Olympicene, process for its preparation and use thereof

Abstract

The application discloses olympicene or a pharmaceutically acceptable salt thereof, which has a structure shown in formula (I). The olympicene or the pharmaceutically acceptable salt thereof shown in formula (I) has low cytotoxicity, can activate the synthesis of glutathione after entering cells, and can strengthen the antioxidant capacity of cells. Meanwhile, the metabolism of the olympicene or the pharmaceutically acceptable salt thereof can up-regulate the expression of genes in an electron transport chain in mitochondria, so that the effect of promoting mitochondrial metabolism is achieved.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of organic synthesis, and in particular to an olympicene and a preparation method and application thereof. BACKGROUND

[0002] Free radicals are open-shell compounds, whose unpaired electrons occupy a single occupied molecular orbital (SOMO). In biological systems, appropriate free radical concentrations help control physiological functions, while abnormal free radical concentrations can cause cell damage. In the past few decades, singlet biradical materials have become a kind of redox-active substances that have been intensively studied. Due to the coexistence of the quinolone closed-shell and the aromatic open-shell resonance structure, single-electron and double-electron redox can be stabilized by restoring aromaticity, thus making the molecule have clear multi-level redox polymorphism. In addition, the redox activity of singlet biradical molecules can be adjusted by modifying the biradical coefficient (y0), the substituent group and the molecular strain.

[0003] Compared with the rapid development of singlet biradical materials in the field of organic semiconductors, little attention has been paid to their application in the biomedical field. This is mainly because the polycyclic aromatic hydrocarbon structure of singlet biradical molecules may have potential carcinogenicity, and it is difficult to be made into a material with water solubility and biocompatibility.

[0004] Therefore, it is necessary to develop a new olympicene compound, which has the functions of activating mitochondrial function and promoting cell antioxidant function. SUMMARY

[0005] The present application aims to at least solve one of the problems in the prior art. To this end, the first aspect of the present application provides an olympicene, which has low cytotoxicity and enhances the antioxidant capacity of cells, and its metabolism can up-regulate the expression of genes in the electron transport chain of mitochondria, achieving the effect of promoting mitochondrial metabolism.

[0006] The second aspect of the present application also provides a preparation method of the olympicene.

[0007] The third aspect of the present application also provides a nanoparticle.

[0008] The third aspect of the present application also provides an application of the olympicene.

[0009] According to the first aspect of the present application, the olympicene or the pharmaceutically acceptable salt thereof has the structure shown in formula (I):

[0010]

[0011] The olympicene according to the embodiments of the present application has at least the following beneficial effects:

[0012] The olympicene shown in the formula (I) or the pharmaceutically acceptable salt thereof provided by the application has low cytotoxicity, and can activate the synthesis of glutathione after entering cells, and strengthen the antioxidant capacity of cells; on the other hand, the metabolism thereof can up-regulate the expression of genes of the electron transport chain in mitochondria, so as to achieve the effect of promoting mitochondrial metabolism.

[0013] The preparation method of olympicene provided by the second aspect of the embodiments of the application comprises the following steps:

[0014] S1, mixing compound 1, phenyllithium and a first organic solvent and reacting at -10℃ to 10℃ for 20 to 50 min; then, the temperature is increased to room temperature for continuous reaction, and the intermediate product is obtained after post-treatment;

[0015] S2, mixing the intermediate product, tin chloride and a second organic solvent under an inert atmosphere to obtain the structure shown in the formula (I);

[0016] The structural formula of the compound 1 is as follows:

[0017]

[0018] According to some embodiments of the application, in step S1, the step of post-treatment is:

[0019] Water is added to the reaction solution in step S1, and extraction, drying and concentration are performed to obtain the intermediate product.

[0020] According to some embodiments of the application, compound 1, phenyllithium and a first organic solvent are mixed and reacted at -5℃ to 5℃. For example, a specific embodiment can be 0℃.

