Use of terazosin or a pharmaceutically acceptable salt thereof for the manufacture of a medicament for the prevention and / or treatment of liver fibrosis

Abstract

This invention provides the use of terazosin or its pharmaceutically acceptable salts in the preparation of medicaments for the prevention and / or treatment of liver fibrosis, relating to the field of biomedical technology. This invention is the first to discover that terazosin or its pharmaceutically acceptable salts can be used to prevent and treat liver fibrosis. It reveals that terazosin or its pharmaceutically acceptable salts can improve the degree of liver fibrosis by improving liver function, reducing the content of α-SMA (a marker of activated hepatic stellate cells) in liver tissue, reducing the content of Col1a1 in liver tissue, and reducing collagen deposition in liver tissue. Furthermore, the liver-targeted sustained-release drug provided by this invention can reduce the peak plasma concentration of terazosin and prolong its plasma half-life, achieving therapeutic effects equivalent to multiple low-dose injections of terazosin, and has broad application prospects.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and more particularly to the use of terazosin or a pharmaceutically acceptable salt thereof in the preparation of medicaments for the prevention and / or treatment of liver fibrosis. Background Technology

[0002] Liver fibrosis is the pathological result of persistent chronic inflammation of the liver and a common outcome of various liver diseases, including viral, toxic, metabolic, and autoimmune liver diseases. The degree of liver fibrosis is a key factor determining the long-term morbidity (such as cirrhosis or liver cancer) and mortality in patients with non-alcoholic fatty liver disease, especially non-alcoholic steatohepatitis. Severe liver fibrosis that progresses to cirrhosis has a certain probability of leading to hepatocellular carcinoma.

[0003] Terazosin's chemical name is 1-(4-amino-6,7-dimethoxy-2-quinazolinyl)-4-[(tetrahydrofuran-2-formyl)]piperazine, and its molecular formula is C2. 19 H 25 N5O4 has the following chemical structural formula:

[0004] .

[0005] Terazosin is a highly effective and selective α1-adrenergic receptor blocker that can act simultaneously on three receptor subtypes: α1A, α1B, and α1D (K-adrenergic receptors). i =3.28nM, 0.689nM, 1.09nM), due to its poor water solubility (30.6mg / L at 22.5℃), the commonly used preparation in clinical treatment is its hydrochloride form, mainly used to treat hypertension, and can be used alone or in combination with other drugs for treating hypertension.

[0006] To date, there are no existing records or reports regarding the preventive and / or therapeutic effects of terazosin on liver fibrosis. Summary of the Invention

[0007] This invention is the first to discover that terazosin or a pharmaceutically acceptable salt thereof can be used for the prevention and treatment of liver fibrosis. Based on this, the following technical solution is proposed:

[0008] A first aspect of the invention provides the use of terazosin or a pharmaceutically acceptable salt thereof in the preparation of a medicament for the prevention and / or treatment of liver fibrosis.

[0009] Furthermore, the terazosin is used to achieve any one or more of the following objectives (a)-(d):

[0010] (a) Improves liver function;

[0011] (b) Reduce the content of α-SMA, a marker of activated hepatic stellate cells, in liver tissue;

[0012] (c) Reduce the content of type I collagen in liver tissue;

[0013] (d) Reduces collagen deposition in liver tissue.

[0014] Furthermore, the drug is prepared in unit dose form, each unit dose containing 0.000001 mg to 20 mg of terazosin or a pharmaceutically acceptable salt thereof.

[0015] Furthermore, the daily dose of terazosin in the drug is 0.0001-1 mg / kg body weight, and / or the administration method of the drug includes, but is not limited to, any one of daily, every 1-2 days, every 3-4 days, every 5-6 days, and every 7-8 days.

[0016] Preferably, the daily dose of terazosin in the drug is 0.001-0.3 mg / kg body weight, more preferably 0.0025-0.2 mg / kg body weight, and even more preferably 0.005-0.1 mg / kg body weight.

[0017] Furthermore, the dosage form of the drug is tablets, capsules, granules, pills, powders, ointments, oral liquids, infusions, or injections.

[0018] A second aspect of the invention provides a liver-targeted sustained-release drug comprising a therapeutically effective amount of terazosin or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier or excipient, said liver-targeted sustained-release drug for the prevention and / or treatment of liver fibrosis.

[0019] Furthermore, the liver-targeted sustained-release drug comprises PEG- b -Plate-like nanoparticles obtained by self-assembly of PLLA (polylactic acid-polyethylene glycol copolymer) and an outer surface modifier that targets and binds to liver cells, wherein the outer surface modifier is attached to the outer surface of the plate-like nanoparticles and the core of the plate-like nanoparticles is loaded with the terazosin.

