Process for the preparation of amino fullerene derivatives

Abstract

The application provides a preparation method of an amino fullerene derivative, which comprises the following steps: adding an amine compound into a chlorinated fullerene solution, stirring the reaction at room temperature, and obtaining the amino fullerene derivative after the reaction is completed; the amine compound is a primary amine compound; no acidic reagent is added in the reaction process of the method; the fullerene is selected from at least one of hollow fullerenes, metal fullerenes, heterocyclic fullerenes and endohedral fullerenes. 60 The method realizes controllable amino modification of the fullerene by adding a secondary amine or a tertiary amine to regulate the nucleophilic substitution reaction of the primary amine compound on C 16, and obtains a series of novel structure tetraamino epoxy fullerene derivatives with clear molecular structures.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of organic synthesis, in particular to a preparation method of a new amino fullerene derivative with a clear molecular structure. BACKGROUND

[0002] Fullerenes are a series of molecules composed of different numbers of carbon atoms with a cage-like fused ring structure. Due to its large conjugated electron structure, fullerenes can be bonded to a variety of groups through chemical reactions to achieve functional modification, thereby obtaining a variety of fullerene derivatives. Among them, amino fullerene derivatives are considered to have great application prospects in the field of biological medicine due to their good tumor inhibition, bacteriostasis, and antiviral properties. Amino fullerene derivatives are mostly prepared by nucleophilic reaction of fullerenes with amino compounds. Among them, the direct reaction of secondary amine compounds with oxidizing agents after being dissolved in chlorobenzene can obtain tetraamino epoxide fullerene derivatives. Such molecules have a clear structure, which is conducive to biomedical applications (DOI: 10.1039 / c39950002023; DOI: 10.1021 / jo050432y; DOI: 10.1246 / cl.2007.20). However, this method is limited to the types of amino compounds substrates and is only suitable for secondary amine derivatives, not for primary amine compounds. 60

[0003] Hexachlorofullerene (C 60 C l6 ) is a well-defined fullerene derivative that is simple and fast to synthesize and can be prepared in large quantities. C 60 Cl6 has stronger reaction selectivity and reactivity than C 60 , and can undergo nucleophilic substitution reactions with amino, thiol, alkoxy, and other groups to obtain corresponding well-defined fullerene derivatives, so it is often used as a precursor in the preparation of fullerene derivatives. C 60 C l6 can directly react with primary amine compounds, but only well-defined pentaamino fullerene derivatives can be prepared (DOI: 10.1039 / c2cc00071g). In the preparation method reported in this document, an acidic reagent needs to be added, and the reaction yield is not more than 80%, which is not conducive to large-scale production of products. Therefore, based on this reaction, a synthesis method for controllable synthesis of other amino fullerene derivatives with different structures is designed, which realizes the regulation of the structure of amino fullerene derivatives and has great significance in the field of organic synthesis and biomedical applications. SUMMARY

[0004] In order to solve the problems existing in the prior art, the purpose of the present application is to provide a preparation method of a new amino fullerene derivative with a clear molecular structure, which is based on C 60 ​C16 is a method for rapidly preparing amino fullerene derivatives by adding secondary amine or tertiary amine to regulate the reaction of primary amine compound with C 60 The nucleophilic substitution reaction of C16 realizes the controllable amino modification of fullerene, and a series of novel structure tetra-amino epoxy fullerene derivatives with clear molecular structure are obtained. Meanwhile, the amino fullerene derivatives prepared by the method of the application can significantly shorten the reaction time and improve the preparation efficiency.

[0005] To solve the above technical problems, the application adopts the following technical solutions:

[0006] In one aspect, the application provides a method for preparing an amino fullerene derivative, comprising the following steps:

[0007] The amine compound is added to the chlorinated fullerene solution, and the reaction is stirred at room temperature. After the reaction is completed, an amino fullerene derivative is obtained. The amine compound is a primary amine compound. No acidic reagent is added during the reaction process of the method. The fullerene is selected from at least one of hollow fullerene, metal fullerene, heterocyclic fullerene and endohedral fullerene.

[0008] In one embodiment, the method further comprises the step of adding a secondary amine compound or a tertiary amine compound to the chlorinated fullerene solution.

[0009] In one embodiment, the amine compound, the secondary amine compound or the tertiary amine compound, and the cumene hydroperoxide are added to the chlorinated fullerene solution, and the reaction is stirred at room temperature. After the reaction is completed, the amino epoxy fullerene derivative is obtained by washing and drying. The epoxy fullerene is a fullerene containing an oxirane structure.

[0010] In one embodiment, the fullerene is selected from any one or mixture of C 2n , M@C 2n , M2@C 2n , MA@C 2n , M3N@C 2n , M2C2@C 2n , M2S@C 2n , M2O@C 2n and M x A 3-x N@C 2n , wherein M and A are both metal elements, and the M and A are both selected from any one of Sc, Y and lanthanide metal elements. Preferably, the fullerene is selected from one or more fullerene molecules containing C 2n , wherein 2n is the number of carbon atoms, 30≤n≤60; preferably, the fullerene is selected from C 60 , C 70 , C 76 , C 78 , C 80 , C84 one or more; more preferably, the fullerene is C 60 .