[0021] According to some embodiments of the application, the first organic solvent comprises at least one of anhydrous tetrahydrofuran and chloroform. For example, a specific embodiment can be anhydrous tetrahydrofuran.

[0022] According to some embodiments of the application, the molar ratio of the compound 1 to phenyllithium is 1:(2 to 5).

[0023] According to some embodiments of the application, the second organic solvent comprises at least one of dichloromethane, ethanol and isopropanol. For example, a specific embodiment can be dichloromethane.

[0024] According to some embodiments of the application, the molar ratio of the intermediate product to tin chloride is 1:(2 to 5).

[0025] According to some embodiments of the application, the inert atmosphere is at least one of an argon atmosphere, a helium atmosphere or a neon atmosphere.

[0026] According to some embodiments of the present application, the temperature of the reaction in step S2 is -10℃ to 10℃.

[0027] The third aspect of the present application provides a nanoparticle formed by self-assembly of olympicene prepared by the above method.

[0028] The olympicene of the present application is adsorbed to each other by intermolecular hydrophobic interaction, and the charges generated by the free radicals increase with the increase of the volume after adsorption, which makes them repel each other, and finally form nanoparticles by self-assembly in a steady state.

[0029] According to some embodiments of the present application, the particle size of the nanoparticle is 100-200 nm.

[0030] According to some embodiments of the present application, the self-assembly is performed as follows:

[0031] Olympicene represented by formula (I), dimethyl sulfoxide and water are ultrasonically mixed, wherein the volume of dimethyl sulfoxide accounts for less than 5% of the total volume.

[0032] According to some embodiments of the present application, the concentration of the nanoparticle in the solution is 10-200 mg / L. For example, it can be specifically 50 mg / L.

[0033] The fourth aspect of the present application provides the use of the above-mentioned olympicene or the above-mentioned nanoparticle in improving the antioxidant activity of cells and promoting mitochondrial metabolism.

[0034] Definitions and general terms

[0035] The term "pharmaceutically acceptable salt" used in the present application refers to organic and inorganic salts of the compounds of the present application

[0036] Salts. Pharmaceutically acceptable salts are well known in the art, for examples, see S. M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19, 1977. Pharmaceutically acceptable non-toxic acid addition salts include those derived from inorganic acids, such as hydrochloric, hydrobromic, phosphoric, sulfuric, perchloric, and organic acids, such as acetic, oxalic, maleic, tartaric, citric, succinic, malonic, and other similar acids. Other pharmaceutically acceptable salts include adipate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxyethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, pivalate, propionate, stearate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Salts derived from an appropriate base include alkali metal, alkaline earth metal, ammonium, and N + (C 1-4 alkyl)4 salts. The present application also contemplates the quaternary ammonium salts of any group containing N. Water or oil-soluble or dispersible products can be obtained by quaternization. Alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Pharmaceutically acceptable salts further include appropriate, nontoxic ammonium, quaternary ammonium, and amine cations formed by the addition of inorganic or organic acids to a free amine group of the present compounds. Such salts are known to be formed by means well known in the art, for example, S. M. Berge et al., describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19, 1977. 1-8Sulfonates and aromatic sulfonates. Amine salts such as, but not limited to, N,N'-dibenzylethylenediamine salts, chloroprocaine salts, choline salts, ammonia salts, isopropylamine salts, benzathine salts, cholinate salts, lysine salts, meglumine salts, piperazine salts, tromethamine salts, diethanolamine salts and other hydroxyalkylamine salts, ethylenediamine salts, N-methylglucamine salts, procaine salts, N-benzylphenethylamine salts, 1 -p-chlorobenzyl-2-pyrrolidine-1 '-ylmethyl-benzimidazole salts and other alkylamine salts, piperazine salts and tris(hydroxymethyl)aminomethane salts; alkaline earth metal salts such as, but not limited to, barium salts, calcium salts and magnesium salts; transition metal salts such as, but not limited to, zinc salts.

