Vincamine derivatives, preparation method therefor and use thereof

Abstract

The present invention relates to vincamine derivatives, a preparation method therefor and a use thereof. The disclosed vincamine derivatives have the following advantages: (i) modification of the tail chain increases the lipid solubility of a vincamine compound without affecting the cerebral blood flow regulation of vincamine itself, which helps a carried drug to penetrate the blood-brain barrier and exert a brain protective effect, improving brain microcirculation disorders; (ii) the tertiary amine group in the parent nucleus structure of the vincamine derivatives has the property of being ionizable under acidic conditions, which can achieve efficient carrying and lysosomal escape of nucleic acid drugs by means of charge adsorption, thereby improving intracellular transport; and (iii) the vincamine derivatives inherit the various pharmacological activities inherent in vincamine and have a high degree of safety. The disclosed vincamine derivatives have good application prospects in brain-targeted delivery of drugs for treating brain diseases.

Description

Vincamine derivatives and their preparation methods and applications Technical Field

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

[0002] The treatment of central nervous system (CNS) diseases, such as neurodegenerative diseases, brain tumors, brain infections, and stroke, is severely limited by the blood-brain barrier (BBB), which prevents most small-molecule drugs and macromolecules (such as peptides, nucleic acid drugs, and protein drugs) from entering the brain. Extensive efforts have been made to improve the efficiency of drug delivery to the brain, including direct CNS administration, BBB disruption, and receptor-mediated delivery methods. However, direct CNS administration is invasive and can cause infection and tissue damage. It is also limited by diffusion distances and the rapid efflux of drugs from the CNS within hours. Techniques such as osmotic disruption, biochemical disruption, and ultrasound-mediated BBB disruption can effectively introduce drugs into the brain; however, these transient BBB 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 BBB, particularly gene and nucleic acid therapeutics, to the CNS via intravenous injection remain to be improved. Currently, lipid nanoparticles (LNPs) composed mainly of ionizable lipids are mainly 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] Furthermore, given the steric hindrance issues during synthesis and preparation, most ionizable lipid molecules are designed to achieve pH sensitivity and endosomal escape by introducing simple linear tertiary amine groups, rather than cyclic tertiary amine head groups. Only a few reports have demonstrated that a single cyclic tertiary amine head group can also exhibit favorable binding interactions with nucleic acids, such as the piperazine-containing C12-200, which can simultaneously carry five liver-targeted siRNAs into cells. Furthermore, aside from promoting RNA encapsulation and cellular uptake, synthetic lipids do not always possess ideal bioactivity. Indeed, potential cytotoxicity and immunogenicity are major drawbacks associated with the use of cationic lipid materials. These materials can activate innate immunity through the complement system and Toll-like receptors, and may also produce proinflammatory cytokines and reactive oxygen species. This is particularly true when long-term, repeated administration is required, necessitating the pre-administration of glucocorticoids and antihistamines in practical LNP applications.

[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 possess vast scaffold diversity and multiple pharmacophores, making them promising lead compounds that dominate drug development. In recent years, some small molecule natural products have also advanced drug delivery from new perspectives. Alkaloids, a class of nitrogenous alkaline organic compounds with significant biological activity, are among the earliest studied natural products. A large number of alkaloid-based drugs have been discovered and approved by the FDA. Representative indole alkaloids include vinpocetine and vincamine, which belong to the vincamine family. They exhibit numerous pharmacological benefits across the brain, cardiovascular system, and circulatory system, including promoting glucose and oxygen uptake and utilization in the brain, increasing ATP, and reducing lactate production during ischemic hypoxia; preventing brain cell excitotoxicity; alleviating brain hypoxic damage and protecting neurons; enhancing dopaminergic, serotonergic, and noradrenergic neuronal function; preventing ischemic damage in the brain, liver, muscle tissue, and other areas; scavenging free radicals and combating lipid peroxidation; alleviating age-related brain dysfunction; and improving blood lipoprotein composition. Their significant anti-inflammatory and antioxidant effects are expected to counteract the potential toxicity of conventional cationic / ionizable lipid materials. Furthermore, the protonated amine structure of the indole heterocycle of the vincamine class is expected to interact electrostatically with nucleic acid drugs, enabling their intracellular entrapment and delivery. However, due to the short half-life and low bioavailability of vinpocetine and other drugs, frequent dosing is required for the long-term treatment of chronic conditions such as cerebrovascular disease. Studies have shown that targeted delivery systems can prolong vinpocetine's circulation time and allow it to penetrate the blood-brain barrier, thereby increasing its reach to lesions. Based on vinpocetine's pentacyclic fused backbone, structural modifications and fatty chain derivatization of intermediates and end products during the synthesis process have revealed that some modified vincamine-derived lipids exhibit 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.

[0006] Summary of the Invention

[0007] 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; and 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.

[0008] In order to achieve the above object, the present invention provides the following technical solutions:

[0009] 1. Vincamine derivatives, the structural formula of the vincamine derivatives is Or any one of AO-NH-C,

[0010] Wherein A is a group formed after vincamine and its derivatives having an indole ring structure lose a hydroxyl group;

[0011] 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 positive integers;

[0012] C is a straight-chain alkyl group with 10 to 18 carbon atoms, a straight-chain alkenyl group with 10 to 18 carbon atoms, a straight-chain alkyl group with 10 to 18 carbon atoms substituted with a hydroxyl group, or a group 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 2'-OH site is carboxylated, or any one of an amino acid chain, wherein x, a, b, d, and e are all positive integers.

