol, which can be formulated as a suspension or solution of the antidepressant composition of the present invention in a suitable propellant, such as a fluorocarbon or hydrocarbon. For ophthalmic administration, the ophthalmic formulation can be formulated with a solution or suspension of the antidepressant composition in a suitable ophthalmic carrier at a suitable weight percentage, thereby maintaining sufficient contact time between the compound and the ocular surface to allow the compound to penetrate into the cornea and internal regions of the eye.

[0068] Useful pharmaceutical dosage forms for administering the compounds of the present invention include, but are not limited to, hard and soft gelatin capsules, tablets, parenteral injections, and oral suspensions.

[0069] When the therapeutic and / or preventive composition for depression of the present invention is administered stepwise or in combination with other therapeutic agents, the same dosage form as described above may be used. When the drugs are administered in a physical combination, the dosage form and route of administration should be selected based on the compatibility of the combined drugs. The therapeutic and / or preventive composition for depression of the present invention may be administered as the sole active ingredient or in combination with a second active ingredient, the second active ingredient including active ingredients known to be effective in the treatment and / or prevention of depression.

[0070] The beneficial effects of this invention are as follows: The present invention has at least the following beneficial effects: 1. Compared with the prior art, the present invention has better technical effects in terms of HT22 cell survival rate, efficacy, latency time of mouse jumping platform test, open arm time of elevated cruciate maze test, immobility time during forced swimming test and tail suspension test.

[0071] 2. Good synergistic effect of components: The three components, namely separable ether glycosides, xanthone glycosides and β-amyrin type pentacyclic triterpenoids, work synergistically with each other. In particular, gentiopicrin, mangiferin and oleanolic acid have a significant synergistic effect at a certain ratio. They can work together from multiple angles to target the pathological links related to depression and enhance the antidepressant effect.

[0072] 3. Improved safety: In addition to the excellent antidepressant effect, no adverse reactions were observed during the testing process. Detailed Implementation

[0073] The following non-limiting embodiments are intended to enable those skilled in the art to gain a more comprehensive understanding of the present invention, but do not limit the invention in any way. The following content is merely an exemplary description of the scope of protection claimed by the present invention, and those skilled in the art can make various changes and modifications to the present invention based on the disclosed content, and such changes should also fall within the scope of protection claimed by the present invention.

[0074] The present invention will be further described below by way of specific embodiments. Unless otherwise specified, all instruments, devices, equipment, reagents, products, etc., used in the embodiments of the present invention are obtained through conventional commercial means.

[0075] Experiment 1 Obtaining Active Ingredients 1. Prepare Artemisia capillaris extract according to the following method (1) Extraction: After crushing or cutting the Tibetan Artemisia capillaris, add 7.5 times the weight of the medicinal material in 85% ethanol solution, soak at 80℃ for 2 hours, and perform hot reflux extraction at 0.15MPa steam pressure for 3 hours. Filter the reflux liquid, add 6 times the weight of the unrecirculated residue in 85% ethanol solution, and repeat the above soaking and extraction steps. Combine the obtained reflux liquids, add 6 times the weight of the unrecirculated residue in 85% ethanol solution, and repeat the above soaking and extraction steps. Combine the obtained reflux liquids to obtain the extract. (2) Concentration: The extracts obtained from each example and comparative example were concentrated under reduced pressure twice according to the reduced pressure concentration times indicated in Table 1. Specifically, the first reduced pressure concentration of Examples 1-4 was 17-20 hours, with a solid content of 10%-15%; the second reduced pressure concentration was 8-12 hours, with a solid content of 23%-40%. (3) Drying: Vacuum dry the concentrate to ensure that the moisture content is ≤5%, and pulverize the dried material through an 80-mesh sieve to obtain the Artemisia capillaris extract.

[0076] Among them, comparative examples 3-4 were extracted according to the extraction method of Example 1 in our patent CN113041276A.

