Vincamine derivatives and preparation methods and applications thereof

Abstract

The present invention relates to vincamine derivatives and preparation methods and applications thereof, and belongs to the technical field of preparation of vincamine derivatives. The vincamine derivatives disclosed in the present invention have the following advantages: (i) modification of the tail chain increases the lipid solubility of vincamine compounds, and does not affect the cerebral blood flow regulation of vincamine itself, which helps the carried drug to penetrate the blood-brain barrier, and exerts a brain protective effect, improving brain microcirculation disorders; (ii) the tertiary amine group in the parent nucleus structure of vincamine derivatives has the property of being ionizable under acidic conditions, and can achieve efficient carrying and lysosomal escape of nucleic acid drugs through charge adsorption, thereby improving intracellular transport; (iii) vincamine derivatives inherit various pharmacological activities inherent in vincamine and have high safety. Therefore, the vincamine derivatives disclosed in the present invention have good application prospects in brain-targeted delivery of drugs for treating brain diseases.

Description

Technical Field

[0001] The invention belongs to the technical field and relates to vincamine derivatives and preparation methods and applications thereof. Background Art

[0002] Treatment of central nervous system diseases, such as neurodegenerative diseases, brain tumors, brain infections, and stroke, is severely limited by the blood-brain barrier because it prevents most small molecule drugs and macromolecules (such as peptides, nucleic acid drugs, and protein drugs) from entering the brain. To date, extensive efforts have been made to improve the efficiency of drug delivery into the brain, including direct CNS administration, disruption of the blood-brain barrier, and receptor-mediated delivery methods. However, direct CNS administration is invasive, can cause infection and tissue damage, and is also limited by diffusion distance and the rapid efflux of drugs out of the central nervous system within hours. Using techniques such as osmotic disruption, biochemical disruption, and ultrasound-mediated disruption of the blood-brain barrier, drugs can be effectively introduced into the brain; however, these transient blood-brain barrier openings also allow plasma proteins to leak into the brain, leading to neurotoxicity, vasculopathy, and chronic neuropathological changes in the brain. Therefore, methods for safely and effectively delivering substances that cannot penetrate the blood-brain barrier, especially gene and nucleic acid therapies, to the central nervous system by intravenous injection remain to be improved. Currently, lipid nanoparticles (LNPs) mainly composed of ionizable lipids are used to encapsulate nucleic acid drugs. However, due to their high positive charge characteristics, they are mostly distributed to the liver after entering the body. Although there is an LNP platform technology called SORT (Selective organ targeting) that achieves non-hepatic delivery of nucleic acid drugs to the spleen and lungs by adjusting the surface charge of the preparation, it still does not solve the problem of nucleic acid drugs entering the brain.

[0003] In addition, considering the steric hindrance during synthesis and preparation, most ionizable lipid molecules are generally designed to achieve pH sensitivity and intracellular escape by introducing simple linear tertiary amine groups rather than cyclic tertiary amine head groups. Only a few reports have shown that a single cyclic tertiary amine head group may also have good binding interactions with nucleic acids, such as piperazine-containing C12-200, which can carry 5 liver-targeted siRNAs into cells at the same time; and, in addition to promoting RNA encapsulation and cellular uptake, synthetic lipids do not always have ideal biological activity. In fact, potential cytotoxicity and immunogenicity are the main defects associated with the use of cationic lipid materials, because they can activate innate immunity through the complement system and Toll-like receptors, and may also produce proinflammatory cytokines and reactive oxygen free radicals, especially when long-term repeated administration is required. Therefore, glucocorticoids and antihistamines are often required to be administered in advance in the actual use of LNPs.

[0004] Natural product libraries provide a rich source of compounds and play a key role in drug discovery. Small molecule natural products, in particular, often have huge compound skeleton diversity and multiple pharmacophores, and are good lead compounds that dominate drug development. In recent years, some small molecule natural products have also promoted the drug delivery process from a new perspective. Among them, alkaloids are a class of nitrogen-containing alkaline organic compounds with significant biological activity. As one of the earliest studied natural products, a large number of alkaloid drugs have been discovered and approved by the FDA. Representative drugs of indole alkaloids are vinpocetine and vincamine, which belong to the vincamine alkaloids. They have a variety of pharmacological effects that are beneficial to the brain, cardiovascular, blood circulation and other systems: promoting the uptake and utilization of glucose and oxygen in the brain, increasing ATP, and reducing the production of lactic acid during ischemia and hypoxia; preventing brain cell excitotoxic death; alleviating brain hypoxia damage and protecting neurons; enhancing the functions of dopaminergic, serotonergic and noradrenergic nerves; preventing ischemic damage to the brain, liver, muscle tissue and other parts; scavenging free radicals and resisting lipid peroxidation; alleviating aging brain dysfunction; improving the composition of lipoproteins in the blood, etc. Among them, the significant anti-inflammatory and antioxidant effects are expected to counteract the potential toxicity of conventional cationic / ionizable lipid materials. In addition, the indole heterocycle of vincamine has a proton amine structure, which is expected to interact electrostatically with nucleic acid drugs, thereby encapsulating and delivering nucleic acid drugs into cells. However, due to the short half-life and low bioavailability of vinpocetine and other drugs, frequent administration is required when used for long-term treatment of chronic diseases such as cerebrovascular diseases. Studies have shown that targeted delivery systems can be used to prolong the circulation time of vinpocetine in the body and penetrate the blood-brain barrier, so that more drugs can reach the lesion site. Based on the five-ring fused skeleton of vinpocetine, intermediates and end products in the synthesis process were selected for structural modification and fatty chain derivatization, and it was found that some modified vinpocetine derivative lipids showed brain targeting.

[0005] Therefore, vincamine-derived lipids can be used as an ionizable lipid to construct a safe and efficient brain-targeted delivery system for nucleic acid drugs. Summary of the invention

[0006] In view of this, one of the objects of the present invention is to provide vincamine derivatives; a second object of the present invention is to provide a method for preparing vincamine derivatives; a third object of the present invention is to provide the use of vincamine derivatives in brain-targeted delivery of drugs for treating brain diseases.

[0007] In order to achieve the above object, the present invention provides the following technical solutions:

[0008] 1. Vincamine derivatives, the structural formula of the vincamine derivatives is Or any one of AO-NH-C,

[0009] Wherein A is a group formed after vincamine and its derivatives having an indole ring structure skeleton lose a hydroxyl group;

[0010] B is -(CH2) n -、-(CH2) n -SS-(CH2) n -、-(CH2) n -TK-(CH2) n -or-(CH2) n -S-Mal-(PEG) m- Any of NHS-, where n and m are both positive integers;

[0011] C is a straight-chain alkyl group having 10 to 18 carbon atoms, a straight-chain alkenyl group having 10 to 18 carbon atoms, a straight-chain alkyl group having 10 to 18 carbon atoms substituted with a hydroxyl group, or a straight-chain alkyl group having 10 to 18 carbon atoms substituted with a chemical formula of C x H 2x+1 OCO(CH2) y -group, (C a H 2a+1 )2N(CH2) b -group, (C d H 2d+1 OCH2CH2)2NCO(CH2) e -group, (C d H 2d+1 COOCH2CH2)2NCO(CH2) e -, DSPE-PEG-, PEG-, a group formed after doxorubicin loses its amino group, a group formed after paclitaxel is carboxylated at the 2'-OH site, or any one of an amino acid chain, wherein x, a, b, d and e are all positive integers.

[0012] Preferably, A is any one of structural formulas A1 to A12:

[0013]

[0014] Preferably, B is any one of B1 to B5, wherein B1 is -(CH2)2-, B2 is -(CH2)6-, B3 is -(CH2)2-SS-(CH2)2-, B4 is -(CH2)2-SC(CH3)2-S-(CH2)2-, and B5 is -(CH2)2-S-Mal-PEG-NHS-.

[0015] Preferably, C is any one of the structural formulas C1 to C14:

[0016]

[0017] n is a positive integer

[0018]

[0019] C also includes C15, wherein C15 is bovine serum albumin.

[0020] Preferably, the vincamine derivatives include any one of Compounds 1 to 38 having the following structural formulas:

[0021]

[0022]

[0023]

[0024]

[0025] n is a positive integer

[0026]

[0027] (A1-B5-C15), where C15 is bovine serum albumin, R is n is a positive integer.

[0028] 2. The preparation method of the above vincamine derivatives, wherein the structural formula of the vincamine derivatives is When the preparation method comprises any one of method 1, method 2, method 3 and method 4,

[0029] Method 1: When A is a group of structural formula A1, B is a group of structural formula B1, and C is a group of structural formula C14, a compound of structural formula A1-OH, a compound of structural formula H2N-B1-NH2 and benzotriazole-1-yl-oxytripyrrolidinophosphorus hexafluorophosphate are dissolved in an organic solvent at a molar ratio of 0.17:0.27:0.28, and stirred at 20-40°C for 12-48h. The product obtained by the reaction is rotary evaporated, redissolved, extracted, dried, and then concentrated under reduced pressure, and then the organic solvent is used to obtain the product. The intermediate compound I is solidified by a solvent; the compound with the structural formula C14-OH, the intermediate compound I and benzotriazole-1-yl-oxytripyrrolidinophosphorus hexafluorophosphate are dissolved in an organic solvent at a molar ratio of 0.31:0.052:0.047, the pH is adjusted to 7-8, and the reaction is stirred at 20-40° C. for 1-12 hours, and the product obtained by the reaction is rotary evaporated, redissolved, extracted, dried, and then concentrated under reduced pressure to obtain a vincamine derivative with the structural formula A1-NH-B1-NH-C14;

[0030] Method 2: When A is a group of structural formula A1, B is a group of structural formula B5, and C is a bovine serum albumin (BSA) sequence, a compound of structural formula A1-OH, cystamine and benzotriazole-1-yl-oxytripyrrolidinophosphine hexafluorophosphate are dissolved in an organic solvent at a molar ratio of 0.15:0.23:0.23, the pH is adjusted to 7-8 with an organic solvent, and the reaction is stirred at 20-40° C. for 1-12 hours, the product obtained by the reaction is solidified with an organic solvent, vacuum dried to obtain a crude product, and then dissolved in an organic solvent, and an aqueous solution of tris(2-carbonylethyl)phosphine hydrochloride (the molar ratio of the compound of structural formula A1-OH to tris(2-carbonylethyl)phosphine hydrochloride is 0.15:0.17) is added dropwise, and the reaction is stirred at 20-40° C. for 1-12 hours, and the intermediate compound II and NHS-PEG are concentrated under reduced pressure, precipitated with water, and then dried to obtain an intermediate compound II; the intermediate compound II and NHS-PEG n -Mal was dissolved in an organic solvent at a molar ratio of 0.02:0.01, the reaction mixture was stirred at 20-40°C for 1-12 hours, then added into a PBS buffer solution containing bovine serum albumin (BSA), stirred at 20-40°C for 1-12 hours, dialyzed with pure water, and freeze-dried to obtain a structural formula of (A1-NH-B5-NH) 11 -C15 vincamine derivatives.

