Perylenequinone compound of alternaria tenuissima and application of perylenequinone compound in preparation of medicine for preventing and / or treating neuroinflammation diseases

Abstract

The invention belongs to the technical field of deep sea microbial materials, and discloses a perylenequinone compound of Alternaria sp. Xiao20, a derivative of the perylenequinone compound, or a pharmaceutically acceptable salt of the perylenequinone compound, a preparation method of the perylenequinone compound, and an application of the perylenequinone compound and the derivative or the pharmaceutically acceptable salt of the perylenequinone compound in preparation of medicines for preventing and / or treating neuroinflammation diseases. The invention provides a perylenequinone compound or a derivative thereof separated from a fermentation product of alternaria alternata, and the perylenequinone compound or the derivative thereof has important significance on development of marine drugs; the perylenequinone compound or the derivative or the pharmaceutically acceptable salt of the perylenequinone compound disclosed by the invention can be used for inhibiting the release of a neuromicroglia inflammatory factor NO; according to the present invention, the release level of the LPS-induced nerve microglial cell inflammatory factor NO is significantly reduced, the technical effect of neuroinflammation disease treatment is achieved, and the LPS-induced nerve microglial cell inflammatory factor NO can be prepared, such that the LPS-induced nerve microglial cell inflammatory factor NO can be used for preparing the neuroinflammation disease prevention and / or treatment drug, and has important significance on neuroinflammation disease treatment and new drug development.

Description

Technical Field

[0001] The present invention belongs to the technical field of deep-sea microbial materials, and particularly relates to perylenequinone compounds, their derivatives or pharmaceutically acceptable salts of Alternaria sp. Xiao20 from the deep sea, and their preparation methods and applications in the preparation of drugs for preventing and / or treating neuroinflammatory diseases. Background Art

[0002] Inflammation is the core mechanism of the body's defense and repair. However, in the central nervous system, the uncontrolled neuroinflammatory response has become the core pathological driving factor of neurodegenerative diseases (such as Alzheimer's disease, multiple sclerosis, Parkinson's disease), post-stroke injury, and autoimmune neurological diseases. Neuroinflammation is triggered by the overactivation of glial cells such as microglia and astrocytes, accompanied by the release of pro-inflammatory factors (IL-1β, TNF-α, CCL2), reactive oxygen species (ROS), and neurotoxic metabolites, leading to blood-brain barrier disruption, neuronal apoptosis, and synaptic dysfunction. Although anti-inflammatory treatment is theoretically promising, existing clinical means still face severe challenges between effectiveness and safety.

[0003] Microglia are key immune cells in the central nervous system and play a crucial role in maintaining neural homeostasis and responding to injury or infection. Similar to macrophages, microglia have the ability to clear dead and damaged nerve cells and participate in the neuroinflammatory response. Recent studies have revealed that regulating microglial polarization (transformation from M1 pro-inflammatory type to M2 anti-inflammatory type) and inhibiting the activation of A1-type astrocytes are key nodes for intervening in neuroinflammation.

[0004] Nitric oxide (NO) is a molecular mediator of many physiological processes, including vasodilation, inflammation, thrombosis, immunity, and neurotransmission. When microglia are stimulated by microbial endotoxins, inflammatory mediators, etc., they will change from the normal monitoring state to M1 pro-inflammatory cells and simultaneously generate a large amount of nitric oxide synthase to generate NO for immune response. Therefore, inhibiting NO generation is a direct indicator of the anti-inflammatory activity of compounds. The anti-inflammatory drug cell activity screening uses mouse microglial cells (BV2) to detect the inhibitory effect of compounds on NO generation. This model uses lipopolysaccharide (LPS) to induce cell inflammatory responses, evaluates the activity of the test substance in inhibiting NO generation at the cell level, and thereby reflects its anti-neuroinflammatory activity. Summary of the Invention

[0005] In order to overcome the above-mentioned disadvantages and deficiencies of the prior art, the primary object of the present invention is to provide a perylenequinone compound, its derivative, or its pharmaceutically acceptable salt.

[0006] The perylenequinone compound, its derivative or its pharmaceutically acceptable salt provided by the present invention can significantly reduce the release level of the inflammatory factor NO and can be used in the preparation of drugs for preventing and / or treating neuroinflammatory diseases.