[0077] Table 1. Extraction conditions for each embodiment and comparative example

[0078] 2. Component analysis of Artemisia capillaris extract. (1) Determination of the content of gentiopicrin and mangiferin According to the detection method of Example 1 in patent CN115541796B, the specific test conditions are as follows: Experimental instruments and reagents: Agilent 1260 high performance liquid chromatograph, Yuexu Ultimate XB-C18 column, KQ-700VDB ultrasonic instrument, chromatographic grade acetonitrile, analytical grade phosphoric acid, analytical grade methanol and ultrapure water.

[0079] Reference standards: gentiopicrin and mangiferin, purchased from the National Institutes for Food and Drug Control.

[0080] Test sample: Take 0.2g of the above extract, accurately weigh it, place it in a 100ml stoppered Erlenmeyer flask, add 25ml of 80% methanol, weigh it, sonicate it (power 270W, frequency 40kHz) for 40 minutes, remove it, let it cool, weigh it again, replenish the lost weight with 80% methanol, filter it with dry filter paper, discard the initial filtrate, take the subsequent filtrate and filter it through a 0.45μm microporous membrane to obtain the test sample solution.

[0081] step (A) Chromatographic detection conditions: Octadecyl-bonded silica gel was used as the stationary phase, acetonitrile as mobile phase A and 0.1% phosphoric acid as mobile phase B. The column temperature was 30℃, the flow rate was 0.7–1 ml / min, and the detection wavelength was 205–240 nm. Gradient elution was performed according to the table below, and the detection wavelength and flow rate were changed according to Table 2. The theoretical plate number, calculated based on the peak of the anthocyanin, should not be less than 10,000. The analysis time was 150 minutes.

[0082] Table 2. Detection Wavelength and Flow Rate Transformation Parameters

[0083] (B) Preparation of reference solution: Take an appropriate amount of gentiopicroside and mangiferin reference standards, weigh them accurately, add methanol to prepare a solution containing 0.5 mg per 1 ml, and the reference solution is obtained. (C) Preparation of test solution: Take 0.1 g of extract, accurately weigh it, place it in a 100 ml stoppered Erlenmeyer flask, add 25 ml of 80% methanol, weigh it, sonicate (power 270 W, frequency 40 kHz) for 40 minutes, take it out, let it cool, weigh it again, make up the weight loss with 80% methanol, filter it with dry filter paper, discard the initial filtrate, take the subsequent filtrate and filter it with a 0.45 μm microporous membrane to obtain the test solution; (D) Content determination: 10 μl of the test solution and the reference solution were injected into the liquid chromatograph and the contents of gentiopicrin and mangiferin were calculated.

[0084] (2) Determination of oleanolic acid content (A) Chromatographic detection conditions: Octadecyl bonded silica gel was used as the stationary phase, methanol-0.1% phosphoric acid (90:10) was used as the mobile phase, flow rate: 0.6 ml / min; detection wavelength: 205 nm; theoretical plate number calculated based on oleanolic acid peak should not be less than 10,000, column temperature 30℃.

[0085] (B) Preparation of reference solution: Take oleanolic acid reference standard, accurately weigh it, add methanol to prepare a solution containing 0.5 mg of oleanolic acid per 1 ml. (C) Preparation of test solution: Take 0.1g of the contents of the preparation example, weigh accurately, place in a 100ml stoppered Erlenmeyer flask, add 20ml of methanol accurately, weigh, sonicate (power 270W, frequency 40kHz) for 40 minutes, take out, cool, weigh again, replenish the lost weight with methanol, filter, discard the initial filtrate, and take the subsequent filtrate to obtain the test solution. (D) Determination: Accurately pipette 20 μl of the above reference solution and 20 μl of the test solution, respectively, and inject them into the liquid chromatograph. The content of oleanolic acid is then calculated.