[0013] Preferably, the A is any one of the structural formulas A1 to A12:

[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, the 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] Where C15 is bovine serum albumin, R is n is a positive integer.

[0026] 2. The preparation method of the above vincamine derivatives, wherein the structural formula of the vincamine derivatives is When the preparation method includes any one of method 1, method 2, method 3 and method 4,

[0027] 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 washed with an organic solvent. The intermediate compound I is solidified by a solvent; a compound with a structural formula of C14-OH, the intermediate compound I and benzotriazole-1-yl-oxytripyrrolidinophosphine 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 mixture is stirred and reacted at 20-40° C. for 1-12 hours. The product obtained by the reaction is rotary evaporated, redissolved, extracted, dried, and then concentrated under reduced pressure to obtain a vincamine derivative with a structural formula of A1-NH-B1-NH-C14;

[0028] 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), 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 is added dropwise (the molar ratio of the compound of structural formula A1-OH and tris(2-carbonylethyl)phosphine hydrochloride is 0.15:0.17), and the reaction is stirred at 20-40° C. for 1-12 hours. After reduced pressure concentration, water precipitation and drying are performed to obtain intermediate compound II; 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 and then added to a PBS buffer solution containing bovine serum albumin (BSA) of the sequence, stirred at 20-40°C for 1-12 hours, dialyzed against pure water, and freeze-dried to obtain a solution having the structural formula (A1-NH-B5-NH) 11 -C15 vincamine derivatives.

[0029] Method 3: The preparation method of vincamine derivatives of other structural formulas comprises the following steps:

[0030] (1) A compound with the structural formula C-Br, a compound with the structural formula HO-B-NH2 or H2N-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 and reacted for 12-48 hours. The product obtained by the reaction was cooled and filtered. The obtained filtrate was extracted with n-hexane and separated by silica gel chromatography to obtain a product with the structural formula 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 linear alkyl group having 10 to 18 carbon atoms substituted by a hydroxyl group;

[0031] Alternatively, a linear alkane having 10 to 18 carbon atoms substituted with an epoxy group and a compound having the structural formula 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 the structural formula An intermediate compound, wherein C is a straight-chain alkyl group having 10 to 18 carbon atoms substituted by a hydroxyl group;

[0032] (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 and reacted 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 structural formula of Vincamine derivatives;

[0033] The preparation method of vincamine derivatives with the structural formula AO-NH-C is as follows:

[0034] 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 benzotriazol-1-yl-oxytripyrrolidinophosphine hexafluorophosphate are dissolved in an organic solvent, stirred at 20-40° C. for 24-96 hours, dialyzed against pure water, and freeze-dried to obtain a vincamine derivative of structural formula A1-O-NH-C12;

[0035] 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-oxytripyrrolidinophosphine hexafluorophosphate are dissolved in an organic solvent, the pH is adjusted to 7-8, and the mixture is stirred and reacted at 20-40°C for 1-12 hours. The reaction product is rotary evaporated, redissolved, extracted, dried, and then concentrated under reduced pressure, and then solidified with an organic solvent to obtain a vincamine derivative of structural formula A1-O-NH-C13.

[0036] 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.

[0037] 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.

[0038] 3. Application of the above-mentioned vincamine derivatives in brain-targeted delivery of drugs for treating brain diseases.

[0039] The present invention has the following beneficial effects: The present invention discloses vincamine derivatives and their preparation methods, which have the following advantages: (i) modification of the tail chain increases the lipid solubility of vincamine compounds without affecting the cerebral blood flow regulation of vincamine itself, thereby facilitating the transport of carried drugs through the blood-brain barrier, exerting a brain-protective effect, and improving cerebral microcirculatory disorders; (ii) the tertiary amine group in the parent nucleus structure of vincamine derivatives is ionizable under acidic conditions, enabling efficient delivery and lysosomal escape of nucleic acid drugs through charge adsorption, thereby improving intracellular transport; and (iii) vincamine derivatives inherit the various pharmacological activities inherent to vincamine and are highly safe. Therefore, the vincamine derivatives disclosed in the present invention have good application prospects in brain-targeted delivery of drugs for treating brain diseases.

[0040] Other advantages, objects, and features of the present invention will be described in part in the following description and, in part, will be apparent to those skilled in the art upon examination of the following description or may be learned from practice of the present invention. The objects and other advantages of the present invention may be realized and obtained through the following description. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] 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 with reference to the accompanying drawings, in which:

[0042] Figure 1 is a diagram of compound 1 in Example 1 1 H NMR spectrum;

[0043] FIG2 is a transmission electron micrograph of lipid nanoparticles (LNP) (compound 2 (A5-B2-C6)-14);

[0044] FIG3 shows the pKa values ​​of lipid nanoparticles prepared from different vincamine derivatives measured by TNS fluorescence method;

[0045] FIG4 is the FAM fluorescence intensity of different drug administration groups (lipid nanoparticles prepared by adding different vincamine derivatives) observed under a fluorescence microscope;