[0007] Another object of the present invention is to provide a preparation method of the above perylenequinone compound, its derivative or its pharmaceutically acceptable salt.

[0008] Another object of the present invention is to provide the application of the above perylenequinone compound, its derivative or its pharmaceutically acceptable salt in the preparation of drugs for preventing and / or treating neuroinflammatory diseases.

[0009] The object of the present invention is achieved by the following solutions:

[0010] In the first aspect of the present invention, a perylenequinone compound, its derivative or its pharmaceutically acceptable salt is provided, and the structural formula is shown as formula (I):

[0011]

[0012] Among them, R1, R2, and R7 are the same or different and are respectively selected from H, halogen, hydroxyl, carboxyl, cyano, nitro, amino, substituted or unsubstituted C1-C20 straight-chain or branched-chain alkyl, substituted or unsubstituted C3-C20 cycloalkyl, substituted or unsubstituted C1-C20 alkoxy, substituted or unsubstituted C1-C20 haloalkyl, C1-C10 carbonyl, C1-C10 ester, substituted or unsubstituted C1-C10 amino, substituted or unsubstituted C6-C20 aryl;

[0013] R3 and R4 are the same or different and are respectively selected from H, halogen, hydroxyl, carboxyl, cyano, nitro, amino, substituted or unsubstituted C1-C20 straight-chain or branched-chain alkyl, substituted or unsubstituted C3-C20 cycloalkyl, substituted or unsubstituted C1-C20 alkoxy, substituted or unsubstituted C1-C20 haloalkyl, C1-C10 carbonyl, C1-C10 ester, substituted or unsubstituted C1-C10 amino, substituted or unsubstituted C6-C20 aryl, or R3 and R4 are connected to form a C2-C10 heterocycle containing one or more of N, O, and S;

[0014] R5 and R6 are the same or different and are respectively selected from H, O, halogen, hydroxyl, carboxyl, cyano, nitro, amino, substituted or unsubstituted C1-C20 straight-chain or branched-chain alkyl, substituted or unsubstituted C3-C20 cycloalkyl, substituted or unsubstituted C1-C20 alkoxy, substituted or unsubstituted C1-C20 haloalkyl, C1-C10 carbonyl, C1-C10 ester, substituted or unsubstituted C1-C10 amino, substituted or unsubstituted C6-C20 aryl.

[0015] One or more hydrogen atoms in the above-mentioned substituents, which are the same or different, in the respective groups may be substituted by groups such as halogen, a straight-chain or branched alkyl group having 1 to 10 carbon atoms, a cycloalkyl group having 3 to 10 carbon atoms, a hydrocarbonoxy group having 1 to 10 carbon atoms, a hydroxyl group, a carboxyl group, a cyano group, a nitro group, an amino group, -CF3, etc.

[0016] Furthermore, the present invention provides a perylenequinone compound or its derivative or its pharmaceutically acceptable salt, the structural formula of which is shown in formula (I), wherein R1, R2, and R7, which are the same or different, are each independently selected from H, halogen, hydroxyl, carboxyl, cyano, nitro, amino, a straight-chain or branched alkyl group having 1 to 20 carbon atoms, a cycloalkyl group having 3 to 20 carbon atoms, an alkoxy group having 1 to 20 carbon atoms, a haloalkyl group having 1 to 20 carbon atoms, a carbonyl group having 1 to 10 carbon atoms, an ester group having 1 to 10 carbon atoms, an amino group having 1 to 10 carbon atoms, an aryl group having 6 to 20 carbon atoms;

[0017] R3 and R4, which are the same or different, are each independently selected from H, halogen, hydroxyl, carboxyl, cyano, nitro, amino, a straight-chain or branched alkyl group having 1 to 20 carbon atoms, a cycloalkyl group having 3 to 20 carbon atoms, an alkoxy group having 1 to 20 carbon atoms, a haloalkyl group having 1 to 20 carbon atoms, a carbonyl group having 1 to 10 carbon atoms, an ester group having 1 to 10 carbon atoms, an amino group having 1 to 10 carbon atoms, an aryl group having 6 to 20 carbon atoms, or R3 and R4 are linked to form a heterocyclic ring containing O and having 2 to 10 carbon atoms;

[0018] R5 and R6, which are the same or different, are each independently selected from H, O, halogen, hydroxyl, carboxyl, cyano, nitro, amino, a straight-chain or branched alkyl group having 1 to 20 carbon atoms, a cycloalkyl group having 3 to 20 carbon atoms, an alkoxy group having 1 to 20 carbon atoms, a haloalkyl group having 1 to 20 carbon atoms, a carbonyl group having 1 to 10 carbon atoms, an ester group having 1 to 10 carbon atoms, an amino group having 1 to 10 carbon atoms, an aryl group having 6 to 20 carbon atoms.