[0011] In an embodiment, the synthetic route of the method is as follows:

[0012] The amine compound and the cumene hydroperoxide are added to the chlorinated fullerene solution, and the reaction is stirred at room temperature until the reaction is completed to obtain the amino fullerene derivative;

[0013] Preferably, the amino fullerene derivative is a tetra-substituted amino-epoxy fullerene derivative;

[0014] Preferably, the molar ratio of the chlorinated fullerene and the primary amine compound is 1:(10-30); more preferably, the molar ratio of the chlorinated fullerene and the primary amine compound is selected from 1:10, 1:15, 1:20, 1:25 or 1:30;

[0015] Preferably, the molar ratio of the chlorinated fullerene, the primary amine compound, the secondary amine compound and the cumene hydroperoxide is (1-3):(10-30):(1-5):(1:5);

[0016] Preferably, the molar ratio of the chlorinated fullerene, the primary amine compound, the secondary amine compound and the cumene hydroperoxide is 1:15:3:3;

[0017] Preferably, the molar ratio of the chlorinated fullerene, the primary amine compound, the tertiary amine compound and the cumene hydroperoxide is (1-3):(10-30):(3-8):(1:5);

[0018] Preferably, the molar ratio of the chlorinated fullerene, the primary amine compound, the tertiary amine compound and the cumene hydroperoxide is 1:15:5:3;

[0019] Preferably, the time of stirring at room temperature is 0.6h-1h; preferably, 1h.

[0020] In an embodiment, the chlorinated fullerene solution is obtained by dissolving chlorinated fullerene in an organic solvent selected from toluene or chlorobenzene.

[0021] In an embodiment, the amine compound is NH2CR 1 R 2 , R 1 , R 2 are each independently selected from -C1-C6alkyl, -C1-C6alkyl-C1-C6alkyl-C6-C8aryl, -C(O)O-C1-C6alkyl, -C6-C8aryl; preferably, the R 1 , R2 each independently selected from the group consisting of -C1-C6alkyl, -C1-C6alkyl-C1-C6alkyl-C6-C8aryl, -C(O)O-C1-C6alkyl, -C6-C8aryl; preferably, the amine compound is selected from one or more of the following:

[0022] In one embodiment, the secondary amine compound is pyrrolidine or piperidine.

[0023] In one embodiment, the tertiary amine compound is NR 5 R 6 R 7 wherein R 5 , R 6 , R 7 each independently selected from the group consisting of -C1-C6alkyl.

[0024] In one embodiment, the tertiary amine is triethylamine.

[0025] In one embodiment, the amino fullerene derivative has the following structure:

[0026]

[0027] In one embodiment, the amino fullerene derivative has the following structure:

[0028]

[0029] In another aspect, the present application provides a compound of the following formula:

[0030]

[0031]

[0032] In another aspect, the present application provides a pharmaceutical composition comprising the amino fullerene derivative prepared by the aforementioned method, the aforementioned compound or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0033] In another aspect, the present application provides a use of the aforementioned method, the aforementioned compound or a pharmaceutically acceptable salt thereof, the aforementioned pharmaceutical composition in the preparation of a medicament for treating cancer.

[0034] In one embodiment, the cancer is selected from one or more of liver cancer, lung cancer, colorectal cancer, kidney cancer, pancreatic cancer, bone cancer, breast cancer, ovarian cancer, prostate cancer, esophageal cancer, stomach cancer, oral cancer, nasal cancer, laryngeal cancer, cholangiocarcinoma, cervical cancer, uterine cancer, testicular cancer, meningioma, skin cancer, melanoma, lymphoma, glioma, leukemia, or sarcoma.

[0035] The beneficial effects of the present application are at least as follows:

[0036] (1) Using chlorinated fullerene and primary amine as raw materials, the nucleophilic substitution reaction of primary amine compound on chlorinated fullerene is regulated by adding secondary amine or tertiary amine, and in the presence of cumene hydroperoxide, the controlled amino modification of fullerene is realized, and tetra-substituted amino-epoxy fullerene derivative is produced;

[0037] (2) Without adding acidic reagents, the protonation of amino group of amine compounds is effectively avoided;

[0038] (3) The purity of the product is relatively high, which is maintained above 95%;

[0039] (4) The accurate preparation of a certain number of amino fullerene derivatives is realized. BRIEF DESCRIPTION OF DRAWINGS

[0040] Figure 1 The molecular structure (left) and high-resolution mass spectrum (right) of C 60 Cl6 prepared in Example 1 are shown.

[0041] Figure 2 The high-performance liquid chromatogram of C 60 Cl6 prepared in Example 1 is shown.

[0042] Figure 3 The nuclear magnetic resonance carbon spectrum of C 60 Cl6 prepared in Example 1 is shown.

[0043] Figure 4 The ultraviolet absorption spectrum of C 60 Cl6 dissolved in chloroform prepared in Example 1 is shown.

[0044] Figure 5 The molecular structure (left) and high-resolution mass spectrum (right) of the amino fullerene TGly-OBu t prepared in Example 2 are shown.

[0045] Figure 6 The high-performance liquid chromatogram of the amino fullerene molecule TGly-OBu t prepared in Example 2 is shown.