[0086] Table 3. Content of each active ingredient and processing characteristics of each group

[0087] Experiment 2: Study on Antidepressant Effect In this experiment, the antidepressant effect of the active ingredient of this application was explored through the experiment on the viability of HT22 cells (mouse hippocampal neuron cells) and the mouse model of depression respectively.

[0088] Table 4. Source of Raw Materials

[0089] Detection of Model 1: Experiment on the Effect of HT22 Cells (Mouse Hippocampal Neuron Cells) Viability (1) Cell Culture DMEM medium supplemented with 10% fetal bovine serum and 1% double antibody was used to culture HT22 cells. The cells were placed in a constant temperature incubator at 37°C with 5% carbon dioxide.

[0090] (2) Determination of Cell Survival Rate by CCK8 Method CCK-8 was used to detect the protective effect of the composition on glutamate-induced oxidative stress injury of HT22. After cell counting, the cells were inoculated into 9-six-well plates at a density of 2×104 cells / mL, 100 μL of cell suspension was added to each well, and after culturing in a constant temperature incubator for 24 h, the medium was discarded for subsequent operations.

[0091] 100 μL of complete medium containing 15 mmol / L glutamate and 100 μmol / L extracts of each group were added to the wells for stimulation for 12 h, with 6 replicates for each concentration.

[0092] The medium in the wells was aspirated, 100 μL of medium containing 10% CCK-8 was added, incubated in a constant temperature incubator for 4 h, and the OD value was measured at a wavelength of 450 nm. The well with only CCK-8 medium without cells was used as the blank well, the well without any stimulation of cells was used as the control well, the well with cells containing glutamate and CCK-8 medium was used as the model well, and the well with cells containing glutamate, extracts of each group and CCK-8 medium was used as the drug-containing well, and the effective rate of each group of compositions was calculated.

[0093] Cell survival rate = (OD of drug-containing well - OD of blank well) / (OD of control well - OD of blank well); Effective rate = (Cell survival rate of drug-containing well - Cell survival rate of model well) / (Cell survival rate of control well - Cell survival rate of model well); Compusyn software was used to calculate the CI combination index (CI < 0.3 indicates strong synergy, 0.3 < CI < 0.7 indicates synergy, 0.7 < CI < 1 indicates slight synergy, CI = 1 indicates addition, CI > 1 indicates antagonism).

[0094] (3) Experimental results and analysis The formulations and test results of each embodiment and comparative example are shown in Table 5 below: Table 5. Formulations and test results for each example and comparative example.

[0095] Analysis of Comparative Examples 1-3 in the table above shows that all three single drugs have a certain protective effect on damaged HT22 cells, but the highest efficacy of each single drug does not exceed 0.5, indicating limited improvement. Analysis of Comparative Examples 4-6 in the table shows that the CI values ​​of the two-drug combination groups are mostly between 0.7 and 1.0, with some showing additive effects, but the overall effect is weaker than that of the three-drug combination group. The two-drug combination is better than the single drug, but significantly lower than the three-drug combination, and the CI values ​​are mostly greater than 0.7, indicating a lack of strong synergistic effect. The three-drug combination has a much better protective effect on HT22 cells than the single drug or the two-drug combination. The CI values ​​of the three-drug combination group are mostly below 0.8, while the CI values ​​of the three-drug combination group using a specific ratio are mostly less than 0.5, showing a strong synergistic effect.

[0096] Model 2: Establishment of a mouse model of depression and its efficacy experiments The corresponding examples and comparative examples were set according to the mass formulations in Table 6, and subsequent efficacy experiments were conducted according to the mass formulations in the table below.

[0097] Table 6. Formulations of Examples and Comparative Examples

[0098] (1) Experimental animals Six- to seven-week-old SPF-grade C57BL / 6 mice (purchased from the Animal Experiment Center of Xi'an Jiaotong University), with an average weight of 18-20g, were housed in a controlled environment with a room temperature maintained at 22±2℃ and humidity at 55%±5%, following a 12-hour light / dark cycle. They were allowed free access to food and water. All experimental procedures involving the mice were performed in accordance with the National Institutes of Health's guidelines for laboratory animal care and use.