[0019] Furthermore, the present invention provides a perylenequinone compound or its derivative or its pharmaceutically acceptable salt, the structural formula of which is shown in formula (I), wherein R1, R2, and R7, which are the same or different, are each independently selected from H, halogen, hydroxyl, carboxyl, cyano, nitro, amino, a straight-chain or branched alkyl group having 1 to 20 carbon atoms, a cycloalkyl group having 3 to 20 carbon atoms, an alkoxy group having 1 to 20 carbon atoms, a haloalkyl group having 1 to 20 carbon atoms;

[0020] R3 and R4, which are the same or different, are each independently selected from H, halogen, hydroxyl, carboxyl, cyano, nitro, amino, a straight-chain or branched alkyl group having 1 to 20 carbon atoms, a cycloalkyl group having 3 to 20 carbon atoms, an alkoxy group having 1 to 20 carbon atoms, a haloalkyl group having 1 to 20 carbon atoms, or R3 and R4 are linked to form a heterocyclic ring containing O and having 2 to 10 carbon atoms;

[0021] R5 and R6 are the same or different and are each independently selected from H, O, halogen, hydroxyl, carboxyl, cyano, nitro, amino, a C1-C20 straight-chain or branched-chain alkyl group, a C3-C20 cycloalkyl group, a C1-C20 alkoxy group, or a C1-C20 haloalkyl group.

[0022] Furthermore, a perylenequinone compound, a derivative thereof, or a pharmaceutically acceptable salt thereof has a structural formula as shown in formula (I), and one or more hydrogen atoms in the structure shown in formula (I) can each be independently substituted by halogen, hydroxyl, carboxyl, cyano, nitro, amino, a C1-C10 straight-chain or branched-chain alkyl group, a C3-C10 cycloalkyl group, a C1-C10 alkoxy group, a C1-C10 haloalkyl group, a C1-C10 carbonyl group, a C1-C10 ester group, or an aryl group.

[0023] Furthermore, the present invention provides a perylenequinone compound, a derivative thereof, or a pharmaceutically acceptable salt thereof, and the structural formula is as

[0024] shown in any one of formula 1 - formula 6:

[0025]

[0026] The perylenequinone compound, a derivative thereof, or a pharmaceutically acceptable salt thereof according to the present invention can be prepared by existing conventional synthesis methods or can be isolated from plants, strains, or their fermentation broths.

[0027] In a second aspect of the present invention, there is provided a method for preparing the above perylenequinone compound, a derivative thereof, or a pharmaceutically acceptable salt thereof, which is isolated from the fermentation product of Alternaria sp.

[0028] Furthermore, the Alternaria sp. (Alternaria sp. Xiao20) refers to CN116286391A, and the deposit number is CCTCC M 2022501, which was deposited with the China Center for Type Culture Collection (CCTCC) on April 27, 2022.

[0029] Furthermore, the fermentation product can be obtained by fermenting Alternaria sp. in a fermentation culture medium.

[0030] Furthermore, the fermentation conditions can be carried out at room temperature (such as 25 °C). The fermentation time can be 21 - 42 days.

[0031] Furthermore, the composition of the fermentation culture medium: per 1 L, 50 g of glucose, 500 g of potatoes (fresh, filtered), and 37.5 g of sea salt, pH 7.5.

[0032] Furthermore, the pH of the fermentation broth can be adjusted using NaOH.

[0033] Furthermore, Alternaria sp. Xiao20 can be cultured on a PDA plate at 25 °C for 1 - 3 days to obtain mycelia; the mycelia are transferred to a culture medium containing PDB, and after culture, a seed solution is obtained; then the seed solution is inoculated into the fermentation broth for fermentation to obtain a fermentation product.

