A mitochondria-targeting compound, and a preparation method and application thereof

Abstract

The application belongs to the technical field of organic synthesis and medicinal chemistry, and particularly relates to a mitochondria-targeting compound and a preparation method and application thereof.The physicochemical properties of the mitochondria-targeting compound can be regulated by the length of a carbon chain and a parent nucleus structure, the mitochondria-targeting compound has excellent mitochondria-targeting capability and a mitochondria bioactivity intensifying effect, can be efficiently enriched on a mitochondrial membrane, and can exert inherent bioactivity of a mitochondria active compound and a mitochondria protection synergistic effect of triphenylphosphine, can be adapted to various preparation forms such as oral administration, injection and external use, and the targeted enrichment characteristics reduce a systemic drug dosage and reduce toxic side effects, and the mitochondria-targeting compound has a good clinical application prospect in mitochondria protection products.

Description

Technical Field

[0001] This invention belongs to the fields of organic synthesis and medicinal chemistry, and specifically relates to a mitochondrial-targeting compound, its preparation method, and its application. Background Technology

[0002] Mitochondria are the core organelles of energy metabolism in eukaryotic cells. They generate ATP through oxidative phosphorylation to power cellular life activities. Their dysfunction is closely related to neurodegenerative diseases (Alzheimer's disease, Parkinson's disease, etc.), cardiovascular diseases (heart failure, myocardial ischemia-reperfusion injury, etc.), metabolic diseases (diabetes, metabolic syndrome, etc.), as well as cellular aging and tissue degeneration. Developing bioactive compounds with mitochondrial-targeting capabilities to achieve precise mitochondrial protection is an important research direction for disease treatment and health maintenance.

[0003] Triphenylphosphine (TPP) and its derivatives are classic mitochondrial-targeting molecules. Their lipophilic cationic properties can be driven by the mitochondrial inner membrane potential to achieve high enrichment in mitochondria. Targeting molecules such as MitoQ and SkQ1 have shown good mitochondrial protective effects. However, the biological activities of these molecules are singular, making it difficult to achieve multi-effect synergistic protection.

[0004] Various compounds in nature contain hydroxyl or amino / amine groups and have protective or functionally improving effects on mitochondria, including vitamins (such as vitamin C, vitamin B1, and vitamin B5), pyridoxine, nicotinamide nucleotides, PQQ, and astaxanthin. These compounds can improve mitochondrial function through antioxidant effects, regulation of energy metabolism, and stabilization of mitochondrial membrane structure. However, these compounds in their natural state generally suffer from poor subcellular targeting, making it difficult to penetrate the mitochondrial membrane and reach effective therapeutic concentrations. Vitamin C, in particular, has multiple hydroxyl groups that are easily oxidized, and direct esterification is difficult, requiring prior protective modification to achieve stable bonding. At the same time, the water-soluble or lipid-soluble single physicochemical properties of various active compounds result in low bioavailability, and high-dose use may also produce systemic toxic side effects, greatly limiting their application in mitochondrial-related diseases.

[0005] Existing mitochondrial protection technologies suffer from the following key shortcomings: ① Lack of targeting: Most mitochondrial active compounds lack specific mitochondrial targeting capabilities, failing to accumulate at the target site and exhibiting weak effects at low doses; ② Narrow range of applicable raw materials: Existing targeted modification technologies have not designed protective strategies for the oxidizability of vitamin C and have not developed universal modification systems for various active compounds such as pyridoxine, nicotinamide nucleotides, PQQ, and astaxanthin; ③ Limited selection of halogenated fatty acid raw materials: Existing technologies lack clear limitations on the medium-to-short chain length of halogenated fatty acids used to prepare triphenylphosphine salts. Long-chain fatty acids can easily lead to decreased membrane permeability, affecting targeting efficiency; ④ Weak synergistic effect: Neither a single targeting molecule nor a single mitochondrial active compound can simultaneously achieve targeted enrichment and multi-effect mitochondrial protection, resulting in limited therapeutic effects.

[0006] Therefore, there is an urgent need in this field to develop a universal mitochondrial targeted modification system that esterifies vitamins with triphenylphosphine salts of short-chain halogenated fatty acids in C5-C12, thereby achieving amidation or esterification modification of various active compounds such as vitamin A, vitamin E, pyridoxine, and nicotinamide nucleotides. This would enable precise targeted delivery and synergistic enhancement of the bioactivity of various mitochondrial active compounds, while simplifying the preparation process and expanding the range of applicable raw materials. Summary of the Invention

[0007] The purpose of this invention is to provide a mitochondrial-targeting compound, its preparation method, and its application, addressing the problems of poor targeting, low bioavailability, and the easy oxidation and difficulty in direct esterification of vitamin C in existing mitochondrial active compounds. By first protecting vitamin C to propylidene vitamin C, and then condensing and linking it with a C5-C12 fatty acid triphenylphosphine salt, while simultaneously binding various active compounds such as pyridoxine and nicotinamide nucleotides to TPP fatty acyl chloride intermediates through amidation or esterification, this invention endows various mitochondrial active compounds with highly efficient mitochondrial targeting capabilities, while simultaneously achieving a synergistic mitochondrial protective effect between triphenylphosphine and the mitochondrial active compounds. This invention also aims to establish a universal targeted modification preparation method, expanding the applicable range of raw materials and reducing the difficulty of industrial production.

[0008] Therefore, the present invention provides the following technical solution.

[0009] One aspect of the present invention provides a mitochondrial-targeting compound having the structure shown in general formula (I): [MO / -NH]-[C(=O)-R1]-PPh3 + X - (I); In formula (I): M is selected from any one of vitamin A, vitamin E, vitamin C, pyridoxine, nicotinamide nucleotide, vitamin B1, vitamin B5, pyrroloquinoline quinone, and astaxanthin; R1 is a straight-chain or branched alkyl group containing 5-12 carbon atoms, or a carbon chain containing unsaturated bonds; X is a halide ion or an organic acid radical ion. PPh3 is a triphenylphosphine group.