[0037] The pharmaceutically acceptable salts of the present application can be synthesized from the parent compound which contains a basic or acidic moiety by conventional chemical methods. Generally, such salts can be prepared by reacting the free acid or base forms of these compounds with a stoichiometric amount of the appropriate base or acid in water or in an organic solvent, or in a mixture of the two. Generally, use of nonaqueous media, such as ether, ethyl acetate, ethanol, isopropanol, or acetonitrile, is desirable, where appropriate. Lists of additional suitable salts can be found, e.g., in "Remington's Pharmaceutical Sciences", 20th ed., Mack Publishing Company, Easton, Pa., (1985); and "Hand book of Pharmaceutical Salts: Properties, Selection, and Use", Stahl and Wermuth (Wiley-VCH, Weinheim, Germany, 2002).

[0038] "Room temperature" in the present application means 20°C ± 5°C.

[0039] Additional features and advantages of the application will be set forth in the description that follows, and in part will become apparent to those skilled in the art upon examination of the following or can be learned by practice of the application. BRIEF DESCRIPTION OF DRAWINGS

[0040] The foregoing and / or additional aspects and advantages of the present application will become apparent and be readily understood upon examination of the following specification, including the drawings in which:

[0041] Figure 1 is a TEM image of nanoparticles prepared according to Example 2 of the present application. DETAILED DESCRIPTION

[0042] The following are specific embodiments of the present application, and the technical solutions of the present application are further described in combination with the embodiments, but the present application is not limited to these embodiments.

[0043] The reagents, methods and equipment used in the present application are all conventional reagents, methods and equipment in the technical field unless otherwise specified.

[0044] Example 1

[0045] Example 1 provides an olympicene, the reaction equation and the preparation method thereof are as follows:

[0046]

[0047] S1, compound 1 (50 mg, 0.20 mmol, commercially available) was dissolved in 3.5 mL of anhydrous tetrahydrofuran, and the solution was cooled to 0°C with an ice bath. A hexane solution of phenyllithium (1.0 mole per liter, 0.60 mL, 0.60 mmol) was added dropwise at 0°C. The reaction mixture was stirred at 0°C for 30 minutes. The reaction mixture was slowly warmed to room temperature and stirred overnight. Then pure water was added, the mixture was extracted with dichloromethane and dried over sodium sulfate, and the solution was concentrated under vacuum to obtain an intermediate product, which was directly used in the next reaction;

[0048] S2, the intermediate product was dissolved in 2 mL of dichloromethane, and tin chloride (113 mg, 0.60 mmol) was added at once. The mixture was stirred for another 15 minutes under the protection of argon, then the reaction solution was filtered, the solvent was removed under reduced pressure, and the crude mixture was purified on a gel permeation chromatograph using polystyrene column and dichloromethane as the eluent to obtain olympicene.

[0049] Structure confirmation: the olympicene prepared was subjected to single crystal confirmation, and single crystal X-ray diffraction study was performed on an XtaLAB Synergy diffractometer (Rigaku) using Cu Kα radiation (λ = 1.5418 A) at 100 K. The diffractometer was equipped with a Hypix6000 HE detector (Rigaku). The carbon atom numbers are as follows. The specific data results are as shown in Table 1:

[0050]

[0051] Table 1

[0052]

[0053]

[0054] Example 2

[0055] Example 2 provides a kind of nanoparticles, specific steps as follows:

[0056] Olympic alkene prepared in Example 1 is dissolved in dimethyl sulfoxide at a concentration of 10 mg / ml, after ultrasonic mixing, the solution is dropped into pure water, fully mixed, when the volume ratio of dimethyl sulfoxide is less than 5%, Olympic alkene self-assembles into uniform and stable nanoparticles.

[0057] The prepared nanoparticles are subjected to TEM detection, and the results are shown in Figure 1 The particle size is between 100-200 nm.