[0034] Furthermore, the separation method includes the following steps:

[0035] The fermentation product of Alternaria sp. is extracted with ethyl acetate, and the obtained extract is separated by normal-phase silica gel column chromatography and gradient eluted with a dichloromethane - methanol system to obtain 5 fractions (Fr.1 - Fr.5); fraction Fr.4 is separated by ODS column chromatography (eluted with a methanol - water system) to obtain 5 sub-fractions (Fr.4 - 1 to Fr.4 - 5); for sub-fraction Fr.4 - 1, it is first separated using a Sephadex dextran gel chromatography column (eluted with methanol), and then separated using high-performance liquid chromatography (gradient elution with acetonitrile - water) to obtain compound 1, compound 2, compound 4, and compound 6; for sub-fraction Fr.4 - 4, it is separated using high-performance liquid chromatography (gradient elution with acetonitrile - water) to obtain compound 3 and compound 5.

[0036] The present invention provides a method for separating perylenequinone compounds or their derivatives from the fermentation product of Alternaria sp., which is of great significance for marine drug development.

[0037] In the third aspect of the present invention, there is provided an application of the above perylenequinone compound or its derivative or its pharmaceutically acceptable salt in the preparation of one of the following products: (1) a drug for preventing and / or treating neuroinflammatory diseases; (2) a drug for inhibiting the release of the inflammatory factor NO in BV-2 cells; (3) a drug for inhibiting the release of the inflammatory factor NO in LPS-induced BV-2 cells.

[0038] Furthermore, the neuroinflammatory diseases include but are not limited to: depression, schizophrenia, Alzheimer's disease (AD), Parkinson's disease, multiple sclerosis (MS), postoperative cognitive dysfunction (POCD), spinal cord injury (SCI), AIDS dementia complex (ADC), ischemia, stroke, traumatic brain injury (TBI), Huntington's disease, amyotrophic lateral sclerosis, brain or central nervous system infections, and brain tumors, etc.

[0039] The perylenequinone compound, its derivative or its pharmaceutically acceptable salt provided by the present invention can reduce the release level of the inflammatory factor NO in microglial cells (BV-2); further, it can significantly reduce the release level of the inflammatory factor NO in microglial cells (BV-2) induced by LPS, thereby achieving the technical effect of treating neuroinflammatory diseases, and can be used for preparing drugs for preventing and / or treating neuroinflammatory diseases, which is of great significance for the treatment of neuroinflammatory diseases and the development of new drugs.

[0040] Further, the drugs, which may be the same or different, respectively include a therapeutically effective amount of a perylenequinone compound, its derivative or its pharmaceutically acceptable salt.

[0041] Further, the drugs, which may be the same or different, respectively can be made into various pharmaceutical dosage forms by conventional methods, and these dosage forms include: tablets, sugar-coated tablets, film-coated tablets, enteric-coated tablets, capsules, hard capsules, soft capsules, oral liquids, buccal tablets, granules, infusion preparations, pills, pellets, suspensions, medicinal wines, tinctures, drops, tube feeding preparations, injections, sprays or ointments.

[0042] Further, the drugs, which may be the same or different, respectively may further contain one or more pharmaceutically acceptable carriers or excipients.

[0043] Further, the carrier or excipient may include diluents, binders, surfactants, wetting agents, adsorption carriers, lubricants, fillers, disintegrants, preservatives, etc.

[0044] In the fourth aspect of the present invention, there is provided a pharmaceutical composition for preventing and / or treating neuroinflammatory diseases, which contains the perylenequinone compound, its derivative or its pharmaceutically acceptable salt.

[0045] In the fifth aspect of the present invention, there is provided an inhibitor for the release of the inflammatory factor NO in BV-2 cells, which contains the perylenequinone compound, its derivative or its pharmaceutically acceptable salt.

[0046] Compared with the prior art, the present invention has the following advantages:

[0047] The present invention provides a perylenequinone compound or its derivative or its pharmaceutically acceptable salt and its new use. The perylenequinone compound or its derivative or its pharmaceutically acceptable salt of the present invention is isolated from the fermentation product of Alternaria sp. from the deep sea. It is verified by experiments that the perylenequinone compound or its derivative or its pharmaceutically acceptable salt has the effect of inhibiting the release of the inflammatory factor NO of microglial cells (BV-2), significantly reducing the release level of the inflammatory factor NO of microglial cells (BV-2) induced by LPS, and further achieving the technical effect of treating neurological inflammatory diseases. It can be used to prepare drugs for preventing and / or treating neurological inflammatory diseases, and is of great significance for the treatment of neurological inflammatory diseases and the development of new drugs. Description of the Drawings

[0048] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings required for use in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention, and therefore should not be regarded as limiting the scope. For those of ordinary skill in the art, other related drawings can be obtained based on these drawings without creative efforts.

