Substituted trifluoroalkyl thioalkyl benzothioate derivative, and preparation method therefor and use thereof

Abstract

Disclosed in the present invention are a substituted trifluoroalkyl thioalkyl benzothioate derivative, and a preparation method therefor and the use thereof. The derivative has a structure as shown in a formula (I). Further disclosed in the present invention is the preparation method for the above-mentioned substituted trifluoroalkyl thioalkyl benzothioate derivative. The substituted trifluoroalkyl thioalkyl benzothioate derivative of the present invention has anti-inflammatory activity and neuroprotective activity, and can be used in the preparation and application of pharmaceuticals, foods or health care products related to anti-inflammation, neuroprotection, and anti-neurodegenerative diseases. The substituted trifluoroalkyl thioalkyl benzothioate derivative can also delay the aging process, prevent aging-related diseases, also has the effect of increasing the rate and density of hair growth, and thus can be used for developing pharmaceuticals, foods, health care products or personal care products, etc. aimed at anti-aging, and the prevention and / or treatment of alopecia.

Description

Substituted benzoic acid trifluoroalkylthioalkylthioester derivatives and preparation methods and applications thereof Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to substituted benzoic acid trifluoroalkylthioalkylthioester derivatives and preparation methods and applications thereof. Background Art

[0002] Inflammation is a major contributor to many diseases and a key factor in age-related disorders. BV-2 microglia are a common cell model for studying inflammation. Upon brain injury, such as ischemia, microglia are the first to respond and become activated. Activated microglia can differentiate into distinct phenotypes, including the neurodegenerative M1 and neuroprotective M2.

[0003] Lipopolysaccharide (LPS) can induce activation of BV-2 microglia to the M1 phenotype and promote the release of various pro-inflammatory mediators, thereby inducing an inflammatory response. BV-2 microglia require simple culture conditions, can be serially subcultured, are easy to operate, and are relatively inexpensive. By establishing an LPS-induced BV-2 microglia activation model, high-throughput screening can be used to identify lead compounds with anti-inflammatory activity. Furthermore, the brains of mice fed a high-fat diet and normal aging mice are also affected by inflammation, which can lead to various neurological diseases.

[0004] Aging is broadly defined as the time-related functional decline that affects most organisms. Aging increases susceptibility to age-related chronic diseases such as cancer, metabolic, cardiovascular, musculoskeletal, and neurodegenerative diseases. The main cause of aging in organisms is cellular senescence, which is a relatively stable state that cells enter after they irreversibly exit the cell cycle and lose their ability to proliferate. Currently, the causes of cellular senescence are mainly divided into three types: replicative senescence, oncogene-induced premature senescence, and stress-induced premature senescence. Various stress stimuli include oxidative stress, carcinogenic stress, and chemotherapeutic drug toxicity stress. Etoposide, as a chemotherapeutic drug, can inhibit topoisomerase, disrupt the reconnection of DNA supercoils after unwinding, cause DNA damage, and lead to cellular senescence.

[0005] One of the most prominent characteristics of human aging is the graying and loss of hair. The morphology of hair depends on the growth and development of hair follicles. In human adulthood, the cycle of hair follicle replacement is irregular and mainly depends on the proliferation and differentiation of hair follicle stem cells. It is generally believed that under the influence of the aging process, the ability of hair follicle stem cells to proliferate, differentiate and maintain their own stable state is impaired, resulting in a progressive decrease and thinning of the number of hair follicles and hair diameter during the growth phase, and a prolonged resting phase of the hair cycle, resulting in sparse hair. Aging-related graying of hair may be related to melanocyte damage induced by ultraviolet rays and reactive oxygen free radicals (ROS). With age, melanocytes age and melanin production is impaired, leading to the occurrence of age-related gray hair.

[0006] Patent document with publication number CN111777588A discloses a phenylpropanoid compound from Pseudomonas aeruginosa and its application. The invention extracts a new phenylpropanoid compound from Pseudomonas aeruginosa. The new phenylpropanoid compound exhibits an inhibitory effect on the inflammatory mediator NO produced by LPS-induced BV-2 cells, has a significant anti-inflammatory effect, and is non-toxic to cells. It can be used to prepare drugs related to inflammation.

[0007] Patent publication number CN111574581A discloses low-toxicity, anti-inflammatory ursolic acid derivatives, their preparation methods, and applications. This invention uses ursolic acid as a lead compound and undergoes structural modification to produce a new low-toxic, anti-inflammatory ursolic acid derivative. This derivative exhibits significant inhibitory activity against lipopolysaccharide-induced NO release in RAW264.7 cells and exhibits extremely low cytotoxicity.

[0008] Patent publication number CN113952362A discloses the use of inducible extracellular vesicles (ICEVs) in the preparation of a formulation for extending mammalian lifespan or treating or preventing aging. The ICEVs are produced from stem cells or by inducing apoptosis in mesenchymal stem cells through the addition of staurosporine, ultraviolet irradiation, starvation, heat stress, or a combination thereof. These ICEVs exhibit anti-aging effects, can extend mammalian lifespan, and alleviate hair loss in older adults.

[0009] Although the existing technology has conducted a lot of research on anti-inflammatory, anti-aging or anti-hair loss products, it is still of great significance to discover some novel structural types of drugs with significant anti-inflammatory, anti-aging or anti-hair loss effects and good safety. Summary of the Invention

[0010] The first objective of the present invention is to provide a substituted benzoic acid trifluoroalkylthioalkyl thioester derivative. The thioester derivative has very significant anti-inflammatory and neuroprotective effects on BV-2 microglia cells. The anti-inflammatory activity is comparable to that of the positive drug butylphthalide, and the active concentration is much lower than that of the positive drug. It can be used for the preparation and application of anti-inflammatory and neuroprotective drugs.

[0011] A substituted benzoic acid trifluoroalkylthioalkylthioester derivative, whose structure is shown in formula (I):

[0012] Wherein, R1 and R2 are OH, OAc or H, and X and Y are linear or branched alkyl groups having 1 to 9 carbon atoms.

[0013] The present invention also provides a method for preparing the above-mentioned substituted benzoic acid trifluoroalkylthioalkylthioester derivative, comprising the following steps:

[0014] (1) NaH (60% dispersed in mineral oil) is added to a thiol-containing solvent at a temperature below 0°C, and the mixture is heated to room temperature and stirred for 10 to 30 minutes. A bromotrifluoroalkyl compound and TBAI are then added at a temperature below 0°C to react. The reaction endpoint is detected by TLC. After the reaction is complete, the reaction product is extracted to obtain the target product.

[0015] (2) The target product obtained in step (1) is dissolved in an organic solvent, EDC·HCl, substituted benzoic acid and DMAP are added, and the reaction is carried out at room temperature. The reaction end point is detected by TLC. After the reaction is completed, the reaction product is separated and purified to obtain the substituted benzoic acid trifluoroalkylthioalkylthioester derivative.

[0016] Preferably, in step (1), the thiol is a dithiol having 1 to 10 carbon atoms.

