Melatonin derivative as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a melatonin derivative as well as a preparation method and application thereof. The melatonin derivative Mito-n-Mel (n = 2, 4, 6, 8, 10) provided by the invention can act on mitochondria in a targeting manner to cause damage to mitochondrial membrane potential in cells, so that active oxygen in the cells is excessively generated, the metabolic function of the mitochondria is destroyed, pyroptosis of the cells is induced, and growth and proliferation of tumor cells such as prostatic cancer are inhibited; therefore, the technical problems that in the prior art, targeted mitochondria is lacked, mitochondrial metabolism is damaged, and diseases are treated are solved.

Description

Technical Field

[0001] This application belongs to the field of pharmaceutical technology, and particularly relates to a melatonin derivative, a preparation method thereof, and an application thereof. Background Art

[0002] Mitochondria are the main sites for cells to perform oxidative phosphorylation and synthesize adenosine triphosphate (ATP), and are the centers of the body's energy metabolism; sugars, fats, proteins, etc. are cleaved into pyruvate and fatty acids in the cell and then enter the mitochondria to be cleaved into acetyl coenzyme A, and enter the tricarboxylic acid cycle to be gradually transmitted through the oxidative respiratory chain, generating end products such as water and carbon dioxide. The energy released in this process oxidizes and phosphorylates adenosine diphosphate (ADP) to generate adenosine triphosphate (ATP) to maintain the normal physiological functions of cells.

[0003] When mitochondria are damaged, a large amount of mitochondrial reactive oxygen species (ROS) are released into the cell, causing inflammation. In addition, the accumulation of reactive oxygen species (ROS) causes oxidative stress, damages vascular endothelial cells, and causes cardiovascular diseases such as atherosclerosis. At the same time, the continuous release of mitochondrial reactive oxygen species (ROS) is the direct inducement of oxidative stress, and oxidative stress is an important cause of neurodegenerative diseases such as Alzheimer's disease; therefore, achieving a healthy state of cells by maintaining the normal function of mitochondria can be used to treat diseases such as inflammation, cardiovascular diseases, and neurodegenerative diseases; in addition, mitochondria are deeply involved in the four major cell death pathways of apoptosis, autophagy, necrosis, and pyroptosis through multiple mechanisms such as dynamically regulating membrane permeability, energy metabolism, and signal molecule release, and play a key role in regulating apoptosis, autophagy, necrosis, and pyroptosis; Pyroptosis is an inflammatory cell death process dependent on caspase-1, which is characterized by the release of a large number of intracellular inflammatory mediators (such as IL-1β and IL-18) and subsequent cell membrane rupture; Oxidative stress and changes in mitochondrial membrane permeability are crucial for regulating pyroptosis. Reactive oxygen species (ROS) produced by mitochondria not only activate the NLRP3 inflammasome, but also promote the cleavage and activation of gasdermin D (GSDMD), resulting in the formation of pores on the cell membrane and pyroptosis; When pyroptosis is induced in tumor cells, the release of inflammatory factors not only promotes local inflammatory responses, but also recruits and activates immune cells such as dendritic cells, macrophages, and T cells, converting the tumor microenvironment into a highly immunogenic state, thereby enhancing the immune system's recognition and attack on tumor cells; Therefore, in addition to maintaining the normal function of mitochondria, another approach can be used, that is, by damaging or regulating mitochondrial function to achieve the killing and pro-apoptotic effects on abnormal cells (such as tumor cells) to treat diseases such as tumors; However, current research on targeting mitochondria and disrupting mitochondrial metabolism to treat diseases such as tumors is not deep enough, and there is a lack of drugs that target mitochondria and disrupt mitochondrial metabolism. Summary of the Invention

[0004] In view of this, the present application provides a melatonin derivative, its preparation method and application, which are used to solve the technical problem of the current lack of drugs that target mitochondria and disrupt mitochondrial metabolism to treat diseases.

[0005] The first aspect of the present application provides a melatonin derivative, the chemical structure of which is shown in Formula I:

[0006] ;

[0007] In Formula I, n = 2, 4, 6, 8, 10.