[0020] Liver fibrosis is the process of fibrotic scarring in liver tissue, characterized by increased extracellular matrix (ECM) content and altered composition. This manifests as excessive deposition of fibrinogens such as type I collagen (COL1A1) in the Disse space, replacing damaged normal tissue. The Disse space contains a low-density basement membrane-like matrix, crucial for maintaining parenchymal cell differentiation and possessing sufficient permeability for blood flow and material exchange and metabolism between hepatocytes. Hepatic stellate cells (HSCs) reside in this space. When ECM is excessively deposited, the normal structure of the Disse space and surrounding tissues is disrupted, leading to functional impairment and hemodynamic changes. Resting HSCs are activated and differentiate into myofibroblasts that highly express α-smooth muscle actin (α-SMA), further secreting large amounts of collagen, ECM degradation inhibitors, pro-inflammatory cytokines, and chemokines, exacerbating liver fibrosis. When severe liver fibrosis progresses to cirrhosis, a large number of intracellular enzymes (alanine aminotransferase and aspartate aminotransferase) are released into the blood, leading to the depletion of free radical scavenging enzymes and further aggravating liver tissue disorder.

[0021] The Disse space is a narrow gap, approximately 0.4 μm wide, located between sinusoidal endothelial cells and hepatocytes. The sinusoidal wall, composed of sinusoidal endothelial cells, has unique fenestrated structures with a diameter of approximately 150-175 nm. These fenestrated structures serve as structural hubs for the exchange of substances between blood and hepatocytes. For drugs to reach HSCs in the Disse space, they must first penetrate the fenestrated structures of the sinusoidal wall, and then specifically bind to markers on the surface of HSCs to achieve targeted regulation. Small molecule and protein drugs have a size advantage in penetrating physical barriers; however, they are also easily and rapidly eliminated and metabolized.

[0022] Therefore, this invention, by employing sheet-like nanoparticles loaded with terazosin for liver-targeted sustained-release, can enhance the liver-targeting of the drug, prolong its half-life, effectively avoid the rapid clearance and metabolism of terazosin when it penetrates physical barriers, and improve pharmacological efficacy. Moreover, compared with spherical nanoparticles of the same mass and diameter of about 100 nm, the sheet-like nanoparticles in this liver-targeted sustained-release drug, when surface modified with N-succinimidyl-S-acetylthioacetate (SATA), have a maximum loading rate of CD44 IgG that is approximately twice that of spherical nanoparticles, and exhibit a stronger enrichment effect in the liver.

[0023] Compared with direct injection of the same dose of terazosin, injection of the liver-targeted sustained-release drug provided by this invention can reduce the peak plasma terazosin concentration by nearly 1 / 3 and prolong the plasma terazosin metabolic endpoint by nearly 5 times.

[0024] Furthermore, in mouse model drug administration, compared with the same dose of terazosin (especially high-dose terazosin) treatment group, the carrier control group, and the model control group, the treatment group internally loaded with terazosin sheet-like nanoparticles (liver-targeted sustained-release drug) effectively improved liver function in mice with liver fibrosis, reduced the content of α-SMA, a marker of activated myofibroblasts, in their liver tissue, and decreased collagen deposition in their liver tissue, exhibiting more significant effects. This indicates that the use of the above-mentioned liver-targeted sustained-release drug can significantly enhance the efficacy of terazosin or its pharmaceutically acceptable salts in the preparation of drugs for the prevention and / or treatment of liver fibrosis.

[0025] Furthermore, the PEG- b -PLLA has a hydrophilic end with exposed maleimide;

[0026] And / or, the number-average molecular weight ratio of PEG to PLLA is 0.02-1, the number-average molecular weight of PEG is 1000-5000, and the number-average molecular weight of PLLA is 1000-50000.

[0027] Furthermore, the liver cells include any one or more of hepatic stellate cells, hepatic sinusoidal endothelial cells, hepatocytes, and Kuffer cells.

[0028] Furthermore, the encapsulation efficiency of the sheet-like nanoparticles for the terazosin is 0-56.22%, and the loading rate is 0-19.08%.

[0029] And / or, the average particle size of the sheet-like nanoparticles is 100-200 nm, and the average thickness is 8-30 nm;

[0030] And / or, the outer surface modification includes at least one of CD44 IgG, FAP IgG, CD44 scFv, FAP scFv, mannose, cRGDyK, hyaluronic acid, and sodium cholate.

[0031] Preferably, the average particle size of the sheet-like nanoparticles is 120-170 nm, more preferably 130-150 nm.

[0032] Preferably, the average thickness of the sheet-like nanoparticles is 10-20 nM, more preferably 10-15 nM.

[0033] Preferably, the outer surface modifier is CD44 IgG, and the loading rate of the sheet-like nanoparticles for the CD44 IgG is 0-38.71%.