[0017] More preferably, the thiol is 1,2-ethanedithiol, 1,4-butanedithiol, 1,6-hexanedithiol, 1,8-octanedithiol or 1,10-decanedithiol.

[0018] Preferably, in step (1), the solvent comprises DMF.

[0019] Preferably, in step (1), the molar ratio of NaH to thiol is 1 to 2:1; the molar ratio of the bromotrifluoroalkyl compound to thiol is 1:1; and the molar ratio of TBAI to thiol is 0.1 to 1:1.

[0020] Preferably, the substituted benzoic acid includes 2,3-dihydroxybenzoic acid, 2,3-diacetoxybenzoic acid, 2-hydroxybenzoic acid, 3-hydroxybenzoic acid, 2-acetoxybenzoic acid or 3-acetoxybenzoic acid.

[0021] Preferably, in step (2), the molar ratio of EDC·HCl to thiol is 1 to 4:1; the molar ratio of substituted benzoic acid to thiol is 1 to 10:1; and the molar ratio of DMAP to thiol is 0.1 to 4:1.

[0022] The present invention also provides the use of the aforementioned substituted benzoic acid trifluoroalkylthioalkyl thioester derivatives in the preparation of drugs, health products, or foods for preventing and / or treating inflammation. The substituted benzoic acid trifluoroalkylthioalkyl thioester derivatives of the present invention exhibit significant anti-inflammatory activity in an in vitro screening model, BV-2 cells, and can reduce the overactivation of microglia and astrocytes. These thioester derivatives can be used as active ingredients, along with pharmaceutically acceptable carriers, diluents, and the like, to prepare drugs for preventing and treating inflammation-related diseases.

[0023] The present invention also provides the use of the aforementioned substituted benzoic acid trifluoroalkylthioalkylthioester derivatives in neuroprotective drugs, health products, or foods. The substituted benzoic acid trifluoroalkylthioalkylthioester derivatives of the present invention have the effect of reducing neuronal damage and death and can be used in the preparation of neuroprotective drugs by adding pharmaceutically acceptable carriers, diluents, etc.

[0024] The present invention also provides the use of the aforementioned substituted benzoic acid trifluoroalkylthioalkylthioester derivatives in the preparation of drugs, health products, or foods for preventing and / or treating neurodegenerative diseases. The substituted benzoic acid trifluoroalkylthioalkylthioester derivatives of the present invention can be used to prevent and / or treat neurodegenerative diseases, particularly Alzheimer's disease, etc.

[0025] The present invention also provides the use of the above-mentioned substituted benzoic acid trifluoroalkylthioalkyl thioester derivatives in the preparation of anti-aging drugs, health products or foods for prevention and / or treatment. The substituted benzoic acid trifluoroalkylthioalkyl thioester derivatives of the present invention have significant activity in alleviating the production of aging markers in etoposide-induced senescent PC12 cells, an in vitro screening model of cell senescence. An effective dose of the substituted benzoic acid trifluoroalkylthioalkyl thioester derivative can be used as an active ingredient, and a pharmaceutically acceptable carrier, diluent, etc. can be added to prepare drugs for preventing and alleviating aging and aging-related diseases.

[0026] The present invention also provides the use of the aforementioned substituted benzoic acid trifluoroalkylthioalkylthioester derivatives in the preparation of pharmaceuticals, health products, or foods for preventing and / or treating hair loss. The substituted benzoic acid trifluoroalkylthioalkylthioester derivatives of the present invention can achieve a hair growth effect similar to that of minoxidil at a dosage several times lower than that of minoxidil. Pharmaceuticals for preventing and / or treating hair loss can be prepared using the substituted benzoic acid trifluoroalkylthioalkylthioester derivatives as active ingredients in combination with pharmaceutically acceptable carriers, diluents, and the like.

[0027] Preferably, the substituted benzoic acid trifluoroalkylthioalkylthioester derivatives, administered via skin application at a dose of 0.25 times that of minoxidil, achieve similar hair growth effects as minoxidil. For example, 0.50% of Compound 3 from the present invention achieved comparable hair growth in length and density to a 2.0% minoxidil dose.

[0028] Preferably, when the substituted benzoic acid trifluoroalkylthioalkyl thioester derivative is used to prevent and / or treat hair loss, the oral dosage of the substituted benzoic acid trifluoroalkylthioalkyl thioester derivative is 1 to 25 mg / kg body weight, where the body weight refers to human body weight.

[0029] The present invention provides a pharmaceutical composition, health product or food for neuroprotection, prevention and / or treatment of inflammation and neurodegenerative diseases. The pharmaceutical composition, health product or food contains the substituted benzoic acid trifluoroalkylthioalkylthioester derivative.

[0030] The present invention also provides an anti-aging and hair loss prevention and / or treatment product, which comprises the substituted benzoic acid trifluoroalkylthioalkylthioester derivative according to claim 1.

[0031] Preferably, the product of the present invention is an anti-aging and hair loss prevention and / or treatment drug, wherein the anti-aging, hair loss prevention and / or treatment drug contains the above-mentioned substituted benzoic acid trifluoroalkylthioalkylthioester derivative as an active ingredient.

[0032] Preferably, the product of the present invention is an anti-aging and hair loss prevention and / or treatment food or health product, wherein the food or health product is composed of the above-mentioned substituted benzoic acid trifluoroalkylthioalkylthioester derivative and a carrier acceptable to the food or health product.

[0033] Preferably, the product of the present invention is a personal care product for preventing and / or treating hair loss, wherein the personal care product is composed of the above-mentioned substituted benzoic acid trifluoroalkylthioalkylthioester derivative and an acceptable carrier in the personal care product.

[0034] Pharmaceutically acceptable carriers refer to conventional pharmaceutical carriers, including fillers such as sucrose, starches, microcrystalline cellulose, and inorganic salts; binders such as cellulose derivatives, starch slurry, povidone, and gelatin; humectants such as distilled water and ethanol; lubricants such as magnesium stearate, micropowdered silica gel, and polyethylene glycols; absorption enhancers such as polysorbate and lecithin; and surfactants such as fatty acids such as sorbitan and poloxamer. Other adjuvants such as sweeteners and flavoring agents may also be added to the pharmaceutical composition.

[0035] The substituted benzoic acid trifluoroalkylthioalkylthioester derivatives of the present invention can be administered in the form of a unit dose, and the administration route is enteral administration or non-enteral administration, including oral administration, intravenous injection, intramuscular injection, subcutaneous injection, skin administration, nasal administration, etc.

[0036] The dosage forms of the drugs of the present invention may be solid preparations, semisolid preparations, liquid preparations, etc., including tablets, pills, powders, dispersible tablets, sachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, soft capsules, hard capsules, sterile injections, liniments, suppositories, etc. The above-mentioned various dosage forms can be prepared by conventional methods, for example, by mixing the active ingredient with one or more carriers and then forming the mixture into the desired dosage form.

