Experimental method of Sanwei sandalwood capsule based on material basis

Abstract

The invention discloses a three-component sandalwood capsule experiment method based on material basis, the three-component sandalwood capsule experiment method is used for preparing Mongolian medicine for treating and / or preventing depression and medicine for promoting interaction between nerve active ligands and receptors and sensitive to stimulation response according to the characteristics of the three-component sandalwood material basis, and the three-component sandalwood capsule experiment method is stable in raw material supply, low in cost, low in preparation cost and suitable for large-scale production. The medicine effect is reliable; the experiment is based on a UHPLC-QTOF / MS (Ultra High Performance Liquid Chromatography-Quadrature Time of Flight / Mass Spectrometry) technology, firstly, the in-vitro chemical substance basis of the Sanwei sandalwood capsule is represented, and then in combination with the general rule of in-vivo biotransformation, the potential active ingredients of the compound playing the efficacy in vivo are screened out from the traditional Chinese medicine ingredients which can be utilized by the body to generate obvious in-vivo exposure after the Sanwei sandalwood capsule is administered. And an effective, accurate and feasible research clue is provided for subsequent selection of traditional Chinese medicine compound quality control markers and pharmacodynamic mechanism research of active components.

Description

Technical Field

[0001] The present invention relates to the technical field of experimental methods for Santalum album capsule with three flavors, and particularly to an experimental method for Santalum album capsule with three flavors based on the material basis. Background Art

[0002] The current situation of the existing process of Santalum album capsule with three flavors: First, there are relevant reports on the treatment of coronary heart disease with Santalum album capsule with three flavors at present, but there is no report in the relevant research on depression; Second, only partial chemical components of Santalum album capsule with three flavors have been studied in terms of chemical components, and there is no systematic research on the material basis, including in vitro compound component research and in vivo metabolic network research.

[0003] Development: The prescription of Santalum album capsule with three flavors is derived from "Four Medical Canons". Santalum album with three flavors is a basic prescription for treating cardiovascular diseases in Mongolian and Tibetan medicines, and is composed of three medicinal materials: wild jujube fruit, nutmeg and sandalwood. Among them, wild jujube fruit is the monarch drug in the prescription; some studies have found that Santalum album capsule with three flavors has an obvious effect of promoting sleep and improving sleep quality. In addition, some people have used the method of network pharmacology to preliminarily reveal the main active components of Santalum album with three flavors and their targets and related pathways for treating depression, providing new ideas for the treatment of depression.

[0004] In summary, the current clinical indications of Santalum album with three flavors have not involved the in vitro and in vivo analysis of the material basis. Therefore, an experimental method for Santalum album capsule with three flavors based on the material basis is proposed. Summary of the Invention

[0005] The purpose of the present invention is to solve the disadvantages existing in the prior art as described in the background art, and to propose an experimental method for Santalum album capsule with three flavors based on the material basis.

[0006] In order to achieve the above purpose, the present invention adopts the following technical scheme:

[0007] An experimental method for Santalum album capsule with three flavors based on the material basis, comprising the following steps:

[0008] S1. Select 72 C57 experimental mice and test to leave the mice that meet the experiment;

[0009] S2. Select the modeling method of subcutaneous injection of corticosterone in the neck;

[0010] S3. All the modeled experimental mice are subcutaneously injected with corticosterone continuously every day between 10:00 and 12:00 for 5 weeks;

[0011] S4. During the 5-week period of subcutaneous injection of corticosterone, the mice treated with the vehicle are used as the control group, and a behavioral experiment is carried out at the end of the second week to determine whether depressive-like behavior occurs;

[0012] S5. To determine whether corticosterone can induce depressive-like behavior in mice, open field test (OFT), tail suspension test (TST), and sucrose preference test (SPT) were conducted on mice.

[0013] As a further step in the method of the present invention, S11. Baseline sucrose preference test was performed on 72 C57 experimental mice.

[0014] As a further step in the method of the present invention, S12. After the test, mice with high-level differences were removed, and the remaining mice were available for experiments.

[0015] As a further step in the method of the present invention, S21. Corticosterone at 20 mg / kg was dissolved in 2% DMSO + 40% PEG300 + 5% Tween-80 + 53% Saline.