[0008] The second aspect of the present application provides a preparation method of a melatonin derivative, which can prepare the melatonin derivative described in Formula I of the first aspect. The preparation method includes the following steps:

[0009] Step S1: Perform a nucleophilic substitution reaction on triphenylphosphine and bromoorganic acid to obtain an intermediate;

[0010] Step S2: Perform an amidation reaction on the intermediate and 5-methoxytryptamine to obtain a melatonin derivative.

[0011] Preferably, in Step S1, the bromoorganic acid used in the nucleophilic substitution reaction is selected from 2-bromoacetic acid, 4-bromobutyric acid, 6-bromohexanoic acid, 8-bromooctanoic acid or 10-bromodecanoic acid.

[0012] Preferably, in Step S1, the organic solvent used in the nucleophilic substitution reaction is selected from acetonitrile.

[0013] Preferably, in Step S2, the reagent used to activate the carboxylic acid in the amidation reaction is 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, the reagent used to prevent racemization is 1-hydroxybenzotriazole, the organic base used for catalysis is N,N-diisopropylethylamine, and the organic solvent is dichloromethane.

[0014] Preferably, in Step S1, the temperature of the nucleophilic substitution reaction is 81°C to 82°C, and the time is 6 h to 12 h.

[0015] Preferably, in Step S2, the temperature of the amidation reaction is room temperature, and the time is 0.5 h to 24 h.

[0016] Preferably, in Step S1, the molar ratio of triphenylphosphine to bromoorganic acid used in the nucleophilic substitution reaction is 1 to 1.2:1 to 1.2.

[0017] Preferably, in Step S2, the molar ratio of the intermediate to 5-methoxytryptamine used in the amidation reaction is 1 to 1.2:1 to 1.2.

[0018] Preferably, after Step S1 and before Step S2, rotary evaporation concentration is also included to purify and crystallize the intermediate;

[0019] After Step S2, vacuum distillation and chromatographic purification are also included to obtain a refined product of the melatonin derivative.

[0020] The third aspect of the present application provides the use of a melatonin derivative as described in Formula I of the first aspect in the preparation of a drug for inducing pyroptosis.

[0021] The fourth aspect of the present application provides the use of a melatonin derivative as described in Formula I of the first aspect in the preparation of a drug for preventing or treating mitochondrial metabolism-related diseases.

[0022] The fifth aspect of the present application provides the use of a melatonin derivative described in Formula I of the first aspect in the preparation of a drug for preventing or treating tumors.

[0023] Preferably, the tumor is a benign tumor, a malignant tumor or a borderline tumor.

[0024] The sixth aspect of the present application provides a pharmaceutical composition, comprising a pharmaceutically active ingredient and a pharmaceutical excipient;

[0025] The pharmaceutically active ingredient is selected from a melatonin derivative described in Formula I of the first aspect or a pharmaceutically acceptable salt, ester, prodrug or hydrate thereof.

[0026] Compared with the prior art, a melatonin derivative provided by the present application has at least the following beneficial effects:

[0027] 1. The melatonin derivative provided by the present application can target mitochondria, cause damage to the mitochondrial membrane potential in cells, lead to excessive generation of reactive oxygen species in cells, disrupt mitochondrial metabolic functions, induce pyroptosis, and inhibit the growth and proliferation of tumor cells such as prostate cancer cells like PC3 and DU145.

[0028] 2. The melatonin derivative provided by the present application can be used to prepare a drug for inducing pyroptosis, a drug for preventing or treating mitochondrial metabolism-related diseases, or a drug for preventing or treating tumors. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] In order to more clearly illustrate the specific embodiments of the present application or the technical solutions in the prior art, the following will briefly introduce the drawings required for use in the description of the specific embodiments or the prior art. Obviously, the drawings in the following description are some embodiments of the present application. For those of ordinary skill in the art, other drawings can be obtained based on these drawings without creative efforts.