[0037] Compared with the prior art, the present invention has at least the following advantages:

[0038] (1) The substituted benzoic acid trifluoroalkylthioalkylthioester derivatives of the present invention contain a thioester group, an alkyl chain sulfur atom and a trifluoromethyl group. The introduction of these groups can increase the stability and metabolic process of the drug in the body and is a candidate compound for improving bioavailability. Among them, the introduction of the trifluoromethyl group is widely used in non-steroidal anti-inflammatory drugs, including the marketed drugs cyclooxygenase (COXs) inhibitors flufenamic acid and celecoxib.

[0039] (2) The substituted benzoic acid trifluoroalkylthioalkylthioester derivatives of the present invention can significantly reduce the LPS-induced BV-2 cell activation ratio at a relatively low concentration (0.03-0.3 μM), and their neuroprotective and anti-inflammatory activities are comparable to those of positive drugs.

[0040] (3) The substituted benzoic acid trifluoroalkylthioalkylthioester derivatives of the present invention can alleviate the oxidative stress level, cell cycle arrest, aging marker SA-β-Gal and lipofuscin production of senescent cells, and are potential anti-aging active compounds that can delay the aging process, prevent aging-related diseases, and prolong the healthy lifespan of living organisms. They have broad application prospects in anti-aging products.

[0041] (4) The substituted benzoic acid trifluoroalkylthioalkylthioate derivatives of the present invention can promote hair growth and increase hair density, and have good application prospects in anti-hair loss drugs, health products and toiletries. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] FIG1 shows the effects of the substituted benzoic acid trifluoroalkylthioalkylthioester derivatives prepared in Examples 1 to 8 on LPS-activated BV-2 microglial cells after 24 hours of action.

[0043] Figure 2 shows the effects of the substituted benzoic acid trifluoroalkylthioalkylthioester derivatives prepared in Example 3 on the activity of microglia and astrocytes in the cerebral cortex and hippocampus of high-fat diet model mice and the expression of inducible nitric oxide synthase (INOS) protein, an inflammation-related indicator, wherein Figures 2A-B show the effects on the activity of microglia in the cerebral cortex and hippocampus; Figures 2C-D show the effects on the activity of astrocytes in the cerebral cortex and hippocampus; and Figures 2E-F show the effects on the expression of INOS protein in the cerebral cortex and hippocampus.

[0044] Figure 3 shows the anti-inflammatory and neuroprotective efficacy evaluation results of the substituted benzoic acid trifluoroalkylthioalkylthioester derivatives prepared in Example 3, wherein Figures 3A-C show the effects on the activity of microglia and astrocytes and the expression of inducible nitric oxide synthase (INOS) protein, an inflammation-related indicator, in the cerebral cortex of naturally aged mice; Figures 3D-E show the effects on the number of mature neurons in the cerebral cortex and hippocampus of naturally aged mice.

[0045] Figure 4 shows the effect of compound 3 on the cognitive function of mice with lipopolysaccharide (LPS)-induced neuroinflammatory disease, wherein Figures 4A-B show the effect of compound 3 on the alternating arm entry rate and total number of arm entries in the Y-maze of mice with neuroinflammatory disease, Figures 4C-D show the effect of compound 3 on target recognition and discrimination indices during the training and testing phases of the novel object recognition test in mice with neuroinflammatory disease, and Figures 4E-G show the effect of compound 3 on the escape latency and number of platform crossings during the training and testing phases of the water maze test in mice with neuroinflammatory disease.

[0046] FIG5 shows the effects of compounds 1 to 5 on etoposide-induced senescent PC12 cells.

[0047] Figure 6 shows the effects of compound 3 on the cell survival rate, oxidative stress level, cell proliferation level, and senescence marker - lipofuscin of etoposide-induced senescent PC12 cells, wherein Figure 6A shows the effect on the cell survival rate of etoposide-induced senescent PC12 cells, Figure 6B shows the effect on the reactive oxygen species level of PC12 cells after etoposide treatment, Figure 6C shows the effect on the proportion of proliferating cells with EdU incorporation, and Figure 6D shows the effect on the production of lipofuscin, a senescence marker induced by etoposide.

[0048] Figure 7 shows the effects of compound 3 on the hair loss area, hair growth length and hair growth density of aged mice, wherein Figure 7A shows the effect on the percentage of hair loss area in aged mice, Figure 7B shows the effect on the hair length of hair loss rats, and Figure 7C shows the effect on the hair growth density of hair loss rats.

[0049] FIG8 shows the effects of compounds 1 and 5 on the length and density of hair growth in rats, wherein FIG8A shows the effect on the hair length of depilated rats, and FIG8B shows the effect on the relative hair growth density of rats.

[0050] Figure 9 shows the effects of compounds 1, 2, 4 and 5 on the hair growth length and hair growth density of mice, wherein Figure 9A shows the effect on the relative growth density of mouse hair, and Figure 9B shows the effect on the hair growth length of mice. DETAILED DESCRIPTION

[0051] The above contents of the present invention are further described in detail below in conjunction with the accompanying drawings and embodiments, but this should not be understood as limiting the scope of the above subject matter of the present invention to the following embodiments. All technologies implemented based on the above contents of the present invention fall within the scope of the present invention.

[0052] Example 1

[0053] The preparation method of the substituted benzoic acid trifluoroalkylthioalkylthioester derivative in this embodiment is as follows:

[0054] (1) 1,2-Ethanedithiol (188.0 mg, 2.0 mmol) was dissolved in 10 ml of dry anhydrous DMF. NaH (60% dispersion in mineral oil, 96.0 mg, 2.4 mmol) was added at 0°C. After warming to room temperature and stirring for 30 min, 1-bromo-4,4,4-trifluorobutane (0.3 mL, 2.0 mmol) and TBAI (74.0 mg, 0.2 mmol) were added at 0°C and allowed to react overnight. The end point of the reaction was detected by TLC (n-hexane:ethyl acetate = 20:1). After the reaction was completed, ethyl acetate was added to dilute the mixture, and the mixture was washed and extracted with 1N HCl solution, water, saturated sodium bicarbonate solution, and NaCl solution. The resulting organic phase was dried over sodium sulfate, filtered, and then concentrated.

[0055] (2) The product obtained in step (1) was dissolved in 10 ml of dry dichloromethane, and EDC·HCl (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride, 382.0 mg, 2.0 mmol), 2,3-dihydroxybenzoic acid (154.0 mg, 1.0 mmol) and DMAP (4-dimethylaminopyridine, 122.0 mg, 1.0 mmol) were added. The reaction was allowed to react at room temperature overnight. The end point of the reaction was detected by TLC (n-hexane:ethyl acetate = 5:1). After the reaction was completed, the product was washed with 1N HCl solution, water, saturated sodium bicarbonate solution and NaCl solution, respectively, and extracted. The organic layer was dried and concentrated, and purified by silica gel column chromatography (n-hexane:ethyl acetate = 80:1). It was then purified by ODS open column chromatography (methanol:water = 75:25) to obtain compound 1.