[0010] In the aforementioned mitochondrial-targeting compounds, the active molecule M comprises mitochondrial active compound residues, including vitamin A, vitamin E, vitamin C, pyridoxine, nicotinamide nucleotide, vitamin B1, vitamin B5, PQQ, and astaxanthin. These provide mitochondrial protective biological activities such as antioxidant activity, regulation of energy metabolism, and membrane structure stabilization, and can be flexibly replaced according to therapeutic needs. Vitamin C, after being protected by a propionyl group, effectively avoids oxidation and enhances bond stability. The R1 chain is a C5-C12 fatty acid carbon chain, serving as a linker arm to regulate the compound's lipophilicity and mitochondrial membrane permeability. The medium-to-short chain structure ensures the compound's transmembrane efficiency; C6, C8, C10, and C12 carbon chains are preferred. PPh3 + The terminal triphenylphosphine cation provides specific mitochondrial targeting capability, driving the enrichment of compounds in mitochondria. - As a counterion, it maintains the electroneutrality of the compound, without affecting its targeting and bioactivity. Regarding the bonding mechanism, ester or amide bonds can be used depending on the functional group characteristics of the M-terminal compound, ensuring bonding stability and biocompatibility.

[0011] In a preferred embodiment of the present invention, the bonding method of M is as follows: When M is vitamin A, vitamin C, nicotinamide nucleotide, vitamin E, or astaxanthin, an ester bond -O-CO- is formed; When M is pyridoxine, vitamin B1, vitamin B5, or pyrroloquinoline quinone, an amide bond -CONH- is formed.

[0012] In a preferred embodiment of the present invention, R1 is a straight-chain or branched alkyl group containing 6, 8, 10 or 12 carbon atoms, or a carbon chain containing unsaturated bonds.

[0013] In a preferred embodiment of the present invention, X is selected from any one of bromide ion, iodide ion, chloride ion, p-toluenesulfonate ion, methanesulfonate ion, and trifluoroacetate ion.

[0014] In a preferred embodiment of the present invention, X is a bromide ion or an iodide ion.

[0015] In a preferred embodiment of the present invention, the mitochondrial targeting compound is selected from one of the following: vitamin C-TPP fatty acid ester, pyridoxine-TPP fatty amide, nicotinamide nucleotide-TPP fatty acid mono / diester, vitamin B1-TPP fatty amide, vitamin B5-TPP fatty amide, pyrroloquinoline quinone-TPP fatty amide, astaxanthin-TPP fatty diester, vitamin E-TPP fatty amide, and vitamin A-TPP fatty diester.

[0016] Another aspect of the present invention provides a method for preparing a mitochondrial-targeting compound as described above, the method comprising the following steps: S1: Triphenylphosphine and halofatty acids are reacted in an anhydrous organic solvent to obtain triphenylphosphine salts of fatty acids; S2: Add an acylating agent to the obtained triphenylphosphine salt of fatty acid to react and obtain TPP fatty acyl chloride (full Chinese name: brominated triphenylphosphine fatty acyl chloride). S3: The active molecule M is reacted with TPP fatty acyl chloride in a solvent to prepare the mitochondrial-targeting compound; The active molecule M is selected from at least one of vitamin A, vitamin E, vitamin C, pyridoxine, vitamin B1, vitamin B5, pyrroloquinoline quinone, nicotinamide nucleotide, or astaxanthin; The reaction conditions are selected based on the type of active groups on the active molecule M: (a) When the active group is an alcohol hydroxyl group, the esterification reaction is carried out at 40-80°C for 6-24 hours in the presence of an esterification catalyst; or (b) When the active group is amino, the amidation reaction is carried out at 0-15°C in the dark for 8-18 hours in the presence of an alkaline acid binder.

[0017] In a preferred embodiment of the present invention, in step S1, the halogenated fatty acid is a C5-C12 terminal halogenated fatty acid, selected from at least one of 5-bromopentanoic acid, 6-bromohexanoic acid, 7-bromoheptanoic acid, 8-bromooctanoic acid, 9-bromononanoic acid, 10-bromodecanoic acid, 11-bromoundecanoic acid, 12-bromododecanoic acid, 5-iodopentanoic acid, and 12-iodododecanoic acid.

[0018] In a preferred embodiment of the present invention, in step S1, the molar ratio of the triphenylphosphine to the halofatty acid is (1:1) to (1.5:1).

[0019] In a preferred embodiment of the present invention, in step S1, the anhydrous organic solvent is selected from at least one of acetonitrile, N,N-dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, and dichloromethane.

[0020] In a preferred embodiment of the present invention, in step S1, the reaction conditions are: temperature 60~100℃, time 4~12 hours.

[0021] In a preferred embodiment of the present invention, in step S2, the acylating agent is selected from one of thionyl chloride, phosphorus trichloride, phosgene, and triphosgene.

[0022] In a preferred embodiment of the present invention, in step S2, the amount of acylating agent added is 1.0-5.0 molar equivalents of the active molecule M.

[0023] In a preferred embodiment of the present invention, in step S2, the amount of acylating agent added is 2-3 molar equivalents.

[0024] In a preferred embodiment of the present invention, in step S2, the reaction conditions are: a temperature of 5-80°C, preferably 50-70°C, and a time of 3-8 hours.

[0025] In a preferred embodiment of the present invention, in step S3, when the active molecule M is vitamin C, the method specifically includes: Vitamin C and TPP fatty acyl chloride were esterified at 40-80°C for 6-24 hours in the presence of an esterification catalyst to obtain vitamin C-TPP fatty acid ester.

[0026] In a preferred embodiment of the present invention, in step S3, when the active molecule M is nicotinamide nucleotide, the method specifically includes: Nicotinamide nucleotides and TPP fatty acyl chlorides were esterified at 40-70°C for 6-12 hours in the presence of an esterification catalyst to obtain monoesters or diesters.

[0027] In a preferred embodiment of the present invention, in step S3, when the active molecule M is astaxanthin, the method specifically includes: Astaxanthin was esterified with 2 molar equivalents of TPP fatty acyl chloride in the presence of an esterification catalyst under an organic base environment for 12-24 hours to obtain astaxanthin diester.

[0028] In a preferred embodiment of the present invention, the organic base is triethylamine or pyridine.

[0029] In a preferred embodiment of the present invention, the esterification catalyst is selected from at least one of N,N'-dicyclohexylcarbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, and 4-dimethylaminopyridine.

[0030] In a preferred embodiment of the present invention, the amount of the esterification catalyst is 5 to 20% of the total molar amount of the reactants.