[0030] Figure 1 It is the experimental result diagram of Experimental Example 1 of the present application;

[0031] Figure 2 It is the experimental result diagram of Experimental Example 2 of the present application;

[0032] Figure 3 It is the experimental result diagram of Experimental Example 3 of the present application;

[0033] Figure 4 It is the experimental result diagram of Experimental Example 4 of the present application;

[0034] Figure 5 It is the experimental result diagram of Experimental Example 5 of the present application. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0035] The present application provides a melatonin derivative, a preparation method and an application thereof, which are used to solve the technical problem that there is currently a lack of a technology for targeting mitochondria and disrupting mitochondrial metabolism to treat diseases.

[0036] The technical solutions of the present application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are part of the embodiments of the present application, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments in the present application without making creative efforts shall fall within the protection scope of the present application.

[0037] In view of the current defect that the research on targeting mitochondria and disrupting mitochondrial metabolism to treat diseases such as tumors is not deep enough, the present application provides a melatonin derivative, and the melatonin derivative provided by the present application is shown in Formula I;

[0038] Formula I.

[0039] In Formula I, n = 2, 4, 6, 8, 10, and they can be respectively named Mito-2-Mel, Mito-4-Mel,

[0040] Mito-6-Mel, Mito-8-Mel, Mito-10-Mel.

[0041] The melatonin derivative shown in Formula I provided by the present application can cause damage to the mitochondrial membrane potential in cells, resulting in the excessive generation of reactive oxygen species in cells. The accumulation of reactive oxygen species will further aggravate the membrane potential damage, disrupt the mitochondrial metabolic function, induce pyroptosis in tumor cells such as prostate cancer, and inhibit the growth and proliferation of tumor cells such as prostate cancer. Therefore, the melatonin derivative shown in Formula I provided by the present application can target mitochondria and disrupt mitochondrial metabolism to treat diseases such as tumors.

[0042] Preferably, among the melatonin derivatives provided by the present application, when n = 10, that is, when the melatonin derivative named Mito-10-Mel is selected, the performance is better than that of melatonin derivatives such as Mito-2-Mel, Mito-4-Mel, Mito-6-Mel, and Mito-8-Mel.

[0043] The present application correspondingly provides a preparation method for the above-mentioned melatonin derivative, and the synthetic route of the preparation method is shown as follows:

[0044] .

[0045] Among them, 2a - 2e are bromoorganic acids such as 2-bromoacetic acid, 4-bromobutyric acid, 6-bromohexanoic acid, 8-bromooctanoic acid, or 10-bromodecanoic acid.

[0046] The present application also provides the use of the above melatonin derivatives, which can be used for preparing drugs for inducing pyroptosis, drugs for preventing or treating mitochondrial metabolism-related diseases, and drugs for preventing or treating tumors; meanwhile, the present application also provides a pharmaceutical composition, which includes a drug active ingredient and a pharmaceutical excipient; the pharmaceutical composition is for inducing pyroptosis, preventing or treating mitochondrial metabolism-related diseases, and preventing or treating tumors; the pharmaceutical composition consists of a drug active ingredient and a pharmaceutical excipient, and the drug active ingredient is selected from the above melatonin derivatives Mito-2-Mel, Mito-4-Mel, Mito-6-Mel, Mito-8-Mel, Mito-10-Mel, while the pharmaceutical excipient is selected from solvents for carrying drug dissolution, solubilizers for increasing drug solubility, preservatives for preventing microbial contamination of drugs during storage and use, flavoring agents for improving the taste and smell of drugs, coloring agents for imparting specific colors to drugs, suspending agents for increasing the suspension of insoluble drugs, disintegrants for promoting the rapid fragmentation of tablets into fine particles to facilitate drug absorption, and other pharmaceutical excipients.

[0047] Example 1

[0048] Example 1 of the present application provides a preparation method of a melatonin derivative, and the preparation method includes the steps of nucleophilic substitution reaction and amidation reaction.