[0046] FIG5 shows the cell survival rate calculated by measuring the absorbance of each well (in which lipid nanoparticles formed by different vincamine derivatives were added) at 570 nm using a microplate reader;

[0047] FIG6 shows the effect of lipid nanoparticles formed from different vincamine derivatives on cerebral microvascular blood flow in mice measured by laser speckle imaging;

[0048] FIG7 shows the N / P ratio of siRNA encapsulated in lipid nanoparticles formed by vincamine derivatives (A1-B1-C5) determined by gel retardation experiment;

[0049] FIG8 is a confocal microscope image of the endosomal escape effect of siRNA loaded on lipid nanoparticles formed by compound 1 (A1-B1-C5), compound 2 (A5-B2-C6) and compound 31 (A11-B3-C11) in Table 2;

[0050] FIG9 shows the effect of lipid nanoparticles formed with compound 2 (A5-B2-C6) on cellular ROS levels detected by a single cell analyzer;

[0051] Figure 10 shows NH2-PEG 2000 -Mass spectra of DSPE (a) and compound 35 (b) prepared in Example 6;

[0052] Figure 11 is a mass spectrum of compound 36 prepared in Example 7;

[0053] FIG12 is a mass spectrum of compound 37 prepared in Example 7;

[0054] FIG13 is the TLC verification result of compound 38 prepared in Example 8;

[0055] FIG14 is a standard curve obtained by measuring the absorbance after the reaction of L-leucine (0-10 mM) with OPA reagent;

[0056] Figure 15 shows fluorescence images collected by the VISQUE in vivo Smart-LF System at predetermined time points (0.5 h, 1 h, 2 h) after Compound 31, Compound 35, Compound 36, Compound 37, and Compound 38 (A1-B5-C15) were injected into mice. DETAILED DESCRIPTION

[0057] The following describes the embodiments of the present invention by means of 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 are only schematic illustrations of the basic concept of the present invention, and the following embodiments and features in the embodiments can be combined with each other without conflict.

[0058] In the following examples, the related groups in the structural formulas and the structural formulas of the compounds are shown below:

[0059] A is any one of the structural formulas A1 to A12:

[0060] 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-;

[0061] C is or C' is any one of the structural formulas C1 to C14:

[0062]

[0063] n is a positive integer

[0064]

[0065] In addition, C also includes C15, where C15 is bovine serum albumin (BSA);

[0066] The structural formulas of compounds 1 to 38 are:

[0067]

[0068]

[0069]

[0070]

[0071] Where C15 is bovine serum albumin (BSA), R is n is a positive integer.

[0072] Example 1

[0073] A vincamine derivative (Compound 1) was prepared, the structural formula of which is:

[0074] The specific preparation method is as follows:

[0075] (1) (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br) (400 mg, 1.21 mmol), 2-aminoethanol (HO-B1-NH2) (35 μL, 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. The reaction mixture was heated and stirred at 65°C for 24 h.

[0076] (2) The mixture was cooled to room temperature, and the filter cake was washed three times with n-hexane. The filtrate was extracted with n-hexane, concentrated under reduced pressure, and separated by silica gel chromatography (methanol / dichloromethane = 15 / 85 (v / v)). The structure of the product was obtained by rotary evaporation. intermediate compounds;

[0077] (3) A1-OH (50 mg, 0.15 mmol), 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;

[0078] (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: Vincamine derivatives (Compound 1), which 1 The H NMR (400 MHz, DMSO-d6) spectrum is shown in Figure 1.

[0079] Example 2

[0080] Prepare a vincamine derivative (Compound 2) with the structural formula:

[0081] The specific preparation method is as follows:

[0082] (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;

[0083] (2) The reaction product was concentrated under reduced pressure and separated by silica gel chromatography (methanol / dichloromethane = 1 / 30 (v / v)), and rotary evaporation was performed to obtain the structural formula: intermediate compounds;

[0084] (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;

[0085] (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 = 8.0Hz, 2H), 7.07 (dd, J = 8.4, 6.9Hz, 1H), 6.99 (dd, J = 8.1, 6.8Hz, 1H), 4.84 (d, J = 10.1Hz, 2H), 4.32 (t, J = 5.1Hz, 1H), 4.27 (d, J = 3.7Hz, 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).

[0086] Example 3

[0087] According to the reaction steps in Example 1 (i.e., replacing certain reactants in the example, and the addition ratio and other reaction conditions are the same as those in Example 1), the following compounds were prepared:

[0088] (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 with the structural formula Compound 3, its 1H NMR (400 MHz, DMSO-d6) δ7.62 (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).

[0089] (2) Compound 4 (A1-B1-C3): (6Z)-12-bromododecan-6-ene (C3-Br) was substituted for (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) δ7.62 (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).

[0090] (3) Compound 5 (A1-B2-C4): (9Z)-16-bromohexadecane-9-ene (C4-Br) was substituted for (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 6-aminohexanol (HO-B2-NH2) was substituted for 2-aminoethanol (HO-B1-NH2), and A2-OH was substituted for A1-OH to obtain the structural formula: Compound 5, its 1H NMR (400 MHz, DMSO-d6) δ7.62 (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).