[0031] In a preferred embodiment of the present invention, in step S3, when the active molecule M is pyridoxine, vitamin B1, vitamin B5, or pyrroloquinoline quinone, the method specifically includes: The active molecule M and TPP fatty acyl chloride were reacted with an amidation reaction at 0-15°C in the dark for 8-18 hours in the presence of an alkaline acid binder to obtain the amidated product.

[0032] In a preferred embodiment of the present invention, the alkaline acid-binding agent is selected from at least one of triethylamine and N,N-diisopropylethylamine.

[0033] In a preferred embodiment of the present invention, the amount of alkaline acid-binding agent is 1.2 to 1.5 times the molar amount of TPP fatty acyl chloride.

[0034] Another aspect of the present invention provides the use of a mitochondrial-targeting compound as described above, or a mitochondrial-targeting compound prepared according to the method described above, in the preparation of a product for the protection of mitochondrial function.

[0035] In a preferred embodiment of the present invention, the product includes: (1) Anti-aging products can improve the decline of mitochondrial function during the cellular aging process; (2) Neuroprotective products for the prevention and treatment of neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease; (3) Cardiovascular protection products used to alleviate myocardial ischemia-reperfusion injury and improve mitochondrial dysfunction in heart failure; (4) Antioxidant products can remove excess reactive oxygen species in mitochondria and reduce oxidative stress damage; or (5) Products for the prevention and treatment of metabolic syndrome, used to regulate abnormal mitochondrial energy metabolism related to glucose and lipid metabolism.

[0036] By employing the above technical solution, the present invention has at least the following advantages: First, the mitochondrial-targeting compound of this invention achieves selective enrichment within mitochondria through its triphenylphosphine group, resulting in a mitochondrial / cytoplasmic concentration ratio of over 15 times, achieving effective therapeutic concentrations even at low doses. The short-chain fatty acid linker arms in the C5-C12 region ensure the compound's transmembrane efficiency, avoiding the membrane permeability reduction issues caused by long-chain fatty acids. The triphenylphosphine group exerts membrane stabilizing and mild antioxidant effects within mitochondria, synergizing with the inherent biological activities of various mitochondrial active compounds at the M-terminus, resulting in a significantly superior overall protective effect compared to using various mitochondrial active compounds or triphenylphosphine salts alone.

[0037] Secondly, this invention establishes a universal targeted modification system in which the M-terminus of the compound structure can be replaced with various active compounds such as vitamin C, pyridoxine, and nicotinamide nucleotide, and the R1 chain can be flexibly adjusted within the C5-C12 range. The physicochemical properties and biological activities of the compound can be optimized according to the treatment needs of different diseases. At the same time, for active compounds with different functional groups, corresponding ester bonds or amide bonds are used to connect them to improve the bonding stability.

[0038] Third, the mitochondrial-targeting compounds of the present invention adopt a strategy of "unified preparation of intermediates + targeted bonding", which is simple to operate and has no harsh reaction conditions; the C5-C12 terminal halogenated fatty acids, triphenylphosphine and various mitochondrial active compounds used are all readily available raw materials with low cost; the overall reaction yield can reach 50-88%, and the purification steps are simple, making it suitable for large-scale industrial production.

[0039] Fourth, the physicochemical properties of the mitochondrial-targeting compound of the present invention can be regulated by carbon chain length and core structure, possessing both excellent mitochondrial targeting ability and mitochondrial bioactivity enhancement effect. It can be efficiently enriched on the mitochondrial membrane, exerting the inherent bioactivity of mitochondrial active compounds and the synergistic effect of triphenylphosphine on mitochondrial protection. It can be adapted to various formulations such as oral, injection, and topical application, and the targeted enrichment characteristics reduce the systemic dosage and reduce toxic side effects, showing good clinical application prospects in mitochondrial protection products.

[0040] Fifth, the preparation method of the mitochondrial-targeting compound of the present invention is simple, the reaction conditions are mild, the raw materials are widely applicable, the yield is high, and it is suitable for industrial production.

[0041] The above description is merely an overview of the technical solution of the present invention. In order to better understand the technical means of the present invention and to implement it in accordance with the contents of the specification, the preferred embodiments of the present invention are described in detail below. Detailed Implementation

[0042] To make the technical means, creative features, achieved objectives, and effects of this invention readily understandable, the technical solutions in the embodiments of this invention will be clearly and completely described below in conjunction with the embodiments of this invention. Obviously, the described embodiments are merely some embodiments of this invention, and not all embodiments. Based on the embodiments of this invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this invention.

[0043] Unless otherwise specified, the preparation method of TPP fatty acyl chloride intermediate -- TPP-octanoyl chloride in the following embodiments of the present invention includes the following steps: Step A: Preparation of fatty acid triphenylphosphine salt – octanoic acid – TPP bromide In a 100 mL three-necked flask, 50 mL of anhydrous acetonitrile, 2.62 g of triphenylphosphine, and 2.23 g of 8-bromooctanoic acid were added. The mixture was then stirred in an oil bath at 75°C for 9 hours under nitrogen protection. After the reaction was complete, the mixture was cooled to room temperature, and the solvent was removed by vacuum evaporation. The residue was washed three times with anhydrous diethyl ether (20 mL each time) and then dried under vacuum to obtain a white solid, triphenylphosphine octanoic acid bromide, abbreviated as octanoic acid-TPP bromide. The yield of the product, octanoic acid-TPP bromide, was calculated to be 88%; MS (ESI+): m / z 423.2 [M-Br]+.

[0044] The chemical reaction formulas involved in the above process are as follows: ; Step B: Preparation of TPP-fatty acyl chloride intermediate – TPP-octanoyl chloride Take 4.37 g of the prepared octanoic acid-TPP bromide, dissolve it in 50 mL of anhydrous dichloromethane, then cool it to 0-5°C in an ice bath, slowly add 2.42 g of thionyl chloride, and stir the reaction in the dark for 3 hours. After the reaction is complete, remove excess thionyl chloride and dichloromethane by rotary evaporation under reduced pressure to obtain TPP-octanoyl chloride for later use.

[0045] It should be noted that all TPP-octanoyl chloride involved in this invention is prepared and used immediately.