[0049] The steps of the nucleophilic substitution reaction include: adding a solution of 3.81 mmol of triphenylphosphine and 3.81 mmol of 2-bromoacetic acid to 30 ml of acetonitrile, heating to reflux for 9 hours, then concentrating the acetonitrile on a rotary evaporator, and purifying and crystallizing to obtain a white solid intermediate; the reaction can be monitored by thin-layer chromatography (TLC) to determine that the reaction has been completed.

[0050] The steps of the amidation reaction include: sequentially adding 0.24 mmol of the intermediate, 0.48 mmol of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide EDCI, 0.24 mmol of 1-hydroxybenzotriazole HOBT, and 10 ml of CH2Cl2 to a dry 50-ml reaction flask at room temperature, stirring the reaction mixture for 1 hour, then adding 0.24 mmol of 5-methoxytryptamine under stirring, and reacting overnight at room temperature with an appropriate amount of organic base N,N-diisopropylethylamine DIPEA as a catalyst; the reaction can be monitored by thin-layer chromatography (TLC) to determine that the reaction has been completed, then washing with 80 ml of water, then drying the organic layer over Na2SO4, distilling the solvent under reduced pressure, and purifying the crude product by flash chromatography on silica gel (CH2Cl2 / CH3OH = 20:1, v / v)) to obtain a white solid powder of Mito-2-Mel respectively.

[0051] The spectral data are as follows:

[0052] Example 2

[0053] Example 2 of the present application provides a method for preparing a melatonin derivative. The difference between the preparation method and that of Example 1 is that 4-bromobutyric acid is used to replace the bromo organic acid, and the method includes the steps of a nucleophilic substitution reaction and an amidation reaction.

[0054] The steps of the nucleophilic substitution reaction include: adding a solution of 3.81 mmol of triphenylphosphine and 3.81 mmol of 4-bromobutyric acid to 30 ml of acetonitrile, heating the mixture to reflux for 9 hours, then concentrating the acetonitrile on a rotary evaporator, and purifying and crystallizing to obtain a white solid intermediate; the reaction can be monitored by thin-layer chromatography (TLC) to determine that the reaction is complete.

[0055] The steps of the amidation reaction include: adding 0.24 mmol of the intermediate, 0.48 mmol of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide EDCI, 0.24 mmol of 1-hydroxybenzotriazole HOBT, and 10 ml of CH2Cl2 to a dry 50-ml reaction flask in sequence at room temperature. The reaction mixture is stirred for 1 hour, then 0.24 mmol of 5-methoxytryptamine is added under stirring, and the reaction is carried out overnight at room temperature with an appropriate amount of organic base N,N-diisopropylethylamine DIPEA as a catalyst; the reaction can be monitored by thin-layer chromatography (TLC) to determine that the reaction is complete, then washed with 80 ml of water, and then the organic layer is dried over Na2SO4, the solvent is distilled under reduced pressure, and the crude product is purified by flash chromatography on silica gel (CH2Cl2 / CH3OH = 20:1, v / v) to obtain a white solid powder of Mito-4-Mel respectively.

[0056] The spectral data are as follows:

[0057] Example 3

[0058] Example 3 of the present application provides a method for preparing a melatonin derivative. The difference between the preparation method and that of Example 1 is that 6-bromohexanoic acid is used to replace the bromo organic acid, and the method includes the steps of a nucleophilic substitution reaction and an amidation reaction.

[0059] The steps of the nucleophilic substitution reaction include: adding a solution of 3.81 mmol of triphenylphosphine and 3.81 mmol of 6-bromohexanoic acid to 30 ml of acetonitrile, heating the mixture to reflux for 9 hours, then concentrating the acetonitrile on a rotary evaporator, and purifying and crystallizing to obtain a white solid intermediate; the reaction can be monitored by thin-layer chromatography (TLC) to determine that the reaction is complete.