[0016] As a further step in the method of the present invention, S41. After the appearance of depressive-like behavior was determined, the mice were randomly assigned to a blank control group, a model group, a low-dose galangin group (0.5625 mg / kg), a medium-dose galangin group (1.125 mg / kg), a high-dose santalum album group (2.25 mg / kg), and a positive drug fluoxetine group (30 mg / kg), with 12 mice in each group.

[0017] 11. As a further step in the method of the present invention, S42. Galangin and fluoxetine treatments were given at the beginning of the third week;

[0018] S43. After the end of the fifth week, behavioral tests were conducted again to evaluate the depressive-like behavior of the mice.

[0019] As a further step in the method of the present invention, S51. In the open field test (OFT) of mice, compared with the control mice, the total distance moved by the corticosterone group decreased, and the galangin group was able to reverse this behavior and significantly increase the movement distance of the mice.

[0020] As a further step in the method of the present invention, S52. Compared with the control mice, the duration of immobility of the mice treated with corticosterone increased in the tail suspension test (TST), and the immobility time of the corticosterone mice was significantly reduced after treatment with santalum album.

[0021] As a further step in the method of the present invention, S53. In the sucrose preference test (SPT), compared with the control group, the preference for sucrose in the corticosterone group decreased significantly, and the preference for sucrose in the mice increased significantly after treatment with galangin.

[0022] Advantages of the present invention

[0023] This experiment is based on the UHPLC-QTOF / MS technology. First, the in vitro chemical substance basis of the Sanwei Tanxiang Capsule is characterized. Then, combined with the general law of in vivo biotransformation, the traditional Chinese medicine components (in the form of component prototypes and / or metabolites) that can be utilized by the body to produce significant in vivo exposure after administration of the Sanwei Tanxiang Capsule are screened out, providing effective, accurate, and feasible research clues for the selection of quality control markers of traditional Chinese medicine compound and the study of the pharmacodynamic mechanism of active components. Description of the Drawings

[0024] Figure 1 It is the result graph of the open field test for mice;

[0025] Figure 2 It is the result graph of the tail suspension test for mice;

[0026] Figure 3 It is the result graph of the sucrose preference test;

[0027] Figure 4 It is the network diagram of the in vivo and in vitro substance basis of the Sanwei Tanxiang Capsule;

[0028] Figure 5 It is the matching schematic diagram of metabolites;

[0029] Figure 6 It is the base peak chromatogram of the Sanwei Tanxiang Capsule;

[0030] Figures 7 - 10 It is the chemical composition characterization graph of the Sanwei Tanxiang Capsule;

[0031] Figure 11 It is the technical route diagram of the substance basis characterization of Sanwei Tanxiang;

[0032] Figure 12 It is the schematic diagram of the effect of Sanwei Tanxiang on the blood drug concentration of paroxetine;

[0033] Figures 13 - 15 It is the schematic diagram of the curative effect of Sanwei Tanxiang combined with paroxetine on depression.

[0034] Among them, the abstract drawing is Figure 5 . Detailed Description of the Invention

[0035] The content of the present invention can be more easily understood by referring to the following detailed description of the preferred embodiments of the present invention and the included examples. Unless otherwise defined, all technical and scientific terms used in this application have the same meaning as commonly understood by those of ordinary skill in the art to which the present invention belongs. When there is a contradiction, the definition in this specification shall prevail.

[0036] As used in this application, the term "composed of" is synonymous with "comprising". The terms "comprising", "including", "having", "containing" or any other variation thereof used in this application are intended to cover non-exclusive inclusion. For example, a composition, step, method, article or device containing the listed elements need not be limited to those elements, but may include other elements not expressly listed or elements inherent to such composition, step, method, article or device.

[0037] The connecting phrase "consisting of" excludes any unrecited element, step or component. If used in a claim, this phrase renders the claim closed-ended, excluding materials other than those described, except for conventional impurities associated therewith. When the phrase "consisting of" appears in a clause within the body of a claim rather than immediately following the subject, it limits only the elements described in that clause; other elements are not excluded from the claim as a whole.

[0038] When an equivalent, concentration, or other value or parameter is expressed as a range, a preferred range, or a range defined by a series of upper preferred values and lower preferred values, this should be understood to specifically disclose all ranges formed by any pairing of any upper range limit or preferred value with any lower range limit or preferred value, whether or not the ranges are separately disclosed. For example, when the range "1 to 5" is disclosed, the described range should be interpreted to include the ranges "1 to 4", "1 to 3", "1 to 2", "1 to 2 and 4 to 5", "1 to 3 and 5", etc. When a numerical range is described in this application, unless otherwise stated, the range is intended to include its end values and all integers and fractions within the range.