[0099] (2) Grouping of animals Animals were placed in a suitable environment for 7 days to acclimatize, and their body weight and sucrose preference rates were tested. Mice with abnormal values ​​were excluded. Animals were randomly assigned to groups of 10 each. The groups were: blank control group, model group, positive control group (Shugan Jieyu capsules, 150 mg / kg), example group, and comparative group.

[0100] (3) Animal modeling A mouse model of depression and anxiety was established by administering chronic unpredictable mild stimuli (CUMS) for 5 weeks to the model group, positive control group, example group, and comparative group. The chronic unpredictable mild stimuli included the following eight different stimuli: S1 Forced Swimming: Fill a 22cm deep glass jar with water at 23±2℃, place the mouse in the jar for 10 minutes, and carefully cover it with a heavy lid to prevent it from escaping. Then put the mouse back in a clean, dry cage and line it with fresh bedding to avoid it getting cold and catching a cold.

[0101] S2 restraint: The mouse is placed in a 50mL plastic tube and adjusted from the outside with plastic tape to prevent the animal from moving. There is a hole at the far end of the tube to allow for normal breathing.

[0102] S3 Water scarcity: Remove the water bottle from the cage within 24 hours and put it back after the time is up.

[0103] S4 Isolation: The mice were placed alone in a new cage for 24 hours, and then returned to the cage with their companions.

[0104] S5 Food Shortage: Remove food from the cage for 24 hours, then put the food back in.

[0105] S6 Wet bedding: Place the mice individually in a new cage with 200 ml of water per 100 g of bedding for 24 hours, then return the mice to their cages with their companions.

[0106] No bedding: The mice were placed alone in a new cage without bedding for 24 hours, and then returned to the cage with their companions.

[0107] S7 Tilting Cage: Tilt the cage at 45° for 24 hours, then restore the cage to its normal state.

[0108] Tail clamping: Clamp a plastic clamp 2 cm from the end of the tail for 3 minutes each time, 3 times a day, and return the animal and its companions to the cage.

[0109] S8 Crowded Space: Place a plastic partition in the cage to reduce the activity area by half. Remove the partition after 24 hours.

[0110] (4) Intervention methods To avoid animal adaptation to specific stimuli, which could affect the accuracy of experimental results, one or two stimuli were randomly applied daily, six days a week. Furthermore, to ensure the unpredictability of the experiment, each stimulus was used no more than three times cumulatively throughout the entire experimental period. This method aims to create a complex and variable stress environment to induce behavioral and physiological responses in mice similar to human depression and anxiety.

[0111] Throughout the experiment, the blank group and the model group were fed in a routine manner, while the positive control group, the example group, and the comparative group were fed in a routine manner and began to be administered medication by gavage on day 36. The dosage was the same as that of the corresponding combination drug in the example and the comparative. The positive control group was given Shugan Jieyu capsules at a dosage of 150 mg / kg·d once a day.

[0112] (5) Behavioral evaluation: Behavioral evaluations of all mice began in week seven, including the jumping platform test, elevated cross maze test, forced swimming test, and tail suspension test.

[0113] S1 Jumping Platform Experiment: Mice were placed on a platform and trained with an electric shock grid (0.2 mA). When a mouse jumped off the platform, it received an electric shock, thus generating pain memory. The training time was three minutes. If a mouse did not jump off the platform within three minutes, it was removed. After the training, all mice returned to their original cages. The memory retrieval phase occurred 24 hours after training: mice were placed on the platform and tested for three minutes without electric shock. The latency time for each group of mice to jump off the platform was recorded, and the differences were statistically significant (P<0.05, P<0.01).

[0114] S2 elevated cross maze experiment: Mice were placed in the central area of ​​the maze, facing the open arms, and allowed to move freely for 6 minutes. The percentage of time spent in the open arms for each group was counted, and the differences were statistically significant (P<0.05, P<0.01).