[0046] Figure 7 The nuclear magnetic resonance hydrogen spectrum of the amino fullerene TGly-OBu t prepared in Example 2 is shown.

[0047] Figure 8 The nuclear magnetic resonance carbon spectrum of the amino fullerene TGly-OBu t prepared in Example 2 is shown.

[0048] Figure 9 The aminofullerene TPhe-OBu prepared in Example 3 is shown. t The molecular structure (left) and high-resolution mass spectra (right).

[0049] Figure 10 The aminofullerene molecule TPhe-OBu prepared in Example 3 is shown. t High performance liquid chromatography.

[0050] Figure 11 The aminofullerene TPhe-OBu prepared in Example 3 is shown. t The proton nuclear magnetic resonance spectrum.

[0051] Figure 12 The aminofullerene TPhe-OBu prepared in Example 3 is shown. t The carbon NMR spectrum.

[0052] Figure 13 The aminofullerene TLeu-OBu prepared in Example 4 is shown. t The molecular structure (left) and high-resolution mass spectra (right).

[0053] Figure 14 The aminofullerene molecule TLeu-OBu prepared in Example 4 is shown. t High performance liquid chromatography.

[0054] Figure 15 The aminofullerene TLeu-OBu prepared in Example 4 is shown. t The proton nuclear magnetic resonance spectrum.

[0055] Figure 16 The aminofullerene TLeu-OBu prepared in Example 4 is shown. t The carbon NMR spectrum.

[0056] Figure 17 The molecular structure (left) and high-resolution mass spectra (right) of the aminofullerene TPhe-OMe prepared in Example 5 are shown.

[0057] Figure 18 The high-performance liquid chromatography of the aminofullerene molecule TPhe-OMe prepared in Example 5 is shown.

[0058] Figure 19 The aminofullerene TAla-OBu prepared in Example 6 is shown. t The molecular structure (left) and high-resolution mass spectra (right).

[0059] Figure 20 The aminofullerene molecule TAla-OBu prepared in Example 6 is shown. t High performance liquid chromatography.

[0060] Figure 21 The molecular structure (left) and high resolution mass spectrum (right) of the aminofullerene molecule TDBMAC prepared in Example 8 are shown.

[0061] Figure 22 The high performance liquid chromatogram of the tetra-substituted aminooxofullerene derivative prepared by reacting C16 with a primary amine compound is shown.

[0062] Figure 23 The high performance liquid chromatogram of the tetra-substituted aminooxofullerene derivative prepared by reacting C16 with a primary amine compound is shown. 60 Cl6 with a primary amine compound. DETAILED DESCRIPTION

[0063] I. DEFINITIONS

[0064] In the present application, unless otherwise specified, the scientific and technical terms used herein have the meanings commonly understood by one of ordinary skill in the art. Also, the relevant terms and laboratory operation steps used herein are the terms and conventional steps widely used in the corresponding field. At the same time, in order to better understand the present application, the definitions and explanations of the relevant terms are provided as follows.

[0065] As used herein and unless otherwise indicated, the term "about" or "approximately" means within 10% of a given value or range. In the case of integers, the term means within 10% of a given value or range, rounded to the nearest integer.

[0066] In the description herein, reference is made to "some embodiments", "some implementations" or "some aspects", which describe a subset of all possible embodiments, but it can be understood that "some embodiments" can be the same subset or different subsets of all possible embodiments, and can be combined with each other without conflict.

[0067] All ranges disclosed herein are to be interpreted as implicitly disclosing all sub-ranges of the same base or distinct end points of the ranges. For example, disclosure of the range 0.5-1 h is to be interpreted to also disclose the sub-ranges 0.6-0.9 h, 0.7-0.8 h, etc., as well as the individual points 0.8, 0.85, 0.9, 0.95, etc. Similarly, disclosure of the range 0.5-1 h is to be interpreted to also disclose the sub-ranges 0.6-1 h, 0.7-1 h, 0.8-1 h, etc., as well as the individual points 0.8, 0.85, 0.9, 0.95, 0.99, 1.0, 1.05, etc.

[0068] The term "alkyl" refers to saturated aliphatic hydrocarbon groups, including straight-chain or branched-chain saturated hydrocarbon groups having the number of carbon atoms indicated. For example, the term "C1-C6 alkyl" includes C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, examples of which include, but are not limited to, methyl, ethyl, n-propyl, i-propyl, n-butyl, i-butyl, t-butyl, n-pentyl, 2-pentyl, 3-pentyl, n-hexyl, 2-hexyl, 3-hexyl, and the like.

[0069] As used herein, the term "fullerene" is a series of spherical-like cluster molecules composed of an even number of carbon atoms, 12 pentagons, and the rest of hexagons. Fullerene includes hollow fullerene, endohedral fullerene, which is a metal or metal cluster encapsulated inside the carbon cage structure of fullerene.

[0070] The term "metal fullerene", "endohedral fullerene" refers to a variety of different metals or metal clusters encapsulated inside the carbon cage structure of fullerene, forming a class of compounds with special structure and properties, such compounds are commonly referred to as endohedral fullerene, generally represented by the form M@C2n, wherein M represents a metal element.