[0039] The singular form includes plural referents unless the context clearly dictates otherwise. "Optional" or "any one" means that the subsequently described matter or event may or may not occur, and the description includes both the case where the event occurs and the case where the event does not occur.

[0040] Approximating language, as used in the specification and claims, is used to modify a quantity, indicating that the present invention is not limited to the specific quantity and also includes modified portions that are close to the quantity and acceptable without causing a change in the relevant basic function. Accordingly, modifying a numerical value with "about", "approximately", etc. means that the present invention is not limited to the exact numerical value. In some instances, the approximating language may correspond to the precision of the instrument for measuring the value. In the specification and claims of this application, range limitations may be combined and / or interchanged, and these ranges include all sub-ranges subsumed therein if not otherwise stated.

[0041] In addition, the indefinite articles "a" and "an" before the elements or components of the present invention do not limit the quantity requirements (i.e., the number of occurrences) of the elements or components. Therefore, "a" or "an" should be construed to include one or at least one, and the singular forms of the elements or components also include the plural forms, unless the quantity clearly refers to the singular form.

[0042] In the description of the present invention, "a plurality of" means two or more, unless otherwise specifically defined.

[0043] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments.

[0044] Example 1

[0045] An experimental method for the Sami Santalum Album Capsule based on the material basis, classifying and attributing unknown components according to the cracking rules of components of different structural types and diagnostic ions. 97 components were identified in the Sami Santalum Album Capsule. Among them, 38 chemical components were identified in Choerospondias axillaris, which were basically various carboxylic acids or phenolic acids and flavonoid compounds, such as ellagic acid and ellagic acid derivatives, etc.; 14 compounds were identified in Santalum album, among which sesquiterpenoids, such as α-Santaldiol, β-Santaldio, etc. were its characteristic components, and there were also a small amount of carboxylic acids and phenylpropanoid compounds; 49 compounds were identified in Myristica fragrans, mainly phenylpropanoids and phenylpropanoid derivatives (lignans), as well as flavonoid compounds; reference can be made to Figure 6 The base peak chromatogram of the labeled extract ((a) negative ion; (b) positive ion), Figures 7 - 10 Schematic diagram of the chemical component characterization of the Sami Santalum Album Capsule.

[0046] The specific process conditions are as follows:

[0047] Sample preparation: Take about 0.1 g of the content of the Sami Santalum Album Capsule, weigh it, add 10 mL of 70% methanol, extract ultrasonically 2 times, 30 minutes each time, combine the supernatants, take 2 mL of the supernatant, blow to dryness with nitrogen until the methanol is completely volatilized, add pure water to 2 mL, load it onto an activated C18 solid-phase extraction small column, wash it with 1 mL of pure water, and then elute it with 2 mL of pure methanol. The eluate is centrifuged at 12000 r / min for 10 minutes, take 400 μL of the supernatant, dry it with nitrogen, re-dissolve it with 400 μL of 70% methanol, centrifuge it at 13000 r / min for 10 minutes, and aspirate 2 μL of the supernatant and inject it into LC-MS.

[0048] Animals and administration dosage: Twelve male Wistar rats (230 g), ordered from Shanghai Slake Laboratory Animal Co., Ltd. (20220004046254), were randomly divided into a blank group and an administration group when their body weights reached 220 - 240 g after 7 days of adaptive feeding (in a room with a temperature of 22 ± 1°C and a 12 / 12-hour light / dark cycle). They were fasted 12 hours before administration, and the administration dosage was 0.252 g / kg.

[0049] Collection and preparation of plasma samples: Blood was collected from the orbital venous plexus at 5 min, 15 min, 30 min, 60 min, 120 min, 240 min, 360 min, 480 min, and 600 min after administration, and placed in EP tubes containing 10 μl of EDTA (0.09 g / ml). Plasma was obtained by centrifugation at 4000 r / min for 5 min. Blank plasma was prepared in the same way.

[0050] Chronic unpredictable mild stress depression model: Mice in the chronic unpredictable mild stress model were subjected to a series of stimuli, including: noise (30 min), tilting the cage (45°, 8 h), wet bedding (8 h), circadian rhythm inversion (24 h), shaking the cage (5 min), restraint (2 h), tail clamping (5 min), cold water swimming (8°C, 6 min), foreign objects (8 h). Mice in the model group, positive drug group, and each administration group were continuously subjected to the above chronic mild stress for 8 weeks, randomly receiving two of the above stimuli every day, and the same stimulus could not be received on adjacent days. Mice in the blank group did not receive any chronic mild stress.