[0115] S3 Forced Swimming Experiment: A 30 cm glass tank filled with water at 23-25°C was used. On the first day, all mice were forced to swim for 15 minutes as a training exercise. On the second day, the mice were placed in the same environment for 10 minutes. Motion activity was monitored using a video tracking system, and the immobility time of each animal during the test was calculated (immobility was defined as inactivity other than the floating action required to keep the head above the water surface). Differences were considered statistically significant (P<0.05, P<0.01).

[0116] S4 Tail Suspension Test: The tip of a mouse's tail (2 cm from the tail tip) was taped to a suspension bar (30 cm above the ground). The mouse was then allowed to touch the suspension. Initially, the mouse instinctively struggled violently to escape the predicament. However, if unable to escape, it became immobile in a state of despair, exhibiting depressive-like behavior. The longer the immobility, the more depressed the mouse became. Movement was monitored using a video tracking system for 6 minutes, and the immobility time for each animal during the test was calculated (immobility was defined as no voluntary or escapist movement). Differences were statistically significant (P < 0.05, P < 0.01).

[0117] The results of the above behavioral tests are expressed as mean ± standard deviation (s), and the test results are shown in Table 7 below.

[0118] Table 7. Results of the behavioral evaluation experiment

[0119] In the table, a vs. blank group, b vs. model group, *P<0.05, **P<0.01, ***P<0.001.

[0120] Verification of technical effectiveness and / or analysis of technical problem solving Experimental Conclusions: As shown in the table above, compared with the model group mice, the latency time in the jumping platform test, immobility time in the forced swimming test, and immobility time in the tail suspension test were significantly shortened in Examples 1-5, while the open arm time in the elevated cruciate maze test was significantly prolonged. Overall, the antidepressant effect was superior to the positive control group. Examples 1-5 (especially 4 and 5) showed the best performance in improving core depressive symptoms (shortened immobility time) and anxiety (prolonged open arm time), with effects superior to the positive control group and approaching the level of the blank group. This indicates strong antidepressant activity, which is statistically significant compared to the model group, proving that this antidepressant composition has an antidepressant effect.

[0121] Compared with Comparative Examples 1-6, Examples 1-5 showed a significantly improved antidepressant effect in mice. This demonstrates that the effect of using gentiopicroside, mangiferin, and oleanolic acid as a single component or in pairs is far less than that of using the three components in combination, proving that these three components have a synergistic antidepressant effect.

[0122] Compared with Comparative Examples 7-10, Examples 1-5 showed a significant improvement in the antidepressant effect in mice. The combination of gentiopicroside, mangiferin and oleanolic acid in a specific ratio showed better antidepressant effect than combinations in other ratios, further demonstrating that the three components in the specific ratio had a significant synergistic effect.

[0123] Preparation Example Weigh the ingredients according to the following prescription: Gentianoside 3g 3g of mangiferin Oleanolic acid 3g 3g of swertiamarin 20g of starch The above raw materials are passed through an 80-mesh sieve and dried. The four compounds other than starch are mixed together, and then starch is added. The mixture is then filled into 100 capsules.

[0124] (I) Analysis of capsule contents According to the testing method in Experiment 1, the calculated content of gentiopicroside was 9.64%, mangiferin was 9.48%, swertiamarin was 9.16%, and oleanolic acid was 9.39%.

[0125] (II) Experiment on the effect of HT22 cell (mouse hippocampal neuron) viability Cell culture and cell viability were performed according to the method described in detection model 1 above. The experimental setup and results are as follows: Table 8. Experimental Results

[0126] The above experimental results show that, in the experiment on the effect of HT22 cell viability, swertiamarin had no effect on the technical efficacy of gentiopicrin, mangiferin, and oleanolic acid.