[0091] (4) Compound 6 (A2-B2-C8): 10-bromooctyl octyl 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) δ7.84 (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).

[0092] (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).

[0093] (6) Compound 8 (A2-B3-C10): Substitute N,N-di(3-oxodecyl)-nonanamide (C10-Br) for (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 6-amino-3,4-dithia-hexanol (HO-B3-NH2) for 2-aminoethanol (HO-B1-NH2), and A2-OH for A1-COOH to obtain the structural formula: Compound 8, its 1H NMR (400 MHz, DMSO-d6) δ7.84 (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).

[0094] (7) Compound 9 (A3-B4-C3): (6Z)-12-bromododecan-6-ene (C3-Br) was substituted for (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 4,4-dimethyl-3,5-thioketal-heptanol (HO-B4-NH2) was substituted for 2-aminoethanol (HO-B1-NH2), and A3-OH was substituted for A1-OH to obtain the structural formula: Compound 9, its 1H NMR (400 MHz, DMSO-d6) δ7.74 (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).

[0095] (8) Compound 10 (A3-B1-C4): (9Z)-16-bromohexadecane-9-ene (C4-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 A3-OH was substituted for A1-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).

[0096] (9) Compound 11 (A3-B1-C5): Substitute ethylenediamine (NH2-B1-NH2) for 2-aminoethanol (HO-B1-NH2) and A3-OH for A1-OH to obtain the structural formula: Compound 11, its 1H NMR (400 MHz, DMSO-d6) δ7.74 (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).

[0097] (10) Compound 13 (A4-B2-C7): 3-Bromooctyl 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) δ7.73 (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).

[0098] (11) Compound 14 (A5-B3-C8): 10-bromooctyl octyl 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) δ9.20 (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).

[0099] (12) Compound 15 (A5-B3-C9): N,N-dioctyl-2-bromo-ethylamine (C9-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 15, its 1H NMR (400 MHz, DMSO-d6) δ9.20 (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).

[0100] (13) Compound 16 (A6-B4-C10): Substitute N,N-di(3-oxodecyl)-nonanamide (C10-Br) for (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 4,4-dimethyl-3,5-thioketal-1,7-heptanediamine (HO-B4-NH2) for 2-aminoethanol (HO-B1-NH2), and A6-OH for A1-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).

[0101] (14) Compound 17 (A6-B4-C11): Substitute N,N-bis(3-nonyl acetate)-propionamide (C11-Br) for (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 4,4-dimethyl-3,5-thioketal-1,7-heptanediamine (HO-B4-NH2) for 2-aminoethanol (HO-B1-NH2), and A6-OH for A1-OH to obtain the structural formula: Compound 17, its 1H NMR (400 MHz, DMSO-d6) δ9.31 (s, 1H), 7.54 (s, 1H), 7.30 (d, 1H), 7.23 (d, 1H), 7.16 (t, 1H), 6.64 (t, 1H), 4.56 (t, 8H), 3.54 (m, 14H), 2.84-2.17 (m, 28H), 1.73-1.26 (m, 66H), 0.89 (m, 12H).

[0102] (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: 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).

[0103] (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) 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, 62H), 0.89 (m, 9H).

[0104] (17) Compound 20 (A8-B2-C3): (6Z)-12-bromododecan-6-ene (C3-Br) was substituted for (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 6-aminohexanol (HO-B2-NH2) was substituted for 2-aminoethanol (HO-B1-NH2), and A8-OH was substituted for A1-OH to obtain the structural formula: Compound 20, 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, 40H), 1.16 (s, 9H), 0.89 (m, 6H).

[0105] (18) Compound 21 (A8-B2-C4): (9Z)-16-bromohexadecane-9-ene (C4-Br) was substituted for (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 6-aminohexanol (HO-B2-NH2) was substituted for 2-aminoethanol (HO-B1-NH2), and A8-OH was substituted for 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).

[0106] (19) Compound 22 (A8-B3-C5): (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br) was substituted for (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 2-aminoethanol (HO-B1-NH2) was substituted for 6-amino-3,4-dithia-hexanol (HO-B3-NH2), and A1-OH was substituted for A8-OH to obtain the structural formula: Compound 22, 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, 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).

[0107] (20) Compound 24 (A9-B4-C7): 3-Bromooctyl 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).

[0108] (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).

[0109] (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).

[0110] (23) Compound 28 (A10-B2-C7): 3-Bromooctyl propionate (C7-Br) was substituted for (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), hexamethylenediamine (NH2-B2-NH2) for 2-aminoethanol (HO-B1-NH2), and A10-OH for 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).

[0111] (24) Compound 29 (A11-B2-C9): N,N-dioctyl-2-bromo-ethylamine (C9-Br) was substituted for (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), hexamethylenediamine (NH2-B2-NH2) was substituted for 2-aminoethanol (HO-B1-NH2), and A11-OH was substituted for A1-OH to obtain the structural formula: Compound 29, its 1H NMR (400 MHz, DMSO-d6) δ9.70 (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).

[0112] (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) δ9.70 (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).

[0113] (26) Compound 31 (A11-B3-C11): N,N-bis(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) δ9.70 (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).

[0114] (27) Compound 32 (A12-B3-C11): N,N-bis(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).