[0060] The steps of the amidation reaction are as follows: In a dry 50 mL reaction flask, 0.24 mmol of the intermediate, 0.48 mmol of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 0.24 mmol of 1-hydroxybenzotriazole (HOBT), and 10 mL of CH2Cl2 are added in sequence at room temperature. The reaction mixture is stirred for 1 hour, then 0.24 mmol of 5-methoxytryptamine is added with stirring, and the reaction is carried out overnight at room temperature using an appropriate amount of organic base N,N-diisopropylethylamine (DIPEA) as a catalyst; the reaction can be monitored by thin-layer chromatography (TLC) until completion, then washed with 80 mL of water, and then the organic layer is dried over Na2SO4. The solvent is distilled under reduced pressure, and the crude product is purified by flash chromatography on silica gel (CH2Cl2 / CH3OH = 20:1, v / v) to obtain the white solid powder of Mito-6-Mel respectively.

[0061] The spectral data are as follows:

[0062] Example 4

[0063] Example 4 of this application provides a preparation method of a melatonin derivative. The difference between the preparation method and that of Example 1 is that 8-bromooctanoic acid is used instead of bromo organic acid, including the steps of nucleophilic substitution reaction and amidation reaction.

[0064] The steps of the nucleophilic substitution reaction are as follows: In 30 mL of acetonitrile, a solution of 3.81 mmol of triphenylphosphine and 3.81 mmol of 8-bromooctanoic acid is heated to reflux and reacted for 9 hours. Then the acetonitrile is concentrated on a rotary evaporator, and the purified crystal is obtained as a white solid intermediate; the reaction can be monitored by thin-layer chromatography (TLC) until completion.

[0065] The steps of the amidation reaction are as follows: In a dry 50 mL reaction flask, 0.24 mmol of the intermediate, 0.48 mmol of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 0.24 mmol of 1-hydroxybenzotriazole (HOBT), and 10 mL of CH2Cl2 are added in sequence at room temperature. The reaction mixture is stirred for 1 hour, then 0.24 mmol of 5-methoxytryptamine is added with stirring, and the reaction is carried out overnight at room temperature using an appropriate amount of organic base N,N-diisopropylethylamine (DIPEA) as a catalyst; the reaction can be monitored by thin-layer chromatography (TLC) until completion, then washed with 80 mL of water, and then the organic layer is dried over Na2SO4. The solvent is distilled under reduced pressure, and the crude product is purified by flash chromatography on silica gel (CH2Cl2 / CH3OH = 20:1, v / v) to obtain the white solid powder of Mito-8-Mel respectively.

[0066] The spectral data are as follows:

[0067] Example 5

[0068] Example 5 of the present application provides a method for preparing a melatonin derivative. The difference between the preparation method and that of Example 1 is that the bromo organic acid is replaced with 10-bromodecanoic acid, and it includes the steps of nucleophilic substitution reaction and amidation reaction.

[0069] The steps of the nucleophilic substitution reaction include: adding a solution of 3.81 mmol of triphenylphosphine and 3.81 mmol of 10-bromodecanoic acid to 30 ml of acetonitrile, heating to reflux for 9 hours, then concentrating the acetonitrile on a rotary evaporator, and purifying and crystallizing to obtain a white solid intermediate; the reaction can be monitored by thin layer chromatography (TLC) to determine that the reaction is complete.

[0070] The steps of the amidation reaction include: adding 0.24 mmol of the intermediate, 0.48 mmol of 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide EDCI, 0.24 mmol of 1-hydroxybenzotriazole HOBT, and 10 ml of CH2Cl2 to a dry 50-ml reaction flask in sequence at room temperature. The reaction mixture is stirred for 1 hour, then 0.24 mmol of 5-methoxytryptamine is added with stirring, and the reaction is carried out overnight at room temperature using an appropriate amount of organic base N,N-diisopropylethylamine DIPEA as a catalyst; the reaction can be monitored by thin layer chromatography (TLC) to determine that the reaction is complete, then washed with 80 ml of water, and then the organic layer is dried over Na2SO4. The solvent is distilled under reduced pressure, and the crude product is purified by flash chromatography on silica gel (CH2Cl2 / CH3OH = 20:1, v / v) to obtain a white solid powder of Mito-10-Mel respectively.