[0056] Analysis of HR ESI-MS and 1 The structure of compound 1 was confirmed by H NMR data, HR ESI-TOF-MS m / z 363.0316, calculated for C 13 H15 F3O3S2Na(M+Na) + 363.0307. 1 H NMR (500 MHz, CDCl3): δ = 11.08 (1H, s), 7.39 (1H, dd, J = 1.3, 8.0 Hz), 7.12 (1H, dd, J = 1.3, 8.0 Hz), 6.83 (1H, t, J = 8.0 Hz), 5.69 (1H, s), 3.27 (2H, m), 2.79 (2H, m), 2.71 (2H, t, J = 7.1 Hz), 2.25 (2H, m), 1.93 (2H, m). The structure of the obtained compound 1 is shown below:

[0057] Example 2

[0058] In this example, 1,4-butanedithiol (244.0 mg, 2.0 mmol) was used to prepare a substituted benzoic acid trifluoroalkylthioalkylthioester derivative. The rest was the same as in Example 1. The obtained product was recorded as Compound 2.

[0059] Analysis of HR ESI-MS and 1 The structure of compound 2 was confirmed by H NMR data, HR ESI-TOF-MS m / z 391.0625, calculated for C 15 H 19 F3O3S2Na(M+Na) + 391.0620. 1 H NMR (500 MHz, CDCl3): δ = 11.19 (1H, s), 7.41 (1H, dd, J = 1.4, 8.0 Hz), 7.11 (1H, dd, J = 1.4, 8.0 Hz), 6.82 (1H, t, J = 8.0 Hz), 5.69 (1H, s), 3.09 (2H, t, J = 7.1 Hz), 2.57 (4H, m), 2.22 (2H, m), 1.83 (4H, m), 1.73 (2H, m). The structure of the obtained compound 2 is shown below:

[0060] Example 3

[0061] In this example, 1,6-hexanedithiol (301.0 mg, 2.0 mmol) was used to prepare a substituted benzoic acid trifluoroalkylthioalkylthioester derivative. The eluent used for its ODS open column purification was MeOH / H2O with a volume ratio of 80:20. The rest was the same as in Example 1. The obtained product was recorded as Compound 3.

[0062] Analysis of HR ESI-MS and 1The structure of compound 3 was confirmed by H NMR data, HR ESI-TOF-MS m / z 419.0933, calculated for C 17 H 23 F3O3S2Na(M+Na) + 419.0933. 1 H NMR (500 MHz, CDCl3): δ = 11.24 (1H, s), 7.41 (1H, dd, J = 1.4, 8.0 Hz), 7.11 (1H, dd, J = 1.4, 8.0 Hz), 6.82 (1H, t, J = 8.0 Hz), 5.67 (1H, s), 3.07 (2H, t, J = 7.3 Hz), 2.50-2.59 (4H, m), 2.22 (2H, m), 1.85 (2H, m), 1.69 (2H, m), 1.61 (2H, m), 1.45 (4H, m). The structure of the obtained compound 3 is shown below:

[0063] Example 4

[0064] In this example, 1,8-octanedithiol (357.0 mg, 2.0 mmol) was used to prepare a substituted benzoic acid trifluoroalkylthioalkylthioester derivative. The rest was the same as in Example 3. The obtained product was recorded as Compound 4.

[0065] Analysis of HR ESI-MS and 1 The structure of compound 4 was confirmed by H NMR data, HR ESI-TOF-MS m / z 447.1257, calculated for C 19 H 27 F3O3S2Na(M+Na) + 447.1246. 1 H NMR (500 MHz, CDCl3): δ = 11.25 (1H, s), 7.41 (1H, dd, J = 1.0, 8.0 Hz), 7.10 (1H, dd, J = 1.0, 8.0 Hz), 6.81 (1H, t, J = 8.0 Hz), 5.68 (1H, s), 3.06 (2H, t, J = 7.3 Hz), 2.54 (4H, m), 2.22 (2H, m), 1.85 (2H, m), 1.68 (2H, m), 1.60 (2H, m), 1.32-1.47 (8H, m). The structure of the obtained compound 4 is shown below:

[0066] Example 5

[0067] In this example, 1,10-decanedithiol (413.0 mg, 2.0 mmol) was used to prepare a substituted benzoic acid trifluoroalkylthioalkylthioester derivative. The rest was the same as in Example 3. The obtained product was recorded as Compound 5.

[0068] Analysis of HR ESI-MS and 1 The structure of compound 5 was confirmed by H NMR data, HR ESI-TOF-MS m / z 475.1544, calculated for C 21 H 31 F3O3S2Na(M+Na) + 475.1559. 1 H NMR (500 MHz, CDCl3): δ = 11.26 (1H, s), 7.41 (1H, dd, J = 1.4, 8.0 Hz,), 7.10 (1H, dd, J = 1.4, 8.0 Hz), 6.81 (1H, t, J = 8.0 Hz), 5.71 (1H, s), 3.06 (2H, t, J = 7.4 Hz), 2.57 (2H, m), 2.50 (2H, m), 2.22 (2H, m), 1.85 (2H, m), 1.68 (2H, m), 1.57 (2H, m), 1.43 (2H, m), 1.38 (2H, m), 1.25-1.45 (8H, m). The structure of the obtained compound 5 is shown below:

[0069] Example 6

[0070] Considering that the two hydroxyl groups on the benzene ring of compound 3 may potentially affect its ability to penetrate the blood-brain barrier, this example acetylated compound 3 based on the prepared compound 3 to synthesize compound 6, a prodrug of compound 3. The steps were as follows: acetic anhydride (0.47 mL, 5.0 mmol) was added to a solution of compound 3 (200.0 mg, 0.5 mmol) in anhydrous pyridine (1.0 mL) at room temperature. After stirring, the reaction was quenched with 1N HCl solution and concentrated in vacuo. The concentrate was extracted with EtOAc, and the organic phase was dried over Na2SO4, filtered, and concentrated. The remaining steps were the same as in Example 3. The resulting product was designated as compound 6.

[0071] Analysis of HR ESI-MS and 1 The structure of compound 6 was confirmed by H NMR data, HR ESI-TOF-MS m / z 481.1327, calculated for C 21 H 28 F3O5S2(M+H) + 481.1325. 1H NMR (500 MHz, CDCl3): δ = 7.79 (1H, dd, J = 1.7, 7.9 Hz), 7.37 (1H, dd, J = 1.7, 7.9 Hz), 7.32 (1H, t, J = 7.9 Hz), 3.01 (2H, t, J = 7.5 Hz), 2.57 (2H, t, J = 7.1 Hz), 2.50 (2H, m), 2.33 (3H, s), 2.30 (3H, s), 2.20 (2H, m), 1.84 (2H, m), 1.66 (2H, m), 1.60 (2H, m), 1.43 (4H, m). The structure of the obtained compound 6 is shown below:

[0072] Example 7

[0073] In this example, 1,10-decanedithiol (413.0 mg, 2.0 mmol) and benzoic acid (122.0 mg, 1.0 mmol) were used to prepare substituted benzoic acid trifluoroalkylthioalkylthioester derivatives. The rest was the same as in Example 3. The obtained product was recorded as Compound 7.