[0034] A third aspect of the present invention provides a method for preparing the liver-targeted sustained-release drug as described above, comprising the following steps:

[0035] (1) Using an organic solvent to treat the PEG- b -PLLA and the terazosin are dissolved to obtain an organic phase;

[0036] (2) The organic phase is dispersed in water, and the organic solvent is removed after cooling to obtain an aqueous solution of drug-loaded sheet-like nanoparticles;

[0037] (3) Add a freeze-drying protectant to the aqueous solution of the drug-loaded sheet nanoparticles to obtain a freeze-dried powder of the drug-loaded sheet nanoparticles;

[0038] (4) Add the N-succinimide-S-acetylthioacetate solution to the modification solution containing the outer surface modifier, mix and incubate, then add hydroxylamine solution, mix and incubate to remove excess small molecules, and obtain a mixed aqueous solution;

[0039] (5) Add the mixed aqueous solution to the lyophilized powder of the drug-loaded nanoparticles, incubate, and then perform solid-liquid separation.

[0040] Preferably, the concentration of the N-succinimide-S-acetyl mercaptoacetate solution is 1.73-34.6 mM, more preferably 17.3 mM.

[0041] Preferably, the concentration of the modified solution is 0.1-4 mg / mL, more preferably 2 mg / mL.

[0042] Preferably, the concentration of the hydroxylamine solution is 20-60 mg / mL, more preferably 50 mg / mL.

[0043] Preferably, the volume ratio of the N-succinimide-S-acetylthioacetate solution to the modified solution is 1:(50-1000), more preferably 1:500.

[0044] Preferably, the volume ratio of the N-succinimide-S-acetylthioacetate solution to the hydroxylamine solution is 1:(5-100), more preferably 1:50.

[0045] Furthermore, in the organic phase, PEG- b - The concentration of PLLA is 0.5 mg / mL-5 mg / mL, and the concentration of terazosin is 0.001 mg / mL-1 mg / mL;

[0046] And / or, the freeze-drying protectant includes at least one of sucrose, trehalose, mannitol, glycerol, and arginine, and its addition amount is 2%-20%.

[0047] Preferably, the PEG- b - The concentration of PLLA was 0.8 mg / mL-2 mg / mL, and the concentration of terazosin was 0.4 mg / mL-0.6 mg / mL.

[0048] Furthermore, the organic solvent is N,N-dimethylformamide (DMF).

[0049] Furthermore, the heating temperature during dissolution in step (1) is 55℃-90℃.

[0050] Furthermore, the water addition ratio in step (2) is 1%-20% (v / v).

[0051] Further, in step (2), water is added to the organic phase and stirred at 1000 rpm-1500 rpm for 0-20 min; then stirred at 1200 rpm-2000 rpm for 5-10 min in a room temperature water bath; and then rapidly cooled and solvent replacement is performed using an ultrafiltration tube to remove the organic solvent.

[0052] Furthermore, the molecular sieve size of the ultrafiltration tube is M. W =50-120kDa, more preferably 100kDa; the volume percentage of the sample solution in ultrapure water for each ultrafiltration is 5%-10%; the number of ultrafiltration cycles is 4-6.

[0053] Furthermore, in step (5), the solid-liquid separation method is centrifugation, and the centrifugation process also includes a washing step with water.

[0054] A fourth aspect of the invention provides the use of the liver-targeted sustained-release drug as described above in the preparation of pharmaceutical compositions for the prevention and / or treatment of liver fibrosis. The sustained-release delivery of low doses of terazosin using the liver-targeted sustained-release drug can exert a low-side-effect effect in preventing and treating liver fibrosis.

[0055] Furthermore, the liver-targeted sustained-release drug is used to achieve any one or more of the following purposes:

[0056] (1) Used to reduce the peak plasma drug concentration;

[0057] (2) Used to prolong the half-life of drugs in plasma.

[0058] Furthermore, when the liver-targeted sustained-release drug is used, the dosage is determined according to its active ingredient terazosin, and the drug is administered to the test mammal via multiple routes of administration at multiple time intervals at a daily dose of 0.0001-1 mg / kg body weight. Preferably, the daily dose is 0.0005-0.5 mg / kg body weight, more preferably 0.001-0.3 mg / kg body weight, even more preferably 0.0025-0.2 mg / kg body weight, and even more preferably 0.005-0.1 mg / kg body weight.

[0059] Furthermore, the various time intervals include, but are not limited to, at least one of daily, 1-2 day intervals, 3-4 day intervals, 5-6 day intervals, and 7-8 day intervals, with a preferred interval of 1-2 days.

[0060] In some embodiments, the test mammals include, but are not limited to, rats, mice, non-human primates, humans, dogs, cats, horses, cattle, sheep, pigs, and goats, preferably humans or mice.