[0115] (28) Compound 33 (A12-B4-C1): 12-bromododecane (C1-Br) was substituted for (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 4,4-dimethyl-3,5-thioketal-1,7-heptanediamine (H2N-B4-NH2) for 2-aminoethanol (HO-B1-NH2), and A12-OH for A1-OH to obtain the structural formula: Compound 33, its 1H NMR (400 MHz, DMSO-d6) δ9.70 (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).

[0116] (29) Compound 34 (A12-B4-C2): 16-bromohexadecane (C2-Br) was substituted for (6Z,9Z)-18-bromooctadecane-6,9-diene (C5-Br), 4,4-dimethyl-3,5-thioketal-1,7-heptanediamine (H2N-B4-NH2) for 2-aminoethanol (HO-B1-NH2), and A12-OH for A1-OH to obtain the structural formula: Compound 34, its 1H NMR (400 MHz, DMSO-d6) δ9.70 (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).

[0117] Example 4

[0118] According to the reaction steps in Example 2, the following compounds were prepared:

[0119] (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) δ7.73 (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).

[0120] (2) Compound 23 (A9-B3-C6): 6-amino-3,4-dithia-hexanol (HO-B3-NH2) was substituted for 6-aminohexanol (HO-B2-NH2), and A9-OH was substituted for A5-OH to obtain the structural formula: Compound 23, 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), 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).

[0121] (3) Compound 27 (A10-B1-C6): 1-ethylenediamine (NH2-B1-NH2) was used to replace 6-aminohexanol (HO-B2-NH2), and A10-OH was used to replace A5-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).

[0122] Example 5

[0123] Compound 1 (A1-B1-C5) was prepared into lipid nanoparticles (LNPs), and its relevant physicochemical properties and in vitro and in vivo activities were tested as follows:

[0124] (1) First, lipid nanoparticles (LNPs) were prepared according to the 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 the corresponding molar ratio and mixed. The mixture was mixed with 6.25 mM sodium acetate buffer (pH = 5) containing siRNA (CUU ACG CUG AGU ACU UCG ATT) at a ratio of v / v = 3:1 (water: organic solvent) using a microfluidic device. The mixture was transferred to a dialysis bag and dialyzed overnight with PBS buffer solution (pH = 7.4). Then, it was concentrated using an ultracentrifugal filter. A1-B1-C5 was made into ionizable lipids and 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). The particle size 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.

[0125] Compounds 1 to 34 prepared in the above examples were prepared according to formulation No. 14 in Table 1 to obtain lipid nanoparticles (LNPs) formed from different ionizable lipids in the same formulation. The particle size and PDI (polymer dispersibility index) are shown in Table 3. As shown in Table 3, the LNPs prepared from some vincamine compounds have a particle size <100 nm and good dispersibility, indicating potential for brain targeting in terms of particle size and suitable for subsequent experiments.

[0126] Table 1 Lipid nanoparticles (LNPs) with different molar ratios

[0127] Table 2 Particle size and PDI of different lipid nanoparticles (LNPs)

[0128] Table 3 Particle size and PDI of lipid nanoparticles (LNPs) formed from different ionizable lipids in the same formulation

[0129] (2) The lipid nanoparticles (LNPs) prepared using Compound 2 (A5-B2-C6) as the ionizable lipid according to the formulation 14 in Table 1 were subjected to transmission electron microscopy. The results are shown in Figure 2. As can be seen from Figure 2, the LNP particle size is approximately 100 nm.

[0130] (3) Lipid nanoparticles (A1-B1-C5-14) were prepared by using A1-B1-C5 as ionizable lipids according to the recipe of label 14 in Table 1; lipid nanoparticles (A5-B2-C6-14) were prepared by using A5-B2-C6 as ionizable lipids according to the recipe of label 14 in Table 1; lipid nanoparticles (A2-B3-C10-14) were prepared by using A2-B3-C10 as ionizable lipids according to the recipe of label 14 in Table 1; lipid nanoparticles (A5-B3-C8-14) were prepared by using A5-B3-C8 as ionizable lipids according to the recipe of label 14 in Table 1; lipid nanoparticles (A5-B3-C8-14) were prepared by using A6-B4-C11 as ionizable lipids according to the recipe of label 14 in Table 1; The lipid nanoparticles (A6-B4-C11-14) were prepared using the formula of , A8-B2-C3 was used as the ionizable lipid and the formula of label 14 in Table 1 was used as the lipid nanoparticles (A8-B2-C3-14), A8-B2-C4 was used as the ionizable lipid and the formula of label 14 in Table 1 was used as the lipid nanoparticles (A8-B2-C4-14), A9-B3-C6 was used as the ionizable lipid and the formula of label 14 in Table 1 was used as the lipid nanoparticles (A9-B3-C6-14), and A11-B3-C10 was used as the ionizable lipid and the formula of label 14 in Table 1 was used as the lipid nanoparticles (A11-B3-C10-14). The pKa of the lipid nanoparticles prepared from the vincamine derivatives was determined by TNS fluorescence assay (the specific method is as follows: the prepared lipid nanoparticles were diluted to 2 mM in PBS buffer solution, a buffer solution containing 130 mM NaCl, 10 mM ammonium acetate, 10 mM Hepes, and 10 mM MES was prepared, and the lipid nanoparticles were diluted to 100 μM. In addition, the buffer solution was divided into 8 groups, and the pH value of each group of buffer solution was adjusted to 3, 4, 5, 6, 7, 8, 9, and 10 using NaOH or HCl solution; the anionic fluorescent dye 2-(p-toluidino)-6-naphthalenesulfonic acid (TNS) was dissolved in distilled water to form a solution with a concentration of 100 μM, and the final concentration of the lipid nanoparticles was adjusted to 1 uM solution. At room temperature, the fluorescence intensity of each group was measured using a multifunctional microplate reader with an excitation wavelength of 321 nm and an emission wavelength of 445 nm, where the pKa was defined as the pH value rising to half of the maximum fluorescence intensity). The final results are shown in Figure 3. As can be seen from FIG3 , the pKa value of the LNP prepared from the vincamine derivatives of the present invention is between 6.2 and 6.5, which is conducive to the efficient encapsulation and lysosomal escape of nucleic acids.