[0051] Example 2

[0052] An experimental method for Santalum album capsule based on the material basis, which is used to prepare Mongolian medicine for treating and / or preventing depression according to the characteristics of the material basis (effective chemical components) of Santalum album, promoting the interaction between neuroactive ligands and receptors, and drugs that are sensitive to stimuli. Its raw material supply is stable, the cost is low, the preparation cost is low, and the drug effect is reliable.

[0053] The in-depth analysis of the material basis research of the effective components of Santalum album capsule is as follows:

[0054] The cleavage of flavonoids is illustrated by taking naringenin in Choerospondias axillaris as an example. Figure 5 It is a display diagram for metabolite matching: Figure 5 In (A), the superimposed extracted ion chromatogram of M60, from top to bottom are the extract group of Santalum album capsule, the administration group (bile, feces, plasma, urine), and the blank group (bile, feces, plasma, urine); Figure 5 In (B), the secondary mass spectrum of M60 in the extract of Santalum album capsule; Figure 5 In (C), the secondary mass spectrum of M60 in the urine of the administration group;Figure 5 The secondary mass spectrum of (D) M60 in the urine of the bile group; Figure 5 The main in vivo metabolic pathway of (E) Scopoletin.

[0055] As Figure 5 As shown in Figure A in, the most classic and characteristic cleavage of flavonoids is RDA cleavage, which occurs by breaking the middle ring to generate the corresponding highest m / z 151 and m / z 119. Additionally, m / z 187 can also be generated through hydrogen rearrangement;

[0056] The cleavage of lignan compounds is illustrated by dehydrodiisoeugenol in nutmeg, as Figure 5 As shown in Figure B in, the loss of the side chain outside the benzene ring can be observed in the secondary mass spectrum, generating fragments such as m / z 286 and m / z 271. Further, the benzene ring can crack under the induction of free radicals to generate m / z 203, and then C3H4 is lost to generate m / z 163 and m / z 164;

[0057] The cleavage of phenylpropanoid compounds is illustrated by eugenol rutinoside in nutmeg, as Figure 5 As shown in Figure C in, for phenylpropanoid compounds containing sugar, fragments m / z 307 and m / z 163 generated after sugar hydrolysis can be observed in the secondary mass spectrum. The sugar can further crack to generate fragments such as m / z 247, m / z 163, and m / z 103. The phenylpropanoid parent nucleus part can also lose a methyl radical to generate m / z 148, etc.;

[0058] The cleavage of polyphenol compounds is illustrated by gallic acid in Choerospondias axillaris, as Figure 5 As shown in Figure D in, gallic acid loses CO2 to generate the characteristic fragment m / z 125. The benzene ring cracks to lose CO to generate the fragment m / z 97, and further loses a molecule of water to generate m / z 79. For polyphenol compounds containing an ellagic acid group, the characteristic fragment is m / z 300 (299.99);

[0059] The cleavage of sesquiterpene compounds is illustrated by santalol in sandalwood. Santalol has a good response in the positive ion mode and rich secondary mass spectrum fragments. The fragments generated by the sequential loss of the carbon chain can be observed. The specific cleavage rules and fragments are shown in Figure 5 Figure E in.

[0060] The analysis of the metabolites of the Sanwei Tanxiang Capsule in vivo is as follows:

[0061] Traditional Chinese medicine compound has complex components. After oral administration, drugs need to undergo a series of biotransformation processes in the body, such as absorption, distribution, metabolism, and excretion. After compounds enter the body, some compounds (prototype and some metabolites) are absorbed in the gastrointestinal tract. The absorbed components are transported to the liver through the portal vein for further metabolism, then enter the systemic blood circulation and are distributed to various tissues and organs to exert their effects, and finally pass through the kidneys and are excreted out of the body through urine. Other components not absorbed by the gastrointestinal tract are excreted out of the body through feces. Therefore, the change process of chemical components in the body after oral administration of traditional Chinese medicine is an important part of studying the material basis of the efficacy of traditional Chinese medicine. See Figure 4 ;

[0062] Figure 4 It is the network diagram of the material basis of Sanweitanxiang Capsule in vitro and in vivo: In the figure, Figure A is the network correlation diagram between the prototype and metabolites; Figure B is the distribution of the prototype and metabolites in tissues; Figure C is the schematic diagram of the chemical structure of the compounds with tissue distribution; green represents the prototype compound, and red represents the metabolite.