[0074] Analysis of HR ESI-MS and 1 The structure of compound 7 was confirmed by H NMR data, HR ESI-TOF-MS m / z 387.1022, calculated for C 17 H 23 F3OS2Na(M+Na) + 387.1035. 1 H NMR (500 MHz, CDCl3): δ = 7.97 (2H, dd, J = 1.2, 7.9 Hz), 7.56 (1H, t, J = 7.9 Hz), 7.45 (2H, t, J = 7.9 Hz), 3.07 (2H, t, J = 7.5 Hz), 2.57 (2H, t, J = 7.0 Hz), 2.51 (2H, t, J = 7.3 Hz), 2.22 (2H, m), 1.85 (2H, m), 1.69 (2H, m), 1.60 (2H, m), 1.45 (4H, m). The structure of the obtained compound 7 is shown below:

[0075] Example 8

[0076] In this example, 1,4-butanedithiol (244.0 mg, 2.0 mmol) and 1-bromo-5,5,5-trifluoropentane (0.3 mL, 2.0 mmol) were used to prepare substituted benzoic acid trifluoroalkylthioalkylthioester derivatives. The rest was the same as in Example 1. The obtained product was recorded as Compound 8.

[0077] Analysis of HR ESI-MS and1 The structure of compound 8 was confirmed by H NMR data, HR ESI-TOF-MS m / z 383.0959, calculated for C 16 H 22 F3O3S2(M+H) + 383.0957. 1 H NMR (500 MHz, CDCl3): δ = 11.20 (1H, s), 7.40 (1H, dd, J = 1.2, 8.0 Hz,), 7.11 (1H, dd, J = 1.2, 8.0 Hz), 6.82 (1H, t, J = 8.0 Hz), 5.70 (1H, s), 3.10 (2H, t, J = 7.0 Hz), 2.55 (4H, m), 2.09 (2H, m), 1.65-1.84 (8H, m). The structure of the obtained compound 8 is shown below:

[0078] Application Example 1

[0079] BV-2 cell activity assay of the substituted benzoic acid trifluoroalkylthioalkylthioester derivatives prepared in Examples 1 to 8

[0080] 1. Culture medium preparation

[0081] RPMI 1640 basal medium: Add 5 mL of dual-antibody (10,000 U / mL penicillin and 10 mg / mL streptomycin) to 500 mL of RPMI 1640 medium, mix well, and store in a refrigerator at 4°C until use.

[0082] RPMI 1640 complete medium: Add 5 mL of double-antibody (10,000 U / mL penicillin and 10 mg / mL streptomycin) to 500 mL of RPMI 1640 medium, mix thoroughly, remove 50 mL of medium as the above RPMI 1640 basal medium, add 50 mL of fetal bovine serum to the remaining 450 mL of medium, mix thoroughly, and store in a refrigerator at 4°C until use.

[0083] 2. BV-2 Cell Anti-inflammatory Activity Assay Procedure

[0084] When BV-2 cells reached approximately 70-80% confluence on the culture dish, subculture was initiated. In a 24-well plate, 50,000 cells were added to each well of the plate with 1 mL of complete RPMI 1640 medium. The cells were cultured in a CO2 incubator for 24 hours before sample addition. Each well was treated with 500 μL of RPMI 1640 basal medium containing the sample for 2 hours, followed by 500 μL of culture medium containing LPS (1 μg / mL) for 24 hours.

[0085] After 24 hours, the culture medium was aspirated, each well was washed 3 times with PBS, and fixed with 4% paraformaldehyde at room temperature for 20 minutes. The fixative was discarded, each well was washed 3 times with PBS, and immunostaining blocking solution was added for 60 minutes. The blocking solution was discarded, and diluted primary antibody anti-Iba1 was added to each well and incubated at 4°C overnight. The next day, the supernatant was discarded, each well was washed 3 times with PBS, each time for 5 minutes, and diluted secondary antibody goat anti-rabbit IgG H&L (Alexa Fluor 5000) was added. 488) and incubated at room temperature for 1 hour. Cell nuclei were then stained with diluted DAPI for 5-10 minutes, and fluorescence was observed under an inverted fluorescence microscope. Three randomly selected locations were photographed, and images were analyzed using Image J software. Data were analyzed using one-way ANOVA using Prism 5.0 statistical software. Statistical results are expressed as mean ± standard error (X + SEM).

[0086] 3. Test results analysis

[0087] Figure 1 shows the LPS-induced cell activation rate of BV-2 cells after 24 hours of treatment with different concentrations of substituted benzoic acid trifluoroalkylthioalkylthioester derivatives (Compounds 1-8) prepared in Examples 1-8. 0.5% DMSO was used as a negative control (C), LPS (1 μg / mL) was used as an experimental control, and butylphthalide (10 μM) was used as a positive control. *P<0.05, **P<0.01, ***P<0.001, ### P<0.001. ### P < 0.001 indicates that BV-2 cells were significantly activated after LPS (1 μg / mL) treatment alone. *** P < 0.001 indicates that compounds 1 to 8 significantly inhibited BV-2 cell activation. The results showed that compounds 1 to 8 were most effective at a concentration of 0.3 μM, with compounds 3, 4, and 5 showing the most significant effects.

[0088] Evaluation of the anti-inflammatory efficacy of substituted benzoic acid trifluoroalkylthioalkylthioester derivatives in animals (taking compound 3 as an example)

[0089] 1. Animal grouping

[0090] Grouping of high-fat diet model mice: 60 ICR white mice, 3-4 weeks old, 10-15 g, male, were randomly divided into six groups, with 10 mice in each group.

[0091] Natural aging model mouse grouping: 40 C57BL / 6J black mice, 10 of which were 8 weeks old, about 25 g, female, were divided into one group (young group); the other 30 were 18 months old, about 30 g, naturally aging, female, and randomly divided into three groups (natural aging groups), with 10 mice in each group.

[0092] 2. Dosage

[0093] All mice were given the drug orally by gavage.

[0094] High-fat diet-induced mice were treated with soybean oil as a blank control (control); the first high-fat diet-fed group was given soybean oil as a negative control (HFD); the second high-fat diet-fed group was given metformin (140 mg / kg), a drug with anti-Alzheimer's disease efficacy, as a positive control (Met); the third, fourth, and fifth high-fat diet-fed groups were given compound 3 at doses of 0.1, 5, and 20 mg / kg, respectively, as experimental groups. Each mouse received 150 μL of the drug daily for two consecutive months.

[0095] Naturally aged mice were treated with 0.5% DMSO as a blank control group; the first group of the natural aged mice was treated with 0.5% DMSO as a negative control group; the second group of the natural aged mice was treated with 3 mg / kg of donepezil, a first-line clinical anti-Alzheimer's disease drug, as a positive control group; and the third group of the natural aged mice was treated with 5 mg / kg of Compound 3 as an experimental group. Each mouse received 150 μL of the drug daily for three consecutive months.