[0071] The term "chlorinated fullerene" refers to a fullerene derivative obtained by combining one or more carbon atoms on the fullerene with chlorine.

[0072] The term "amino fullerene derivative" refers to the amino modification of fullerene, and the modified fullerene includes one or more same or different amino-containing groups outside.

[0073] The term "pharmaceutically acceptable salt" refers to a salt that retains the biological effectiveness of the free acids and bases of a particular compound without biological adverse effects. For example, acid (including organic acid and inorganic acid) addition salt or base addition salt (including organic base and inorganic base).

[0074] The pharmaceutically acceptable salt of the present application can be synthesized from the parent compound containing an acid group or a base group by conventional chemical methods. Generally, the salt of such a compound is prepared by reacting the free acid or base form of the compound in water or in an organic solvent, or in a mixture of both, with a stoichiometric amount of the appropriate base or acid.

[0075] II. Examples

[0076] In order to make the purpose, technical scheme and advantages of the present application more clear, the present application will be further described in detail below, and the described examples should not be regarded as limitation of the present application, all other examples obtained by those skilled in the art without making creative efforts shall fall within the scope of protection of the present application.

[0077] Before further detailed description of the examples of the present application, the terms and phrases involved in the examples of the present application are explained, and the terms and phrases involved in the examples of the present application are applicable to the following explanations.

[0078] The raw materials and equipment used in the specific embodiments of the present application are known products, which are obtained by purchasing commercially available products.

[0079] Example 1: Preparation of hexachlorofullerene C 60 Cl6

[0080]

[0081] Synthesis method: 1 g of C 60 was dissolved in 100 mL of chlorobenzene, 4 g of iodine monochloride was added dropwise, after stirring at room temperature for 1 h, the product was filtered through a solvent filter (nylon filter membrane, 0.22 μm), precipitated by adding 500 mL of ethanol, the precipitate was collected by filtration, washed with ethanol for 3 times, and dried at 60 °C under vacuum for 24 h to obtain red solid product C 60 Cl6.

[0082] Structural characterization: The molecular weight was detected by electrospray mass spectrometry (ESI-MS, Thermo Scientific Exactive, negative ion mode), and the results are shown in Figure 1 ; its purity was detected by high performance liquid chromatography (HPLC, LC-2030C 3D, Shimadzu), C18 chromatographic column (Agilent, Eclipse XDB-C18, 4.6*250mm, filler particle size 5 μm), toluene-acetonitrile as mobile phase, specific gradient method is shown in Table 1, ultraviolet detector (detection wavelength: 310 nm), flow rate is 1 mL / min, and the purity detection results are shown in Figure 2 ; the detection results of elemental analysis are shown in Table 2; nuclear magnetic resonance (Bruker Avance III 400HD, 400 MHz) was used to analyze the structure of the prepared C 60 Cl6sample was dissolved in carbon disulfide and deuterated chloroform for carbon spectrum detection, and the structure of the product was analyzed, and the detection results are shown in Figure 3 ; the product was dissolved in chloroform, and ultraviolet visible spectrometer (Hitachi, UH4150) was used to detect ultraviolet absorption, and the detection results are shown in Figure 4 .

[0083] Conclusion: The mass spectrometry signal in the literature is 897.5 (C 60 Cl5 - mass spectrometry signal) and 968.5 (C 60 Cl7 - mass spectrometry signal), the molecular weight of the product detected by ESI-MS is 897.8 and 968.8, which is basically the same as the mass spectrometry data reported in the literature, and the isotopic distribution of the molecular mass spectrometry signal of the product is completely consistent with the theoretical distribution (literature: Seven-Minute Synthesis of Pure Cs-C 60 Cl6from

[60] Fullerene and Iodine Monochloride: First IR, Raman, and Mass Spectra of 99 mol% C 60Cl6); HPLC purity > 91%; elemental analysis showed the mass percentage of medium carbon was 76.2%, which was very close to the theoretical value (77.2%); Figure 3 The chemical shift of the nuclear magnetic resonance carbon spectrum was C 60 Cl6 structure was consistent; Figure 4 Characteristic ultraviolet absorption at 214, 255, 278, 388 nm, which was basically consistent with the reported data (211, 257, 280, 386 nm) (literature: Preparation and 13 C NMRSpectroscopic Characterisation of C 60 C16).

[0084] Table 1 shows the high performance liquid chromatography detection of C 60 Cl6 toluene-acetonitrile gradient method

[0085]

[0086] Table 2 shows the C 60 Cl6 elemental analysis detection results

[0087]

[0088] Example 2: Preparation of tetra-substituted amino epoxy fullerene molecule TGly-OBu t

[0089]

[0090] Synthesis method: 1g C 60 Cl6 was dissolved in 1000 mL of toluene, 600 μL of hydrogen peroxide isopropylbenzene (80%) (3 equivalents) was added, 2.19 mL of glycine tert-butyl ester (15 equivalents) was mixed with 745 μL of triethylamine (5 equivalents) and added to the above reaction solution, stirred (1500 rpm) at room temperature for 1 h, the reaction solution was taken for HPLC detection, the reaction was complete, the reaction was stopped, the reaction solution was washed with 500 mL of saturated ammonium chloride solution and 500 mL of saturated sodium bicarbonate solution in turn, the organic layer was dried with anhydrous sodium sulfate, concentrated and evaporated at 40°C to obtain a red-brown solid.