[0049] Figure 1 Effect of the perylenequinone compound of the present invention on the release of NO by LPS-induced BV-2 microglial cells. Among them, *p<0.05, **p<0.01, ***p<0.001, vs LPS.

[0050] Figure 2 Half inhibitory concentration of the perylenequinone compound of the present invention on the release of NO by LPS-induced BV-2 microglial cells.

[0051] Figure 3 Effect diagram of the perylenequinone compound of the present invention on dopaminergic neurons in PD mice. Among them, *p<0.05, **p<0.01, ***p<0.001. Detailed Description of the Embodiments

[0052] The present invention will be further described in detail below in conjunction with the embodiments, but the embodiments of the present invention are not limited thereto. The materials involved in the following embodiments can be obtained from commercial channels without special instructions. The methods are conventional methods without special instructions.

[0053] The preservation information of the strain is as follows:

[0054] The Alternaria sp. Xiao20 mentioned above refers to CN116286391A, with the preservation number of CCTCC M2022501, and was preserved in the China Center for Type Culture Collection (CCTCC) on April 27, 2022.

[0055] Example 1: Preparation of the fermentation broth and fermentation compounds of Alternaria sp. Xiao20

[0056] 1. Preparation of the Alternaria sp. fermentation broth: The Alternaria sp. Xiao20 (preservation number CCTCC M2022501) was cultured on a PDA plate at 25 °C for 2 days; the fresh mycelium was inoculated into a culture solution containing 1000 mL of PDB. After 24 h, 20 mL of the seed solution was inoculated into 1 L Erlenmeyer flasks (200 flasks) and statically fermented at 25 °C for 31 days. The Erlenmeyer flasks contained 20 g of glucose, 200 g of potatoes (fresh, filtered), and 15 g of sea salt. The above substances were dissolved in 400 mL of deionized water and the pH was adjusted to 7.5 with NaOH to obtain the fermentation broth.

[0057] 2. Separation of perylenequinone compounds: The obtained fermentation broth was extracted three times with ethyl acetate, and the organic solvent was evaporated under reduced pressure to obtain a crude extract; the obtained crude extract was separated by normal-phase silica gel column chromatography, and gradient elution was performed with a dichloromethane-methanol system (50:1, 30:1, 20:1, 10:1, 5:1, 3:1) to obtain 5 fractions (Fr.1 - Fr.5); the crude fraction Fr.4 was separated by ODS column chromatography, and gradient elution was performed with a methanol-water system (5% to 100%) to obtain 5 sub-fractions (Fr.4-1 - Fr.4-5); for sub-fraction Fr.4-1, it was first separated using a Sephadex dextran gel chromatography column (methanol), and then separated using high-performance liquid chromatography (gradient elution with acetonitrile-water, 30% to 75%) to obtain Compound 1, Compound 2, Compound 4, and Compound 6; for sub-fraction Fr.4-4, it was separated using high-performance liquid chromatography (gradient elution with acetonitrile-water, 30% to 60%) to obtain Compound 3 and Compound 5.

[0058] The NMR data analysis of the above-obtained compounds confirmed that the chemical structural formulas are shown in Formulas 1 - 6 respectively. The 1 H (400 MHz) and 13 C (100 MHz) NMR data (CD3OD) are shown in Table 1.

[0059]

[0060] Table 1

[0061] NO. <![CDATA[δ C ,type]]> <![CDATA[δ H (mult,JinHz)]]> 1 133.9d 8.03(d,8.9) 2 119.5d 7.01(d,8.9) 3 162.2s 3a 115.3s 3b 138.6s 4 204.6s 5 37.7t 3.18 (dd, 17.5, 3.2) 3.36m 6 80.6d 4.34s 6a 71.5s 6b 41.3d 3.62s 7 51.0d 3.75m 8 56.7d 3.56m 9 62.1d 5.26s 9a 123.4s 9b 128.7s 10 157.9s 11 115.1d 6.84(d,8.4) 12 125.5d 7.59(d,8.4) 12a 125.3s 12b 126.8s OMe 57.1q