[0071] The spectral data are as follows:

[0072] Experimental Example 1

[0073] In Experimental Example 1 of the present application, performance tests were carried out on the melatonin derivatives Mito-2-Mel, Mito-4-Mel, Mito-6-Mel, Mito-8-Mel, and Mito-10-Mel (yield 52.7% - 69.8%) provided in Examples 1 - 5.

[0074] The performance tests included inhibiting the growth and proliferation of prostate cancer cell lines PC3 and DU145. The instruments used included a microplate reader (BioTek Synergy HT), the cell lines included PC3 and DU145, and the reagents included a CCK-8 kit, DMEM and MEM complete media, DMSO, FBS, PBS, and melatonin (Mel).

[0075] The experimental procedure for inhibiting the growth and proliferation of prostate cancer cell lines PC3 and DU145 was as follows: The cells were cultured in a medium containing different concentrations of the experimental drug at a density of 2000 cells per well in a 96-well plate, with a volume of 100 μL, at a culture temperature of 37 °C for 24 hours. Then, the complete medium was replaced with a medium containing different Mito-Mel analogs and cultured for 24 hours. Then, 10 μL of the cell counting kit (CCK-8) was added to each well, and the culture plate was further cultured in a humidified CO2 incubator at 37 °C and 5% CO2 for 2 - 4 hours. Finally, the absorbance at 450 nm was measured using a microplate reader (BioTek Synergy HT).

[0076] The experimental results of inhibiting the growth and proliferation of prostate cancer cell lines PC3 and DU145 were as Figure 1 shown. It was found that the compound Mito-10-Mel could significantly inhibit the proliferation of tumor cells. In addition, Mito-10-Mel significantly inhibited the proliferation of PC3 and DU145 cells at a concentration 100 times lower than that of melatonin Mel. This indicated that Mito-10-Mel in the present invention had good activity in inhibiting the proliferation of tumor cells.

[0077] The performance tests also included the effect on the growth and proliferation of normal prostate cells WPMY-1. The instruments used included an inverted microscope, the cell lines included PC3, DU145, and WPMY-1, and the reagents included DMEM and MEM complete media, 4% paraformaldehyde, 0.1% crystal violet, DMSO, Mito-10-Mel, and melatonin (Mel).

[0078] The experimental procedure for the effect on the growth and proliferation of normal prostate cells WPMY-1 was as follows: The cells were seeded in a 6-well plate (1000 cells per well) and treated with Mel (2 mM) and Mito-Mel (2 μM), and then cultured at 37 °C for 5 days. Subsequently, the medium was replaced with fresh drug-free medium and the cells were cultured for another 10 days. Finally, the cells were fixed with 4% formaldehyde, stained with crystal violet, and photographed.

[0079] The experimental results of the effect on the growth and proliferation of normal prostate cells WPMY-1 were as Figure 1As shown, it was found that compound Mito-10-Mel could significantly inhibit the growth of PC3 and DU145 cells without affecting the growth of normal prostate cells WPMY-1.

[0080] Experimental Example 2

[0081] In Experimental Example 2 of this application, performance tests were carried out on the melatonin derivative Mito-10-Mel provided in Example 5; the performance tests included: inducing mitochondrial membrane potential damage, reactive oxygen species (ROS) accumulation and mitochondrial structure damage in prostate cancer cells, and the results were as Figure 2 shown.

[0082] To induce mitochondrial membrane potential damage in prostate cancer cells, JC-1 was used to detect mitochondrial membrane potential damage. The instruments used included a flow cytometer (BECKMAN COULTER), and the reagents included JC-1 (BD). The experimental process included incubating PC3 cells with the compound for 24 h, collecting the cells and washing them with PBS, then staining with JC-1 for 30 min at room temperature in the dark, and performing flow cytometry analysis.

[0083] To detect the accumulation of reactive oxygen species (ROS), a ROS detection kit was used to detect ROS. The instruments used included a flow cytometer (BECKMAN COULTER), and the reagents included Reactive Oxygen Species Assay Kit (Yeasen). The experimental process included incubating PC3 cells with the compound for 24 h, staining with ROS dye for 30 min, collecting the cells and washing them with PBS, and performing flow cytometry analysis.