[0091] The reaction product was dissolved in toluene, and a rapid preparation liquid chromatograph (Sephaven TM machine) was used for purification. The standard type rapid separation column S-8101-0120 was pre-filled with normal phase silica gel, the particle size was 40-63 μm, and the pore size was 60 A. The obtained product was concentrated and evaporated at 40°C to obtain a red-brown solid.

[0092] Structural characterization: The molecular weight was detected by electrospray mass spectrometry (ESI-MS, Thermo Scientific Exactive, positive ion mode), and the detection results are shown in FIG. 1. Figure 5 ; high performance liquid chromatography (HPLC, LC-2030C 3D, Shimadzu) was used, a C18 chromatographic column (Agilent, Eclipse XDB-C18, 4.6*250mm, filler particle size 5μm) was used, toluene-acetonitrile was used as the mobile phase, the specific gradient method is shown in Table 3, and an ultraviolet detector (detection wavelength: 310nm) was used, the flow rate was 1mL / min, and the purity was detected, and the detection results are shown in FIG. 2. Figure 6 ; nuclear magnetic resonance (Bruker Avance III 400HD, 400MHz) was used, the sample was dissolved in deuterated chloroform for hydrogen spectrum and carbon spectrum detection, and the product structure was analyzed, and the hydrogen spectrum and carbon spectrum detection results are shown in FIG. 3 and FIG. 4, respectively. Figure 7 Figure 8 .

[0093] Conclusion: The molecular weight detected by ESI-MS is completely consistent with the theoretical molecular weight of the glycine tert-butyl ester four-addition epoxy fullerene (TGly-OBu t ), the purity detected by HPLC is more than 95%, and the hydrogen atom shift and integral area of the hydrogen spectrum and the carbon atom shift of the carbon spectrum of the nuclear magnetic resonance are consistent with the structure of TGly-OBu t . It can be seen that the method can efficiently prepare the four-substituted amino fullerene derivative TGly-OBu t , and realize accurate preparation of a certain number of amino fullerene derivatives.

[0094] Table 3 shows the toluene-acetonitrile gradient method of high performance liquid chromatography

[0095]

[0096] Example 3: Preparation of four-substituted amino epoxy fullerene molecule TPhe-OBu t

[0097]

[0098] Synthetic method: 1g C 60 ​C16 was dissolved in 1000 mL of toluene, 600 μL of cumene hydroperoxide (80%) (3 equivalents) was added, 3 mL of phenylalanine tert-butyl ester (15 equivalents) was mixed with 745 μL of triethylamine (5 equivalents) and then added to the above reaction solution, stirred at room temperature (1500 rpm) for 1 h, the reaction solution was taken for HPLC detection, the reaction was complete, the reaction was stopped, the reaction solution was washed with 500 mL of saturated ammonium chloride solution and 500 mL of saturated sodium bicarbonate solution in turn, the organic layer was dried with anhydrous sodium sulfate, concentrated and evaporated at 40°C to obtain a red-brown solid.

[0099] The reaction product was dissolved in toluene and column chromatography was performed using a fast preparation liquid chromatograph (SepaBean TM machine), normal phase silica gel column (SepaBean standard fast separation column S-8101-0120, pre-packed normal phase silica gel, particle size 40-63 μm, pore size 120 g). The eluent was ethyl acetate / toluene, the volume fraction of ethyl acetate was 1-5%, the flow rate was 60 mL / min, and the ultraviolet detector (detection wavelength 1: 310 nm, detection wavelength 2: 320 nm). The obtained product was concentrated and evaporated at 40°C to obtain a red-brown solid.

[0100] Structure characterization: the molecular weight was detected by electrospray mass spectrometry (ESI-MS, Thermo Scientific Exactive, positive ion mode); high performance liquid chromatography (HPLC, LC-2030, Shimadzu) was used, C18 chromatographic column (Agilent, Eclipse XDB-C18, 4.6*250 mm, filler particle size 5 μm), toluene-acetonitrile as mobile phase (Table 3), ultraviolet detector (detection wavelength: 310 nm), flow rate 1 mL / min, to detect the purity; nuclear magnetic resonance (Bruker Avance III400HD, 400 MHz) was used, the sample was dissolved in deuterated o-dichlorobenzene for hydrogen spectrum detection and carbon spectrum detection to analyze the structure of the product. Figure 9 Figure 10 Figure 11 Figure 12

[0101] Conclusion: the molecular weight detected by ESI-MS is completely consistent with the theoretical molecular weight of phenylalanine tert-butyl ester four-substituted epoxy fullerene (TPhe-OBu t ), the purity detected by HPLC is more than 95%, and the hydrogen atom shift and integral area of the hydrogen spectrum and the carbon atom shift of the carbon spectrum are consistent with the structure of TPhe-OBu t . It can be seen that the four-substituted amino fullerene derivative TGly-OBu t ​​​​, to achieve accurate preparation of a determined number of amino fullerene derivatives.