[0063] Example 3

[0064] An experimental method for Sanweitanxiang Capsule based on the material basis includes the following content:

[0065] Before the experiment started, in order to exclude individual differences, a baseline sucrose preference test was conducted on 72 C57 experimental mice, and mice with high-level differences were removed. According to previous research 41, the modeling method of subcutaneous injection of corticosterone in the neck was selected. Corticosterone (20 mg / kg) was dissolved in 2% dimethyl sulfoxide + 40% polyethylene glycol 300 + 5% emulsifier T-80 + 53% saline solution. Specifically, all the modeling experimental mice were subcutaneously injected with corticosterone between 10:00 and 12:00 every day for 5 weeks. Mice treated with the vehicle during the same period were used as the control group. At the end of the second week, a behavioral experiment was conducted to determine whether depressive-like behaviors occurred. After determining the occurrence of depressive-like behaviors, the mice were randomly assigned to a blank control group, a model group, a low-dose galangin group (0.5625 mg / kg), a medium-dose galangin group (1.125 mg / kg), a high-dose Sanweitanxiang group (2.25 mg / kg), and a positive drug fluoxetine group (30 mg / kg). Each group contained 12 mice. Galangin and fluoxetine treatments were given at the beginning of the third week. After the end of the fifth week, a behavioral test was conducted again to evaluate the depressive-like behaviors of the mice.

[0066] Results: In order to determine whether corticosterone can induce depressive-like behaviors in mice, we conducted the open field test (OFT) for mice, the tail suspension test (TST) for mice, and the sucrose preference test (SPT) for mice;

[0067] In the open field test (OFT) of mice, compared with the control mice, the total distance moved by the corticosterone group decreased, and the galangin group was able to reverse this behavior and significantly increase the movement distance of the mice.

[0068] Compared with the control mice, the mice treated with corticosterone showed an increased duration of immobility in the tail suspension test (TST), and the administration of Santali Albi Radix could significantly reduce the immobility time of the corticosterone-treated mice.

[0069] In the sucrose preference test (SPT), it was observed that compared with the control group, the preference for sucrose in the corticosterone group decreased significantly, and the treatment with Santali Albi Radix could significantly increase the preference of mice for sucrose.

[0070] Example 4

[0071] Refer to Figure 11 , the technical roadmap for the characterization of the material basis of Santali Albi Radix, specifically described as follows: The serum, urine, and fecal samples before and after administration were analyzed using the UPLC-QTOF / MS method; after normalizing the data collected by UPLC-QTOF / MS, the data was imported into the software for OPLS-DA analysis; the OPLS-DA analysis of the data before and after administration could distinguish endogenous substances from exogenous substances; based on the results of the previous characterization of the decoction components, phase I and phase II metabolic formulas were established based on metabolic reactions. Identification of metabolite structures: The similarity values of the in vivo metabolites of Santali Albi Radix and their matching prototype compounds were arranged, and the structures of the metabolites were identified by combining the structures of the corresponding prototype components and the analysis of characteristic fragments.

[0072] Example 5

[0073] Refer to Figures 12 - 15 , to explore the changes in blood drug concentration, efficacy, and safety when Santali Albi Radix capsules are combined with paroxetine in the treatment of depression patients.

[0074] Recruit 70 patients aged 18 - 65 years old who met the diagnostic criteria for depression according to the 10th edition of the International Classification of Diseases (ICD - 10) in Inner Mongolia Mental Health Center from December 2022 to March 2024. They were divided into an experimental group using paroxetine combined with Sanwei Tanxiang Capsule and a control group using paroxetine alone according to the random number table method. The concentration of paroxetine was measured at the 2nd, 4th, and 8th weeks of treatment in both groups. The 24 - item Hamilton Rating Scale for Depression (HAMD - 24) and Montgomery and Asberg Depression Rating Scale (MADRS) were used to evaluate the depressive symptoms of patients at baseline and the 2nd, 4th, and 8th weeks of treatment. The Treatment Emergent Symptom Scale (TESS) was used to evaluate the safety of the drugs after treatment. The scores and reduction rates of HAMD - 24 and the scores of MADRS were used as the main evaluation indicators, and adverse reactions were used as the secondary evaluation indicators.