[0131] (4) The encapsulation of siRNA by lipid nanoparticles formed by different vincamine derivatives in Table 2 was tested, as shown below:

[0132] The Quant-iT RiboGreen RNA Assay kit was used to quantitatively measure the siRNA concentration in lipid nanoparticles formed from different vincamine derivatives listed in Table 2. The method was as follows: the dialyzed lipid nanoparticles were diluted into Tris-EDTA (TE) buffer in a 96-well plate. Simultaneously, the dialyzed lipid nanoparticles were diluted into TE buffer containing 2% Triton X-100 for 30 minutes. RiboGreen working solution was then added to the corresponding wells. Fluorescence intensity at an excitation wavelength of 485 nm and an emission wavelength of 530 nm was measured using a microplate reader to measure the amount of free siRNA and dialyzed siRNA. The siRNA concentration was calculated using an siRNA standard curve, and the encapsulation efficiency was then calculated. The results are shown in Table 4. As can be seen from Table 4, the LNPs prepared from the vincamine derivatives of the present invention had a good nucleic acid encapsulation efficiency exceeding 90%.

[0133] Table 4 Encapsulation efficiency of siRNA by lipid nanoparticles prepared from different vincamine derivatives

[0134] (5) Fluorescence microscopy of the in vitro transfection effect of siRNA loaded on lipid nanoparticles prepared from vincamine derivatives:

[0135] bEnd.3 cells were seeded in 6-well plates (30% confluence) and cultured in 1 mL of complete medium without anti-IL-6 antibody for 12 hours. The medium was then replaced with serum-free and anti-IL-6 antibody-free medium, and lipid nanoparticles loaded with FAM fluorescent-tagged siRNA (compounds 3 (A1-B1-C2), compound 8 (A2-B3-C10), compound 10 (A3-B1-C4), compound 13 (A4-B2-C7), compound 14 (A5-B3-C8), and compound 17 (A6-B4-C11)) were added to the cells. , 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 an example), incubated at 37 ° C, 5% CO2 environment for 6 hours, the culture medium was replaced with complete medium without double antibody, and the culture was continued for 18 hours. The FAM fluorescence intensity of different dosing groups was observed under a fluorescence microscope, and the results are shown in Figure 4. As can be seen from Figure 4, compared with the gold standard cell transfection reagent Lipo2000, some vincamine compounds prepared LNPs can better carry siRNA for transfection into cells.

[0136] (6) MTT assay of the cytotoxicity of lipid nanoparticles formed by different vincamine derivatives in Table 2:

[0137] 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 listed 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-C10), and Compound 20 (A8-B1-C11). 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) were incubated at 37°C in a 5% CO2 environment for 48 hours. At the end of the experiment, 3-(4,5)-dimethylthiazolylazo(-z-y1)-3,5-diphenyltetrazolium (MTT) solution (5 mg / ml, 20 μL) was added and incubated at 37°C for a further 4 hours. Cell viability was calculated by measuring the absorbance of each well at 570 nm using a microplate reader, and the results are shown in Figure 5. As can be seen from Figure 5, the LNPs prepared from the vincamine derivatives of the present invention have low cytotoxicity and high biocompatibility.

[0138] (7) Laser speckle imaging was used to measure the effects of lipid nanoparticles formed from different vincamine derivatives in Table 2 on brain microvascular blood flow in mice:

[0139] Mice were anesthetized with isoflurane and fixed in a brain stereotactic apparatus. The skull and skin were incised along the midline, and cerebral microvascular blood flow was measured using a laser speckle imager. After the mice stabilized for 30 minutes, 200 μL of lipid nanoparticles prepared from different vincamine derivatives listed in Table 2 were administered via the tail vein at a dose of 30 mg / kg. Laser speckle blood flow images were recorded for 60 minutes after administration to identify cerebral microvascular blood perfusion (ROIs). The results are shown in Figure 6. As can be seen from Figure 6, LNPs prepared from some vincamine compounds have the effect of increasing cerebral microvascular blood flow in mice.