[0031] Method 3: The preparation method of vincamine derivatives of other structural formulas comprises the following steps:

[0032] (1) A compound with a structural formula of C-Br, a compound with a structural formula of HO-B-NH2 or H2N-B-NH2, potassium carbonate and potassium iodide are dissolved in an organic solvent at a molar ratio of 1.21:0.552:2.43:0.552, and the mixture is heated to 45-65° C. and reacted for 12-48 hours. The product obtained by the reaction is cooled and filtered, and the obtained filtrate is extracted with n-hexane and separated by silica gel chromatography to obtain a compound with a structural formula of An intermediate compound wherein B is -(CH2) n -、-(CH2) n -SS-(CH2) n -, or -(CH2) n -TK-(CH2) n -, C is a group other than a straight-chain alkyl group having 10 to 18 carbon atoms substituted by a hydroxyl group;

[0033] Alternatively, a linear alkane having 10 to 18 carbon atoms substituted by an epoxy group and a compound having a structural formula of HO-B-NH2 or H2N-B-NH2 are dissolved in an organic solvent at a molar ratio of 1.26:0.57, and the mixture is heated to 55 to 75°C and reacted for 12 to 48 hours. The product obtained by the reaction is concentrated under reduced pressure and separated by silica gel chromatography to obtain a compound having a structural formula of An intermediate compound, wherein C is a straight-chain alkyl group having 10 to 18 carbon atoms substituted by a hydroxyl group;

[0034] (2) a compound with the structural formula A-OH and a compound with the structural formula The intermediate compound, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and 4-dimethylaminopyridine are dissolved in an organic solvent in a molar ratio of 0.15:1.59:0.183:0.0081, heated to 20-40°C for reaction for 24-96 hours, the product obtained by the reaction is diluted, extracted and dried, concentrated under reduced pressure and separated by silica gel chromatography to obtain a product with the structural formula Vincamine derivatives;

[0035] The preparation method of vincamine derivatives with the structural formula AO-NH-C is as follows:

[0036] When A is a group of structural formula A1 and C is a group of structural formula C12, a compound of structural formula A1-OH, a compound of structural formula C12-NH2(DSPE-PEG-NH2), and benzotriazole-1-yl-oxytripyrrolidinophosphine hexafluorophosphate are dissolved in an organic solvent, stirred and reacted at 20-40° C. for 24-96 hours, dialyzed with pure water, and freeze-dried to obtain a vincamine derivative of structural formula A1-O-NH-C12;

[0037] When A is a group of structural formula A1 and C is a group of structural formula C13, a compound of structural formula A1-OH, a compound of structural formula C13-NH2 (DOX-NH2) and benzotriazole-1-yl-oxytripyrrolidinophosphorus hexafluorophosphate are dissolved in an organic solvent, the pH is adjusted to 7-8 and then stirred to react at 20-40° C. for 1-12 hours, the product obtained by the reaction is rotary evaporated, redissolved, extracted, dried and concentrated under reduced pressure, and then solidified with an organic solvent to obtain a vincamine derivative with structural formula A1-O-NH-C13.

[0038] Preferably, the eluent used in the silica gel chromatography separation in step (1) is a mixed solution composed of methanol and dichloromethane in a volume ratio of 10:90 to 90:10, and the eluent used in the silica gel chromatography separation in step (2) is a mixed solution formed by mixing acetone and n-hexane in a volume ratio of 20:80 to 80:20.

[0039] Preferably, the organic solvent is any one of acetonitrile, ethanol, methanol, N,N-dimethylformamide, N,N-diisopropylethylamine, ethyl acetate, methyl tert-butyl ether, triethylamine, pyridine or dichloromethane.

[0040] 3. Application of the above vincamine derivatives in brain-targeted delivery of drugs for the treatment of brain diseases.

[0041] The beneficial effects of the present invention are as follows: the present invention discloses vincamine derivatives and a preparation method thereof, and the vincamine derivatives have the following advantages: (i) modification of the tail chain increases the lipid solubility of vincamine compounds, and does not affect the cerebral blood flow regulation of vincamine itself, which helps the carried drug to penetrate the blood-brain barrier, and exerts a brain protective effect, and improves brain microcirculation disorders; (ii) the tertiary amine group in the parent nucleus structure of the vincamine derivative has the property of being ionizable under acidic conditions, and can achieve efficient carrying and lysosomal escape of nucleic acid drugs through charge adsorption, thereby improving intracellular transport; (iii) the vincamine derivatives inherit the various pharmacological activities inherent in vincamine and have a high degree of safety. Therefore, the vincamine derivatives disclosed in the present invention have good application prospects in brain-targeted delivery of drugs for treating brain diseases.

[0042] Other advantages, objectives and features of the present invention will be described in the following description to some extent, and to some extent, will be obvious to those skilled in the art based on the following examination and study, or can be taught from the practice of the present invention. The objectives and other advantages of the present invention can be realized and obtained through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention will be described in detail below in conjunction with the accompanying drawings, wherein:

[0044] Figure 1 is the compound 1 in Example 1 1 H NMR spectrum;

[0045] Figure 2 is a transmission electron microscopy image of lipid nanoparticles (LNP) (compound 2 (A5-B2-C6)-14);

[0046] Figure 3 The pKa values ​​of lipid nanoparticles prepared from different vincamine derivatives were determined by TNS fluorescence method;

[0047] Figure 4 To observe the FAM fluorescence intensity of different drug administration groups (lipid nanoparticles prepared by adding different vincamine derivatives) under fluorescence microscope;

[0048] Figure 5The cell survival rate was calculated by measuring the absorbance of each well (in which lipid nanoparticles formed by different vincamine derivatives were added) at 570 nm using an ELISA instrument;

[0049] Figure 6 Laser speckle imaging was used to measure the effects of lipid nanoparticles formed by different vincamine derivatives on brain microvascular blood flow in mice;

[0050] Figure 7 To determine the N / P ratio of siRNA encapsulated in lipid nanoparticles formed by vincamine derivatives (A1-B1-C5) using gel retardation assay;

[0051] Figure 8 Table 2 Endosomal escape effect of lipid nanoparticles loaded with siRNA formed by compound 1 (A1-B1-C5), compound 2 (A5-B2-C6) and compound 31 (A11-B3-C11) was taken with a confocal microscope;

[0052] Fig. 9 To detect the effect of lipid nanoparticles formed with compound 2 (A5-B2-C6) on cellular ROS levels using a single cell analyzer;

[0053] Fig.10 NH2-PEG 2000 -Mass spectra of DSPE (a) and compound 35 (b) prepared in Example 6;

[0054] Fig.11 Mass spectrum of compound 36 prepared in Example 7;

[0055] Fig.12 The mass spectrum of compound 37 prepared in Example 7;

[0056] Fig.13 TLC verification result of compound 38 prepared in Example 8;

[0057] Fig.14 A standard curve was obtained to measure the absorbance after the reaction of L-leucine (0-10 mM) with OPA reagent;

[0058] Fig.15 Fluorescence images collected at predetermined time points (0.5h, 1h, 2h) by the VISQUE in vivo Smart-LF System after Compound 31, Compound 35, Compound 36, Compound 37 and Compound 38 (A1-B5-C15)) were injected into mice. DETAILED DESCRIPTION

[0059] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the illustrations provided in the following embodiments only illustrate the basic concept of the present invention in a schematic manner, and the following embodiments and features in the embodiments can be combined with each other without conflict.

[0060] In the following embodiments, the related groups in the structural formula and the structural formula of the compound involved are as follows:

[0061] A is any one of the structural formulas A1 to A12:

[0062]

[0063] B is any one of B1 to B5, wherein B1 is -(CH2)2-, B2 is -(CH2)6-, B3 is -(CH2)2-SS-(CH2)2-, B4 is -(CH2)2-SC(CH3)2-S-(CH2)2-, and B5 is -(CH2)2-S-Mal-PEG-NHS-;

[0064] C is or C' is any one of the structural formulas C1 to C14:

[0065]

[0066] n is a positive integer

[0067]

[0068] In addition, C also includes C15, wherein C15 is bovine serum albumin (BSA);

[0069] The structural formulas of Compound 1 to Compound 38 are:

[0070]

[0071]

[0072]

[0073]

[0074] n is a positive integer

[0075]

[0076] (A1-B5-C15), where C15 is bovine serum albumin (BSA), R is n is a positive integer.