[0075] Bivariate correlation analysis was used to evaluate the effect of Sanwei Tanxiang Capsule on the blood drug concentration of paroxetine, and independent - sample t - test, sample t - test analysis, etc. were used to compare the efficacy differences. At the 4th week, the blood drug concentration of the control group was higher than that of the experimental group, and the difference was statistically significant (P < 0.05). There was no statistically significant difference in the baseline HAMD - 24 and MADRS scores between the two groups (P > 0.05). At the 2nd, 4th, and 8th weeks, the HAMD scores of the experimental group were lower than those of the control group, and the differences were statistically significant (P < 0.05). The reduction rate of HAMD in the experimental group was higher than that in the control group, and the difference was statistically significant (P < 0.05). At the 8th week, the MADRS score of the experimental group was lower than that of the control group, and the difference was statistically significant (P < 0.05). Long - term observation found that the drug side effects in the experimental group decreased over time, and the side effects were milder in the experimental group than in the control group in the TESS assessment.

[0076] Combining Sanwei Tanxiang with paroxetine can maintain the blood drug concentration of paroxetine within an optimal effective range. And compared with using paroxetine alone in the treatment of depression, it can more effectively relieve depressive symptoms. This combination therapy also reduces adverse reactions and enhances the safety of treatment.

[0077] Example Six

[0078] The treatment results of the traditional Chinese medicine preparation of the present invention are as follows:

[0079] Typical case: 1

[0080] Wang Moumou, female, 57 years old, with poor sleep, low mood, restlessness, anxiety, and general discomfort; HMAD score: 31 points.

[0081] Treatment method: Santalum album L. Capsule 0.6g orally twice a day, Paroxetine Hydrochloride 25mg starting from daily and gradually increasing the dose to 50mg orally daily, for 14 days of treatment. Palpitation disappeared, sleep improved, mood relieved, HAMD score: 24 points; on the 30th day, the patient's mood was basically fine, physical discomfort disappeared, HAMD score: 14 points.

[0082] Typical case: 2

[0083] Gao Moumou, female, 41 years old, with poor sleep, restlessness, anxiety, palpitation, fatigue, listlessness, hot and cold sensations on the body, HMAD score: 31 points;

[0084] Treatment method: Santalum album L. Capsule 0.6g orally twice a day, Paroxetine Hydrochloride 20mg starting from daily and gradually increasing the dose to 40mg orally daily, for 14 days of treatment. Symptoms such as palpitation and restlessness disappeared, sleep improved, HAMD score: 28 points; on the 30th day, the patient's condition basically returned to normal, HAMD score: 14 points.

[0085] Typical case: 3

[0086] Zhang Moumou, female, 56 years old, with low mood, restlessness, anxiety, fidgeting, palpitation, and repeated general discomfort, HMAD score: 33 points;

[0087] Treatment method: Santalum album L. Capsule 0.6g orally twice a day, Paroxetine Hydrochloride 20mg starting from daily and gradually increasing the dose to 40mg orally daily, for 14 days of treatment. Palpitation disappeared, HAMD score: 30 points; on the 30th day, the patient's mood basically returned to normal, and general discomfort was basically relieved, HAMD score: 20 points.

[0088] Typical case: 4

[0089] Xu Moumou, female, 56 years old, with intermittent low mood, restlessness, anxiety, and a sense of suicidal ideation, palpitations and sweating in the morning, hot and cold sensations on the body, HMAD score: 30 points;

[0090] Treatment method: Santalum album L. Capsule 0.6g orally twice a day, Paroxetine Hydrochloride 25mg starting from daily and gradually increasing the dose to 50mg orally daily, for 14 days of treatment. Restlessness and suicidal ideation decreased compared to before, mood relieved, HAMD score: 22 points; on the 30th day, the patient's mood basically returned to normal, and physical discomfort symptoms such as palpitation disappeared, HAMD score: 12 points.

[0091] Typical case: 5

[0092] Ge Moumou, female, 42 years old, with poor sleep, low mood, anxiety, self-abasement and self-blame, reduced interest, accompanied by headache and head fullness, HMAD score: 35 points;

[0093] Treatment method: Santalum album capsule 0.6g orally twice a day, paroxetine hydrochloride 25mg starting from daily and gradually increasing the dose to 50mg orally daily, treated for 14 days, the symptoms of headache and head fullness disappeared, sleep improved compared with before, mood relieved, HAMD score: 24 points; On the 30th day, the patient's condition basically returned to normal, HAMD score: 16 points.

