Application of inula flower lactone HA in preparation of medicine for preventing or treating depression

Abstract

The invention belongs to the technical field of medicines, and particularly relates to application of inula flower lactone HA in preparation of a medicine for preventing or treating depression, and the structure of the inula flower lactone HA is as shown in a formula I. A chronic social contusion stress induced model mouse test proves that the inula flower lactone HA has an obvious improvement effect on depression-like behaviors and has anti-depression activity, and a thought and a scheme are provided for developing the compound into an anti-depression drug in the future. And the formula I of the # imgabs0 # is shown in the specification.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and specifically relates to the application of inula lactone HA in the preparation of a drug for preventing or treating depression. Background Art

[0002] Depression, also known as depressive disorder, is a mental disorder with high morbidity, high clinical cure rate, low treatment acceptance rate and high recurrence rate. Its main feature is significant and persistent low mood. Some patients may have self-harm, suicidal behavior, and may even be accompanied by psychotic symptoms such as delusions and hallucinations. Clinically, depressive disorders can be divided into mild, moderate and severe according to the number, type and severity of symptoms. They are respectively targeted at different groups such as the elderly, children, and pregnant women, and are called elderly depression, children's depression, postpartum depression, etc. The treatment of depression mainly includes drug therapy, psychotherapy and physical therapy. Drug therapy is the main treatment method. Research on depression is ongoing, and its exact cause and pathogenesis are still unclear.

[0003] Inula japonica is a plant of the genus Inula in the Asteraceae family. It is traditionally used to reduce qi, eliminate phlegm, promote fluid secretion and stop vomiting. Currently, a variety of active ingredients including 1-O-acetylbritannilactone (ABL) and britannilactone (BL) have been extracted from Inula japonica. Xing P et al. found that the sesquiterpene 1β-hydroxyalantolactone from Inula britannica in Yuncheng, Shanxi Province has the strongest cytotoxic effect on HEp2 (laryngeal cancer), SGC-7901 (gastric cancer) and HCT116 (colorectal cancer) cells. Its cytotoxic effect is equivalent to that of the positive control etoposide, and the killing effect on HEp2 cells is achieved through the pro-apoptotic mechanism related to the cleavage of proaspartase-3 and PARP; the study showed that the compound 1β-hydroxyalantolactone has potential anticancer activity against HEp2 cells (Xiang P, Guo X, Han YY, et al. Cytotoxic and Pro-apoptotic Activities of Sesquiterpene Lactones from [J]. Natural Product Communications, 2016, 11 (1): 7-10). In 2019, Zhou Qiang et al. found that 1,6-O,O-diacetyl inula lactone (OODBL) can promote apoptosis of HepG2 cells and block the cells in the G2-M phase; among them, the Bcl-2 family is involved in the apoptosis process of HepG2 cells through Bcl-2 phosphorylation and endoplasmic reticulum stress (Zhou Qiang. Study on the pharmacokinetics of 1,6-O,O-diacetyl inula lactone and its mechanism of inducing apoptosis of HepG2 liver cancer cells [J]. 2019).

[0004] However, there is no report on the application of HA component of Inula japonica to the preparation of drugs for preventing or treating depression. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide an application of inula lactone HA in the preparation of a drug for preventing or treating depression.

[0006] To solve the above problems, the technical solution adopted by the present invention is: Use of inula lactone HA, and its pharmaceutically acceptable salts or isomers in the preparation of drugs for preventing or treating depression, wherein the structural formula of the inula lactone HA is as shown in Formula I, Formula I.

[0007] As a further improvement of the present invention, the depression includes major depression, bipolar depression, psychotic depression, reactive depression, menopausal depression, secondary depression, postpartum depression and / or seasonal depression.

[0008] The beneficial effects of adopting the above technical solution are: The present invention uses a mouse model experiment induced by chronic social frustration (CSDS) stress to show that the model mice treated with HA inula lactone have significantly improved depressive-like behaviors in behavioral experiments compared with the model mice in the negative control group. The model mice treated with HA inula lactone have significantly higher activity levels and responses to pleasant stimuli in forced swimming experiments and sugar water experiments than the mice in the negative control group. This indicates that HA inula lactone has a significant improvement effect on depressive-like behaviors and has antidepressant activity, which provides new ideas and new solutions for the preparation and research of antidepressant drugs today and has important practical value and research significance. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 It is an experimental flow chart of Example 1 of the present invention; Figure 2 is a graph showing weight changes of mice in Example 1 of the present invention before and after chronic social frustration; Figure 3 This is a graph showing the results of an open field test of the effect of inula lactone HA on the behavior of model mice in Example 1 of the present invention; Figure 4 This is a diagram showing the results of a forced swimming test of the behavioral effects of inula lactone HA on model mice in Example 1 of the present invention; Figure 5 This is a graph showing the results of a sucrose preference experiment of the behavioral effects of Inula lactone HA on model mice in Example 1 of the present invention ( # P<0.05). DETAILED DESCRIPTION