[0127] (III) Establishment of a mouse model of depression and drug efficacy experiments Modeling and efficacy testing were conducted according to the method described in detection model 2 above. The experimental setup and results are as follows: Table 9. Experimental Setup and Results

[0128] In the table, a vs. blank group, b vs. model group, *P<0.05, **P<0.01, ***P<0.001 The above experimental results show that, in the mouse depression model experiment, the addition of swertiamarin to gentiopicrin, mangiferin and oleanolic acid did not affect the therapeutic effect on mice. Therefore, swertiamarin has no effect on the technical efficacy of gentiopicrin, mangiferin and oleanolic acid.

[0129] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A pharmaceutical composition for the treatment and / or prevention of depression, characterized by, The pharmaceutical composition comprises active ingredients and one or more pharmaceutically acceptable excipients, wherein the active ingredients comprise secoiridoid glycosides, xanthone glycosides and β-amyrin-type pentacyclic triterpenes in a mass ratio of 1:0.25-6:0.1-5.

2. The pharmaceutical composition of claim 1, wherein, The secoiridoid glycosides are at least one of gentalpicroside, loganin acid; the xanthone glycosides are at least one of mangiferin, 7-O-methyl mangiferin, senegin XI, gambogic acid; and the β-amyrin-type pentacyclic triterpenes are at least one of oleanolic acid, glycyrrhizic acid, and maslinic acid.

3. The pharmaceutical composition of claim 2, wherein, The secoiridoid glycosides are gentalpicroside, the xanthone glycosides are mangiferin, and the β-amyrin-type pentacyclic triterpenes are oleanolic acid.

4. The pharmaceutical composition of claim 3, wherein, The active ingredients comprise gentalpicroside, mangiferin, and oleanolic acid in a mass ratio of 1:0.25-6:0.1-5.

5. The pharmaceutical composition of claim 4, wherein, The active ingredients are obtained by extraction from plants of the genus Rhynchosia, and the preparation method comprises the following steps: (1) extraction: after the Rhynchosia plants are crushed or cut, 6-8 times the mass of the plants of 80-90% ethanol solution is added, the mixture is soaked at 75-85°C for 1-3 hours, and then hot reflux extraction is performed at a steam pressure of 0.12-0.17 MPa for 2-3 hours; the refluxed liquid is filtered, and the remaining residue that is not refluxed is added with 5-7 times 80-90% ethanol solution; the above-mentioned soaking and extraction steps are repeated, and the obtained refluxed liquids are combined; the remaining residue that is not refluxed is added with 4-6 times 80-90% ethanol solution, and the above-mentioned soaking and extraction steps are repeated; the obtained refluxed liquids are combined to obtain an extract; the Rhynchosia plants are Rhynchosia delavayi or Rhynchosia henryi; (2) concentration: the extract is concentrated twice under reduced pressure, and the solid content is 10%-15% in the first time of concentration and 23%-40% in the second time of concentration; (3) drying: the concentrated liquid is vacuum dried to ensure that the moisture content is less than or equal to 5%, and the dried material is crushed to pass through an 80-mesh sieve to obtain the Rhynchosia extract.

6. The pharmaceutical composition of claim 1, wherein, The pharmaceutical composition can further comprise other active ingredients, wherein the other active ingredients comprise glutamate receptor modulators, neuropeptide modulators, inflammatory pathway inhibitors, and natural products and active components derived therefrom.

7. The pharmaceutical composition of claim 1, wherein, The excipients comprise pharmaceutically acceptable carriers, diluents, excipients, or other additives, and the dosage form of the pharmaceutical composition comprises solid preparations, liquid preparations, or semi-solid preparations.

8. Use of the pharmaceutical composition according to any one of claims 1-7 in the preparation of a medicament for treating and / or preventing depression.

9. Use according to claim 8, characterized in that, The manifestations of depression include a core symptom group, a psychological symptom group, and a somatic symptom group.

10. Use according to claim 8, characterized in that, The manifestations of depression include symptoms of low mood, anxiety, slow thinking, sleep disorders, and loss of energy.

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