[0061] The beneficial effects of terazosin or its pharmaceutically acceptable salts provided by this invention in the preparation of medicaments for the prevention and / or treatment of liver fibrosis are as follows: This invention reveals for the first time the therapeutic mechanism of terazosin, which can improve the degree of liver fibrosis by improving liver function, reducing the content of α-SMA, a marker of activated hepatic stellate cells in liver tissue, reducing the content of Col1a1 in liver tissue, and reducing collagen deposition in liver tissue; and the use of the liver-targeted sustained-release drug provided by this invention can reduce the risk of rapid clearance and metabolism of terazosin, effectively reduce the dosage of terazosin, reduce its side effects, and further improve its application effect, thus having broad application prospects. Attached Figure Description

[0062] To more clearly illustrate the technical solutions in this invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0063] Figure 1 This is a schematic diagram of the CCl4-induced mouse liver fibrosis model and the terazosin treatment process.

[0064] Figure 2 These are the results of the detection of liver function indicators—alanine aminotransferase (ALT) and aspartate aminotransferase (AST)—in mice with liver fibrosis after terazosin treatment.

[0065] Figure 3These are the results of Western blot analysis of Col1a1 and α-SMA in liver tissue homogenates from mice with liver fibrosis after terazosin treatment.

[0066] Figure 4 These are the results of marson staining on liver tissue sections from mice with liver fibrosis after terazosin treatment.

[0067] Figure 5 This is an electron microscope image of terazosin sustained-release nanoparticles.

[0068] Figure 6 This is an electron microscope image of liver-targeted terazosin sustained-release nanoparticles.

[0069] Figure 7 This is an atomic force microscope image of liver-targeted terazosin sustained-release nanoparticles.

[0070] Figure 8 This is a thickness analysis diagram obtained using an atomic force microscope.

[0071] Figure 9 This is a particle size distribution diagram of liver-targeted terazosin sustained-release nanoparticles.

[0072] Figure 10 This is a graph showing the zeta potential analysis before and after loading terazosin nanoparticles and modifying the surface of CD44 IgG.

[0073] Figure 11 This is the encapsulation / drug loading rate of terazosin in liver-targeted sustained-release nanoparticles versus the preparation concentration curve.

[0074] Figure 12 This is the in vitro release time curve of liver-targeted terazosin sustained-release nanoparticles.

[0075] Figure 13 This is a scanning electron microscope image of spherical nanoparticles with surface-modified CD44 IgG.

[0076] Figure 14 The curves show the drug loading rate-preparation concentration of CD44 IgG modified on the surface of sheet-like nanoparticles and spherical nanoparticles.

[0077] Figure 15 These are fluorescence imaging images of liver enrichment after the surface of sheet-like and spherical nanoparticles was modified with CD44 IgG.

[0078] Figure 16 This is the plasma terazosin concentration-time curve corresponding to liver-targeted terazosin sustained-release nanoparticles and free terazosin.

[0079] Figure 17 This is a schematic diagram of the treatment process using a CCl4-induced mouse liver fibrosis model and liver-targeted terazosin sustained-release nanoparticles.

[0080] Figure 18 The results show the detection of liver function indicators—alanine aminotransferase (ALT) and aspartate aminotransferase (AST)—in mice with liver fibrosis after treatment with liver-targeted terazosin sustained-release nanoparticles.

[0081] Figure 19 The results are obtained by immunoblotting of Col1a1 and α-SMA in liver tissue homogenate from mice with liver fibrosis after treatment with liver-targeted terazosin sustained-release nanoparticles.

[0082] Figure 20 These are the results of Masson staining and Sirius red staining of liver tissue sections from mice with liver fibrosis after treatment with liver-targeted terazosin sustained-release nanoparticles.

[0083] Figure 21 These are blood biochemical indicators in mice after intravenous injection of terazosin, sheet-like nanoparticles, and liver-targeted terazosin sustained-release nanoparticles.

[0084] In the picture: "express p <0.05, "express p <0.01, "express p <0.001; “ns” indicates no significant difference. Detailed Implementation

[0085] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.

[0086] Where specific techniques or conditions are not specified in the examples, they shall be performed in accordance with the techniques or conditions described in the literature in this field, or in accordance with the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased through legitimate channels.

[0087] The raw material information for the following embodiments is as follows:

[0088] PEG- b -PLLA (number average molecular weight of PEG is 2000, number average molecular weight of PLLA is 10000): purchased from Xi'an Ruixi Biotechnology Co., Ltd., catalog number R-PLLA-PEG-MAL-2K.

[0089] Terazosin Hydrochloride: Zhejiang Xinsai Pharmaceutical Co., Ltd., batch number 140190307.

[0090] DMF: Purchased from Sinopharm Chemical Reagent Co., Ltd., item number 13026.

[0091] SATA: Purchased from Thermo, part number 26102.