[0010] Terms and Definitions The terms used in the description of the present invention herein are only for describing specific embodiments and are not intended to limit the present invention. The nomenclature used herein and the laboratory operations in the organic chemistry, medicinal chemistry, and biology described herein are well known and commonly used in the art. Unless otherwise mentioned, all technical and scientific terms used herein have the same meaning as those generally understood by those skilled in the art to which this art belongs.

[0011] It should be noted that for the structural formula and chemical formula description in the embodiments or implementation schemes of the present invention, the present invention is intended to cover all replacements, modifications and equivalent technical solutions, which should be deemed to be included in the scope of the present invention. It should be understood by those skilled in the art that many similar or equivalent methods and materials can be used to implement the present invention, so the present invention is not limited to the specific methods and materials described. In the event that the cited literature, patents or similar materials are different or contradictory from the present application (including but not limited to term definitions, term applications, described technologies, etc.), the content of the present invention shall prevail.

[0012] As used in the description of the embodiments of the present invention and the appended claims, the singular forms "the", "the" and "its" refer to both the singular and the plural, unless the context clearly dictates otherwise. For example, a compound includes one or more than one compound.

[0013] The term "treatment" generally refers to the use of drugs to improve, alleviate or cure the state of the disease, and refers to partial or complete relief, improvement, delay of its onset, inhibition of its progression, reduction of its severity and / or reduction of the incidence of one or more symptoms or characteristics. Specific infections, diseases, disorders and / or conditions. For example, "treating" cancer can refer to inhibiting the survival, growth and / or spread of tumors. In order to reduce risk, subjects who do not show diseases, disorders and / or conditions and / or subjects who only show early signs of diseases, disorders and / or conditions can be treated. Pathological development conditions associated with diseases, disorders and / or conditions.

[0014] As used herein, "pharmaceutically acceptable salts" refer to salts that retain the desired biological activity of the target compound and exhibit minimal undesirable toxicological effects. When the compounds of the present invention contain relatively acidic functional groups, base addition salts can be obtained by contacting the neutral form of the compounds of the present invention with a sufficient amount of base in a pure solution or a suitable inert solvent. Pharmaceutically acceptable bases include salts prepared from inorganic bases and organic bases, and the salts of the inorganic bases include aluminum salts, ammonium salts, calcium salts, copper salts, iron salts, ferrous salts, lithium salts, magnesium salts, manganic salts, manganous salts, potassium salts, sodium salts, zinc salts, etc. The salts of the organic non-toxic bases include salts of primary amines, secondary amines and tertiary amines, including substituted amines and cyclic amines. For example: N,N'-dibenzylethylenediamine, diethylamine, 2-diethylaminoethanol, 2-dimethylaminoethanol, aminoethanol, ethanolamine, ethylenediamine, N-ethylmorpholine, N-ethylpiperidine, glucosamine, glucosamine, histidine, hydroxocobalamin, isopropylamine, lysine, methylglucosamine, morpholine, piperazine, piperidine, guanidine, polyamine resin, procaine, purine, theobromine, triethylamine, trimethylamine, tripropylamine, etc. When the compounds of the present invention contain relatively basic functional groups, acid addition salts can be obtained by contacting the neutral form of such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent. Examples of pharmaceutically acceptable acid addition salts include inorganic acid salts, such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid or nitric acid, and the like; organic acids such as formic acid, acetic acid, acetoacetic acid, pyruvic acid, trifluoroacetic acid, propionic acid, butyric acid, hexanoic acid, heptanoic acid, undecanoic acid, lauric acid, benzoic acid, salicylic acid, 2-(4-hydroxybenzoyl)-benzoic acid, camphoric acid, cinnamic acid, cyclopentanepropionic acid, digluconic acid, 3-hydroxy-2-naphthoic acid, nicotinic acid, pamoic acid, pectinic acid, 3-phenylpropionic acid, picric acid, pivalic acid, 2-hydroxyethanesulfonic acid, itaconic acid, aminosulfonic acid, trifluoromethanesulfonic acid, dodecylsulfuric acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, naphthalene disulfonic acid, camphorsulfonic acid, citric acid, tartaric acid, stearic acid, lactic acid, oxalic acid, malonic acid, succinic acid, malic acid, adipic acid, alginic acid, maleic acid, fumaric acid, D-gluconic acid, mandelic acid, ascorbic acid, glucoheptonic acid, glycerophosphate, aspartic acid, sulfosalicylic acid, etc., including salts formed by sodium, potassium, magnesium, lithium, aluminum, calcium, zinc, N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, N-methylglucamine and procaine.