[0046] The chemical reaction formulas involved in the above process are as follows: ; It should be noted that the preparation methods of other intermediates involved in the examples, such as TPP-decanoyl chloride, TPP-hexanoyl chloride, and TPP-lauroyl chloride (C12), are roughly the same as the preparation method of TPP-octanoyl chloride described above. The only difference is that the 8-bromooctanoic acid used in step A is replaced with 10-bromodecanoic acid, 6-bromohexanoic acid, and 12-bromododecanoic acid, respectively. The rest is consistent with the preparation method of TPP-octanoyl chloride described above.

[0047] Example 1: Preparation of ascorbic acid-C8-TPP fatty acid ester In a 500 mL round-bottom flask, 200 mL of anhydrous N,N-dimethylformamide (DMF), 2.33 g of vitamin C, and 0.12 g of 4-dimethylaminopyridine (DMAP) were added. The flask was cooled to 0–5°C in an ice bath, and then 4.65 g of TPP-octanoyl chloride was slowly added dropwise. After the addition was complete, the mixture was heated to 60°C and stirred for 12 hours. After the reaction was complete, the mixture was poured into ice water and extracted three times with ethyl acetate (100 mL each time). The organic layers were combined. The mixture was then washed with water and sodium bicarbonate, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give a pale yellow solid compound, ascorbic acid-C8-TPP fatty acid ester.

[0048] The yield of the obtained product, ascorbic acid-C8-TPP fatty acid ester, was calculated to be 43%; MS (ESI+): m / z 557.3 [M]+.

[0049] The chemical reaction formulas involved in the above process are as follows: ; Example 2: Preparation of carnosine-C10-TPP fatty acid amide In a 500 mL round-bottom flask, 200 mL of anhydrous N,N-dimethylformamide (DMF), 2.44 g of carnosine, and 1.21 g of triethylamine were added. The mixture was cooled to 0–5°C in an ice bath, and then 4.93 g of TPP-decanoyl chloride was slowly added dropwise. After the addition was complete, the mixture was stirred at room temperature in the dark for 16 hours. After the reaction was complete, 150 mL of ice water was added, and the mixture was extracted three times with ethyl acetate (100 mL each time). The organic layers were combined. The mixture was then washed with water and sodium bicarbonate, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give a pale yellow solid compound, carnosine-C10-TPP fatty amide.

[0050] The yield of the obtained product, carnosine-C10-TPP fatty amide, was calculated to be 72%; MS (ESI+): m / z 664.4[M]+.

[0051] The chemical reaction formulas involved in the above process are as follows: ; Example 3: Preparation of pyridoxine-C8-TPP fatty acid amide In a 50 mL round-bottom flask, 20 mL of anhydrous N,N-dimethylformamide (DMF), 1.68 g of pyridoxine, and 1.21 g of triethylamine were added. The mixture was cooled to 0–5°C in an ice bath, and then 4.65 g of TPP-octanoyl chloride was slowly added dropwise. After the addition was complete, the amidation reaction was carried out at room temperature in the dark for 15 hours. After the reaction was completed, 100 mL of ice water was added to quench the reaction, and the mixture was extracted three times with dichloromethane (50 mL each time). The organic layers were combined. The residue was then washed with water to remove salts, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give a pale yellow solid compound, pyridoxine-C8-TPP fatty amide, in 75.2% yield; MS (ESI+): m / z 548.3 [M]+.

[0052] The chemical reaction formulas involved in the above process are as follows: ; Example 4: Preparation of nicotinamide nucleoside-C10-TPP fatty acid ester In a 50 mL round-bottom flask, 20 mL of anhydrous pyridine, 3.34 g of nicotinamide nucleoside (NR), and 0.12 g of 4-dimethylaminopyridine (DMAP) were added. The mixture was cooled to 0-5°C in an ice bath, and then 4.93 g of TPP-decanoyl chloride was slowly added dropwise. After the addition was complete, the temperature was raised to 50°C and the esterification reaction was carried out under controlled temperature for 10 hours. After the reaction was completed, 50 mL of ice water was added for neutralization, and the mixture was extracted three times with dichloromethane (50 mL each time). The organic layers were combined. The mixture was then dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a white solid compound, nicotinamide nucleoside-C10-TPP fatty acid ester.

[0053] The yield of the obtained product, nicotinamide nucleoside-C10-TPP fatty acid ester, was calculated to be 70%; MS (ESI+): m / z 731.4 [M]+.

[0054] The chemical reaction formulas involved in the above process are as follows: ; Example 5: Preparation of Vitamin B1-C6-TPP Amide In a 500 mL round-bottom flask, 250 mL of anhydrous dichloromethane (DCM), 3.00 g of vitamin B1, and 1.55 g of N,N-diisopropylethylamine (DIPEA) were added. The flask was cooled to 0–5°C in an ice bath, and then 4.37 g of TPP-hexanoyl chloride was slowly added dropwise. After the addition was complete, the amide condensation reaction was carried out at room temperature in the dark for 12 hours. After the reaction was completed, the product was washed three times with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a pale yellow solid vitamin B1-C6-TPP amide.

[0055] The yield of the obtained product, vitamin B1-C6-TPP amide, was calculated to be 33%; MS (ESI+): m / z 582.3[M]+.

[0056] The chemical reaction formulas involved in the above process are as follows: ; Example 6: Preparation of pantothenic acid-C12-TPP amidation In a 500 mL round-bottom flask, 200 mL of anhydrous N,N-dimethylformamide (DMF), 2.19 g of pantothenic acid, and 1.21 g of triethylamine were added. The mixture was cooled to 0–5°C in an ice bath, and then 5.21 g of TPP-lauroyl chloride (C12) was slowly added dropwise. After the addition was complete, the amidation reaction was carried out at room temperature in the dark for 16 hours. After the reaction was completed, 100 mL of ice water was added to quench the reaction, and the mixture was extracted three times with ethyl acetate (50 mL each time). The organic layers were combined. The mixture was then washed with water to remove salts, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give a white solid compound, pantothenic acid-C12-TPP amidation.

[0057] The yield of the obtained pantothenic acid-C12-TPP amidate was 31%; MS (ESI+): m / z 618.4 [M]+.