[0140] (8) Taking the lipid nanoparticles containing siRNA formed by vincamine derivatives (A1-B1-C5) in Table 2 as an example, the N / P ratio was determined by gel retardation experiment:

[0141] siRNA (1 OD) was dissolved in 125 μL of DEPC water. Lipid nanoparticles (LNPs) prepared from vincamine derivative compound 1 (A1-B1-C5) listed in Table 2 were mixed with the 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. The mixtures were incubated at room temperature for 30 minutes. The siRNA binding ability of the LNPs was assessed by agarose gel electrophoresis. The LNPs were then mixed with 6× loading buffer and loaded onto a 2% agarose gel containing 0.01% Glodview gel stain. Electrophoresis was performed in 1× TAE buffer at 90 V for 30 minutes. The results were recorded using a gel image analysis system (Tanon, China) at a UV wavelength of 320 nm. The results are shown in Figure 7. As shown in Figure 7, the LNPs prepared from the vincamine derivatives of the present invention can completely encapsulate siRNA when the siRNA / LNP mass ratio is greater than 1:10.

[0142] (9) Confocal microscopy was used to capture the endosomal escape effects of siRNA loaded on lipid nanoparticles formed by different vincamine derivatives in Table 2 (using compound 1 (A1-B1-C5), compound 2 (A5-B2-C6), and compound 31 (A11-B3-C11) as examples):

[0143] bEnd.3 cells were seeded in a 12-well plate (20,000 cells per well) and cultured in 500 μL of culture medium for 12 h. 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 culture medium containing 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)) was discarded, the cells were washed three times with PBS buffer solution, and the cells were stained with Lysotracker. Red (1 uM) was incubated in the dark at 37°C for 2 h. After the incubation, the supernatant was discarded and the cells were washed three times with PBS buffer solution. The cells were fixed with 4% paraformaldehyde fixative at room temperature for 30 min. After discarding the fixative, the cells were washed three times with PBS buffer solution. Hoechst 33342 nuclear stain was added and incubated in the dark for 10 min. After washing three times with PBS buffer solution, fluorescence co-localization was performed using a high-content imaging system in confocal mode. The results are shown in Figure 8. As can be seen from Figure 8, the LNPs prepared from the vincamine derivatives of the present invention can carry siRNA and efficiently escape lysosomes.

[0144] (10) 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:

[0145] To measure cellular oxidative stress and intracellular reactive oxygen species (ROS) levels, a DCFH diacetate reactive oxygen species assay kit was used to detect the green fluorescence signal of ROS. bEnd.3 cells were seeded in cell culture dishes (50,000 cells per dish) and cultured in 1 mL of culture medium for 12 hours. The medium was then discarded and replaced with fresh complete culture medium. Lipid nanoparticles formed from a vincamine derivative (compound 2 (A5-B2-C6)) were added to the cells and incubated in the dark at 37°C in a 5% CO2 atmosphere for 2 hours. Each group of cells was incubated in PBS buffer containing 10 μM DCHF-DA at 37°C for 30 minutes. The cells were then washed three times in PBS buffer containing 0.1% BSA and mounted on glass slides. ROS levels were measured in individual cells using a fiber optic probe with a tip of approximately 10 μm. After the tip was inserted into the cells fixed in PBS, photon counts were continuously detected and calculated for 150 seconds. The results are shown in Figure 9. As can be seen from Figure 9, 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 LNP prepared by vincamine compounds has low cytotoxicity and high biocompatibility.

[0146] Example 6

[0147] Preparation of a vincamine derivative (Compound 35 (A1-C12)) comprises the following steps:

[0148] (1) NH2-PEG 2000 -DSPE (60 mg, 0.02 mmol) (whose mass spectrum is shown in FIG10 a), A1 (13 mg, 0.04 mmol), and benzotriazol-1-yl-oxytripyrrolidinophosphonium 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;

[0149] (2) The reaction solution was placed in a dialysis bag and dialyzed with pure water for 24 h. 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 FIG10 b).

[0150] Example 7

[0151] 1. Preparation of a vincamine derivative (Compound 36 (A1-C13)), specifically comprising the following steps:

[0152] (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) (its 1H NMR (400 MHz, DMSO-d6) δ 12.04 (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, the mass spectrum is shown in Figure 11).

[0153] 2. Preparation of a vincamine derivative (Compound 37 (A1-B5-C14)), specifically comprising the following steps:

[0154] (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 concentrating under reduced pressure, the mixture was dissolved in dichloromethane (5 mL), extracted with 10% anhydrous citric acid and saturated brine in sequence, dried over anhydrous sodium sulfate, and concentrated under reduced pressure to remove the organic phase. The mixture 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. 30 mL of methyl tert-butyl ether was added for precipitation, and dried to obtain A1-B5-NH2.

[0155] (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. After drying, PTX-COOH was obtained;

[0156] (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) and extracted three times with 10% anhydrous citric acid (4 mL) and saturated brine. The mixture was 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) (its 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), the mass spectrum is shown in Figure 12).