[0062] Example 2: Inhibitory Effect of Perylenequinone Compounds on Neuroinflammation

[0063] When immune cells are stimulated by microbial endotoxins, inflammatory mediators, etc., a large amount of nitric oxide synthase will be generated, and NO will be generated for immune response. Therefore, inhibiting NO generation is a direct indicator of the anti-inflammatory activity of compounds. In this example model, LPS was used to induce the inflammatory response of microglia (BV-2), and the Griess diazotization reaction was used to detect nitrite spontaneously oxidized from NO under physiological conditions by spectrophotometry.

[0064] 1. Experimental grouping:

[0065] Solvent control group (DMSO): Equal volume of culture medium, 0.1% dimethyl sulfoxide, without adding compounds;

[0066] Inducer control group (LPS): Equal volume of culture medium, without adding compounds + 1 μg / mL LPS;

[0067] Positive drug control group (Dex): Equal volume of culture medium, 0.1% dimethyl sulfoxide + 1 μM Dex (dexamethasone) + 1 μg / mL LPS;

[0068] Experimental group (6 compounds): Equal volume of culture medium, 0.1% dimethyl sulfoxide + 1 μM of the compound + 1 μg / mL LPS;

[0069] 2. Test content

[0070] Detection was carried out using a Griess kit (G7921, Invitrogen).

[0071] 2.1 Preparation of Griess reagent:

[0072] Volume of H2O required: 65 μL × number of wells;

[0073] Volume of Component A required: 5 μL × number of wells;

[0074] Volume of Component B required: 5 μL × number of wells;

[0075] Mix the Griess reagent, 75 μL / well.

[0076] 2.2 Preparation of standard curve:

[0077] After diluting the NO standard in the kit (1 mM) to 100 μM (the highest concentration of the standard curve), six gradients were serially diluted two-fold with the culture medium, and pure culture medium was used as the blank well. 75 μL of each was transferred to a 96-well plate. Add Griess reagent and mix well, 75 μL / well; incubate at room temperature in the dark for 30 min. Measure its absorbance at 548 nm, and establish a standard curve based on its absorbance.

[0078] 2.31 μM concentration test for inhibitory activity against LPS-induced neuroinflammation: Inoculate cell suspension (200,000 cells / well, 300 μL / well) in a 24-well plate, pre-culture in an incubator for 24 h, add the test drug to the culture plate, add LPS induction 1 h later, set up a DMSO control group and an LPS model group (1 μg / mL), and place in the incubator for 24 h. Take 75 μL of the supernatant culture medium from each well and transfer it to a 96-well plate, add Griess reagent and mix well, 75 μL / well; incubate at room temperature in the dark for 30 min. Measure its absorbance at 548 nm, and perform spectrophotometric quantification according to its absorbance against the standard curve; the results are as Figure 1 .

[0079] 2.4 Inhibitory rate test for NO at different concentrations:

[0080] Inoculate cell suspension (200,000 cells / well, 300 μL / well) in a 24-well plate, pre-culture in an incubator for 24 h, add different concentrations of the test drug to the culture plate, add LPS induction 1 h later, and place in the incubator for 24 h. Take 75 μL of the supernatant culture medium from each well and transfer it to a 96-well plate, add Griess reagent and mix well, 75 μL / well; incubate at room temperature in the dark for 30 min. Measure its absorbance at 548 nm, and perform spectrophotometric quantification according to its absorbance against the standard curve; the results are as Figure 2 , and the curves of Compound 1 - Compound 3 are shown in the figure.

[0081] As can be seen from the figure, the perylenequinone compounds of the present invention have an inhibitory activity against LPS-induced neuroinflammation exceeding 50% at a concentration as low as 1 μM; measure the inhibitory rate of NO at different concentration gradients and calculate its IC50. The results show that the perylenequinone compounds of the present invention effectively reduce the NO release induced by LPS, showing an anti-inflammatory effect on LPS-stimulated BV-2 cells, and can be applied to the preparation of drugs for preventing and / or treating neuroinflammatory diseases.