[0102] Example 4: Preparation of tetra-substituted amino-epoxy fullerene molecule TLeu-OBu t

[0103]

[0104] Synthetic method: 1 g of C 60 C16 was dissolved in 1000 mL of toluene, 600 μL of cumene hydroperoxide (80%) (3 equivalents) was added, 3 mL of leucine tert-butyl ester (15 equivalents) was mixed with 745 μL of triethylamine (5 equivalents) and added to the above reaction solution, stirred at room temperature (1500 rpm) for 1 h, the reaction solution was taken for HPLC detection, the reaction was complete, the reaction was stopped, the reaction solution was washed with 500 mL of saturated ammonium chloride solution and 500 mL of saturated sodium bicarbonate solution in turn, the organic layer was dried with anhydrous sodium sulfate, concentrated and evaporated at 40°C to obtain a red-brown solid.

[0105] The reaction product was dissolved in toluene and column chromatography was performed using a fast preparation liquid chromatograph (SepaBean TM machine), normal phase silica gel column (SepaBean standard fast separation column S-8101-0120, pre-packed normal phase silica gel, particle size 40-63 μm, pore size 120 g). The eluent was ethyl acetate / toluene, the volume fraction of ethyl acetate was 1-5%, the flow rate was 60 mL / min, and the ultraviolet detector (detection wavelength 1: 310 nm, detection wavelength 2: 320 nm). The obtained product was concentrated and evaporated at 40°C to obtain a red-brown solid.

[0106] Structural characterization: the molecular weight was detected by electrospray mass spectrometry (ESI-MS, Thermo Scientific Exactive, positive ion mode), and the detection results are shown in Figure 13 ; high performance liquid chromatography (HPLC, LC-2030, Shimadzu) was used, C18 chromatographic column (Agilent, Eclipse XDB-C18, 4.6*250 mm, filler particle size 5 μm), toluene-acetonitrile as mobile phase, the specific proportion gradient method is shown in Table 3, ultraviolet detector (detection wavelength: 310 nm), flow rate is 1 mL / min, purity detection, the detection results are shown in Figure 14 ; nuclear magnetic resonance (Bruker Avance III 400HD, 400 MHz) was used, the sample was dissolved in deuterated chloroform for hydrogen spectrum and carbon spectrum detection, and the product structure was analyzed, the hydrogen spectrum and carbon spectrum detection results are shown in Figure 15 and Figure 16 .

[0107] Conclusion: The molecular weight detected by ESI-MS is related to the tetraaddition of leucine tert-butyl ester with epoxy fullerene (TLeu-OBu). t The theoretical molecular weight is completely consistent, and the purity detected by HPLC is above 95%. The hydrogen atom shifts and integral areas in the proton NMR spectrum and the carbon atom shifts in the carbon NMR spectrum are consistent with those in the TLeu-OBu spectrum. t The structures are consistent. Therefore, this method can efficiently prepare the tetrasubstituted aminofullerene derivative TLeu-OBu. t This enables the accurate preparation of a defined number of aminofullerene derivatives.

[0108] Example 5: Preparation of tetrasubstituted aminoepoxyfullerene molecule TPhe-OMe

[0109]

[0110] Synthesis method: 1g C 60 Cl6 was dissolved in 1000 mL of toluene, and 600 μL of cumene hydroperoxide (80%) (3 equivalents) was added. 2.8 mL of phenylalanine methyl ester (15 equivalents) and 745 μL of triethylamine (5 equivalents) were mixed and added to the above reaction solution. The mixture was stirred at room temperature (1500 rpm) for 1 h. The reaction solution was analyzed by HPLC. The reaction was complete, and the reaction was stopped. The reaction solution was washed successively with 500 mL of saturated ammonium chloride solution and 500 mL of saturated sodium bicarbonate solution. The organic layer was dried with anhydrous sodium sulfate and concentrated at 40 °C to obtain a reddish-brown solid.

[0111] The reaction product was dissolved in toluene and then processed using a rapid preparative liquid chromatograph (SepaBean). TM machine), positive phase silicone column (Sante Technology) Standard rapid separation column S-8101-0120, pre-filled with normal-phase silica gel, particle size 40-63μm, pore size The packing material (120 g) was used for column chromatography separation. The eluent was ethyl acetate / toluene, with ethyl acetate accounting for 1-5% by volume. The flow rate was 60 mL / min, and a UV detector was used (detection wavelength 1: 310 nm, detection wavelength 2: 320 nm). The obtained product was concentrated and evaporated to dryness at 40 °C to obtain a reddish-brown solid.

[0112] Structural characterization: Molecular weight was determined using electrospray ionization mass spectrometry (ESI-MS, Thermo Scientific Exactive, positive ion mode). Results are shown below. Figure 17; using high performance liquid chromatography (HPLC, LC-2030, Shimadzu), C18 column (Agilent, Eclipse XDB-C18, 4.6*250mm, packing particle size 5 μm), toluene-acetonitrile as mobile phase, the specific proportion gradient method is shown in Table 3, UV detector (detection wavelength: 310 nm), flow rate of 1 mL / min, purity detection, the detection results are shown in Table 4. Figure 18 .