[0015] As used herein, "isomers" refers to the ability of compounds of the invention to exist as racemates, racemic mixtures, single enantiomers, diastereomeric mixtures, single diastereomers, geometric isomers, etc., where the compounds of Formula I contain one or more asymmetric centers and / or double bonds. These compounds may be represented by the symbols "R" or "S," depending on the configuration of substituents around the stereogenic carbon atom, and may also be represented by the symbols "Z" or "E," depending on the arrangement of substituents around the carbon-carbon double bond, or the substituents around the carbon-carbon double bond may be referred to as "cis" or "trans." The compounds disclosed herein may exist as tautomers, and both tautomeric forms are intended to be included within the scope of the invention, even if only one tautomeric structure is depicted, such as keto-enol tautomers, phenol-keto tautomers, nitroso-oxime tautomers, imine-enamine tautomers, etc. In the present invention, the compounds and stereoisomers thereof described are within the scope of protection, including but not limited to enantiomers, diastereomers and cis-trans isomers. Therefore, the compounds protected by the present invention include not only all possible single stereoisomers, but also optically active mixtures and racemates thereof. In addition, it should be understood by those skilled in the art that different stereoisomers may have significant differences in biological activity, efficacy and toxicity, so selecting a specific stereoisomer or a combination thereof may optimize the therapeutic effect. Stereoisomers can be obtained by known chemical or physical methods, such as chiral catalysis, chiral synthesis or chiral separation by chromatography or chemical methods. The present invention also includes stereoisomers obtained by these methods and pharmaceutically acceptable salts thereof.

[0016] In order to make the objectives, technical solutions and advantages of the present invention more clear, the present invention is clearly and completely described below in conjunction with specific embodiments.

[0017] Preparation Example 1 Preparation of Inula lactone HA (5β-hydroxyasperilin (HA)) Inula lactone HA is a monomer separated and extracted from Inula japonica, and its purity has been identified to be 98%.

[0018] (1) Take 7.5 kg of crushed Inula japonica, extract with 95% ethanol for 7 days, repeat the extraction 3 times, and combine the extracts; (2) filtering the obtained extract, and concentrating the filtrate under reduced pressure to a paste to obtain an inula lactone HA extract; (3) suspending the extract in water, extracting with petroleum ether, dichloromethane and ethyl acetate respectively, and concentrating the extract under reduced pressure to obtain a petroleum ether extract, a dichloromethane extract and an ethyl acetate extract; (4) The dichloromethane extract was separated by silica gel column chromatography, dextran gel, preparative thin layer chromatography and preparative high performance liquid chromatography, and the obtained HA monomer compound was a light yellow oil.

[0019] (5) Adoption 1 HNMR and 13 CNMR was used to identify the structure of the compound.

[0020] The data of its NMR spectrum are as follows: 1 H-NMR(600MHz,Methanol-d4)δ:6.22(d,J=3.7Hz,1H),5.77(d,J=3.3Hz,1H),5.10(t,J=1.4Hz,1H), 4.94(ddd,J=11.3,8.6,5.3Hz,2H),3.85(dd,J=11.3,4.5Hz,1H),3.30(m,1H),2.48(dd,J=15.7,7.8H z,1H),2.40-2.34(m,1H),2.32(ddt,J=14.4,5.3,1.7Hz,1H),2.11(dd,J=13.3,5.8Hz,1H),1.95(dd, J=15.7,1.8Hz,1H),1.78(dq,J=12.7,4.3Hz,1H),1.61-1.51(m,1H),1.50-1.43(m,1H),0.77(s,3H); 13 C-NMR(151MHz,Methanol-d4)δ:173.15(C-12),151.69(C-4),140.12(C-11),121.79(C-13),110.31(C-15),77.37(C-8) ,76.75(C-5),70.36(C-1),45.86(C-10),38.77(C-7),34.93(C-9),33.57(C-6),31.40(C-2),30.68(C-3),15.25(C-14).