[0096] 3. Test result analysis

[0097] High-fat diet model mice: As shown in Figure 2A-F, the results showed that compared with the negative control HFD group, the 5 mg / kg and 20 mg / kg compound 3 experimental groups had significantly reduced inflammation-related indicators (Iba1 protein, GFAP protein, INOS protein); compared with the blank control group (Control), the activation ratio of microglia (Iba1 protein) and astrocytes (GFAP protein) in the cerebral cortex and hippocampus and the expression of INOS protein were comparable.

[0098] Naturally aging model mice: As shown in Figure 3A-C, the results showed that the experimental group (aging group + compound administration dose of 5 mg / kg) had significantly reduced inflammation-related indicators (Iba1 protein, GFAP protein, INOS protein) compared with the negative control group (aging group); compared with the blank control group (young group), the activation ratio of microglia (Iba1 protein) and astrocytes (GFAP protein) in the cerebral cortex and the expression of INOS protein were comparable.

[0099] As shown in Figure 3D and E, the results showed that the number of mature neurons (NeuN protein) in the experimental group (aging group + compound dosage of 5 mg / kg) was significantly increased compared with the negative control group (aging group); compared with the blank control group (young group), the number of mature neurons in the cerebral cortex and hippocampus in the brain was comparable, that is, compound 3 has significant neuroprotective activity.

[0100] The present invention provides substituted benzoic acid trifluoroalkylthioalkylthioester derivatives and their preparation methods. Evaluation in a BV-2 cell bioactivity system revealed significant anti-inflammatory activity, and further animal experiments confirmed their anti-inflammatory and neuroprotective efficacy. These derivatives can be used in the preparation and application of anti-inflammatory and neuroprotective drugs. This research provides a basis for the development of new drugs and basic research for inflammatory diseases, and is of great significance.

[0101] Application Example 2

[0102] Evaluation of the cognitive function activity of substituted benzoic acid trifluoroalkylthioalkylthioester derivatives (Compound 3) on lipopolysaccharide (LPS)-induced neuroinflammation mice

[0103] The effect of compound 3 on the cognitive function of mice with LPS-induced neuroinflammation was studied through animal behavioral experiments. Experimental method: 50 of 60 C57BL / 6 mice were randomly selected and continuously injected intraperitoneally with LPS (1 mg / kg) for 30 days, and the remaining 10 mice were in the blank group (injected with the same volume of PBS). Compound 3 and the positive control galanthamine (GA) were dissolved in soybean oil. Subsequently, the LPS model mice were randomly divided into 5 groups and given compound 3 (1 mg / kg, 5 mg / kg, 10 mg / kg), galanthamine (4 mg / kg), and soybean oil (control group) by gavage for 2 months. The mice in each group were subjected to Y-maze and novel object recognition test experiments to evaluate the working memory and short-term memory of the mice. In addition, impaired spatial memory and long-term memory are prominent features of cognitive decline. Therefore, the water maze experiment was used to evaluate the spatial memory and long-term memory of mice, thereby comprehensively evaluating the improvement effect of compound 3 on the cognitive function of mice with LPS-induced neuroinflammation.

[0104] As shown in Figures 4A and 4B, in the Y-maze, the alternating arm entry rate and total arm entries of LPS-induced mice were significantly reduced compared to the blank control group. In contrast, the arm entry rate and total arm entries of mice treated with compound 3 (10 mg / kg) and GA were higher than those of the LPS control group. As shown in Figures 4C and 4D, in the novel object recognition test, there were no significant differences in the target recognition and discrimination indices among the groups during the training phase. However, the recognition and discrimination indices of the LPS control group were lower, while those of the compound 3 (5 mg / kg) and GA-treated groups were higher than those of the LPS control group. These results indicate that both compound 3 and GA can improve working memory and short-term memory in mice with LPS-induced neuroinflammation. As shown in Figures 4E-G, during the four-day training phase, the escape latency of the LPS control group was significantly greater than that of the blank control group, while the escape latency of the GA and compound 3 groups was significantly shorter than that of the LPS control group. During the fifth-day testing phase, similar changes in escape latency were observed in the LPS control, GA, and compound 3-treated groups as in the training phase. In addition, the number of platform crossings in the LPS control group was significantly lower than that in the blank control group. However, these parameters returned to normal levels in the GA and compound 3 (10 mg / kg) groups, thus confirming that compound 3 can improve cognitive dysfunction in mice with LPS-induced neuroinflammation and has anti-Alzheimer's disease efficacy.

[0105] Application Example 3

[0106] In vitro anti-aging biological activity evaluation of substituted benzoic acid trifluoroalkylthioalkylthioester derivatives (Compounds 1-5) prepared in Examples 1-5

[0107] Determination of the activity of compounds 1 to 5 in alleviating senescence markers in etoposide-induced senescent NIH3T3 cells: (1) Effects of compounds 1 to 5 on the activity of senescence-related β-galactosidase in etoposide-induced senescent NIH3T3 cells. Experimental method:

[0108] Place a 12 mm round glass slide in each well of a 24-well cell plate and coat with 250 μL of 100 μg / mL L-polylysine. After overnight incubation, rinse three times with PBS and incubate in a CO2 incubator for 24 hours before loading. Cell loading and treatment: Prepare the sample to be tested with DMSO to the desired concentration. Add the prepared sample to EM to create 1 mL of the desired concentration. Replace the CM in the 24-well plate with the sample solution. The positive control is 500 nM rapamycin (Rapa), and the negative control is 0.5% DMSO (C). After 24 hours, remove the culture medium and add 1 mL of CM containing 0.3 μM etoposide (Eto) to each well, except for the negative control. The negative control is treated with CM containing an equal volume of DMSO. Incubate in a CO2 incubator for an additional 48 hours. Staining treatment: The cells were treated with cell senescence β-galactosidase staining reagent (Shanghai Biyuntian Biotechnology Co., Ltd., product number C0602), then washed with 70% ethanol solution, sliced ​​and sealed, and observed under an ordinary optical microscope. The proportion of cells stained blue (i.e., cells positive for cell senescence β-galactosidase) in the field of view was calculated.

[0109] As shown in Figure 5, etoposide significantly caused NIH3T3 cell senescence. After adding compounds 1 to 5 at different concentrations (0.1 μM, 0.3 μM, 1 μM), the proportion of positive cells decreased significantly, indicating that pretreatment with compounds 1 to 5 can alleviate etoposide-induced cell senescence, and the alleviating effect is stronger as the chain length of the compound increases.

[0110] (2) Effect of compound 3 on cell survival in etoposide-induced senescent PC12 cells

[0111] Experimental Method: 7,000 cells were seeded with 200 μL of CM per well of a 96-well cell plate and incubated in a CO2 incubator for 24 hours before sample loading. The cell loading method in this application example is the same as in Application Example 1. Each well was then replaced with 100 μL of EM solution containing 200 μg / mL thiazolyl blue tetrazolium bromide (MTT). After a 2-hour incubation, 100 μL of DMSO was added to each well. The plates were shaken at room temperature for 10 minutes, and the absorbance of each well was measured at 570 nm using a microplate reader.