[0058] The chemical reaction formulas involved in the above process are as follows: ; Example 7: Preparation of PQQ-C10-TPP fatty acid amide In a 500 mL round-bottom flask, 200 mL of anhydrous N,N-dimethylformamide (DMF), 2.14 g of pyrroloquinoline quinone (PQQ), and 1.21 g of triethylamine were added. The mixture was cooled to 0–5 °C in an ice bath under nitrogen protection. Then, 4.65 g of TPP-octanoyl chloride was slowly added dropwise to induce acylation and generate an amide. The reaction was carried out at low temperature (0–5 °C) in the dark for 18 hours (to prevent oxidative degradation of PQQ). After the reaction was completed, the mixture was extracted three times with 100 mL of ethyl acetate each time, and the organic layers were combined. The mixture was then washed twice with saturated sodium bicarbonate solution, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a pale yellow solid, PQQ-C10-TPP aliphatic amidate.

[0059] The yield of the obtained product PQQ-C10-TPP fatty acid amidation was 54%; MS (ESI+): m / z 567.3 [M]+.

[0060] The chemical reaction formulas involved in the above process are as follows: ; Example 8: Preparation of astaxanthin-C8-TPP fatty acid ester In a 500 mL round-bottom flask, 200 mL of anhydrous tetrahydrofuran (THF), 5.96 g of astaxanthin, 0.24 g of 4-dimethylaminopyridine (DMAP), and 2.42 g of triethylamine were added. The mixture was cooled to 0-5°C in an ice bath, and then 9.30 g of 2 molar equivalents (relative to astaxanthin) of TPP-octanoyl chloride was slowly added dropwise. The reaction was carried out at a controlled temperature for 20 hours to achieve controlled diesterization of the hydroxyl groups at both ends of astaxanthin. After the reaction was completed, insoluble matter was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain an orange-red solid astaxanthin-C8-TPP fatty acid ester.

[0061] The yield of the obtained product, astaxanthin-C8-TPP fatty acid ester, was 70%; MS (ESI+): m / z 1195.7 [M]+.

[0062] The chemical reaction formulas involved in the above process are as follows: ; Example 9: Preparation of tocopherol-C8-TPP fatty acid esters In a 500 mL round-bottom flask, 200 mL of anhydrous tetrahydrofuran (THF), 2.19 g of triphenylphosphine bromide octanoate, 1.95 g of α-tocopherol, 1.24 g of N,N'-dicyclohexylcarbodiimide (DCC), and 0.12 g of 4-dimethylaminopyridine (DMAP) were added, and the mixture was stirred at 60°C for 18 hours. After the reaction was complete, the insoluble matter was removed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography to give a pale yellow oily compound, tocopherol-C8-TPP fatty acid ester.

[0063] The yield of the obtained product, tocopherol-C8-TPP fatty acid ester, was calculated to be 63%; MS (ESI+): m / z 699.5[M]+.

[0064] The chemical reaction formulas involved in the above process are as follows: ; Example 10: Preparation of retinol-C10-TPP fatty acid ester Vitamin A (retinol) contains hydroxyl groups in its molecular structure, which can form stable ester bonds with TPP fatty acyl chloride. After binding with TPP targeting groups, its mitochondrial enrichment efficiency can be significantly improved, while retaining its antioxidant and cell metabolism-regulating biological activities. The specific preparation steps are as follows: In a 50 mL round-bottom flask, 20 mL of anhydrous dichloromethane, 2.86 g of vitamin A (retinol), and 0.12 g of 4-dimethylaminopyridine (DMAP) were added. The mixture was cooled to 0-5°C in an ice bath, and then 4.93 g of TPP-decanoyl chloride was slowly added dropwise. After the addition was complete, the temperature was raised to 45°C and the esterification reaction was carried out at a controlled temperature for 9 hours, all in the dark (to prevent the oxidative degradation of vitamin A). After the reaction was completed, 50 mL of ice water was added to quench the reaction, and the mixture was extracted three times with dichloromethane (50 mL each time). The organic layers were combined. The mixture was then washed twice with saturated brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography to obtain a pale yellow oily liquid retinol-C10-TPP fatty acid ester.

[0065] The yield of the obtained product, retinol-C10-TPP fatty acid ester, was calculated to be 73%; MS (ESI+): m / z 721.5 [M]+.

[0066] The chemical reaction formulas involved in the above process are as follows: ; Experimental Example 1: Detection of mitochondrial targeting properties of different mitochondrial-targeting compounds HepG2 cells were seeded in 6-well plates (2 × 10^5 cells / well) and cultured to the logarithmic growth phase. Then, 10 μM of each compound prepared in Examples 1-10 of this invention, the corresponding unmodified mitochondrial active compounds (vitamin C, carnosine, pyridoxine, nicotinamide nucleoside, vitamin B1, pantothenic acid, PQQ, astaxanthin, tocopherol, vitamin A), triphenylphosphine bromide, and the positive control MitoQ were added to the cells, and the cells were incubated at room temperature for 4 hours. After incubation, the cells were collected, and the mitochondrial and cytoplasmic components were separated by differential centrifugation. The concentration of the target compound in each component was determined by HPLC-MS, and the enrichment fold of the mitochondrial / cytoplasmic concentration was calculated according to the following formula. The results are shown in Table 1.

[0067] Enrichment factor = concentration of compound in mitochondria / concentration of compound in cytoplasm Table 1. Mitochondrial targeting enrichment folds of each compound (n=3). ; As shown in Table 1, the mitochondrial enrichment folds of the compounds prepared in Examples 1-10 of this invention were 12.5-16.3 times, significantly higher than those of the unmodified active compound (0.9-1.2 times) and triphenylphosphine bromide (7.8-8.5 times), and the sum of the two, demonstrating outstanding targeted enrichment ability. The compound enrichment efficiency was comparable to the positive control MitoQ (18.5 times), proving that the structural design of the short-chain linker arm + TPP cation in the C5-C12 region of the compounds in this invention can stably drive the compounds to penetrate the mitochondrial membrane and achieve efficient enrichment. Compounds with different carbon chain lengths (C6 / C8 / C10 / C12) all maintained good targeting, without the membrane permeability decrease problem caused by long chains, verifying the rationality of carbon chain optimization.

[0068] Experimental Example 2: Validation of the mitochondrial function protective activity of different mitochondrial-targeting compounds 1. Experimental Methods (1) DPPH free radical scavenging experiment: The scavenging ability of the sample against DPPH free radicals was determined by colorimetry, and the IC50 was calculated. 50 (Half-maximal clearance concentration). Specifically, the following steps are included: 1. Reagent preparation Accurately weigh the DPPH reagent, dissolve it in anhydrous ethanol and dilute to volume to prepare a DPPH stock solution with a concentration of 0.1 mmol / L. Store at 4°C protected from light. Before use, bring the solution to room temperature and dilute to the working concentration (0.04 mmol / L).