[0077] Example 1

[0078] A vincamine derivative (Compound 1) was prepared, the structural formula of which is:

[0079] The specific preparation method is as follows:

[0080] (1) (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br) (400 mg, 1.21 mmol), 2-aminoethanol (HO-B1-NH2) (35 uL, 0.552 mmol), potassium carbonate (330 mg, 2.43 mmol), and potassium iodide (9 mg, 0.552 mmol) were dissolved in 5 mL of acetonitrile at a molar ratio of 1.21:0.552:2.43:0.552, and the reaction mixture was heated and stirred at 65°C for 24 h;

[0081] (2) Cool to room temperature, wash the filter cake obtained by filtration with n-hexane three times, extract the filtrate obtained by filtration with n-hexane, concentrate under reduced pressure, separate by silica gel chromatography (methanol / dichloromethane=15 / 85 (v / v)), and obtain the structural formula by rotary evaporation: intermediate compounds;

[0082] (3) A1-OH (50 mg, 0.15 mmol) and the structural formula The intermediate compound (88 mg, 1.59 mmol), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl, 35 mg, 0.183 mmol) and 4-dimethylaminopyridine (DMAP, 1 mg, 0.0081 mmol) were dissolved in 6 mL of pyridine / dichloromethane (1 / 1, v / v), and the reaction mixture was stirred at room temperature for 24 h;

[0083] (4) The organic phase obtained by separation was diluted with dichloromethane (6 mL), washed 4 times with 5 mL of 2M hydrochloric acid solution and saturated brine, then dried over sodium sulfate, concentrated under reduced pressure, and separated by silica gel chromatography (acetone / n-hexane = 1 / 1 (v / v); dichloromethane / methanol = 1 / 3 (v / v)). The final product was obtained by rotary evaporation with the structural formula: A vincamine derivative (compound 1), which 1 H NMR (400 MHz, DMSO-d6) spectrum is shown in Figure 1 shown.

[0084] Example 2

[0085] A vincamine derivative (Compound 2) was prepared, the structural formula of which is:

[0086] The specific preparation method is as follows:

[0087] (1) 1,2-epoxydodecane (C6-epoxy 233 mg, 1.26 mmol) and 6-aminohexanol (HO-B2-NH2, 202 mg, 0.57 mmol) were dissolved in 2 mL of ethanol at a molar ratio of 1.26:0.57, and the reaction mixture was heated and stirred at 75°C for 24 h;

[0088] (2) The reaction product was concentrated under reduced pressure and separated by silica gel chromatography (methanol / dichloromethane = 1 / 30 (v / v)), and rotary evaporated to obtain a product having the structural formula: intermediate compounds;

[0089] (3) A5-OH (50 mg, 0.15 mmol) and the structural formula The intermediate compound (88 mg, 1.59 mmol), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (EDC·HCl, 35 mg, 0.183 mmol) and 4-dimethylaminopyridine (DMAP, 1 mg, 0.0081 mmol) were dissolved in 6 mL of pyridine / dichloromethane (1 / 1, v / v), and the reaction mixture was stirred at room temperature for 24 h;

[0090] (4) The organic phase was diluted with dichloromethane (6 mL), washed four times with 2M hydrochloric acid solution (4×5 mL) and saturated brine, then dried over sodium sulfate, concentrated under reduced pressure, and separated by silica gel chromatography (acetone / n-hexane = 1 / 1 (v / v); dichloromethane / methanol = 1 / 3 (v / v)). The final product was obtained by rotary evaporation with the structural formula: Vincamine derivatives (Compound 2), its 1H NMR (400MHz, DMSO-d6) δ10.32 (s, 1H), 7.42 (d, J = 8.0 Hz, 2H), 7.07 (dd, J = 8.4, 6.9 Hz, 1H), 6.99 (dd, J = 8.1, 6.8 Hz, 1H), 4.84 (d, J = 10.1 Hz, 2H), 4.32 (t, J = 5.1 Hz, 1H), 4.27 (d, J = 3.7 Hz, 1H), 4.21(d,J=3.9Hz,1H),4.18–4.09(m,2H),2.09–1.98(m,2H),1.82(ddd,J=13.7,10.3,6.1Hz, 1H), 1.39 (q, J = 9.7Hz, 10H), 1.29–1.18 (m, 50H), 0.86 (t, J = 6.7Hz, 9H), 0.64 (t, J = 7.5Hz, 3H).

[0091] Example 3

[0092] According to the reaction steps in Example 1 (i.e., replacing certain reactants in the example, the addition ratio and other reaction conditions are the same as those in Example 1), the following compounds were prepared:

[0093] (1) Compound 3 (A1-B1-C2): (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br) was replaced by 16-bromohexadecane (C2-Br) and 2-aminoethanol (HO-B1-NH2) was replaced by 2-aminoethanol (HO-B1-NH2) to prepare a compound having the structural formula: Compound 3, its 1H NMR (400 MHz, DMSO-d6) δ 7.62 (d, 1H), 7.51 (d, 1H), 7.46 (t, 1H), 7.17 (t, 1H), 6.74 (s, 1H), 4.27 (t, 2H), 3.63 (s, 1H), 2.90 (m, 6H), 2.68 (m, 6H), 1.64 (m, 2H), 1.48-1.33 (m, 60H), 0.89 (m, 3H), 0.84 (m, 6H).

[0094] (2) Compound 4 (A1-B1-C3): (6Z)-12-bromododecan-6-ene (C3-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br) to prepare a compound having the structural formula: Compound 4, its 1H NMR (400 MHz, DMSO-d6) δ 7.62 (d, 1H), 7.52 (d, 1H), 7.46 (t, 1H), 7.17 (t, 1H), 6.71 (s, 1H), 5.39 (m, 4H), 4.26 (t, 2H), 3.59 (s, 1H), 2.90 (m, 6H), 2.68 (m, 6H), 2.00 (m, 8H), 1.51 (m, 4H), 1.48-1.33 (m, 26H), 0.89 (m, 9H).

[0095] (3) Compound 5 (A1-B2-C4): (9Z)-16-bromohexadecane-9-ene (C4-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 6-aminohexanol (HO-B2-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A2-OH was used to replace A1-OH to obtain the structural formula: Compound 5, its 1H NMR (400 MHz, DMSO-d6) δ 7.62 (d, 1H), 7.52 (d, 1H), 7.46 (t, 1H), 7.17 (t, 1H), 6.71 (s, 1H), 5.39 (m, 4H), 4.20 (t, 2H), 3.59 (s, 1H), 2.68 (m, 4H), 2.68 (m, 6H), 1.99 (m, 8H), 1.49 (m, 6H), 1.49-1.29 (m, 50H), 0.89 (m, 9H).

[0096] (4) Compound 6 (A2-B2-C8): 10-bromooctyl ester (C8-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 6-aminohexanol (HO-B2-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A2-OH was used to replace A1-OH to obtain the structural formula: Compound 6, its 1H NMR (400 MHz, DMSO-d6) δ 7.84 (s, 1H), 7.54 (d, 1H), 7.47 (d, 1H), 6.74 (s, 1H), 4.20 (t, 4H), 4.02 (t, 2H), 3.62 (s, 1H), 2.90 (t, 6H), 2.36 (m, 10H), 1.69-1.29 (m, 66H), 0.89 (m, 9H).

[0097] (5) Compound 7 (A2-B3-C9): Replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br) with N,N-dioctyl-2-bromo-ethylamine (C9-Br), replace 2-aminoethanol (HO-B1-NH2) with 6-amino-3,4-dithia-hexanol (HO-B3-NH2), and replace A1-OH with A2-OH to obtain the structural formula: Compound 7, its 1H NMR (400 MHz, DMSO-d6) δ7.84 (s, 1H), 7.54 (d, 1H), 7.47 (d, 1H), 6.74 (s, 1H), 4.31 (t, 2H), 3.80 (d, 1H), 3.07 (t, 12H), 2.87 (t, 2H), 2.73-2.59 (m, 10H), 2.53 (m, 8H), 1.70-1.32 (m, 50H), 0.89 (m, 15H).

[0098] (6) Compound 8 (A2-B3-C10): Replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br) with N,N-di(3-oxodecyl)-nonanamide (C10-Br), replace 2-aminoethanol (HO-B1-NH2) with 6-amino-3,4-dithia-hexanol (HO-B3-NH2), and replace A1-COOH with A2-OH to obtain the structural formula: Compound 8, its 1H NMR (400 MHz, DMSO-d6) δ 7.84 (s, 1H), 7.54 (d, 1H), 7.47 (d, 1H), 6.74 (s, 1H), 4.01 (t, 2H), 3.80 (t, 8H), 3.54 (s, 1H), 3.28 (m, 16H), 3.07 (t, 4H), 2.87 (t, 2H), 2.73-2.54 (m, 10H), 2.35 (t, 4H), 1.52-1.32 (m, 70H), 0.89 (m, 15H).

[0099] (7) Compound 9 (A3-B4-C3): (6Z)-12-bromododecan-6-ene (C3-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 4,4-dimethyl-3,5-thioketal-heptanol (HO-B4-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A3-OH was used to replace A1-OH to obtain the structural formula: Compound 9, its 1H NMR (400 MHz, DMSO-d6) δ 7.74 (d, 1H), 7.61 (d, 1H), 7.44 (t, 1H), 7.16 (t, 1H), 5.48 (m, 4H), 4.30 (t, 2H), 4.15 (s, 1H), 3.04 (t, 4H), 2.77-2.02 (m, 26H), 1.70-1.29 (m, 32H), 0.89 (m, 6H).

[0100] (8) Compound 10 (A3-B1-C4): (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br) was replaced by (9Z)-16-bromohexadecane-9-ene (C4-Br), 2-aminoethanol (HO-B1-NH2) was replaced by ethylenediamine (NH2-B1-NH2), and A1-OH was replaced by A3-OH to obtain the structural formula: Compound 10, its 1H NMR (400 MHz, DMSO-d6) δ 7.74 (s, 1H), 7.61 (d, 1H), 7.44 (d, 1H), 7.16 (t, 1H), 6.89 (t, 1H), 5.48 (m, 4H), 4.18 (s, 1H), 3.20 (t, 2H), 3.08 (t, 4H), 2.74-1.94 (m, 20H), 1.57-1.28 (m, 44H), 0.89 (m, 6H).