[0094] Typical case: 6

[0095] Pa Moumou, female, 65 years old, with poor sleep, low mood, anxiety, slow brain response, inattentiveness, few words, little activity, feeling unwell all over, HMAD score: 32 points.

[0096] Treatment method: Santalum album capsule 0.6g orally twice a day, paroxetine hydrochloride 20mg starting from daily and gradually increasing the dose to 40mg orally daily, treated for 14 days, sleep improved, mood and physical discomfort relieved compared with before, HAMD score: 27 points; On the 30th day, the patient's anxiety and physical discomfort disappeared, mood returned to normal, HAMD score: 15 points.

[0097] The examples involved in this application are only illustrative, used to explain some features of the method described in the present invention. The appended claims are intended to claim the broadest possible scope that can be conceived, and the embodiments presented in this application are only illustrative of the selected embodiments according to the combination of all possible embodiments. Therefore, the applicant's intention is that the appended claims are not limited by the selection of examples illustrating the features of the present invention. Some numerical ranges used in the claims also include sub-ranges within them, and changes within these ranges should also be interpreted as being covered by the appended claims where possible.

Claims

1. An experimental method for the Three-Flavour Santalum Album Capsule based on the material basis, characterized in that, It includes the following steps: S1. Select 72 C57 experimental mice and test them to leave the mice that meet the experiment requirements; S2. Select the modeling method of subcutaneous injection of corticosterone in the neck; S3. All the modeled experimental mice are subcutaneously injected with corticosterone continuously every day between 10:00 and 12:00 for 5 weeks; S4. During the 5 weeks of subcutaneous injection of corticosterone, the mice treated with the vehicle are used as the control group, and a behavioral experiment is carried out at the end of the second week to determine whether depressive-like behaviors occur; S5. To determine whether corticosterone can induce depressive-like behaviors in mice, the open field test (OFT), tail suspension test (TST) and sucrose preference test (SPT) of mice are carried out.

2. The experimental method of the Santalum album L. capsule based on the material basis according to claim 1, wherein S11. Perform a baseline sucrose preference test on 72 C57 experimental mice.

3. The experimental method of the Santalum album L. three - flavor capsule based on the material basis according to claim 1, characterized in that, S12. After the test, remove the mice with high-level differences and leave the mice available for experiments.

4. A method for the experiment of the Three-Flavour Santalum Album Capsule based on the material basis according to claim 1, characterized in that, S21. Corticosterone at 20 mg / kg is dissolved in 2% DMSO + 40% PEG300 + 5% Tween-80 + 53% Saline.

5. A method for the experiment of the Three-Flavour Sandalwood Capsule based on the material basis according to claim 1, characterized in that, S41. After determining the occurrence of depressive-like behaviors, the mice are randomly assigned to a blank control group, a model group, a low-dose galangin group (0.5625 mg / kg), a medium-dose galangin group (1.125 mg / kg), a high-dose santalum album group (2.25 mg / kg) and a positive drug fluoxetine group (30 mg / kg), with 12 mice in each group.

6. The experimental method of the Santalum album L. three - flavor capsule based on material basis according to claim 5, characterized in that, S42. Administer galangin and fluoxetine treatments at the beginning of the third week; S43. After the end of the fifth week, perform a behavioral test again to evaluate the depressive-like behaviors of the mice.

7. An experimental method for the Three-Flavour Sandalwood Capsule based on the material basis according to claim 1, characterized in that, S51. In the open field test (OFT) of mice, compared with the control mice, the total distance moved by the corticosterone group is reduced, and the galangin group can reverse this behavior and significantly increase the movement distance of the mice.

8. An experimental method for the Three-Flavour Sandalwood Capsule based on the material basis according to claim 7, characterized in that, S52. Compared with the control mice, the mice treated with corticosterone show an increase in the duration of immobility in the tail suspension test (TST), and the immobility time of the corticosterone mice can be significantly reduced after administration of santalum album.

9. The experimental method of the Santalum album capsule with three flavors based on the material basis according to claim 8, characterized in that S53. In the sucrose preference test (SPT), compared with the control group, the preference for sucrose in the corticosterone group is significantly decreased, and the preference for sucrose in the mice can be significantly increased after treatment with galangin.

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