[0021] Example 1 1. Construction of the mouse CSDS model 1. Experimental animals: (1) Twenty 8-week-old SPF male C57BL / 6J mice, 20 10-week-old male CD1 mice, and 20 10-week-old female CD1 mice were selected. After adaptive training, baseline tests of sugar water and body weight were performed to exclude mice that were naturally too small or had no preference for sugar water.

[0022] (2) Screening of highly aggressive CD1 mice: One female CD1 mouse and one male CD1 mouse were placed in each cage and raised at a temperature of 22±1°C, a humidity of 60%, and a 12h light / 12h dark cycle (lights turned off at 7:00 am). Food and water were freely available. After 7 days of breeding, the male mice developed territorial awareness and separated the female mice from the cages. A strange C57BL / 6J male mouse was placed in each cage as a territorial intruder. For the screening of a single highly aggressive CD1 mouse: a male CD1 mouse attacked a C57BL / 6J male mouse within 1 min after the C57BL / 6J male mouse was placed in the cage, and the male CD1 mouse attacked the C57BL / 6J male mouse no less than 5 times every 3 minutes. This male CD1 mouse was considered to be a highly aggressive CD1 mouse. 2. Establishment of the Chronic Social Defeat Stress (CSDS) Model in Mice: (1) Model construction: 18 C57BL / 6J mice were randomly divided into a normal group and a model group; Normal group: 6 C57BL / 6J mice, each individually housed in a cage, housed under the conditions of 22±1°C, 60% humidity, and 12 h light / 12 h dark cycle (lights off at 7:00 a.m.), free access to food and water, and conventional feeding for 10 d; Model group: 12 C57BL / 6J mice, each of which was raised individually in a cage at a temperature of 22±1℃, a humidity of 60%, and a 12h light / 12h dark cycle (lights turned off at 7:00 am), with free access to food and water; every day, the model group mice were placed in the cage of aggressive CD1 mice, and the two mice were allowed to engage in 10-minute social confrontation, and then the model group mice were taken out and put back for feeding, and the feeding was repeated for 10 days; (2) Social interaction experiment: On day 10, C57BL / 6J mice were placed in the behavior chamber to acclimate for approximately 3 h before testing; Behavioral detection in the absence of a social target: a transparent plastic cage (10 cm × 10 cm × 20 cm) was placed in the middle of one side of an open field (50 cm × 50 cm × 50 cm), and a C57BL / 6J mouse to be tested was placed in a corner of the open field; the behavioral characteristics of the mouse were monitored, recorded and analyzed for 2.5 minutes as a baseline for exploratory behavior in the absence of a social target, after which the C57BL / 6J mouse was moved back to its original cage; Behavioral testing in the presence of a social target: After placing an unfamiliar CD-1 mouse in a transparent plastic cage, the same C57BL / 6J mouse was placed in the same corner in the open field for 2.5 minutes of monitoring and recording. After cleaning the open field with 75% alcohol, the next C57BL / 6J mouse was tested; The experimenter remained quiet during the whole process. When changing mice, he / she should be gentle to avoid over-stimulating the mice and causing stress. During the video recording, the partition was pulled up to put the mice in a relatively independent environment to avoid external interference. The head-body-tail three-point method was used to analyze the movement trajectory of the mice. The open field environment was divided into 10×10 squares, and the social area was 20cm×15cm above. The activity time of the mice in this area was the social time. After the experiment, the social experimental platform was cleaned up.

[0023] By comparing and calculating the SIR value, we can judge whether the model group is depressed and whether the model is successfully established. If the SIR value is less than 0.1, the model is successfully established. SIR = cumulative time with strangers in the social area / cumulative time without strangers in the social area.

[0024] 2. Effect of 5β-hydroxyasperilin (HA) on CSDS model mice 1. The CSDS model mice obtained in Example 1 were randomly divided into two groups: HA group (n=6) and negative control group (Neg group, n=6); normal C57BL / 6J male mice were selected as blank control group (Con group, n=6).