[0113] Conclusion: The molecular weight detected by ESI-MS is completely consistent with the theoretical molecular weight of phenylalanine methyl tetra-addition epoxy fullerene (TPhe-OMe), and the purity detected by HPLC is more than 95%. It can be seen that the method can efficiently prepare the tetra-substituted amino fullerene derivative TPhe-OMe, and realize the accurate preparation of a certain number of amino fullerene derivatives.

[0114] Example 6: Preparation of tetra-substituted amino epoxy fullerene molecule TAla-OBu t

[0115]

[0116] Synthetic method: 1g C 60 Cl6 was dissolved in 1000 mL of toluene, 600 μL of hydrogen peroxide isopropylbenzene (80%) (3 equivalents) was added, 2.5 mL of propionyl alanine tert-butyl ester (15 equivalents) was mixed with 745 μL of triethylamine (5 equivalents) and then added to the above reaction solution, stirred (1500 rpm) at room temperature for 1 h, the reaction solution was detected by HPLC, the reaction was complete, the reaction was stopped, and the reaction solution was washed with 500 mL of saturated ammonium chloride solution and 500 mL of saturated sodium bicarbonate solution in sequence, the organic layer was dried with anhydrous sodium sulfate, and concentrated and evaporated at 40°C to obtain a red-brown solid.

[0117] The reaction product was dissolved in toluene and column chromatography was performed using a fast preparation liquid chromatograph (SepaBean TM machine), normal phase silica gel column (SepaBean standard type fast separation column S-8101-0120, pre-packed normal phase silica gel, particle size 40-63 μm, pore size filler amount 120 g) with ethyl acetate / toluene as eluent, the volume fraction of ethyl acetate was 10-30%, the flow rate was 60 mL / min, and the UV detector (detection wavelength 1: 310 nm, detection wavelength 2: 320 nm). The obtained product was concentrated and evaporated at 40°C to obtain a red-brown solid.

[0118] Structural characterization: the molecular weight was detected by electrospray mass spectrometry (ESI-MS, Thermo Scientific Exactive, positive ion mode), and the detection results are shown inFigure 19 High-performance liquid chromatography (HPLC, LC-2030, Shimadzu) was used with a C18 column (Agilent Eclipse XDB-C18, 4.6*250mm, packing particle size 5μm), toluene-acetonitrile as the mobile phase (Table 3), and a UV detector (detection wavelength: 310nm) at a flow rate of 1mL / min to determine the purity. The results are shown in the table below. Figure 20 .

[0119] Conclusion: The molecular weight detected by ESI-MS is related to the tetraaddition of alanine tert-butyl ester with epoxy fullerene (TAla-OBu). t The molecular weights are completely consistent with the theoretical values, and the purity, as determined by HPLC, is above 95%. Therefore, this method can efficiently prepare the tetrasubstituted aminofullerene derivative TAla-OBu. t This enables the accurate preparation of a defined number of aminofullerene derivatives.

[0120] Example 7: Preparation of tetrasubstituted aminoepoxyfullerene molecule TGly-OBu by secondary amine regulation t

[0121]

[0122] Synthesis method: 1g C 60 Cl6 was dissolved in 1000 mL of toluene, and 600 μL of cumene hydroperoxide (80%) (3 equivalents) was added. 2.19 mL of glycine tert-butyl ester (15 equivalents) and 320 μL of piperidine (3 equivalents) were mixed and added to the above reaction solution. The mixture was stirred at room temperature (1500 rpm) for 1 h. The reaction solution was analyzed by HPLC. The reaction was complete, and the reaction was stopped. The reaction solution was washed successively with 500 mL of saturated ammonium chloride solution and 500 mL of saturated sodium bicarbonate solution. The organic layer was dried with anhydrous sodium sulfate and concentrated at 40 °C to obtain a reddish-brown solid.

[0123] The reaction product was dissolved in toluene and then processed using a rapid preparative liquid chromatograph (SepaBean). TM machine), positive phase silicone column (Sante Technology) Standard rapid separation column S-8101-0120, pre-filled with normal-phase silica gel, particle size 40-63μm, pore size The sample was separated by column chromatography (120 g packing material) using ethyl acetate / toluene as the eluent (ethyl acetate volume percentage 10-20%), at a flow rate of 60 mL / min, and with a UV detector (detection wavelength 1: 310 nm, detection wavelength 2: 320 nm). The product was concentrated and evaporated to dryness at 40 °C to obtain a reddish-brown solid.

[0124] Structural characterization: The molecular weight was detected by electrospray mass spectrometry (ESI-MS, Thermo Scientific Exactive, positive ion mode), and the detection results were the same as those of Figure 5 ; the purity was detected by high performance liquid chromatography (HPLC, LC-2030C 3D, Shimadzu), C18 column (Agilent, Eclipse XDB-C18, 4.6*250mm, packing particle size 5um), toluene-acetonitrile as mobile phase, the specific gradient method was shown in Table 3, UV detector (detection wavelength: 310nm), flow rate was 1mL / min, and the detection results were the same as those of Figure 6 ; the structure of the product was analyzed by nuclear magnetic resonance (Bruker Avance III 400HD, 400MHz) by dissolving the sample in deuterated chloroform for hydrogen spectrum and carbon spectrum detection, and the detection results of hydrogen spectrum and carbon spectrum were substantially the same as those of Figure 7 and Figure 8 .