[0101] (9) Compound 11 (A3-B1-C5): 2-aminoethanol (HO-B1-NH2) was replaced by ethylenediamine (NH2-B1-NH2) and A1-OH was replaced by A3-OH to obtain the structural formula: Compound 11, its 1H NMR (400 MHz, DMSO-d6) δ 7.74 (s, 1H), 7.61 (d, 1H), 7.44 (d, 1H), 7.16 (t, 1H), 6.89 (t, 1H), 5.55-5.11 (m, 8H), 4.18 (s, 1H), 3.31 (t, 2H), 3.08 (t, 4H), 2.82 (m, 4H), 2.70-1.94 (m, 20H), 1.53-1.26 (m, 40H), 0.89 (m, 6H).

[0102] (10) Compound 13 (A4-B2-C7): 3-bromo-octyl propionate (C7-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), hexamethylenediamine (NH2-B2-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A4-OH was used to replace A1-OH to obtain the structural formula: Compound 13, its 1H NMR (400 MHz, DMSO-d6) δ 7.73 (s, 1H), 7.61 (d, 1H), 7.51 (d, 1H), 7.44 (t, 1H), 7.16 (t, 1H), 6.75 (t, 1H), 4.38 (t, 4H), 4.15 (t, 1H), 3.72 (t, 4H), 3.22 (t, 2H), 3.03 (t, 2H), 2.82-2.53 (m, 12H), 1.54-1.27 (m, 32H), 1.14 (t, 3H), 0.89 (m, 6H).

[0103] (11) Compound 14 (A5-B3-C8): 10-bromooctyl ester (C8-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 3,4-dithia-1,6-hexanediamine (HO-B3-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A5-OH was used to replace A1-OH to obtain the structural formula: Compound 14, its 1H NMR (400 MHz, DMSO-d6) δ 9.20 (s, 1H), 7.54 (s, 1H), 7.30 (d, 1H), 7.23 (d, 1H), 7.16 (t, 1H), 6.79 (t, 1H), 4.38 (t, 4H), 4.14 (s, 1H), 3.40 (t, 2H), 3.05 (t, 4H), 2.79-2.28 (m, 16H), 2.02 (m, 2H), 1.62-1.27 (m, 60H), 0.89 (m, 6H), 0.85 (m, 3H).

[0104] (12) Compound 15 (A5-B3-C9): Replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br) with N,N-dioctyl-2-bromo-ethylamine (C9-Br), replace 2-aminoethanol (HO-B1-NH2) with 3,4-dithia-1,6-hexanediamine (HO-B3-NH2), and replace A1-OH with A5-OH to obtain the structural formula: Compound 15, its 1H NMR (400 MHz, DMSO-d6) δ 9.20 (s, 1H), 7.54 (s, 1H), 7.30 (d, 1H), 7.23 (d, 1H), 7.16 (t, 1H), 6.79 (t, 1H), 3.90 (t, 2H), 3.40 (s, 1H), 3.05 (t, 8H), 2.79-2.30 (m, 20H), 2.02 (t, 2H), 1.77-1.26 (m, 56H), 0.89 (m, 12H), 0.85 (m, 3H).

[0105] (13) Compound 16 (A6-B4-C10): Replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br) with N,N-di(3-oxodecyl)-nonanamide (C10-Br), replace 2-aminoethanol (HO-B1-NH2) with 4,4-dimethyl-3,5-thioketal-1,7-heptanediamine (HO-B4-NH2), and replace A1-OH with A6-OH to obtain the structural formula The 1H NMR (400 MHz, DMSO-d6) of compound 16 was δ9.31 (s, 1H), 7.54 (s, 1H), 7.30 (d, 1H), 7.23 (d, 1H), 7.16 (t, 1H), 6.64 (t, 1H), 4.26 (t, 8H), 3.54 (m, 3H), 3.28 (m, 16H), 3.07 (t, 4H), 2.77-2.17 (m, 19H), 1.73-1.27 (m, 74H), 0.89 (m, 12H).

[0106] (14) Compound 17 (A6-B4-C11): N,N-di(3-nonyl acetate)-propionamide (C11-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 4,4-dimethyl-3,5-thioketal-1,7-heptanediamine (HO-B4-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A6-OH was used to replace A1-OH to obtain the structural formula: Compound 17, its 1H NMR (400 MHz, DMSO-d6) δ 9.31 (s, 1H), 7.54 (s, 1H), 7.30 (d, 1H), 7.23 (d, 1H), 7.16 (t, 1H), 6.64 (t, 1H), 4.56 (t, 8H), 3.54 (m, 14H), 2.84-2.17 (m, 28H), 1.73-1.26 (m, 66H), 0.89 (m, 12H).

[0107] (15) Compound 18 (A7-B1-C1): Replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br) with 12-bromododecane (C1-Br) and replace A1-OH with A7-OH to obtain the structural formula: The compound 18, its 1H NMR (400MHz, DMSO-d6) δ9.38 (s, 1H), 7.54 (d, 1H), 7.30 (d, 1H), 7.23 (t, 1H), 7.16 (t, 1H), 4.27 (t, 2H), 3.59 (d, 1H), 3.09 (m, 6H), 2.65-2.13 (m, 8H), 1.97 (m, 2H), 1.57-1.21 (m, 46H), 0.89 (m, 9H).

[0108] (16) Compound 19 (A7-B1-C2): Replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br) with 16-bromohexadecane (C2-Br) and replace A1-OH with A7-OH to obtain the structural formula: Compound 19, its 1H NMR (400 MHz, DMSO-d6) δ 9.38 (s, 1H), 7.54 (d, 1H), 7.30 (d, 1H), 7.23 (t, 1H), 7.16 (t, 1H), 4.27 (t, 2H), 3.59 (d, 1H), 3.09 (m, 6H), 2.65-2.13 (m, 8H), 1.97 (m, 2H), 1.57-1.21 (m, 62H), 0.89 (m, 9H).

[0109] (17) Compound 20 (A8-B2-C3): (6Z)-12-bromododecan-6-ene (C3-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 6-aminohexanol (HO-B2-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A8-OH was used to replace A1-OH to obtain the structural formula: Compound 20, its 1H NMR (400 MHz, DMSO-d6) δ 9.27 (s, 1H), 7.54 (d, 1H), 7.30 (d, 1H), 7.23 (t, 1H), 7.16 (t, 1H), 5.48 (m, 4H), 4.14 (t, 2H), 3.90 (s, 1H), 3.48 (t, 2H), 3.05 (t, 6H), 2.74-2.18 (m, 16H), 1.78-1.29 (m, 40H), 1.16 (s, 9H), 0.89 (m, 6H).

[0110] (18) Compound 21 (A8-B2-C4): (9Z)-16-bromohexadecane-9-ene (C4-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 6-aminohexanol (HO-B2-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A8-OH was used to replace A1-OH to obtain the structural formula: Compound 21, its 1H NMR (400 MHz, DMSO-d6) δ9.27 (s, 1H), 7.54 (d, 1H), 7.30 (d, 1H), 7.23 (t, 1H), 7.16 (t, 1H), 5.48 (m, 4H), 4.14 (t, 2H), 3.90 (s, 1H), 3.48 (t, 2H), 3.05 (t, 6H), 2.74-2.18 (m, 16H), 1.78-1.29 (m, 56H), 1.16 (s, 9H), 0.89 (m, 6H).

[0111] (19) Compound 22 (A8-B3-C5): (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 6-amino-3,4-dithia-hexanol (HO-B3-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A8-OH was used to replace A1-OH to obtain the structural formula: Compound 22, its 1H NMR (400 MHz, DMSO-d6) δ 9.27 (s, 1H), 7.54 (d, 1H), 7.30 (d, 1H), 7.23 (t, 1H), 7.16 (t, 1H), 5.48 (m, 4H), 4.14 (t, 2H), 3.90 (s, 1H), 3.48 (t, 8H), 3.05 (t, 6H), 2.74-2.18 (m, 16H), 1.78-1.29 (m, 50H), 1.16 (s, 9H), 0.89 (m, 6H).

[0112] (20) Compound 24 (A9-B4-C7): 3-bromo-octyl propionate (C7-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 4,4-dimethyl-3,5-thioketal-heptanol (HO-B4-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A9-OH was used to replace A1-OH to obtain the structural formula: Compound 24, its 1H NMR (400 MHz, DMSO-d6) δ 9.25 (s, 1H), 7.54 (d, 1H), 7.30 (d, 1H), 7.23 (t, 1H), 7.16 (t, 1H), 4.15 (t, 4H), 4.04 (t, 2H), 3.77 (m, 6H), 3.63 (m, 2H), 2.74-2.30 (m, 18H), 1.77-1.27 (m, 36H), 0.89 (m, 6H).

[0113] (21) Compound 25 (A9-B4-C8): 10-bromooctyl octyl ester (C8-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 4,4-dimethyl-3,5-thioketal-heptanol (HO-B4-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A9-OH was used to replace A1-OH to obtain the structural formula: Compound 25, its 1H NMR (400 MHz, DMSO-d6) δ9.25 (s, 1H), 7.54 (d, 1H), 7.30 (d, 1H), 7.23 (t, 1H), 7.16 (t, 1H), 4.15 (t, 4H), 4.04 (t, 2H), 3.77 (m, 6H), 3.63 (m, 2H), 2.74-2.30 (m, 18H), 1.77-1.27 (m, 64H), 0.89 (m, 6H).

[0114] (22) Compound 26 (A10-B1-C5): (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br) was substituted for (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), ethylenediamine (NH2-B1-NH2) was substituted for 2-aminoethanol (HO-B1-NH2), and A10-OH was substituted for A1-OH to obtain the structural formula: Compound 26, its 1H NMR (400 MHz, DMSO-d6) δ 8.42 (s, 1H), 7.54 (s, 1H), 7.30 (d, 1H), 7.23 (d, 1H), 7.16 (t, 1H), 6.71 (t, 1H), 5.51 (m, 8H), 4.55 (d, 1H), 4.03 (m, 2H), 3.32 (t, 2H), 2.91 (m, 4H), 2.88 (m, 6H), 2.57 (t, 2H), 2.32-1.69 (m, 17H), 1.36-1.26 (m, 36H), 0.89 (m, 6H).