[0092] CD44 IgG: Purchased from Biolegend, catalog number 103002.

[0093] Hydroxylamine hydrochloride: purchased from Thermo, catalog number 26103.

[0094] PVA: Purchased from Kuraray Co., Ltd., Japan, item number PVA217.

[0095] Application Example 1: Application of low-dose, frequent injections of terazosin in the prevention and treatment of liver fibrosis

[0096] 1. Experimental Methods

[0097] Twelve male C57BL / 6J mice, aged 8 weeks and weighing 24.5-25.5g, were selected and divided into four groups of three mice each as biological replicates.

[0098] Terazosin hydrochloride dihydrate was dissolved in physiological saline and administered via tail vein injection.

[0099] A mouse liver fibrosis model was induced by intraperitoneal injection of CCl4 with corn oil as the solvent, at a dose of 1 mL / kg body weight.

[0100] The specific treatment methods and dosages for each group are as follows:

[0101] [Corn oil] Solvent control group (ip corn oil 4 mL / kg, iv physiological saline 0.1 mL);

[0102] [CCl4] model group (ip 25% CCl4 solution 4mL / kg, iv 0.1mL physiological saline);

[0103] [CCl4 / TZ-High Dose] High-dose, high-frequency terazosin treatment group (ip 25% CCl4 solution 4 mL / kg, iv 0.075 mg / mL terazosin aqueous solution 0.1 mL);

[0104] [CCl4 / TZ-Low Dose] Terazosin low-dose high-frequency treatment group (ip 25% CCl4 solution 4mL / kg, iv 0.0075mg / mL terazosin aqueous solution 0.1mL);

[0105] A one-week cycle was followed by intraperitoneal injections for model initiation on Mondays, Wednesdays, and Fridays, and intravenous administration of the drug on Tuesdays, Thursdays, Saturdays, and Sundays for four consecutive weeks. Mice were fasted for 12 hours after the last administration and then sacrificed by cervical dislocation. The timing and procedure for model initiation, drug administration, and sample collection are as follows: Figure 1 As shown.

[0106] 2. Experimental Results

[0107] 2.1 Liver function index testing

[0108] Those skilled in the art will understand that ALT and AST are core indicators of liver function testing, and abnormally elevated levels indicate hepatocellular damage or abnormal liver function. Twenty-four hours after the last administration, blood was collected from the infraorbital venous plexus into anticoagulant tubes. After plasma separation, the ALT and AST levels in the plasma of the mice were measured using an ALT and AST detection kit (Changchun Huili, China).

[0109] Test results as follows Figure 2 As shown, compared with the [CCl4] model group, both the [CCl4 / TZ-high dose] terazosin high-dose high-frequency treatment group and the [CCl4 / TZ-low dose] terazosin low-dose high-frequency treatment group effectively reduced the ALT and AST levels in mouse plasma. Among them, the [CCl4 / TZ-low dose] terazosin low-dose high-frequency treatment group was significantly better than the [CCl4 / TZ-high dose] terazosin high-dose high-frequency treatment group, and had a stronger therapeutic effect.

[0110] 2.2 Molecular immunoblotting detection of characteristic progression of liver fibrosis

[0111] Those skilled in the art will understand that the levels of Col1a1 and α-SMA, a marker protein for activated HSCs, in homogenates of fibrotic liver tissue are characteristic molecules reflecting the degree of liver fibrosis progression. Twenty-four hours after the last administration, mice were euthanized by cervical dislocation and cardiac perfusion was performed, followed by liver tissue sampling. Liver samples from each group were flash-frozen in liquid nitrogen and homogenized using a tissue homogenizer. The levels of Col1a1 and α-SMA were detected by Western blot analysis.

[0112] Test results as follows Figure 3 As shown, compared with the [CCl4] model group, both the [CCl4 / TZ-high dose] terazosin high-dose high-frequency treatment group and the [CCl4 / TZ-low dose] terazosin low-dose high-frequency treatment group effectively reduced the protein content of Col1a1 and α-SMA in mouse liver tissue. Among them, the [CCl4 / TZ-low dose] terazosin low-dose high-frequency treatment group was superior to the [CCl4 / TZ-high dose] terazosin high-dose high-frequency treatment group and had a stronger therapeutic effect.

[0113] 2.3 Detection of collagen fiber deposition in liver tissue

[0114] Furthermore, the inventors investigated collagen fiber deposition in mouse liver tissue using marshmallow staining. Twenty-four hours after the last administration, mice were euthanized by cervical dislocation and their hearts perfused before liver tissue samples were taken. Liver samples from each group were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned. Staining was then performed using a marshmallow staining kit (Solepro, China).