[0084] To detect the damage of mitochondrial structure by immunofluorescence, a high-sensitivity laser confocal microscope was used. The reagents included Mito-Tracker Deep Red FM and DAPI. The experimental process included inoculating PC3 cells on a 24-well plate, and after the cells adhered, culturing them with the compound for 12 h. Then the cells were fixed with 4% paraformaldehyde, washed several times with PBS, and sealed with 4′,6-diamidino-2-phenylindole dihydrochloride (DAPI) (1:10000), and the images were analyzed by a laser scanning confocal microscope (Leica).

[0085] The experimental results were as Figure 2 shown. From Figure 2 it can be seen that the melatonin derivative Mito-10-Mel could induce damage to the mitochondrial membrane potential in PC3 cells in a concentration gradient, and cause the accumulation of reactive oxygen species in tumor cells, affecting the function of tumor cell mitochondria and inducing damage and destruction of the mitochondrial structure of tumor cells.

[0086] Experimental Example 3

[0087] In Experimental Example 3 of this application, the performance of the melatonin derivative Mito-10-Mel provided in Example 5 was tested; the performance tests included: the effect on the mitochondrial metabolic function of prostate cancer cell lines.

[0088] The effect on the mitochondrial metabolic function of prostate cancer cell lines was evaluated by measuring the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR) of the cells to assess mitochondrial metabolic function; the instruments used included the Agilent Seahorse XFe96 analyzer, and the reagents included oligomycin, FCC, rotenone, antimycin A, Glucose, and 2-DG (2-deoxy-glucose).

[0089] The experimental procedure included: inoculating the cells at 4×10 ^4 cells per well in a specific 96-well plate and culturing overnight; then replacing the complete medium with a medium supplemented with the required complexes and continuing to culture for 24 hours; then adding XF analysis medium containing 25 mM glucose, 1 mM sodium propionate, and 2 mM glutamine, as well as the complexes to maintain the stimulating environment, and equilibrating the cells in a CO2-free incubator for 1 hour; transferring the cells to a Seahorse 96-well plate and measuring the oxygen consumption every 10 seconds continuously; then treating the cells sequentially with oligomycin (ATP synthase inhibitor, 1.0 μM), FCCP (mitochondrial uncoupler, 1.0 μM), and rotenone / antimycin (inhibitor of electron transport chain complex I / III, 1.0 μM) to determine the change in mitochondrial oxygen consumption rate (OCR); in addition, the cells were treated sequentially with glucose, oligomycin (ATP synthase inhibitor, 1.0 μM), and 2-DG (glycolysis inhibitor, 1.0 μM) to measure the change in extracellular acidification rate (ECAR).

[0090] The experimental results are as Figure 3 shown. It can be seen from Figure 3 that the melatonin derivative Mito-10-Mel can significantly inhibit the glycolysis of PC3, DU145, and TRAMP-C1 cells at a concentration of 5 μM, and reduce the disruption of the oxidative phosphorylation process of the cells, thereby disrupting the mitochondrial metabolic function of prostate cancer cell lines.

[0091] Experimental Example 4

[0092] In Experimental Example 4 of this application, the performance of the melatonin derivative Mito-10-Mel provided in Example 5 was tested; the performance tests included: the effect on pyroptosis.

[0093] The instruments used in the experiment on the effect of pyroptosis include the transmission electron microscope Hitachi HT-7800, and the reagents include glutaraldehyde.

[0094] The process of the experiment on the effect of pyroptosis includes: inoculating cells in a 6-cm diameter culture dish, treating the cells with the compound after they adhere to the wall and culturing for 3 h; then digesting and centrifuging the cells, adding the glutaraldehyde working solution along the tube wall, and fixing the cells overnight at 4°C; the next day, washing the cells three times with 0.1 M PB solution, and finally analyzing the images by transmission electron microscope.