[0115] (23) Compound 28 (A10-B2-C7): 3-bromo-octyl propionate (C7-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), hexamethylenediamine (NH2-B2-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A10-OH was used to replace A1-OH to obtain the structural formula: Compound 28, its 1H NMR (400 MHz, DMSO-d6) δ 8.42 (s, 1H), 7.54 (s, 1H), 7.30 (d, 1H), 7.23 (d, 1H), 7.16 (t, 1H), 6.71 (t, 1H), 4.55 (d, 1H), 4.16 (t, 4H), 4.03 (t, 4H), 3.14 (m, 2H), 2.94 (m, 4H), 2.88 (m, 2H), 2.58 (m, 4H), 2.32-1.94 (m, 7H), 1.69-1.27 (m, 34H), 0.89 (m, 6H).

[0116] (24) Compound 29 (A11-B2-C9): N,N-dioctyl-2-bromo-ethylamine (C9-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), hexamethylenediamine (NH2-B2-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A11-OH was used to replace A1-OH to obtain the structural formula: Compound 29, its 1H NMR (400 MHz, DMSO-d6) δ 9.70 (s, 1H), 8.03 (t, 2H), 7.58 (d, 1H), 7.39 (d, 1H), 7.12 (d, 2H), 3.71 (s, 1H), 3.11 (t, 24H), 2.45-2.33 (t, 20H), 2.28 (t, 2H), 2.16 (s, 3H), 1.99-1.87 (t, 8H), 1.53-1.26 (m, 112H), 0.92-0.86 (t, 27H).

[0117] (25) Compound 30 (A11-B3-C10): N,N-di(3-oxodecyl)-nonanamide (C10-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 3,4-dithia-1,6-hexanediamine (H2N-B3-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A11-OH was used to replace A1-OH to obtain the structural formula: Compound 30, its 1H NMR (400 MHz, DMSO-d6) δ 9.70 (s, 1H), 8.03 (t, 2H), 7.58 (d, 1H), 7.39 (d, 1H), 7.12 (d, 2H), 3.71 (s, 1H), 3.58 (t, 16H), 3.46 (q, 4H), 3.40 (t, 32H), 3.25 (t, 8H), 2.81-2.64 (t, 12H), 2.48-2.27 (t, 14H), 2.16 (s, 3H), 1.57-1.25 (m, 136H), 0.92-0.88 (t, 27H).

[0118] (26) Compound 31 (A11-B3-C11): N,N-di(3-nonyl acetate)-propionamide (C11-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 3,4-dithia-1,6-hexanediamine (H2N-B3-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A11-OH was used to replace A1-OH to obtain the structural formula: Compound 31, its 1H NMR (400 MHz, DMSO-d6) δ 9.70 (s, 1H), 8.03 (t, 2H), 7.58 (d, 1H), 7.39 (d, 1H), 7.12 (d, 2H), 4.25 (t, 16H), 3.96 (t, 8H), 3.71 (s, 1H), 3.54 (t, 20H), 2.84-2.28 (t, 40H), 2.16 (s, 3H), 1.63-1.56 (t, 24H), 1.40-1.26 (m, 96H), 0.92-0.88 (t, 27H).

[0119] (27) Compound 32 (A12-B3-C11): N,N-di(3-nonyl acetate)-propionamide (C11-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 3,4-dithia-1,6-hexanediamine (H2N-B3-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A12-OH was used to replace A1-OH to obtain the structural formula: Compound 32, its 1H NMR (400 MHz, DMSO-d6) δ9.70 (s, 1H), 8.03 (t, 2H), 7.52 (d, 1H), 7.39 (d, 1H), 7.10 (d, 2H), 4.25 (t, 16H), 3.71 (s, 1H), 3.54 (t, 8H), 3.48-3.25 (t, 22H), 2.83-2.64 (t, 8H), 2.44-2.29 (t, 32H), 2.16 (s, 3H), 1.98 (m, 16H), 1.63-1.56 (t, 8H), 1.39-1.26 (m, 96H), 1.20 (s, 9H), 0.92-0.88 (t, 24H).

[0120] (28) Compound 33 (A12-B4-C1): 12-bromododecane (C1-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 4,4-dimethyl-3,5-thioacetal-1,7-heptanediamine (H2N-B4-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A12-OH was used to replace A1-OH to obtain the structural formula: Compound 33, its 1H NMR (400 MHz, DMSO-d6) δ 9.70 (s, 1H), 8.03 (t, 2H), 7.56 (d, 2H), 7.12 (d, 2H), 3.96 (m, 4H), 3.71 (s, 1H), 3.33 (t, 2H), 3.01 (t, 8H), 2.78-2.51 (t, 12H), 2.31-2.00 (t, 9H), 1.54 (s, 12H), 1.51-1.41 (t, 8H), 1.34-1.27 (m, 80H), 1.20 (s, 9H), 0.88 (t, 12H).

[0121] (29) Compound 34 (A12-B4-C2): 16-bromohexadecane (C2-Br) was used to replace (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 4,4-dimethyl-3,5-thioketal-1,7-heptanediamine (H2N-B4-NH2) was used to replace 2-aminoethanol (HO-B1-NH2), and A12-OH was used to replace A1-OH to obtain the structural formula: Compound 34, its 1H NMR (400 MHz, DMSO-d6) δ 9.70 (s, 1H), 8.03 (t, 2H), 7.54 (d, 2H), 7.09 (d, 2H), 3.95 (m, 4H), 3.71 (s, 1H), 3.33 (t, 2H), 3.01 (t, 8H), 2.79-2.51 (t, 12H), 2.31-2.00 (t, 9H), 1.54 (s, 12H), 1.51-1.41 (t, 8H), 1.36-1.26 (m, 112H), 1.19 (s, 9H), 0.88 (t, 12H).

[0122] Example 4

[0123] According to the reaction steps in Example 2, the following compounds were prepared:

[0124] (1) Compound 12 (A4-B2-C6): Replace 6-aminohexanol (HO-B2-NH2) with hexamethylenediamine (NH2-B2-NH2) and replace A5-OH with A4-OH to obtain the structural formula: Compound 12, its 1H NMR (400 MHz, DMSO-d6) δ 7.73 (s, 1H), 7.61 (d, 1H), 7.51 (d, 1H), 7.44 (t, 1H), 7.16 (t, 1H), 6.75 (t, 1H), 5.37 (s, 2H), 4.38 (t, 1H), 3.45 (m, 2H), 3.22 (t, 2H), 2.68-2.41 (m, 14H), 1.51-1.26 (m, 52H), 1.14 (t, 3H), 0.89 (m, 6H).

[0125] (2) Compound 23 (A9-B3-C6): 6-amino-3,4-dithia-hexanol (HO-B3-NH2) was used to replace 6-aminohexanol (HO-B2-NH2), and A9-OH was used to replace A5-OH to obtain a compound of the formula: Compound 23, its 1H NMR (400 MHz, DMSO-d6) δ 9.27 (s, 1H), 7.54 (d, 1H), 7.30 (d, 1H), 7.23 (t, 1H), 7.16 (t, 1H), 4.31 (s, 2H), 4.04 (t, 2H), 3.84 (m, 2H), 3.63 (m, 4H), 2.85 (t, 2H), 2.71-2.30 (m, 16H), 1.74-1.23 (m, 50H), 0.89 (m, 6H).

[0126] (3) Compound 27 (A10-B1-C6): 6-aminohexanol (HO-B2-NH2) was replaced by 1-ethylenediamine (NH2-B1-NH2), and A5-OH was replaced by A10-OH to obtain the structural formula: Compound 27, its 1H NMR (400 MHz, DMSO-d6) δ 8.42 (s, 1H), 7.54 (s, 1H), 7.30 (d, 1H), 7.23 (d, 1H), 7.16 (t, 1H), 6.71 (t, 1H), 5.37 (s, 2H), 4.55 (d, 1H), 3.84 (m, 6H), 3.34 (t, 2H), 2.82-1.69 (m, 15H), 1.38-1.23 (m, 44H), 0.89 (m, 6H).

[0127] Example 5

[0128] Compound 1 (A1-B1-C5) was prepared into lipid nanoparticles (LNPs), and then its relevant physicochemical properties and in vitro and in vivo activities were tested, as follows:

[0129] (1) First, lipid nanoparticles (LNPs) were prepared according to different formulations in Table 1. The specific preparation method was as follows: ionizable lipids, auxiliary lipids (including any one of DOPE, DOPC or DSPC), cholesterol and PEG-DMG were dissolved in an organic solvent according to corresponding molar ratios and mixed, and the mixture was mixed with 6.25 mM sodium acetate buffer (pH = 5) containing siRNA (CUUACG CUGAGUACUUCGATT) at a ratio of v / v = 3:1 (water: organic solvent) using a microfluidic device, and the mixture was transferred to a dialysis bag and dialyzed with PBS buffer solution (pH = 7.4) overnight, and then concentrated using an ultracentrifugal filter, and A1-B1-C5 was made into ionizable lipids, which were combined with auxiliary lipids, cholesterol and PEG-DMG to form different lipid nanoparticles (LNPs) based on A1-B1-C5 (respectively A1-B1-C5-1 to A1-B1-C5-27), and their particle sizes and PDI (polymer dispersibility index) are shown in Table 2. As can be seen from Table 2, the LNP particle size prepared under the formulation No. 14 is <100 nm, and the dispersion is good. It has brain targeting potential in terms of particle size and can be used for subsequent experiments.

[0130] Compounds 1 to 34 prepared in the above examples were prepared according to the formulation No. 14 in Table 1 to obtain lipid nanoparticles (LNPs) formed by different ionizable lipids in the same formulation, and the particle size and PDI (polymer dispersibility index) are shown in Table 3. As can be seen from Table 3, the LNP particle size prepared by some vincamine compounds is <100 nm, and the dispersion is good, which has brain targeting potential in terms of particle size, and can be used for subsequent experiments.