[0115] Test results as follows Figure 4 As shown, compared with the [CCl4] model group, both the [CCl4 / TZ-high dose] terazosin high-dose high-frequency treatment group and the [CCl4 / TZ-low dose] terazosin low-dose high-frequency treatment group effectively reduced collagen fiber deposition in mouse liver tissue. Among them, the [CCl4 / TZ-low dose] terazosin low-dose high-frequency treatment group was significantly better than the [CCl4 / TZ-high dose] terazosin high-dose high-frequency treatment group, and had a stronger therapeutic effect.

[0116] Example 1: Preparation of liver-targeted terazosin sustained-release nanoparticles (liver-targeted sustained-release drug)

[0117] Weigh 2 mg of PEG- with maleimide hydrophilic end groups. b - PLLA (with a number-average molecular weight of 2000 for PEG and 10000 for PLLA) and 0.5 mg terazosin were added to a glass bottle, along with 1 mL of DMF. The bottle was sealed and heated in a 75°C water bath with magnetic stirring at 3000 rpm until dissolved and clear, yielding an organic phase with a terazosin concentration of 0.5 mg / mL. The stirring speed was reduced to 1500 rpm, and 50 μL of ultrapure water was added to the organic phase in the glass bottle. The heating was maintained at 75°C with continuous stirring for 15 min. Separately, a room-temperature water bath was prepared, and the stirring speed was increased to 2000 rpm. The glass bottle was placed in the bath and stirred for 5 min. The solution was then rapidly cooled to room temperature to obtain a DMF solution containing sheet-like nanoparticles. This solution contained sheet-like nanoparticles loaded with terazosin (terazosin sustained-release nanoparticles). An electron microscope image of these terazosin sustained-release nanoparticles is shown below. Figure 5 .

[0118] Add 9 mL of ultrapure water to a beaker and, while stirring magnetically at 1000 rpm, quickly add 1 mL of DMF solution containing sheet-like nanoparticles. Then transfer the solution to a 100 kDa ultrafiltration tube. Centrifuge at 5000 g at 4 °C until the volume is less than or equal to 1 mL. Add 9 mL of ultrapure water again and repeat the washing process four times to obtain an aqueous solution loaded with terazosin sheet-like nanoparticles.

[0119] Weigh the vial beforehand, add an aqueous solution containing 5% sucrose (w / v), 5% trehalose (w / v), and 10% glycerol (v / v) loaded with terazosin nanoparticles, pre-freeze at -80°C, and freeze-dry overnight to obtain a lyophilized powder of terazosin-loaded nanoparticles. Weigh the vial again, and calculate the mass of the terazosin-loaded nanoparticles by subtracting the mass of the added freeze-drying preservative from the difference in mass of the vial before and after freeze-drying.

[0120] Add 500 µL of DMF to a glass vial containing 2 mg of SATA and mix thoroughly until dissolved to obtain a 17.3 mM SATA solution. Dilute 10 mg / mL CD44 IgG stock solution with PBS to prepare a 2 mg / mL CD44 IgG modified solution. Mix 1 µL of SATA solution with 500 µL of CD44 IgG modified solution and incubate at room temperature for 1 hour. Then, add 50 µL of 50 mg / mL hydroxylamine solution to deprotect the thiol group at the other end of the SATA. After mixing, incubate at room temperature for 2 hours and then pass the solution through a desalting column to obtain a CD44 IgG-SATA-thiol solution.

[0121] CD44 IgG-SATA-thiol solution was added to lyophilized powder of terazosin-loaded flake nanoparticles, incubated at room temperature for 2 hours, centrifuged at 5000g for 10 minutes, and washed three times with ultrapure water to obtain liver-targeted terazosin sustained-release nanoparticles. Figure 6 This refers to liver-targeted sustained-release drugs. Their morphology and structure were characterized using transmission electron microscopy. Figures 5-6 Its thickness was characterized by atomic force microscopy. Figures 7-8 The hydrated particle size and zeta potential were detected by a particle size analyzer. Figures 9-10 ). Figure 10 Medium nanosheets refer to nanoparticles that do not load terazosin and do not modify CD44 IgG. Nanosheets + TZ refer to nanoparticles loaded with terazosin and not modified with CD44 IgG. CD44 IgG-nanosheets + TZ refer to nanoparticles loaded with terazosin and modified with CD44 IgG (i.e., liver-targeted terazosin sustained-release nanoparticles).

[0122] Example 2: Encapsulation efficiency / drug loading rate of terazosin in liver-targeted sustained-release nanoparticles (liver-targeted sustained-release drugs) - preparation concentration curve

[0123] Liver-targeted terazosin sustained-release nanoparticles were prepared according to the scheme described in Example 1. Six concentrations of terazosin (i.e., the concentration of terazosin in the organic phase) were set in a gradient, namely 0 mg / mL, 0.0625 mg / mL, 0.125 mg / mL, 0.25 mg / mL, 0.5 mg / mL, and 1 mg / mL.