[0125] Conclusion: The molecular weight detected by ESI-MS was completely consistent with the theoretical molecular weight of the four-glycine tert-butyl ester tetra-substituted epoxy fullerene (TGly-OBu t ), the purity detected by HPLC was above 95%, and the hydrogen atom shift and integral area of hydrogen spectrum and the carbon atom shift of carbon spectrum were consistent with the structure of TGly-OBu t . It can be seen that the four-substituted amino fullerene derivative TGly-OBu t can be efficiently prepared by the method, and the accurate preparation of a certain number of amino fullerene derivatives is realized.

[0126] Example 8: Preparation of four-substituted amino epoxy fullerene molecule TDBMAC

[0127]

[0128] Synthetic method: 1g C 60 Cl6 was dissolved in 1000mL toluene, 600μL of hydrogen peroxide isopropylbenzene (80%) (3 equivalents) was added, 2.77mL of diphenylamine (15 equivalents) was mixed with 745μL of triethylamine (5 equivalents) and then added to the above reaction solution, and stirred (1500rpm) at room temperature for 1h. The reaction solution was detected by HPLC, and the reaction was complete. The reaction was stopped, and the reaction solution was washed with 500mL of saturated ammonium chloride solution and 500mL of saturated sodium bicarbonate solution in sequence. The organic layer was dried with anhydrous sodium sulfate, and concentrated and evaporated at 40℃ to obtain a red-brown solid.

[0129] The reaction product was dissolved in toluene, and a fast preparation liquid chromatograph (Sepharmachine) was used to purify the product by using a normal phase silica gel column (Sepharmachine TM ). Standard fast separation column S-8101-0120, pre-packed normal phase silica gel, particle size 40-63 μm, pore size 60 A Column chromatography separation was carried out on the column (120 g of filler) with toluene as eluent at a flow rate of 60 mL / min, and an ultraviolet detector (detection wavelength 1: 310 nm, detection wavelength 2: 320 nm). The obtained product was concentrated and evaporated at 40°C to obtain a red-brown solid.

[0130] Structural characterization: The molecular weight was detected by electrospray mass spectrometry (ESI-MS, Thermo Scientific Exactive, positive ion mode), and the detection results are shown in Table 1. Figure 21 ; high performance liquid chromatography (HPLC, LC-2030, Shimadzu) was used, a C18 chromatographic column (Agilent, Eclipse XDB-C18, 4.6*250 mm, filler particle size 5 μm) was used, toluene-acetonitrile was used as the mobile phase (Table 3), an ultraviolet detector (detection wavelength: 310 nm) was used, and the flow rate was 1 mL / min. The purity was detected, and the detection results are shown in Table 2. Figure 22 .

[0131] Conclusion: The molecular weight detected by ESI-MS is completely consistent with the theoretical molecular weight of the tetra-substituted diphenylamine tetra-adduct epoxy fullerene (TDBMAC), and the purity detected by HPLC is more than 95%. It can be seen that the method can efficiently prepare the tetra-substituted amino fullerene derivative TAla-OBu t , and realize the accurate preparation of a certain number of amino fullerene derivatives. It can be seen that the method can efficiently prepare the tetra-substituted amino fullerene derivative TDBMAC, and realize the accurate preparation of a certain number of amino fullerene derivatives.

[0132] The above-described embodiments only express several embodiments of the present application, and the description is more specific and detailed, but it should not be understood as limiting the scope of the patent. It should be noted that for ordinary skilled in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which are within the scope of protection of the present application. Therefore, the scope of protection of the present application patent should be subject to the appended claims.

Claims

1. A method for preparing an aminofullerene derivative, comprising the following steps: An amine compound and cumene hydroperoxide were added to a solution of fullerene chloride and stirred at room temperature. After the reaction was completed, an aminofullerene derivative was obtained. The amine compound is NH2R; The NH2R is , , , , or ; The R is , , , , or ; No acidic reagents are added during the reaction process of the method described above; The fullerene in the chlorinated fullerene is C. 60 ; The method further includes the step of adding a secondary or tertiary amine compound to a fullerene chloride solution; The secondary amine compound is piperidine; The tertiary amine compound is triethylamine; The aminofullerene derivative has the structure shown in the following formula: ； The molar ratio of the fullerene chloride, the amine compound, the secondary amine compound, and the cumene hydroperoxide is (1-3):(10-30):(1-5):(1:5); The molar ratio of the fullerene chloride, the amine compound, the tertiary amine compound and the cumene hydroperoxide is (1-3):(10-30):(3-8):(1:5).

2. The method according to claim 1, characterized in that, The stirring time at room temperature is 0.6 h to 1 h.

3. The method according to claim 2, characterized in that, The stirring time at room temperature is 1 hour.

4. The method according to claim 1, characterized in that, The chlorinated fullerene solution is obtained by dissolving chlorinated fullerene in an organic solvent, wherein the organic solvent is selected from toluene or chlorobenzene.

5. The method according to any one of claims 1 to 4, characterized in that, The aminofullerene derivative has the following structure: 、 、 、 、 、 。

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