[0131] Table 1 Lipid nanoparticles (LNP) with different molar ratios

[0132]

[0133] Table 2 Particle size and PDI of different lipid nanoparticles (LNP)

[0134]

[0135]

[0136] Table 3 Particle size and PDI of lipid nanoparticles (LNPs) formed by different ionizable lipids in the same formulation

[0137] Preparation No. Size(nm) PDI Compound 1(A1-B1-C5)-14 92.6 0.225 Compound 2(A5-B2-C6)-14 89.3 0.213 Compound 3(A1-B1-C2)-14 114.6 0.243 Compound 4(A1-B1-C3)-14 171.3 0.136 Compound 5(A1-B2-C4)-14 164.2 0.228 Compound 6(A2-B2-C8)-14 198.5 0.189 Compound 7(A2-B3-C9)-14 >1um - Compound 8(A2-B3-C10)-14 90.3 0.115 Compound 9(A3-B4-C3)-14 273.6 0.212 Compound 10(A3-B1-C4)-14 165.4 0.315 Compound 11(A3-B1-C5)-14 >1um - Compound 12(A4-B2-C6)-14 167.7 0.305 Compound 13(A4-B2-C7)-14 112.9 0.214 Compound 14 (A5-B3-C8)-14 100.3 0.127 Compound 15(A5-B3-C9)-14 126.9 0.411 Compound 16(A6-B4-C10)-14 >1um - Compound 17(A6-B4-C11)-14 93.2 0.165 Compound 18(A7-B1-C1)-14 115.9 0.247 Compound 19(A7-B1-C2)-14 108.4 0.175 Compound 20(A8-B2-C3)-14 91.3 0.198 Compound 21(A8-B2-C4)-14 88.5 0.166 Compound 22(A8-B3-C5)-14 >1um - Compound 23(A9-B3-C6)-14 86.6 0.201 Compound 24(A9-B4-C7)-14 105.6 0.141 Compound 25(A9-B4-C8)-14 143.6 0.315 Compound 26(A10-B1-C5)-14 150.4 0.251 Compound 27(A10-B1-C6)-14 >1um - Compound 28(A10-B2-C7)-14 116.1 0.196 Compound 29(A11-B2-C9)-14 104.7 0.170 Compound 30(A11-B3-C10)-14 98.3 0.157 Compound 31(A11-B3-C11)-14 110.3 0.249 Compound 32(A12-B3-C11)-14 137.6 0.334 Compound 33(A12-B4-C1)-14 179.3 0.225 Compound 34(A12-B4-C2)-14 158.4 0.114

[0138] (2) The lipid nanoparticles (LNPs) prepared by using compound 2 (A5-B2-C6) as the ionizable lipid according to the formulation 14 in Table 1 were subjected to transmission electron microscopy scanning. The results are as follows: Figure 2 As shown. Figure 2 It can be seen that the LNP particle size is about 100 nm.

[0139] (3) Lipid nanoparticles (A1-B1-C5-14) prepared by using A1-B1-C5 as ionizable lipids and according to the recipe of number 14 in Table 1, lipid nanoparticles (A5-B2-C6-14) prepared by using A5-B2-C6 as ionizable lipids and according to the recipe of number 14 in Table 1, lipid nanoparticles (A2-B3-C10-14) prepared by using A2-B3-C10 as ionizable lipids and according to the recipe of number 14 in Table 1, lipid nanoparticles (A5-B3-C8-14) prepared by using A5-B3-C8 as ionizable lipids and according to the recipe of number 14 in Table 1, and lipid nanoparticles (A5-B3-C8-14) prepared by using A6-B4-C11 as ionizable lipids and according to the recipe of number 14 in Table 1. The lipid nanoparticles (A6-B4-C11-14) were prepared using the formula of A8-B2-C3 as the ionizable lipid and the formula of label 14 in Table 1 (A6-B4-C11-14), the lipid nanoparticles (A8-B2-C3-14) were prepared using A8-B2-C3 as the ionizable lipid and the formula of label 14 in Table 1 (A8-B2-C4-14), the lipid nanoparticles (A9-B3-C6-14) were prepared using A9-B3-C6 as the ionizable lipid and the formula of label 14 in Table 1 (A9-B3-C6-14), and the lipid nanoparticles (A11-B3-C10-14) were prepared using A11-B3-C10 as the ionizable lipid and the formula of label 14 in Table 1 (A11-B3-C10-14). The pka of the lipid nanoparticles prepared by the vincamine derivatives was determined by TNS fluorescence method (specific method: dilute the prepared lipid nanoparticles to 2 mM in PBS buffer solution, prepare a buffer solution containing 130 mM NaCl, 10 mM ammonium acetate, 10 mM Hepes and 10 mM MES, dilute the lipid nanoparticles to 100 μM, divide the buffer solution into 8 groups, and adjust the pH value of each group of buffer solution to 3, 4, 5, 6, 7, 8, 9, 10 using NaOH or HCl solution; dissolve the anionic fluorescent dye 2-(p-toluidino)-6-naphthalenesulfonic acid (TNS) in distilled water to form a solution with a concentration of 100 μM, and adjust the final concentration of the lipid nanoparticles to 1 uM solution. At room temperature, with 321 nm as the excitation wavelength and 445 nm as the emission wavelength, the fluorescence intensity of each group was determined using a multifunctional microplate reader, wherein pKa is defined as the pH value rising to half of the maximum fluorescence intensity). The final results are as follows: Figure 3 As shown. Figure 3 It can be seen that the pka value of LNP prepared from the vincamine derivatives of the present invention is 6.2-6.5, which is conducive to the efficient encapsulation and lysosomal escape of nucleic acids.

[0140] (4) The encapsulation of siRNA by lipid nanoparticles formed by different vincamine derivatives in Table 2 was tested, as shown below:

[0141] The quantitative Quant-iT RiboGreen RNAAssay kit was used to detect the concentration of siRNA in the lipid nanoparticles formed by different vincamine derivatives in Table 2, and the method was as follows: the dialyzed lipid nanoparticles were diluted into the Tris-EDTA (TE) buffer in the 96-well plate, and the dialyzed lipid nanoparticles were diluted into the TE buffer containing 2% Triton X-100 for 30 minutes, and then the RiboGreen working solution was added to the corresponding wells, and the fluorescence intensity at an excitation wavelength of 485nm and an emission wavelength of 530nm was detected by an ELISA instrument to reflect the amount of free siRNA and dialyzed siRNA; the siRNA concentration was calculated using the siRNA standard curve and then the encapsulation rate was calculated, and the results are shown in Table 4. As can be seen from Table 4, the encapsulation rate of nucleic acid prepared by the vincamine derivatives of the present invention is good, higher than 90%.

[0142] Table 4 Encapsulation efficiency of siRNA by lipid nanoparticles prepared from different vincamine derivatives

[0143] Preparation No. Encapsulation rate (%) Compound 1(A1-B1-C5)-14 90.1 Compound 2(A5-B2-C6)-14 87.6 Compound 6(A2-B2-C8)-14 75.3 Compound 9(A3-B4-C3)-14 62.4 Compound 12(A4-B2-C6)-14 83.8 Compound 16(A6-B4-C10)-14 91.4 Compound 19(A7-B1-C2)-14 78.6 Compound 22(A8-B3-C5)-14 83.7 Compound 24(A9-B4-C7)-14 93.5 Compound 26(A10-B1-C5)-14 80..4 Compound 30(A11-B3-C10)-14 95.9 Compound 32(A12-B3-C11)-14 62.1

[0144] (5) Fluorescence microscopy of the in vitro transfection effect of siRNA loaded on lipid nanoparticles prepared from vincamine derivatives:

[0145] bEnd.3 cells were seeded in 6-well plates (confluence of 30%) and cultured in 1 mL of complete medium without dual antibodies for 12 hours. The medium was then replaced with serum-free medium without dual antibodies, and lipid nanoparticles loaded with siRNA with FAM fluorescent labels (compound 3 (A1-B1-C2), compound 8 (A2-B3-C10), compound 10 (A3-B1-C4), compound 13 (A4-B2-C7), compound 14 (A5-B3-C8), compound 17 (A6-B4-C10) were added to the cells. 1), compound 19 (A7-B1-C2), compound 20 (A8-B2-C3), compound 23 (A9-B3-C6), compound 28 (A10-B2-C7), compound 31 (A11-B3-C11), compound 33 (A12-B4-C1) and Lipo2000 (control group) as examples), incubated at 37°C, 5% CO2 for 6 hours, replaced the culture medium with complete culture medium without double antibody, and continued to culture for 18 hours. The FAM fluorescence intensity of different dosing groups was observed under a fluorescence microscope. The results are as follows Figure 4 As shown. Figure 4It can be seen that compared with the gold standard cell transfection reagent lipo2000, LNPs made from some vincamine compounds can better carry siRNA for transfection into cells.

[0146] (6) MTT assay of the cytotoxicity of lipid nanoparticles formed by different vincamine derivatives in Table 2:

[0147] bEnd.3 cells were seeded in 96-well plates (6000 cells per well) and cultured in 100 μL of culture medium for 12 h. Afterwards, lipid nanoparticles formed by different vincamine derivatives in Table 2 (compound 3 (A1-B1-C2), compound 8 (A2-B3-C10), compound 10 (A3-B1-C4), compound 13 (A4-B2-C7), compound 14 (A5-B3-C8), compound 17 (A6-B4-C11), compound 19 (A7-B1-C4), compound 20 (A8-B2-C7), compound 21 (A9-B3-C8), compound 22 (A10-B3-C9), compound 23 (A11-B4-C10), compound 24 (A12-B4-C11), compound 25 (A13-B1-C12), compound 26 (A14-B2-C13), compound 27 (A15-B3-C14), compound 28 (A16-B4-C15), compound 29 (A17-B1-C16), compound 30 (A18-B1 C2), compound 20 (A8-B2-C3), compound 23 (A9-B3-C6), compound 28 (A10-B2-C7), compound 31 (A11-B3-C11) and compound 33 (A12-B4-C1)), incubated at 37°C, 5% CO2 for 48h; at the end of the experiment, 3-(4,5)-dimethylthiazolylazo (-z-y1)-3,5-diphenyltetrazolium urea (MTT) solution (5mg / ml, 20μL) was added and further cultured at 37°C for 4h. The cell viability was calculated by measuring the absorbance of each well at 570nm using an ELISA reader. The results are shown in Figure 5 As shown. Figure 5 It can be seen that the LNP prepared from the vincamine derivatives of the present invention has low cytotoxicity and high biocompatibility.