[0095] The results of the experiment on the effect of pyroptosis are as Figure 4 shown. From Figure 4 it can be seen that at concentrations of 2.5 μM and 5 μM of Mito-10-Mel and an action time of 3 hours, the cells showed changes such as cell membrane swelling, cell membrane rupture, and release of cell contents, resulting in pyroptosis of the cells.

[0096] Experimental Example 5

[0097] In Experimental Example 5 of this application, performance tests were carried out on the melatonin derivative Mito-10-Mel provided in Example 5; the performance tests included: the effect on the occurrence and development of prostate cancer (PC3) tumors and pyroptosis.

[0098] In the process of the performance test, BALB / c(nu / nu) nude mice (weighing 18 -20 g, raised in a SPF-class animal room) were selected to construct an in vitro mouse xenograft tumor model. The mice were grouped and inoculated with human prostate cancer cells (PC3). When the tumor volume in the mice grew to 100 -300 mm3, intraperitoneal injection was given. Each mouse in the administration group was given Mel (200 mg / kg) once a day according to its body weight, , and each mouse in the control group was given the same volume of PBS buffer. At the same time, the volume of each mouse was measured, and the administration was continued for 15 days; after the administration was completed, the mice were sacrificed by cervical dislocation, the tumors of the mice were dissected, photographed, and weighed; the tumor size and volume data were statistically analyzed.

[0099] The experimental results are as Figure 5 shown. From Figure 5 it can be seen that the melatonin derivative can well inhibit human prostate cancer xenografts in mice and induce mitochondrial function damage and pyroptosis in tumor tissue cells; in addition, the HE& staining results of the heart, liver, spleen, lung, and kidney of the administration group and the control group were measured, indicating that the melatonin derivative has no obvious toxic and side effects.

[0100] The above embodiments are only used to illustrate the technical solutions of the present application, rather than to limit them; although the present application has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: they can still modify the technical solutions described in the foregoing embodiments, or perform equivalent replacements on some or all of the technical features; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application.

Claims

1. A melatonin derivative, characterized in that, Its chemical structure is shown in Formula I: Formula I; In Formula I, n = 2, 4, 6, 8, 10.

2. The preparation method of a melatonin derivative according to claim 1, characterized in that, It includes the following steps: Step S1: Perform a nucleophilic substitution reaction on triphenylphosphine and bromo organic acid to obtain an intermediate; Step S1: Perform an amidation reaction on the intermediate and 5-methoxytryptamine to obtain a melatonin derivative.

3. The preparation method of a melatonin derivative according to claim 2, characterized in that, In Step S1, the bromo organic acid used in the nucleophilic substitution reaction is selected from 2-bromoacetic acid, 4-bromobutyric acid, 6-bromohexanoic acid, 8-bromooctanoic acid or 10-bromodecanoic acid.

4. The preparation method of a melatonin derivative according to claim 2, characterized in that, In Step S2, the reagent used for activating the carboxylic acid in the amidation reaction is 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, the reagent used for preventing racemization is 1-hydroxybenzotriazole, the organic base used for catalysis is N,N-diisopropylethylamine, and the organic solvent is dichloromethane.

5. The preparation method of a melatonin derivative according to claim 2, wherein The temperature of the nucleophilic substitution reaction is 81°C to 82°C, and the time is 6 h to 12 h; The temperature of the amidation reaction is room temperature, and the time is 0.5 h to 2 h.

6. Use of a melatonin derivative according to claim 1 in the preparation of a drug for inducing pyroptosis.

7. Use of a melatonin derivative according to claim 1 in the preparation of a drug for preventing or treating mitochondrial metabolism-related diseases.

8. Use of a melatonin derivative according to claim 1 in the preparation of a drug for preventing or treating tumors.

9. The application according to claim 8, wherein The tumor is a benign tumor, a malignant tumor or a borderline tumor.

10. A pharmaceutical composition, characterized in that, It includes a drug active ingredient and a pharmaceutical excipient; The drug active ingredient is selected from a melatonin derivative according to claim 1 or a pharmaceutically acceptable salt, ester, prodrug or hydrate thereof.

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