[0082] Example 3: Protective effect of perylenequinone compounds on dopaminergic neurons in subacute Parkinson's disease mice

[0083] The main pathological feature of Parkinson's disease (PD) is the degeneration of dopaminergic (TH) neurons in the substantia nigra. This neuronal damage is one of the important hallmarks of PD, and PD can be visually identified and analyzed by observing the degeneration of dopaminergic neurons in the substantia nigra. The onset of PD is often accompanied by an inflammatory response. 1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) is a neurotoxin that can selectively damage dopaminergic neurons, thus mimicking the pathological features of PD and is often used as a brain injury modeling reagent to verify anti-neuroinflammatory activity.

[0084] MPTP-induced subacute Parkinson's model:

[0085] 1. Experimental grouping:

[0086] Control group (0.9% NaCl): An equal volume of an aqueous solution containing 1% dimethyl sulfoxide + PBS;

[0087] Model group (MPTP): An equal volume of an aqueous solution containing 1% dimethyl sulfoxide + PBS + 30 mg / kg MPTP;

[0088] Drug administration group: An equal volume of an aqueous solution containing 1% dimethyl sulfoxide + PBS + 30 mg / kg MPTP + 30 mg / kg compound 1;

[0089] 2. The specific steps include

[0090] Fifteen SPF-grade C57 healthy male mice were randomly divided into 3 groups, with 5 mice in each group, namely the control group, the model group, and the drug administration group. The adaptation period was 7 days. The drug administration group was intraperitoneally injected with compound 1, and the other groups were injected with an equal volume of normal saline, and the drug was administered continuously for 7 days. Modeling began on the 3rd day of drug administration. 60 minutes after drug administration, except for the control group, the other groups were intraperitoneally injected with a single dose of 30 mg / kg MPTP solution, and the control group was injected with an equal volume of normal saline. Modeling was carried out continuously for 5 days. Samples were collected on the 8th day, and the complete brains of the mice were collected.

[0091] The immunohistochemical staining images of TH neurons showed that the perylenequinone compounds of the present invention could significantly protect dopaminergic neurons damaged by MPTP, manifested as an increase in TH-positive neurons ( Figure 3 ). The research results showed that the perylenequinone compounds of the present invention showed the potential to effectively prevent Parkinson's disease.

[0092] The above embodiments are the preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention shall be equivalent replacement methods and are all included in the protection scope of the present invention.

Claims

1. A perylenequinone compound or a derivative thereof or a pharmaceutically acceptable salt thereof, characterized in that The structural formula is shown in formula (I): Wherein, R1, R2, and R7 are the same or different and are selected from H, halogen, hydroxyl, carboxyl, cyano, nitro, amino, substituted or unsubstituted C1-C20 straight or branched alkyl, substituted or unsubstituted C3-C20 cycloalkyl, substituted or unsubstituted C1-C20 alkoxy, substituted or unsubstituted C1-C20 haloalkyl, C1-C10 carbonyl, C1-C10 ester, substituted or unsubstituted C1-C10 amine, substituted or unsubstituted C6-C20 aryl; R3 and R4 are the same or different and are selected from H, halogen, hydroxyl, carboxyl, cyano, nitro, amino, substituted or unsubstituted C1-C20 straight or branched alkyl, substituted or unsubstituted C3-C20 cycloalkyl, substituted or unsubstituted C1-C20 alkoxy, substituted or unsubstituted C1-C20 haloalkyl, C1-C10 carbonyl, C1-C10 ester, substituted or unsubstituted C1-C10 amine, substituted or unsubstituted C6-C20 aryl, or R3 and R4 are connected to form a C2-C10 heterocycle containing one or more of N, O and S; R5 and R6 are the same or different and are selected from H, O, halogen, hydroxyl, carboxyl, cyano, nitro, amino, substituted or unsubstituted C1-C20 straight or branched alkyl, substituted or unsubstituted C3-C20 cycloalkyl, substituted or unsubstituted C1-C20 alkoxy, substituted or unsubstituted C1-C20 haloalkyl, C1-C10 carbonyl, C1-C10 ester, substituted or unsubstituted C1-C10 amine, substituted or unsubstituted C6-C20 aryl.

2. The perylenequinone compound or its derivative or its pharmaceutically acceptable salt according to claim 1, characterized in that: The substitutions described are the same or different and refer to that one or more hydrogen atoms in the group are replaced by halogen, C1-C10 straight or branched alkyl, C3-C10 cycloalkyl, C1-C10 alkoxy, hydroxyl, carboxyl, cyano, nitro, amino, or -CF3 groups.