[0148] (7) Laser speckle imaging was used to measure the effects of lipid nanoparticles formed by different vincamine derivatives in Table 2 on the cerebral microvascular blood flow in mice:

[0149] After isoflurane anesthesia, the mice were fixed on a brain stereoscope, the skull skin was cut along the midline, and the mouse brain microvascular blood flow was detected using a laser speckle imager. After the mice were stable for 30 minutes, 200uL of lipid nanoparticles formed by different vincamine derivatives in Table 2 were administered through the tail vein at a dose of 30mg / kg. The laser speckle blood flow images were recorded within 60 minutes after administration to identify the brain microvascular blood perfusion (ROIs). The results are shown in Figure 6 As shown. Figure 6 It can be seen that the LNPs prepared from some vincamine compounds have the effect of enhancing the blood flow in the brain microvessels of mice.

[0150] (8) Taking the lipid nanoparticles encapsulating siRNA formed by vincamine derivatives (A1-B1-C5) in Table 2 as an example, the N / P ratio was determined by gel retardation experiment:

[0151] Dissolve siRNA (1OD) in 125μL DEPC water, mix lipid nanoparticles (LNP) formed by vincamine derivative compound 1 (A1-B1-C5) in Table 2 and siRNA solution at siRNA / LNP mass ratios of 5:1, 1:1, 1:2, 10:1, 1:5, 1:10, 1:15, and 1:20, respectively, and incubate the mixture at room temperature for 30min; examine the siRNA binding ability of LNP by agarose gel electrophoresis; mix with 6× loading buffer, add to 2% agarose gel containing 0.01% gel stain Glodview after mixing; run electrophoresis in 1×TAE buffer, voltage 90V, electrophoresis for 30min. Record the results at 320nm UV wavelength using a gel image analysis system (Tanon, China), as shown in the figure. Figure 7 As shown. Figure 7 It can be seen that the LNP prepared from the vincamine derivatives of the present invention can completely encapsulate siRNA when the siRNA / LNP mass ratio is greater than 1:10.

[0152] (9) Confocal microscopy was used to photograph the endosomal escape effect of siRNA loaded on lipid nanoparticles formed by different vincamine derivatives in Table 2 (compound 1 (A1-B1-C5), compound 2 (A5-B2-C6) and compound 31 (A11-B3-C11) as examples):

[0153] bEnd.3 cells were seeded in 12-well plates (20,000 cells per well), cultured in 500 uL of culture medium for 12 h, then the culture medium in the wells was discarded and replaced with new complete culture medium. LNP@FAM-siRNA (lipid nanoparticles formed by different vincamine derivatives in Table 2) was added to the cells and incubated in the dark at 37°C and 5% CO2 for 2, 4, 6, and 8 h. After the uptake was completed, the LNP (lipid nanoparticles formed by different vincamine derivatives in Table 2 (compound 1 (A1-B1-C5), compound 2 (A5-B2-C6) and compound 31 (A11-B3-C11) as examples)) medium, washed 3 times with PBS buffer solution, incubated with LysotrackerRed (1uM) at 37°C in the dark for 2h, discarded the supernatant after the incubation, added PBS buffer solution to rinse 3 times, fixed the cells with 4% paraformaldehyde fixative at room temperature for 30min, discarded the fixative and added PBS buffer solution to rinse 3 times, added Hoechst 33342 cell nuclear stain solution to incubate in the dark for 10min, washed 3 times with PBS buffer solution, and then used a high-content imaging system to perform fluorescence co-localization photography in confocal mode. The results are shown in FIG. Figure 8 As shown. Figure 8 It can be seen that the LNP prepared from the vincamine derivatives of the present invention can carry siRNA and efficiently escape from lysosomes.

[0154] (10) The single cell analyzer was used to detect the effect of lipid nanoparticles formed by vincamine derivatives (compound 2 (A5-B2-C6) as an example) on cellular ROS levels:

[0155] In order to detect the oxidative stress / intracellular reactive oxygen species (ROS) level of cells, the DCFH diacetate reactive oxygen species analysis kit was used to detect the green fluorescence signal of ROS: bEnd.3 cells were inoculated in cell culture dishes (50,000 cells per dish) and cultured in 1 mL of culture medium for 12 hours. The culture medium in the wells was discarded and replaced with new complete culture medium. Lipid nanoparticles formed by vincamine derivatives (compound 2 (A5-B2-C6)) were added to the cells and incubated in the dark at 37°C and 5% CO2 for 2 hours; the cells of each group were incubated in a PBS buffer solution containing 10 μM DCHF-DA at 37°C for 30 minutes; then, the cells were washed three times in a PBS buffer solution containing 0.1% BSA and then fixed on a glass slide. The ROS level was detected in a single cell using an optical fiber probe with a tip of about 10 μm. The tip was inserted into the cells fixed in PBS, and the photon count was continuously detected and calculated within 150 seconds. The results are shown in Figure 2. Fig. 9 As shown. Fig. 9It can be seen that compared with the positive control of the kit and the commercially available cationic lipid DOTAP and ionizable lipid Dlin-MC3-DMA (MC3), which induce cells to produce high ROS levels and lead to cytotoxicity, the LNPs prepared from vincamine compounds have low cytotoxicity and high biocompatibility.

[0156] Example 6

[0157] Preparation of a vincamine derivative (compound 35 (A1-C12)), specifically comprising the following steps:

[0158] (1) NH2-PEG 2000 -DSPE (60 mg, 0.02 mmol) (mass spectrum as shown Fig.10 As shown in a), A1 (13 mg, 0.04 mmol), benzotriazol-1-yl-oxytripyrrolidinophosphine hexafluorophosphate (PyBop, 31.2 mg, 0.06 mmol) were dissolved in 3 mL of N,N-dimethylformamide, and the reaction mixture was stirred at room temperature for 24 h;

[0159] (2) The reaction solution was placed in a dialysis bag and dialyzed with pure water for 24 hours. The aqueous solution in the dialysis bag was taken out and freeze-dried to obtain a vincamine derivative with the structural formula A1-PEG2000-DSPE, namely compound 35 (A1-C12) (its mass spectrum is shown in Fig.10 (as shown in b).

[0160] Example 7

[0161] 1. Preparation of a vincamine derivative (compound 36 (A1-C13)), comprising the following steps:

[0162] (1) Compound C13-H2 (doxorubicin, DOX-NH2) (20 mg, 0.037 mmol) and compound A1-OH (16.8 mg, 0.052 mmol) were dissolved in 2 mL of N,N-dimethylformamide and stirred to dissolve. Then, benzotriazole-1-yl-oxytripyrrolidinophosphine hexafluorophosphate (27 mg, 0.052 mmol) was added and the pH was adjusted to 7-8 with N,N-diisopropylethylamine. The mixture was reacted at room temperature for 1 h. After removing N,N-dimethylformamide by rotary evaporation, the mixture was dissolved in ethyl acetate (20 mL), washed with 0.10 M hydrochloric acid solution, and then washed with saturated sodium chloride solution until neutral. After drying over anhydrous sodium sulfate, the mixture was filtered under reduced pressure, the organic phase was removed by rotary evaporation, and further purified by washing with methyl tert-butyl ether to obtain a vincamine derivative with the structural formula A1-DOX, namely compound 36 (A1-C13) (1H NMR (400MHz, DMSO-d6) δ12.04 (s, 2H), 8.32 (s, 1H), 7.90-7.40 (m, 6H), 6.90 (t, 1H), 6.70 (d, 1H), 5.37 (s, 1H), 4.90 (d, 2H), 4.70-4.65 (m, 4H), 4.22 (t, 1H), 3.90 (s, 3H), 3.65 (m, 3H), 3.37 (d, 2H), 2.65-1.42 (m, 16H), 1.11 (d, 3H), 0.99 (m, 3H, mass spectrum as shown Fig.11 shown).

[0163] 2. Preparation of a vincamine derivative (compound 37 (A1-B5-C14)), comprising the following steps:

[0164] (1) Compound A1-OH (60 mg, 0.17 mmol) and compound H2N-B1-NH2 (44.7 mg, 0.27 mmol) were dissolved in 2 mL of N,N-dimethylformamide and stirred to dissolve. Then, benzotriazole-1-yl-oxytripyrrolidinophosphine hexafluorophosphate (147 mg, 0.28 mmol) was added and reacted at room temperature overnight. After vacuum concentration, the mixture was dissolved in dichloromethane (5 mL). The mixture was extracted with 10% anhydrous citric acid and saturated brine in sequence. After drying with anhydrous sodium sulfate, the organic phase was concentrated under reduced pressure to remove the organic phase. The organic phase was dissolved in a small amount of methanol (0.5 mL) and washed three times with 15 mL of methyl tert-butyl ether. After dissolving in dichloromethane (1.6 mL), trifluoroacetic acid (0.4 mL) was added and reacted at room temperature with stirring for 1 h. After precipitation with 30 mL of methyl tert-butyl ether, the organic phase was dried to obtain A1-B5-NH2.