[0112] As shown in Figure 6A, etoposide significantly reduced the cell survival rate of PC12 cells. Pretreatment with the positive drug 500nM rapamycin and 0.003-0.3μM compound 3 for 24h had no significant effect on the cell survival rate of etoposide-induced senescent PC12 cells, while pretreatment with 1μM compound 3 could reduce the cell survival rate of etoposide-induced senescent PC12 cells (P=0.02).

[0113] (3) Effect of compound 3 on reactive oxygen species in etoposide-induced senescent PC12 cells

[0114] Experimental Methods: The cell culture and sample loading methods in this application example are the same as those in Application Example 1. After loading cells into a 24-well plate, 200 μL of 10 μM reactive oxygen species probe DCFH-DA (Shanghai Beyotime Biotechnology Co., Ltd., Product No. S0033) was added to each well. The cells were incubated in the dark for 20 minutes, then washed three times with EM solution and photographed using an inverted fluorescence microscope. Fluorescence intensity values ​​in the photographs were quantified using Image J software.

[0115] As shown in Figure 6B, etoposide treatment can significantly increase the level of reactive oxygen species in PC12 cells, while pretreatment with rapamycin and 0.003-1 μM compound 3 can significantly reduce the abnormal reactive oxygen species level induced by etoposide, and as the concentration of compound 3 increases, the ability to reduce the abnormal reactive oxygen species level becomes stronger.

[0116] (4) Effect of compound 3 on cell proliferation in etoposide-induced senescent PC12 cells

[0117] Experimental Methods: The cell culture and sample loading methods in this application example are the same as those in Application Example 1. After loading cells into a 24-well plate, the cells were treated with EdU Cell Proliferation Detection Reagent (Shanghai Beyotime Biotechnology Co., Ltd., Product No. C0071S). Finally, slides were prepared, mounted, and photographed using an upright fluorescence microscope. The ratio of green-fluorescent cells (i.e., proliferating cells with EdU incorporated into their DNA) to blue-fluorescent cells (all cells) in the field of view was calculated.

[0118] As shown in Figure 6C, etoposide significantly reduced the proportion of proliferating cells with EdU incorporation, while pretreatment with rapamycin and 0.1-1 μM compound 3 significantly increased the proportion of proliferating cells with EdU incorporation. Moreover, as the concentration of compound 3 increased, the ability to increase the proportion of proliferating cells with EdU incorporation became stronger, indicating that compound 3 can alleviate the cell cycle arrest of senescent cells.

[0119] (5) Effect of compound 3 on lipofuscin in etoposide-induced senescent PC12 cells

[0120] Experimental Methods: The cell culture and sample loading methods in this application example are the same as those in Application Example 1. After loading and fixing cells in a 24-well plate, wash three times with PBS and stain with 0.2 ml of Sudan Black stain. After washing, counterstain with Nuclear Fast Red stain. After washing, slides are prepared and mounted. Cells are observed and photographed under a standard light microscope. The proportion of cells stained blue-black (i.e., lipofuscin-positive cells) in the field of view is calculated.

[0121] As shown in Figure 6D, etoposide significantly induced the production of lipofuscin, a marker of aging. After adding different concentrations of compound 3, the proportion of cells containing lipofuscin decreased to varying degrees, with 0.3 μM compound 3 having the best effect.

[0122] Application Example 4

[0123] Evaluation of the in vivo hair growth promoting activity of substituted benzoic acid trifluoroalkylthioalkylthioester derivatives 1 to 5 prepared in Examples 1 to 5

[0124] (1) Study on the effect of compound 3 on hair loss in aged mice

[0125] Animal grouping: 20 C57BL / 6J black mice, 18 months old, about 30 g, naturally aged, female, were randomly divided into two groups; 10 C57BL / 6J black mice, 8 weeks old, about 25 g, female, were divided into one group.

[0126] Dosing: All three groups of mice were administered oral gavage. The first group in the natural aging group received 0.5% DMSO and served as a blank control. The second group in the natural aging group received 5 mg / kg of compound 3 as an experimental group. The young group received 0.5% DMSO. Each mouse received 150 μL of the compound daily for three consecutive months.

[0127] As shown in Figure 7A, the blank control group had more severe hair loss than the young group; the experimental group had significantly improved hair loss compared to the blank control group, and the area and length of new hair grown were comparable to those of the young group. (2) Study on the effect of different doses of compound 3 on promoting hair growth in rats

[0128] Experimental method: To establish a rat hair removal experimental animal model: 20 white SD rats, 6-8 weeks old, weighing approximately 250 g, female; anesthesia was performed with 1.0% sodium pentobarbital, i.e., 1 g of sodium pentobarbital was dissolved in 100 mL of 0.9% saline, at a dose of 30 mg / kg. The hair was shaved with an electric shaver. The model was considered successful when no hair remained in a 4 cm × 5 cm area on the back of the rat.

[0129] Animal grouping: The SD rats with successful modeling were randomly divided into 5 groups, with 4 rats in each group.

[0130] Administration: The four groups of rats were sprayed on the hair removal area with the following: 0.02% compound 3 (i.e., 0.02 g of compound 3 in 100 mL of carrier solvent; 0.4 mg / kg), 0.10% compound 3 (2.0 mg / kg), 0.50% compound 3 (10.0 mg / kg), and 2.0% minoxidil (40.0 mg / kg). The fifth group of rats served as blank controls and were sprayed with an equal volume of mixed solvent (water: ethanol: glycerol monoacetate = 0.4:0.2:0.4). The volume of each administration was 0.5 mL, and the administration was done once a day for four weeks.

[0131] Experimental results: a. Hair growth length

[0132] Four areas were randomly selected from the rat dosing area each week, and five hairs were randomly plucked from each area. The hair length was measured and recorded. The results are shown in Figure 7B. Compared with the blank control group, the hair length of the experimental groups treated with different concentrations of compound 3 increased. The hair growth length of the experimental group treated with 0.50% compound 3 and the 2.0% minoxidil group was comparable, and both increased significantly compared with the blank control group.

[0133] b. Hair growth density

[0134] Image J was used to analyze the hair density in the administration area of ​​the rats every week. The results are shown in Figure 7C. Compared with the blank control group, the hair growth density of the experimental groups with different concentrations of compound 3 increased. The hair growth density of the experimental group with 0.50% compound 3 was equivalent to that of the 2.0% minoxidil group, and was significantly increased compared with the blank control group. After 4 weeks, the hair growth density of the experimental groups with different concentrations of compound 3 was significantly increased compared with the blank control group.

[0135] (3) Study on the effect of compounds 1 and 5 on promoting hair growth in rats

[0136] Experimental method: To establish a rat hair removal experimental animal model: 16 white SD rats, 6-8 weeks old, weighing approximately 250 g, female; anesthesia was performed with 1.0% sodium pentobarbital, dose of 30 mg / kg, and hair was shaved with an electric shaver. The model was successfully established when no hair remained in a 4 cm × 5 cm area on the back of the rat.