[0124] After preparation, the mass of each group of liver-targeted terazosin sustained-release nanoparticles was obtained by lyophilization and precise weighing. The 50% methanol aqueous solution of each group of nanoparticles was then disrupted using an ultrasonic disruptor at 30% power. The disrupted solution was then suspended at 37℃ for 48 hours to accelerate PEG- b Hydrolysis of the PLLA polymer promotes the release of terazosin. After centrifugation at 12000 rpm for 20 min, the supernatant was collected and its volume accurately measured. The terazosin content in the supernatant of each sample was determined by high-performance liquid chromatography (HPLC) with a fluorescence detector. The HPLC conditions were as follows: Agilent C18 column was selected. 18 (4.6 × 250 mM, 5 μm), mobile phase A was 30 mMol / L potassium dihydrogen phosphate-4 mMol / L sodium octane sulfonate monohydrate buffer, mobile phase B was 30 mMol / L potassium dihydrogen phosphate-4 mMol / L sodium octane sulfonate monohydrate buffer-acetonitrile (50:50), pH was adjusted to 3.5 with phosphoric acid solution, flow rate was 1.0 mL / min, injection volume was 20 μL, column temperature was 30 ℃, and absorbance at 246 nM was detected by fluorescence detector.

[0125] The mass of terazosin loaded in each group of samples prepared with different concentrations of terazosin was measured using a standard curve. The encapsulation efficiency and drug loading rate of terazosin in liver-targeted sustained-release nanoparticles at different concentrations were calculated according to the following formula, and the encapsulation efficiency / drug loading rate-preparation concentration curve was plotted. Figure 11 The curve shows that when the initial concentration of terazosin is 0.5 mg / mL, the maximum encapsulation efficiency and loading rate under the current preparation conditions can be obtained, which are 56.22% and 19.08%, respectively.

[0126] Formula 1;

[0127] In Equation 1, m 装载特拉唑嗪 Mass of terazosin released after ultrasonic disruption of liver-targeted sustained-release nanoparticles; m 投入载特拉唑嗪 : The feed mass of terazosin during the preparation of liver-targeted sustained-release nanoparticles; m 纳米颗粒 The quality of liver-targeted terazosin sustained-release nanoparticles.

[0128] Example 3: In vitro release time curve of liver-targeted terazosin sustained-release nanoparticles (liver-targeted sustained-release drug)

[0129] The liver-targeted terazosin sustained-release nanoparticles were prepared according to the scheme described in Example 1. Three parallel 10mg sample groups were weighed and placed in 50mL centrifuge tubes. 20mL of DMEM culture medium was added to simulate the physiological environment, and 0.05% (w / v) sodium azide was added to inhibit microbial growth. The samples were continuously suspended in a suspension apparatus at 37℃. After centrifugation at 5000g for 5 min at 0h, 6h, 12h, 24h, 2d, and 4d, 1mL of supernatant was collected, and 1mL of DMEM culture medium was added simultaneously. The samples were then returned to the incubator for continuous suspension. The collected supernatant was stored at -80℃. At the experimental endpoint, the terazosin content in the supernatant at each time point was determined by high-performance liquid chromatography (HPLC) with a fluorescence detector, using the same HPLC conditions as described in Example 2. The release rate was calculated based on the ratio of the mass of terazosin in the supernatant to the total mass of terazosin in the sample at each time point, and an in vitro release time curve of the liver-targeted terazosin sustained-release nanoparticles was plotted. Figure 12 As shown by the curve, under in vitro conditions, the time for complete release of terazosin from liver-targeted sustained-release nanoparticles is approximately 4 days.

[0130] Comparative Example 1: Preparation of spherical nanoparticles with surface-modified CD44 IgG

[0131] 200 mg of PEG-PLLA with maleimide end-group modification was dissolved in 2 mL of ethyl acetate (oil phase, O) to obtain the oil phase. The oil phase was poured into 15 mL of an aqueous solution containing 1.5% PVA (aqueous phase, W), and homogenized at 8000 rpm for 2 min to obtain an emulsion (oil-in-water, O / W). After suspension at room temperature for 15 min, the emulsion was poured into 1000 mL of ultrapure water and magnetically stirred at room temperature for 3 h to allow solvent evaporation and microsphere solidification. The microspheres were collected by centrifugation at 5000 g for 10 min and then lyophilized. The lyophilization method and nanoparticle mass calculation method were the same as in Example 1, and the method for surface modification of CD44 IgG was the same as in Example 1. Spherical nanoparticles with surface modification of CD44 IgG were obtained, and their morphology and structure were characterized by scanning electron microscopy. Figure 13 ).

[0132] Comparative Example 2: Comparison of drug loading rates of CD44 IgG modified on the surfaces of sheet-like nanoparticles and spherical nanoparticles.