[0165] (2) Paclitaxel PTX (25.1 mg, 0.029 mmol) and succinic anhydride (37.5 mg, 0.37 mmol) were added to 0.6 mL of pyridine and stirred at room temperature for 3 hours. The solvent was removed in vacuo, and the residue was washed with 1 mL of water and dried. The precipitate was then dissolved in acetone, and ice water was added dropwise to the solution to induce crystallization. PTX-COOH was obtained after drying;

[0166] (3) PTX-COOH (30 mg, 0.031 mmol) and A1-B5-NH2 (16.8 mg, 0.052 mmol) were dissolved in 2 mL of N,N-dimethylformamide and stirred to dissolve. Then, benzotriazole-1-yl-oxytripyrrolidinophosphine hexafluorophosphate (24.6 mg, 0.047 mmol) was added, and the pH was adjusted to 7-8 with N,N-diisopropylethylamine. The mixture was reacted at room temperature for 2 h. After concentrating under reduced pressure to remove N,N-dimethylformamide, the mixture was dissolved in dichloromethane (5 mL), extracted three times with 10% anhydrous citric acid (4 mL) and saturated brine in sequence, dried with anhydrous sodium sulfate, and rotary evaporated to obtain a vincamine derivative with the structural formula A1-B5-PTX, namely compound 37 (A1-B1-C14) (1H NMR (400MHz, DMSO-d6) δ8.87 (s, 1H), 8.41 (s, 1H), 8.05-7.89 (m, 5H), 7.68-7.27 (m, 15H), 6.77 (s, 1H), 6.70 (s, 1H), 6.50 (d, 1H), 6.21 (s, 1H), 6.10 (d, 1H), 5.13-5.03 (m, 4H), 4.27 (d, 1H), 3.73-3.66 (m, 4H), 3.55 (s, 1H), 3.42 (m, 2H), 2.68-2.41 (m, 11H), 2.19 (s, 6H), 2.11-1.25 (m, 16H), 1.01 (s, 6H), 0.89 (s, 3H), mass spectrum as shown Fig.12 shown).

[0167] Example 8

[0168] Preparation of a vincamine derivative (compound 38 (A1-B5-C15)), specifically comprising the following steps:

[0169] (1) Compound A1-OH (50 mg, 0.15 mmol), cystamine dihydrochloride (52 mg, 0.23 mmol), and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBop, 120 mg, 0.23 mmol) were dissolved in 6 mL of N,N-dimethylformamide / methanol (5 / 1, v / v), and 50 μL of triethylamine was added dropwise. The reaction mixture was stirred at room temperature for 2 h, and the reaction mixture was added dropwise into 30 mL of methyl tert-butyl ether (a large amount of yellow semi-solid precipitate was produced). The precipitate was washed twice with methyl tert-butyl ether and dried under vacuum to obtain a crude product; the obtained crude product was dissolved in 3 mL of N,N-dimethylformamide, tri(2-carbonylethyl)phosphonium hydrochloride (50 mg, 0.17 mmol) was dissolved in 200 μL of water and added dropwise into the solution of compound 1, the reaction mixture was stirred at room temperature for 2 h, concentrated under reduced pressure, precipitated with water, and the precipitate was dried to obtain an intermediate with a structure of A1-NH-(CH2)2-SH;

[0170] (2) The intermediate with the structure A1-NH-(CH2)2-SH (7.8 mg, 0.02 mmol) and NHS-PEG8-Mal (6.9 mg, 0.01 mmol) were dissolved in 100 μL N,N-dimethylformamide and stirred at room temperature for 1 h. The reaction mixture was added dropwise into a PBS solution of BSA (68 mg, 0.001 mmol) and stirred at room temperature for 3 h. The reaction solution was placed in a 8000-14000 Da dialysis bag and dialyzed with pure water for 24 h. Finally, the aqueous solution in the dialysis bag was taken out and freeze-dried to obtain a vincamine derivative compound 38 with the structural formula A1-B5-C15.

[0171] (3) TLC verification of the product (such as Fig.13 As shown in the figure, chloroform and methanol in a volume ratio of 3:1 were used as the developing solvent, and ultraviolet and fluorescent ammonia were used for color development. Compared with the BSA point, the compound 38 (A1-B5-C15) point showed ultraviolet signal, indicating that the coupling was successful.

[0172] (4) Use OPA to determine the free amino group content of BSA and A1-B9-BSA. Dissolve 40 mg OPA in 1 mL methanol, then add 25 mL 0.1 M sodium borate, 200 mg DMA and 5 mL 10% SDS, and add water to make up to 50 mL to obtain OPA reagent. Mix 65 μL of sample (1% protein) with 3 mL OPA reagent solution. After 2 minutes, use an ultraviolet spectrophotometer to measure the absorbance at 340 nm to measure the alkyl isoindole derivatives formed after the reaction of OPA with free amino groups. The standard curve (such as Fig.14The free amino group content of the protein sample was calculated according to the standard curve (as shown in Table 5), and the coupling rate of the intermediate with the structure of A1-NH-(CH2)2-SH and BSA was estimated. Compared with BSA, the free amino group of A1-B5-C15 was reduced by about 10, indicating that on average each BSA was coupled with about 11 small molecules.

[0173] Table 5A1-B5-BSA free amino group number

[0174]

[0175] Example 9

[0176] In vivo imaging was used to capture the in vivo brain targeting of vincamine derivatives (compound 31 (A11-B3-C11), compound 35 (A1-C12), compound 36 (A1-C13), compound 37 (A1-BA-C14), and compound 38 (A1-B5-C15) as examples):

[0177] Preparations loaded with DiD or compounds labeled with Cy5 (compound 31 (A11-B3-C11), compound 35 (A1-C12), compound 36 (A1-C13), compound 37 (A1-BA-C14), and compound 38 (A1-B5-C15)) were injected into mice via the tail vein, and fluorescence images were collected at predetermined time points (0.5 h, 1 h, and 2 h) using the VISQUE in vivo Smart-LF System. The results are shown in Figure 2. Fig.15 As shown. Fig.15 It can be seen that compared with the control group (Control), the vincamine derivatives of the present invention have brain targeting properties.

[0178] In summary, the present invention discloses vincamine derivatives and a preparation method thereof, and the vincamine derivatives have the following advantages: (i) the modification of the tail chain increases the lipid solubility of vincamine compounds, and does not affect the cerebral blood flow regulation of vincamine itself, which helps the carried drug to penetrate the blood-brain barrier, and exerts a brain protective effect, and improves brain microcirculation disorders; (ii) the tertiary amine group in the parent nucleus structure of vincamine derivatives has the property of being ionizable under acidic conditions, and can achieve efficient carrying and lysosomal escape of nucleic acid drugs through charge adsorption, thereby improving intracellular transport; (iii) vincamine derivatives inherit various inherent pharmacological activities of vincamine and have high safety. Therefore, the vincamine derivatives disclosed in the present invention have good application prospects in brain-targeted delivery of drugs for treating brain diseases.

[0179] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solution, which should be included in the scope of the claims of the present invention.

Claims

1. A vincamine derivative, characterized in that: The structural formula of the vincamine derivatives is , Wherein A is any one of the structural formulas A1 to A10: A1 A2 A3 A4 A5 A6 A7 A8 A9 A10 B is any one of B1 to B4, wherein B1 is -(CH2)2-, B2 is -(CH2)6-, B3 is -(CH2)2-SS-(CH2)2-, and B4 is -(CH2)2-SC(CH3)2-S-(CH2)2-; The C is a straight-chain alkyl group having 10 to 18 carbon atoms, a straight-chain alkenyl group having 10 to 18 carbon atoms, a straight-chain alkyl group having 10 to 18 carbon atoms substituted with a hydroxyl group, or any one of the structural formulas C7 to C11: C7 C8 C9 C10 C11.

2. The vincamine derivative according to claim 1, characterized in that: The C is any one of the structural formulas C1 to C6: C1 C2 C3 C4 C5 C6.

3. The vincamine derivative according to claim 1, characterized in that: The vincamine derivatives include any one of Compound 1 to Compound 9 and Compound 18 to Compound 25 with the following structural formula: Compound 1 Compound 2 Compound 3 Compound 4 Compound 5 Compound 6 Compound 7 Compound 8 Compound 9 Compound 18 Compound 19 Compound 20 Compound 21 Compound 22 Compound 23 Compound 24 Compound 25.

4. The method for preparing the vincamine derivatives according to any one of claims 1 to 3, characterized in that: The structural formula of the vincamine derivatives is When, the preparation method comprises the following steps: (1) A compound with the structural formula C-Br, a compound with the structural formula HO-B-NH2, potassium carbonate and potassium iodide were dissolved in an organic solvent at a molar ratio of 1.21:0.552:2.43:0.552, heated to 45-65°C for reaction for 12-48 hours, the product obtained by the reaction was cooled and filtered, the filtrate was extracted with n-hexane and separated by silica gel chromatography to obtain a compound with the structural formula The intermediate compound Alternatively, a linear alkane having 10 to 18 carbon atoms substituted by an epoxy group and a compound having a structural formula of HO-B-NH2 are dissolved in an organic solvent at a molar ratio of 1.26:0.57, and the mixture is heated to 55 to 75°C for reaction for 12 to 48 hours. The product obtained by the reaction is concentrated under reduced pressure and separated by silica gel chromatography to obtain a compound having a structural formula of The intermediate compound (2) Compounds with the structural formula A-OH and The intermediate compound, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride and 4-dimethylaminopyridine are dissolved in an organic solvent at a molar ratio of 0.15:1.59:0.183:0.0081, heated to 20-40°C for reaction for 24-96 hours, the product obtained by the reaction is diluted, extracted and dried, concentrated under reduced pressure and separated by silica gel chromatography to obtain a product with the structural formula Vincamine derivatives.

5. The preparation method according to claim 4, characterized in that: The eluent used in the silica gel chromatography separation in step (1) is a mixed solution composed of methanol and dichloromethane in a volume ratio of 10:90 to 90:10, and the eluent used in the silica gel chromatography separation in step (2) is a mixed solution formed by mixing acetone and n-hexane in a volume ratio of 20:80 to 80:

20.

6. The preparation method according to claim 5, characterized in that: The organic solvent is any one of acetonitrile, ethanol, methanol, N,N-dimethylformamide, N,N-diisopropylethylamine, ethyl acetate, methyl tert-butyl ether, triethylamine, pyridine or dichloromethane.

7. Use of the vincamine derivatives according to any one of claims 1 to 3 in the preparation of drugs for brain-targeted delivery for the treatment of brain diseases.

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