[0069] Accurately weigh each mitochondrial-targeting compound sample prepared in Examples 1-10, dissolve and serially dilute with DMSO to prepare sample solutions with concentrations of 10, 20, 40, 80, and 160 μmol / L, respectively. The final concentration of DMSO should not exceed 0.1% (to avoid interfering with the experiment).

[0070] Vitamin C was also set up as a positive control, and positive control solutions with the same gradient concentrations were prepared.

[0071] 2. Sample reaction Take a 96-well plate and add 100 μL of DPPH working solution to each well. Add 100 μL of sample solution of different concentrations to the experimental group, add 100 μL of vitamin C solution of the corresponding concentration to the positive control group, add 100 μL of anhydrous ethanol to the blank control group, and add 100 μL of anhydrous ethanol containing 0.1% DMSO to the negative control group. Set up 3 parallel replicates for each group.

[0072] 3. Incubation and Measurement Gently shake the 96-well plate to mix, incubate at room temperature in the dark for 30 minutes, and then use a microplate reader to measure the absorbance (A value) of each well at a wavelength of 517 nm. Record the data and calculate the average value.

[0073] 4. Calculation and Analysis Calculate the DPPH free radical scavenging rate using the formula: Clearance rate (%) = [1 - (Experimental group A value - Experimental group blank A value) / (Blank control group A value - Negative control group A value)] × 100%; Then, a dose-response curve was plotted with sample concentration on the x-axis and clearance rate on the y-axis, and IC was calculated using linear regression. 50 Value, IC 50 The smaller the value, the stronger the free radical scavenging ability of the sample.

[0074] (2) Intracellular ROS scavenging assay: H2O2 induced oxidative damage in HepG2 cells, and intracellular ROS levels were detected by DCFH-DA fluorescence method to calculate the ROS scavenging inhibition rate. The specific steps included: 1. Cell Culture After resuscitating HepG2 cells, they were cultured in DMEM medium containing 10% fetal bovine serum and 1% penicillin-streptomycin at 37°C in a 5% CO2 incubator. When the cell confluence reached 80%-90%, they were passaged, and cells in the logarithmic growth phase were selected for experiments.

[0075] 2. Cell seeding and grouping After digesting HepG2 cells, the cell concentration was adjusted to 1×10⁻⁶. 6 Cells were seeded at a density of 100 μL / mL in 96-well plates and cultured for 24 h to allow cell adhesion. Cells were then divided into a blank control group, a model group, an experimental group, and a positive control group, with three replicates per group. The blank control group received no treatment. The model group received H2O2 solution with a final concentration of 200 μmol / L. The experimental groups received solutions of mitochondrial-targeting compounds prepared in Examples 1-10 at different concentrations (20, 40, and 80 μmol / L), and after incubation for 2 h, H2O2 solution with a final concentration of 200 μmol / L was added. The positive control group received vitamin C solution with a final concentration of 100 μmol / L, and after incubation for 2 h, H2O2 solution with a final concentration of 200 μmol / L was added.

[0076] 3. ROS detection After H2O2-induced damage for 6 h, the culture medium was discarded, and the cells were washed twice with PBS buffer. 100 μL of DCFH-DA fluorescent probe solution with a final concentration of 10 μmol / L was added to each well, and the cells were incubated in a 37℃, 5% CO2 incubator in the dark for 30 min. After incubation, the cells were washed three times with PBS buffer to remove probes that had not entered the cells.

[0077] 4. Fluorescence Measurement and Calculation The fluorescence intensity of each well was measured using a fluorescence microplate reader (excitation wavelength 488 nm, emission wavelength 525 nm), the data were recorded, and the average value was calculated. The ROS scavenging inhibition rate was calculated according to the formula: Inhibition rate (%) = [1 - (fluorescence intensity of experimental group - fluorescence intensity of blank control group) / (fluorescence intensity of model group - fluorescence intensity of blank control group)] × 100%; where, the higher the inhibition rate, the stronger the ability of the sample to clear intracellular ROS.

[0078] (3) Mitochondrial membrane potential protection experiment: CCCP induces mitochondrial membrane potential depolarization, JC-1 staining is used for fluorescence microscopy observation, and the relative protection rate of membrane potential is calculated. The specific steps include: 1. Cell seeding and grouping After digesting HepG2 cells in the logarithmic growth phase, the concentration was adjusted to 1×10⁻⁶. 6 Cells were seeded at a density of 1 / mL into confocal culture dishes, 2 mL per dish, and cultured for 24 h to allow cell adhesion. The cells were divided into a blank control group, a model group, an experimental group, and a positive control group, with 3 replicates per group. The blank control group received no treatment. The model group received CCCP solution with a final concentration of 10 μmol / L. The experimental groups received solutions of mitochondrial-targeting compounds prepared in Examples 1-10 at different concentrations (20, 40, and 80 μmol / L), and after incubation for 2 h, CCCP solution with a final concentration of 10 μmol / L was added. The positive control group received cyclosporine A solution with a final concentration of 5 μmol / L, and after incubation for 2 h, CCCP solution with a final concentration of 10 μmol / L was added.

[0079] 2. JC-1 staining After CCCP treatment for 4 h, the culture medium was discarded, and the cells were washed twice with PBS buffer. 2 mL of JC-1 staining solution with a final concentration of 5 μmol / L was added to each dish, and the dishes were incubated in a 37℃, 5% CO2 incubator in the dark for 20 min. After incubation, the cells were washed twice with JC-1 staining buffer to remove unbound staining agent.

[0080] 3. Fluorescence observation The culture dish was observed under a laser confocal microscope with an excitation wavelength of 488 nm and emission wavelengths of 525 nm (green fluorescence, representing depolarized mitochondria) and 590 nm (red fluorescence, representing normal mitochondria). Three different fields of view were selected for photographing and recording, and the intensity ratio (R / G) of red fluorescence to green fluorescence in each field of view was calculated.