[0157] Example 8

[0158] Preparation of a vincamine derivative (Compound 38 (A1-B5-C15)) comprises the following steps:

[0159] (1) Compound A1-OH (50 mg, 0.15 mmol), cystamine dihydrochloride (52 mg, 0.23 mmol), and benzotriazole-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBop, 120 mg, 0.23 mmol) were dissolved in 6 mL 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. 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. Tris(2-carbonylethyl)phosphonium hydrochloride (50 mg, 0.17 mmol) was dissolved in 200 μL of water and added dropwise to the solution of compound 1. The reaction mixture was stirred at room temperature for 2 h, concentrated under reduced pressure, precipitated with water, and dried to obtain an intermediate with the structure of A1-NH-(CH2)2-SH.

[0160] (2) The intermediate with the structure of 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 an 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.

[0161] (3) TLC confirmed the product (as shown in Figure 13). Chloroform and methanol were used as the developing solvent in a volume ratio of 3:1. UV and fluorescent ammonia were used for color development. Compared with the BSA spot, the compound 38 (A1-B5-C15) spot showed UV signal, indicating that the coupling was successful.

[0162] (4) Determination of the free amino group content of BSA and A1-B9-BSA using OPA. Dissolve 40 mg of OPA in 1 mL of methanol, then add 25 mL of 0.1 M sodium borate, 200 mg of DMA, and 5 mL of 10% SDS. Add water to make the volume 50 mL to obtain the OPA reagent. Mix 65 μL of sample (1% protein) with 3 mL of the OPA reagent solution. After 2 minutes, measure the absorbance at 340 nm using a UV spectrophotometer to measure the alkyl isoindole derivatives formed after the reaction of OPA with the free amino groups. A standard curve was obtained by measuring the absorbance after the reaction of L-leucine (0-10 mM) with the OPA reagent (as shown in Figure 14). The free amino group content of the protein sample was calculated based on 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 number of free amino groups in A1-B5-C15 decreased by about 10, indicating that on average each BSA was coupled to about 11 small molecules.

[0163] Table 5 Number of free amino groups in A1-B5-BSA

[0164] Example 9

[0165] In vivo imaging was used to capture the brain targeting properties 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):

[0166] DiD-loaded formulations or Cy5-labeled compounds (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. 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 15 . As can be seen from Figure 15 , the vincamine derivatives of the present invention exhibit brain targeting compared to the control group.

[0167] In summary, the present invention discloses vincamine derivatives and methods for their preparation. These vincamine derivatives have the following advantages: (i) modification of the tail chain increases the lipid solubility of vincamine compounds without affecting the cerebral blood flow regulation of vincamine itself, thereby facilitating drug delivery through the blood-brain barrier, exerting a brain-protective effect, and improving cerebral microcirculatory disorders; (ii) the tertiary amine group in the parent nucleus structure of vincamine derivatives is ionizable under acidic conditions, enabling efficient delivery and lysosomal escape of nucleic acid drugs through charge adsorption, thereby improving intracellular transport; and (iii) vincamine derivatives inherit the various pharmacological activities inherent in vincamine and are highly safe. Therefore, the vincamine derivatives disclosed in the present invention have promising application prospects in brain-targeted delivery of drugs for treating brain diseases.

[0168] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention and are not limiting. 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 solutions of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solutions, which should all be included in the scope of the claims of the present invention.

Claims

1. [Corrected 07.09.2023 in accordance with Rule 91] Vincamine derivatives, characterized in that: The structural formula of the vincamine derivatives is Or any one of AO-NH-C, Wherein A is a group formed after vincamine and its derivatives having an indole ring structure skeleton lose a hydroxyl group; 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 positive integers; 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.

2. [Corrected 07.09.2023 according to Rule 91] The vincamine derivative according to claim 1, characterized in that: The A is any one of the structural formulas A1 to A12:

3. [Corrected 07.09.2023 according to Rule 91] The vincamine derivative according to claim 1, characterized in that: The 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-.

4. [Corrected 07.09.2023 according to Rule 91] The vincamine derivative according to claim 1, characterized in that: The C is any one of the structural formulas C1 to C14: C also includes C15, wherein C15 is bovine serum albumin.

5. [Corrected 07.09.2023 according to Rule 91] The vincamine derivative according to claim 1, characterized in that: The vincamine derivatives include any one of Compounds 1 to 38 with the following structural formulas: Among them, C15 bovine serum albumin, R n is a positive integer.

6. [Corrected 07.09.2023 according to Rule 91] A method for preparing a vincamine derivative according to any one of claims 1 to 5, characterized in that: The structural formula of the vincamine derivatives is When the preparation method comprises any one of method 1, method 2, method 3 or method 4, 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; Method 2: When A is a group of structural formula A1, B is a group of structural formula B5, and C is bovine serum albumin, 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, 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; Method 3: The preparation method of vincamine derivatives of other structural formulas comprises the following steps: (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; 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; (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; The preparation method of vincamine derivatives with the structural formula AO-NH-C is as follows: 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 DSPE-PEG-NH2 and benzotriazole-1-yl-oxytripyrrolidinophosphine hexafluorophosphate are dissolved in an organic solvent, stirred 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; 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 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.

7. [Corrected 07.09.2023 according to Rule 91] The method according to claim 6, 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.

8. [Corrected 07.09.2023 according to Rule 91] The method according to claim 6, 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.

9. [Corrected 07.09.2023 according to Rule 91] Use of the vincamine derivatives according to any one of claims 1 to 5 in brain-targeted delivery of drugs for the treatment of brain diseases.

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