[0137] Animal grouping: The SD rats with successful modeling were randomly divided into 4 groups, with 4 rats in each group.

[0138] Administration: Three groups of rats were sprayed on the hair removal area with the following: 0.50% compound 1 (10.0 mg / kg), 0.50% compound 5 (10.0 mg / kg), and 2.0% minoxidil (40.0 mg / kg). The fourth group of rats served as blank controls and were smeared with an equal volume of a mixed solvent (water: ethanol: glycerol monoacetate = 0.4:0.2:0.4). The volume of each administration was 0.5 mL, and the administration was done once a day for four weeks.

[0139] Experimental results: a. Hair growth length

[0140] The results are shown in FIG8A . From the second week onwards, the hair of the rats in the compound 1 group showed significant growth compared with the blank control group. From the third week onwards, the hair of the rats in the compound 5 group showed significant growth compared with the blank control group.

[0141] b. Hair growth density

[0142] The results are shown in FIG8B . From the first week onwards, the hair density of the rats in the compound 1 group increased significantly compared with the blank control group. From the third week onwards, the hair density of the rats in the compound 5 group increased significantly compared with the blank control group.

[0143] (4) Study on the effect of compounds 1, 2, 4 and 5 on promoting hair growth in mice

[0144] Experimental method: To establish a mouse hair removal experimental animal model: 50 black C57 mice, 6-8 weeks old, weighing approximately 20 g, male; anesthetized with 0.3% sodium pentobarbital, dose of 40 mg / kg, and depilated with depilatory cream. The model was successfully established when no hair remained in a 3 cm × 4 cm area on the back of the mouse.

[0145] Animal grouping: The successfully modeled C57 mice were randomly divided into 10 groups, with 5 mice in each group.

[0146] Dosage: 9 groups of mice were sprayed on the hair removal area and administered: 2.0% minoxidil (100.0 mg / kg), 0.08% compound 1 (4.0 mg / kg), 0.25% compound 1 (12.5 mg / kg), 0.08% compound 2 (4.0 mg / kg), 0.25% compound 2 (12.5 mg / kg), 0.08% compound 4 (4.0 mg / kg), 0.25% compound 4 (12.5 mg / kg), 0.08% compound 5 (4.0 mg / kg), 0.25% compound 5 (12.5 mg / kg), and the 10th group of mice was used as a blank control and was smeared with an equal volume of mixed solvent (water: ethanol: monoacetin = 0.4: 0.2: 0.4). The volume of each administration was 0.1 mL, and the administration was done once a day for three weeks.

[0147] Experimental results: a. Hair growth density

[0148] The results are shown in FIG9A . Starting from the second week, the hair of mice in compound 2, 4 and 5 groups showed significant growth compared with the blank control group. Starting from the third week, the hair of mice in compound 1 group showed significant growth compared with the blank control group.

[0149] b. Hair growth length

[0150] The results are shown in Figure 9B. Starting from the first week, the hair density of mice in the compound 5 group increased significantly compared with the blank control group. Starting from the second week, the hair density of mice in the compound 1, 2 and 4 groups increased significantly compared with the blank control group.

Claims

1. A substituted benzoic acid trifluoroalkylthioalkylthioester derivative having a structure shown in formula (I): in, R1 and R2 are OH, OAc or H, and X and Y are straight-chain or branched-chain alkyl groups having 1 to 9 carbon atoms.

2. The method for preparing a substituted benzoic acid trifluoroalkylthioalkylthioester derivative according to claim 1, characterized in that: The following steps are involved: (1) adding NaH to a thiol-containing solvent at a temperature below 0° C., heating the mixture to room temperature and stirring the mixture for 10 to 30 minutes, then adding a bromotrifluoroalkyl compound and TBAI at a temperature below 0° C. to react, detecting the reaction end point by TLC, and extracting the reaction product after the reaction is completed to obtain the target product; (2) dissolving the target product obtained in step (1) in an organic solvent, adding EDC·HCl, substituted benzoic acid and DMAP, reacting at room temperature, detecting the reaction end point by TLC, and separating and purifying the reaction product after the reaction is completed to obtain the substituted benzoic acid trifluoroalkylthioalkylthioester derivative.

3. The method for preparing a substituted benzoic acid trifluoroalkylthioalkylthioester derivative according to claim 2, characterized in that: In step (1), the thiol is a dithiol having 1 to 10 carbon atoms, including but not limited to 1,2-ethanedithiol, 1,4-butanedithiol, 1,6-hexanedithiol, 1,8-octanedithiol or 1,10-decanedithiol; In step (2), the substituted benzoic acid includes but is not limited to 2,3-dihydroxybenzoic acid, 2,3-diacetoxybenzoic acid, 2-hydroxybenzoic acid, 3-hydroxybenzoic acid, 2-acetoxybenzoic acid or 3-acetoxybenzoic acid; In step (1), the solvent is DMF; the molar ratio of NaH, bromotrifluoroalkyl compound, TBAI and thiol is 1-2:1:0.1-1:1; In step (2), the molar ratio of EDC·HCl, substituted benzoic acid, DMAP and thiol is 1-4:1-10:0.1-4:

1.

4. Use of the substituted benzoic acid trifluoroalkylthioalkylthioester derivative according to claim 1 in the preparation of medicines, health products or foods for preventing and / or treating inflammation.

5. Use of the substituted benzoic acid trifluoroalkylthioalkylthioester derivative according to claim 1 in the preparation of drugs, health products or foods for neuroprotection, prevention and / or treatment.

6. Use of the substituted benzoic acid trifluoroalkylthioalkylthioester derivative according to claim 1 in the preparation of medicines, health products or foods for preventing and / or treating neurodegenerative diseases.

7. Use of the substituted benzoic acid trifluoroalkylthioalkylthioester derivative according to claim 1 in the preparation of drugs, health products or foods for preventing and / or treating anti-aging.

8. Use of the substituted benzoic acid trifluoroalkylthioalkylthioester derivative according to claim 1 in the preparation of medicines, health products, foods or toiletries for preventing and / or treating hair loss.

9. The use according to claim 8, characterized in that: The substituted benzoic acid trifluoroalkylthioalkylthioester derivative can achieve a hair growth effect similar to that of minoxidil by applying the drug to the skin in an amount of 0.25 times that of minoxidil.

10. The use according to claim 8, characterized in that: When used, the oral dosage of the substituted benzoic acid trifluoroalkylthioalkylthioester derivative is 1 to 25 mg / kg body weight.

11. A pharmaceutical composition, health product or food for neuroprotection, prevention and / or treatment of inflammation and neurodegenerative diseases, characterized in that: The pharmaceutical composition, health product or food contains the substituted benzoic acid trifluoroalkylthioalkylthioester derivative according to claim 1.

12. An anti-aging and hair loss prevention and / or treatment product, characterized in that: The anti-aging and hair loss prevention and / or treatment product comprises the substituted benzoic acid trifluoroalkylthioalkylthioester derivative according to claim 1.

Images