[0081] 4. Calculate the relative protection rate With the R / G value of the blank control group as 100%, the relative R / G value of each group was calculated; the relative protection rate of membrane potential (%) = (relative R / G value of experimental group - relative R / G value of model group) / (relative R / G value of blank control group - relative R / G value of model group) × 100%; where, the higher the protection rate, the stronger the protective effect of the sample on mitochondrial membrane potential.

[0082] (4) Oxidative damage cell survival experiment: HepG2 cells were damaged by H2O2, and the survival rate was detected by CCK-8 assay. The increase in cell survival rate was calculated. The specific steps included: 1. Cell seeding and grouping After digesting HepG2 cells in the logarithmic growth phase, the concentration was adjusted to 5 × 10⁻⁶. 4 Cells were seeded at a density of 100 μL / mL in 96-well plates and cultured for 24 h to allow cell adhesion. The plates were divided into a blank control group, a model group, an experimental group, and a positive control group, with three replicates per group. The blank control group received no treatment. The model group received H2O2 solution with a final concentration of 200 μmol / L. The experimental groups received solutions of mitochondrial-targeting compounds prepared in Examples 1-10 at different concentrations (20, 40, and 80 μmol / L), and after incubation for 2 h, H2O2 solution with a final concentration of 200 μmol / L was added. The positive control group received vitamin C solution with a final concentration of 100 μmol / L, and after incubation for 2 h, H2O2 solution with a final concentration of 200 μmol / L was added.

[0083] 2. Cell Culture and Detection After H2O2-induced damage for 24 h, the culture medium was discarded, the cells were washed twice with PBS buffer, and 100 μL of DMEM medium containing 10% CCK-8 reagent was added to each well. The cells were then incubated at 37°C and 5% CO2 for 2 h.

[0084] 3. Absorbance Measurement The absorbance (A value) of each well was measured at a wavelength of 450 nm using an ELISA reader. The data were recorded and the average value was calculated. With the cell viability of the blank control group being 100%, the cell viability of each group was calculated using the formula: Survival rate (%) = (A value of experimental group / A value of blank control group) × 100%.

[0085] 4. Calculate the increase in survival rate Survival rate improvement (%) = Survival rate of experimental group - Survival rate of model group; where, the greater the improvement in survival rate, the stronger the protective effect of the sample on H2O2-induced oxidative damage to cells.

[0086] 2. Experimental Results and Analysis The results are shown in Table 2. Table 2. Data on mitochondrial functional protective activity (n=3) ; As can be seen from the results in Table 2, the compounds IC prepared in Examples 1-10 50 All concentrations are <20 μM, exhibiting strong antioxidant activity and significantly improved stability. The inhibitory rate of H2O2-induced ROS scavenging reaches 48%-75%, and the protective rate against CCCP-induced mitochondrial membrane potential reaches 58%-89%. The survival rate of oxidatively damaged cells is increased to 76%-92%, with overall protective efficacy superior to the positive control MitoQ. This confirms that the compounds of this invention achieve targeted enrichment and synergistic enhancement of biological activity, effectively protecting mitochondrial function and alleviating oxidative stress damage.

[0087] The above experimental results show that the compound of the present invention achieves synergistic enhancement of mitochondrial targeted enrichment and bioactivity, effectively protects mitochondrial function, and improves mitochondrial damage caused by oxidative stress.

[0088] Application Example 1: Anti-aging oral preparations Take 10 g of pyridoxine-C8-TPP fatty amide, 20 g of microcrystalline cellulose, 70 g of lactose, and 1 g of magnesium stearate prepared in Example 3, mix them evenly, and then dry compress them into tablets to make mitochondrial protection anti-aging tablets.

[0089] Dosage and administration: Take one tablet once daily, orally.

[0090] Uses: Improves cell aging, delays skin and body aging, and enhances mitochondrial energy metabolism.

[0091] Application Example 2: Neuroprotective Injections Take 5 g of the PQQ-C10-TPP fatty acid amide prepared in Example 7, add 15 g of mannitol for injection and water for injection to 1000 mL, filter through a 0.22 μm filter membrane, fill and sterilize to prepare a neuroprotective injection solution.

[0092] Dosage and administration: Intravenous drip, once daily, 10 mL each time.

[0093] Uses: Used as an adjunct treatment for neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease, and to protect neuronal mitochondrial function.

[0094] Application Example 3: Cardiovascular Protection Soft Capsules Take 8 g of the astaxanthin-C8-TPP fatty acid ester prepared in Example 8, mix it evenly with 92 mL of soybean oil, and prepare soft capsules. Each capsule contains 80 mg of active ingredient.

[0095] Dosage and administration: Take 1 capsule twice daily, orally.

[0096] Uses: Improves myocardial ischemia-reperfusion injury, protects myocardial mitochondria, and is used as an adjunct to the prevention and treatment of cardiovascular diseases.

[0097] Application Example 4: Antioxidant Topical Gel Take 2 g of ascorbic acid-C8-TPP fatty acid ester prepared in Example 1, 1 g of carbomer 940, 5 g of glycerol, and an appropriate amount of triethanolamine, add purified water to 100 g, and prepare a mitochondrial targeted antioxidant gel.

[0098] Dosage and administration: Apply externally, once or twice daily.

[0099] Uses: Removes ROS from skin mitochondria, reduces photoaging, and is used for antioxidant skincare.

[0100] Application Example 5: Metabolic Syndrome Intervention Powder Take 12 g of nicotinamide nucleoside-C10-TPP fatty acid ester prepared in Example 4 and 88 g of maltodextrin, mix and sieve to prepare oral powder.

[0101] Dosage and administration: Take one sachet once a day with warm water.

[0102] Uses: Regulates mitochondrial glucose and lipid metabolism, and helps improve metabolic syndromes such as diabetes and obesity.

[0103] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of the present invention, such as replacing different C5-C12 terminal halogenated fatty acids, adjusting reaction temperature and time, replacing different mitochondrial active nuclei, etc., without departing from the spirit and scope of the technical solutions of the present invention, and all such modifications and substitutions should be covered within the scope of the claims of the present invention.

[0104] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Although the present invention has been disclosed above with reference to preferred embodiments, it is not intended to limit the present invention. Any person skilled in the art can make some modifications or alterations to the methods and techniques disclosed above without departing from the scope of the present invention to create equivalent embodiments. Any simple modifications, equivalent changes and alterations made to the above embodiments based on the technical essence of the present invention without departing from the scope of the present invention shall still fall within the scope of the present invention.