[0025] 2. Inula lactone HA intragastric solution: Prepare it before use. Add the Inula lactone HA in Preparation Example 1 into normal saline to prepare an Inula lactone HA intragastric solution with a concentration of 1 mg / ml; 3. Dosage regimen: HA group: On days 11-14, the CSDS model mice obtained in Example 1 were intragastrically administered with 0.1 ml / g of 1 mg / ml HA solution of inula lactone every 24 hours; the mice were raised under conditions of 22±1°C, 60% humidity, and 12h light / 12h dark cycle (lights turned off at 7:00 am), with free access to food and water; Negative control group: On days 11-14, the CSDS model mice obtained in Example 1 were gavaged with 0.1 ml / g of normal saline every 24 h; they were raised under the conditions of temperature 22±1°C, humidity 60%, and 12 h light / 12 h dark cycle (lights turned off at 7:00 a.m.), with free access to food and water; Blank control group: On days 11-14, normal C57BL / 6J male mice were gavaged with 0.1 ml / g of normal saline every 24 h; they were housed at 22±1°C, 60% humidity, and a 12 h light / 12 h dark cycle (lights turned off at 7:00 am), with free access to food and water.

[0026] 4. Open field test (OFT): On the 12th day, 60 minutes after gavage with inula lactone HA solution, 6 mice in each of the HA group, negative control group and blank control group were placed into the white open field one by one. The total distance moved by each mouse in the white open field within 60 minutes was recorded, and the average movement distance per 5 minutes was calculated to evaluate the mice's exercise capacity. The experimental results are as follows Figure 3 As shown, the amount of exercise of CSDS model mice in the HA group treated with inula lactone HA had no statistical difference with that of normal C57BL / 6J male mice, but was higher than that of CSDS model mice in the negative control group that were not treated with inula lactone HA.

[0027] 5. Forced swimming test (FST): On the 13th day, 60 minutes after gavage with HA solution, 2 / 3 of the volume of water was poured into a cylindrical transparent container with a diameter of 10 cm and a height of 25 cm. The water temperature was (23±1)℃. Six mice in each of the HA group, negative control group and blank control group were placed in the cylindrical transparent container for 6 minutes, and the cumulative immobility time of the mice was recorded for 2-6 minutes. The experimental results are as follows Figure 4 As shown, the swimming and activity time of the CSDS model mice in the HA group had no statistical difference with that of the normal group mice, but was significantly higher than that of the CSDS model mice in the negative control group that were not treated with inula lactone HA. This result indicates that the compound inula lactone HA has a significant improvement effect on CSDS-induced depressive-like behavior and has antidepressant activity.

[0028] 6. Sugar Preference Test (SPT): (1) Adaptation phase: On days 12-13, each of the six mice in the HA group, negative control group, and blank control group was fed the same amount of water and 1% sucrose solution using the same container at the same time. The sucrose solution and water were swapped every 12 h. (2) Experimental phase: On the 14th day, the mice in the HA group, negative control group, and blank control group were first prohibited from eating and drinking for 4 hours. The mass of the sucrose solution and normal water before the experiment was weighed and recorded. Then, 1% sucrose solution and normal water were placed in the cage of each mouse at the same time. After 1 hour, the mass of the sucrose solution and normal water after the experiment was weighed and recorded. The amount of sucrose solution and water consumed by each mouse was calculated, and the sucrose preference rate formula was used to determine the mouse's preference for sucrose.

[0029] The experimental results are as follows Figure 5 As shown, the response degree of CSDS model mice in the HA group treated with inula lactone HA to pleasant stimulation was significantly higher than that of CSDS model mice in the negative control group not treated with inula lactone HA. This result indicates that the compound inula lactone HA has a significant improvement effect on CSDS-induced depressive-like behavior and has antidepressant activity.

[0030] Based on the above results, it can be seen that the compound HA has a significant improvement effect on the depressive symptoms of model mice and has antidepressant activity, indicating that HA, an inula lactone, has a positive effect in preventing or treating depressive symptoms and depression.

[0031] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of inula lactone HA, and its pharmaceutically acceptable salts or isomers in the preparation of a drug for preventing or treating depression, characterized in that: The structural formula of the inula lactone HA is shown in Formula I: Formula I.

2. Use of the inula lactone HA, and its pharmaceutically acceptable salts or isomers according to claim 1 in the preparation of a drug for preventing or treating depression, characterized in that: The depression includes major depression, bipolar depression, psychotic depression, reactive depression, menopausal depression, secondary depression, postpartum depression and / or seasonal depression.

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