3. The perylenequinone compound or its derivative or its pharmaceutically acceptable salt according to claim 1, characterized in that: The structural formula is shown in formula (I), wherein R1, R2, and R7 are the same or different and are selected from H, halogen, hydroxyl, carboxyl, cyano, nitro, amino, C1-C20 straight or branched alkyl, C3-C20 cycloalkyl, C1-C20 alkoxy, C1-C20 haloalkyl, C1-C10 carbonyl, C1-C10 ester, C1-C10 amine, and C6-C20 aryl; R3 and R4 are the same or different and are selected from H, halogen, hydroxyl, carboxyl, cyano, nitro, amino, C1-C20 straight or branched alkyl, C3-C20 cycloalkyl, C1-C20 alkoxy, C1-C20 haloalkyl, C1-C10 carbonyl, C1-C10 ester, C1-C10 amine, C6-C20 aryl, or R3 and R4 are connected to form a C2-C10 heterocycle containing O; R5 and R6 are the same or different and are selected from H, O, halogen, hydroxyl, carboxyl, cyano, nitro, amino, C1-C20 straight or branched alkyl, C3-C20 cycloalkyl, C1-C20 alkoxy, C1-C20 haloalkyl, C1-C10 carbonyl, C1-C10 ester, C1-C10 amine, and C6-C20 aryl.

4. The perylenequinone compound or its derivative or its pharmaceutically acceptable salt according to claim 1, characterized in that: The structural formula is shown in formula (I), wherein R1, R2, and R7 are the same or different and are selected from H, halogen, hydroxyl, carboxyl, cyano, nitro, amino, C1-C20 straight or branched alkyl, C3-C20 cycloalkyl, C1-C20 alkoxy, and C1-C20 haloalkyl; R3 and R4 are the same or different and are selected from H, halogen, hydroxyl, carboxyl, cyano, nitro, amino, C1-C20 straight or branched alkyl, C3-C20 cycloalkyl, C1-C20 alkoxy, C1-C20 haloalkyl, or R3 and R4 are connected to form a C2-C10 heterocycle containing O; R5 and R6 are the same or different and are selected from H, O, halogen, hydroxyl, carboxyl, cyano, nitro, amino, C1-C20 straight or branched alkyl, C3-C20 cycloalkyl, C1-C20 alkoxy, C1-C20 haloalkyl.

5. The perylenequinone compound or its derivative or its pharmaceutically acceptable salt according to claim 1, characterized in that: The structural formula is shown in formula (I), and one or more hydrogen atoms in the structure shown in formula (I) are respectively replaced by halogen, hydroxyl, carboxyl, cyano, nitro, amino, C1-C10 straight or branched alkyl, C3-C10 cycloalkyl, C1-C10 alkoxy, C1-C10 haloalkyl, C1-C10 carbonyl, C1-C10 ester or aryl.

6. The perylenequinone compound or its derivative or its pharmaceutically acceptable salt according to claim 1, characterized in that The structural formula is one of Formula 1 to Formula 6:

7. A method for preparing the perylenquinone compound or its derivative or its pharmaceutically acceptable salt according to any one of claims 1 to 6, characterized in that It was isolated from the fermentation product of Alternaria sp. Xiao20. The Alternaria sp. (Alternaria sp. Xiao20) has a preservation number of CCTCC M 2022501 and was preserved in the China Center for Type Culture Collection on April 27, 2022.

8. Use of the perylenequinone compound or its derivative or its pharmaceutically acceptable salt according to any one of claims 1 to 6 in the preparation of the following products, characterized in that: (1) Drugs for preventing and / or treating neuroinflammatory diseases; (2) Drugs for inhibiting the release of inflammatory factor NO from BV-2 cells; (3) Drugs for inhibiting the release of inflammatory factor NO from BV-2 cells induced by LPS.

9. A pharmaceutical composition for preventing and / or treating neuroinflammatory diseases, characterized in that The invention comprises the perylequinone compound or its derivative or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6.

10. An inhibitor of the release of inflammatory factor NO in BV-2 cells, characterized in that The invention comprises the perylequinone compound or its derivative or pharmaceutically acceptable salt thereof according to any one of claims 1 to 6.

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