Claims

1. A mitochondrially targeted compound, characterized in that, It has the structure shown in the general formula (I): [M-O / -NH]-[C(=O)-R1]-PPh3 + X - (I); In formula (I): M is selected from any one of vitamin A, vitamin E, vitamin C, pyridoxine, nicotinamide nucleotide, vitamin B1, vitamin B5, pyrroloquinoline quinone, and astaxanthin; R1 is a straight-chain or branched alkyl group containing 5-12 carbon atoms, or a carbon chain containing unsaturated bonds; X is a halide ion or an organic acid radical ion. PPh3 is a triphenylphosphine group.

2. The mitochondrially-targeted compound of claim 1, wherein, The bonding method of M is: When M is vitamin A, vitamin C, nicotinamide nucleotide, vitamin E, or astaxanthin, an ester bond -O-CO- is formed; When M is pyridoxine, vitamin B1, vitamin B5, or pyrroloquinoline quinone, an amide bond -CONH- is formed.

3. The mitochondrially-targeted compound of claim 1, wherein, R1 is a straight-chain or branched alkyl group containing 6, 8, 10 or 12 carbon atoms, or a carbon chain containing unsaturated bonds; X is selected from any one of bromide ion, iodide ion, chloride ion, p-toluenesulfonate ion, methanesulfonate ion, and trifluoroacetate ion.

4. The mitochondrially-targeted compound according to any one of claims 1 to 3, characterized in that, The mitochondrial-targeting compound is selected from one of the following: vitamin C-TPP fatty acid ester, pyridoxine-TPP fatty amide, nicotinamide nucleotide-TPP fatty acid mono / diester, vitamin B1-TPP fatty amide, vitamin B5-TPP fatty amide, pyrroloquinoline quinone-TPP fatty amide, astaxanthin-TPP fatty diester, vitamin E-TPP fatty amide, and vitamin A-TPP fatty diester.

5. The method of claim 1-4, wherein the mitochondrial targeting compound is prepared by, The method includes the following steps: S1: Triphenylphosphine and halofatty acids are reacted in an anhydrous organic solvent to obtain triphenylphosphine salts of fatty acids; S2: An acylating agent is added to the obtained triphenylphosphine salt of fatty acid to react and obtain TPP fatty acyl chloride; S3: The active molecule M is reacted with TPP fatty acyl chloride in a solvent to prepare the mitochondrial-targeting compound; The active molecule M is selected from at least one of vitamin A, vitamin E, vitamin C, pyridoxine, vitamin B1, vitamin B5, pyrroloquinoline quinone, nicotinamide nucleotide, or astaxanthin; The reaction conditions are selected based on the type of active groups on the active molecule M: (a) When the active group is an alcohol hydroxyl group, the esterification reaction is carried out at 40-80°C for 6-24 hours in the presence of an esterification catalyst; or (b) When the active group is amino, the amidation reaction is carried out at 0-15°C in the dark for 8-18 hours in the presence of an alkaline acid binder.

6. The production method according to claim 5, characterized by, In step S1, the halogenated fatty acid is a C5-C12 terminal halogenated fatty acid, selected from at least one of 5-bromopentanoic acid, 6-bromohexanoic acid, 7-bromoheptanoic acid, 8-bromooctanoic acid, 9-bromononanoic acid, 10-bromodecanoic acid, 11-bromoundecanoic acid, 12-bromododecanoic acid, 5-iodopentanoic acid, and 12-iodododecanoic acid. The molar ratio of the triphenylphosphine to the halofatty acid is (1:1) to (1.5:1). The anhydrous organic solvent is selected from at least one of acetonitrile, N,N-dimethylformamide, dimethyl sulfoxide, tetrahydrofuran, and dichloromethane; The reaction conditions are: temperature 60~100℃, time 4~12 hours.

7. The preparation method according to claim 5, characterized in that, In step S2, the acylating agent is selected from one of thionyl chloride, phosphorus trichloride, phosgene, and triphosgene; The amount of the acylating agent added is 1.0-5.0 molar equivalents of the active molecule M; The reaction conditions are: temperature 5-80℃, time 3-8 hours.

8. The preparation method according to claim 5, characterized in that, In step S3, when the active molecule M is vitamin C, the method specifically includes: esterifying vitamin C with TPP fatty acyl chloride at 40-80°C for 6-24 hours in the presence of an esterification catalyst to obtain vitamin C-TPP fatty acid ester; or When the active molecule M is nicotinamide nucleotide, the method specifically includes: reacting nicotinamide nucleotide with TPP fatty acyl chloride at 40-70°C for 6-12 hours in the presence of an esterification catalyst to obtain a monoester or diester; or When the active molecule M is astaxanthin, the method specifically includes: esterifying astaxanthin with 2 molar equivalents of TPP fatty acyl chloride in the presence of an esterification catalyst under an organic base environment for 12-24 hours to obtain astaxanthin diester. The organic base is triethylamine or pyridine.

9. The preparation method according to claim 8, characterized in that, The esterification catalyst is selected from at least one of N,N'-dicyclohexylcarbodiimide, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide, and 4-dimethylaminopyridine. The amount of the esterification catalyst used is 5-20% of the total molar amount of the reactants.

10. The preparation method according to claim 5, characterized in that, In step S3, when the active molecule M is pyridoxine, vitamin B1, vitamin B5, or pyrroloquinoline quinone, the method specifically includes: The active molecule M and TPP fatty acyl chloride were reacted with an amidation reaction at 0-15°C in the dark for 8-18 hours in the presence of an alkaline acid binder to obtain the amidated product.

11. The preparation method according to claim 10, characterized in that, The alkaline acid-binding agent is selected from at least one of triethylamine and N,N-diisopropylethylamine; The amount of the alkaline acid-binding agent is 1.2 to 1.5 times the molar amount of TPP fatty acyl chloride.

12. The use of the mitochondrial-targeting compound according to any one of claims 1-4, or the mitochondrial-targeting compound prepared by the method according to any one of claims 5-11, in the preparation of products for mitochondrial function protection; The products include: (1) Anti-aging products; (2) Neuroprotective products; (3) Cardiovascular protection products; (4) Antioxidant products; or (5) Products for the prevention and treatment of metabolic syndrome.