[0058] Performance test

[0059] 1. Cytotoxicity test:

[0060] Mouse kidney primary cells are plated in 96-well plates and incubated overnight, then different concentrations of Olympic alkene prepared in Example 1 are added and incubated for 24 hours. ATP concentration is detected using CellTiter-Glo luminescent cell viability assay. Quantitative results are read on a multifunctional enzyme marker (Bio-Rad), and the cytotoxicity IC 50 > 2500 μg / mL.

[0061] 2. Cell metabolism test:

[0062] The prepared nanoparticle solution (50 mg / L) is added to the cell culture medium (DMEM / F12 purchased from Thermo Fisher Scientific (Catalog No. 11320033)), and incubated for 24 hours. Reduced glutathione (GSH) content detection kit (Solarbio, Catalog No. BC1175) is used to detect the glutathione content of the control group (phosphate buffer) and the drug group (nanoparticle solution), and it is found that the glutathione level of the drug group is significantly improved (n=3). At the same time, RNA sequencing by second-generation sequencing technology (DNBSEQ sequencer (Huada Gene)) finds that the expression of genes related to mitochondrial electron transport chain in the drug group is significantly improved (n=3). Finally, high-throughput targeted metabolomics detection analysis of metabolites by liquid chromatography tandem mass spectrometry technology (metabolite test by Huada Technology) finds that the key metabolites related to carnitine cycle and organic acid products in the tricarboxylic acid cycle are also significantly improved (n=10), and the specific data are listed in Table 2.

[0063] Table 2

[0064]

[0065] As can be seen from Table 2, the metabolism of the nanoparticles prepared in Example 2 of the present application in cells can activate the synthesis of glutathione by cells, increase the content of glutathione, and further enhance the antioxidant capacity of cells; meanwhile, under the enhancement of carnitine cycle and tricarboxylic acid cycle, the function of mitochondria can be restored or activated, and finally the oxidative phosphorylation process in mitochondria is promoted, so that the cell metabolism is more active.

[0066] The above is described in detail in combination with the embodiments of the present application, but the present application is not limited to the above embodiments, and various changes can be made within the knowledge range possessed by those skilled in the art without departing from the purpose of the present application.

Claims

1. An Olympicene, characterized in that, It has the structure shown in equation (Ⅰ): 。 2. The method for preparing Olympicene according to claim 1, characterized in that, Includes the following steps: S1. Mix compound 1, phenyllithium and the first organic solvent and react at -10℃ to 10℃ for 20 to 50 min; then raise the temperature to room temperature and continue the reaction, and the intermediate product is obtained by post-treatment. S2. Under an inert atmosphere, the intermediate product, tin chloride, and the second organic solvent are mixed and reacted to obtain the structure shown in formula (Ⅰ); The structural formula of compound 1 is as follows: 。 3. The method for preparing Olympicene according to claim 2, characterized in that, In step S1, the post-processing steps are as follows: Water is added to the reaction solution in step S1, and the intermediate is obtained by extraction, drying and concentration.

4. The method for preparing Olympicene according to claim 2, characterized in that, The first organic solvent is at least one of anhydrous tetrahydrofuran and chloroform.

5. The method for preparing Olympicene according to claim 2, characterized in that, The molar ratio of compound 1 to phenyllithium is 1:(2~5).

6. The method for preparing Olympicene according to claim 2, characterized in that, The second organic solvent is at least one of dichloromethane, ethanol, and isopropanol.

7. A nanoparticle, characterized in that, Olympicene, prepared by the method described in claim 1 or by any one of claims 2 to 6, is formed by self-assembly.

8. The nanoparticles according to claim 7, characterized in that, The nanoparticles have a particle size of 100~200nm.

9. The nanoparticles according to claim 7, characterized in that, The self-assembly steps are as follows: The Olympicene, dimethyl sulfoxide, and water described in claim 1 are ultrasonically mixed, wherein the volume of dimethyl sulfoxide accounts for less than 5% of the total volume.

10. The use of the Olympicene of claim 1 or the nanoparticles of any one of claims 7 to 9 in the preparation of drugs that enhance cellular antioxidant capacity and promote mitochondrial metabolism.

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