[0133] The preparation method of the sheet-like nanoparticles differs from that in Example 1 in that terazosin is not added.

[0134] Plate-like nanoparticles with a particle size of approximately 150 nm and spherical nanoparticles prepared in Example 1 were surface-loaded with CD44 IgG according to the surface modification method described in Example 1. Six concentrations of CD44 IgG modification solutions were set in a gradient: 0 mg / mL, 0.25 mg / mL, 0.5 mg / mL, 1 mg / mL, 2 mg / mL, and 4 mg / mL. After modification and thorough washing, the nanoparticles were then lyophilized using the same method as in Example 1, including the lyophilization method and nanoparticle mass calculation method.

[0135] 2 mg of lyophilized nanoparticles were accurately weighed from each group and dissolved in 1 mL of PBS to prepare a 2 mg / mL nanoparticle suspension. After 10-fold dilution, the CD44 IgG content of each group's samples was detected using a Micro BCA kit. The loading rate of CD44 IgG was calculated with reference to the loading rate of terazosin in Formula 1. The loading rate-concentration curves of CD44 IgG modified on the surface of sheet-like nanoparticles and spherical nanoparticles were plotted. Figure 14 As shown in the figure, when the concentration of the CD44 IgG modified solution is 2 mg / mL, the loading rates of both nanosheets (sheet-like nanoparticles with surface modified CD44 IgG) and nanospheres (spherical nanoparticles with surface modified CD44 IgG) tend to reach their maximum values.

[0136] Comparative Example 3: Liver enrichment effect of CD44 IgG modified on the surface of sheet-like nanoparticles and spherical nanoparticles.

[0137] Following the preparation method of sheet-like nanoparticles described in Example 1 (wherein 0.1% Cy7 is added to the organic phase), the preparation method of spherical nanoparticles described in Comparative Example 1 (wherein 0.1% Cy7 is added to the oil phase), and the surface modification method described in Example 1, the concentration of the CD44 IgG modification solution was set to 2 mg / mL, and sheet-like nanoparticles and spherical nanoparticles with a particle size of approximately 150 nm, surface-modified with CD44 IgG, and capable of fluorescence traceability were prepared, respectively corresponding to... Figure 15 The CD44 IgG nanosheets and CD44 IgG nanospheres are included.

[0138] Four SPF-grade 8-week-old male C57BL / 6J mice were randomly selected. Two groups of fluorescent nanoparticles were injected into the mice via the tail vein (n=2), with a nanoparticle concentration of 0.5 mg / mL and an injection volume of 100 μL. Fluorescence was quantified before injection to ensure consistent fluorescence intensity between the two nanoparticle suspensions. Twenty-four hours after injection, the mice were euthanized by cervical dislocation, and the major organs (heart, liver, spleen, lung, and kidney) were dissected. Fluorescence intensity images of Cy7 (excitation wavelength 750 nm, emission wavelength 770 nm) in the major organs were captured using the BIOVIVO in vivo imaging platform. Figure 15As shown in the figure, the liver enrichment effect of the nanosheets after surface modification with CD44 IgG is better than that of the nanospheres.

[0139] Comparative Example 4: Comparison of blood concentration-time curves of liver-targeted terazosin sustained-release nanoparticles (liver-targeted sustained-release drug) and free terazosin.

[0140] Following the preparation method described in Example 1, the concentration of terazosin was set to 0.5 mg / mL, and the concentration of the CD44 IgG modified solution was set to 2 mg / mL, to prepare liver-targeted terazosin sustained-release nanoparticles with a particle size of approximately 150 nm. Terazosin was diluted with physiological saline to prepare an aqueous solution of nanoparticles with a concentration of 0.075 mg / mL.

[0141] Terazosin hydrochloride dihydrate was dissolved in physiological saline to prepare an aqueous solution with a terazosin concentration of 0.075 mg / mL, which was used as the free terazosin group.

[0142] Six SPF-grade 8-week-old male C57BL / 6J mice, weighing approximately 25g each, were randomly selected and administered the drug at a dose of 0.3mg / kg. Both groups of drugs were injected into the mice via the tail vein (n=3), with an injection volume of 100μL. Blood was collected from the mice via the infraorbital venous plexus at 0h, 2h, 4h, 6h, 8h, 10h, 12h, 18h, 24h, 30h, 36h, 42h, and 48h after injection. The blood was placed in anticoagulant tubes containing EDTA and centrifuged at 3500rpm for 10min to separate the plasma.

[0143] The terazosin content in plasma was determined by LC-MS / MS under the following chromatographic conditions: Agilent C18 column was selected. 18(2.1 × 50 mM, 1.8 μm), mobile phase: methanol: 5 mmol / L ammonium acetate (containing 0.15% formic acid) = 35:65, flow rate: 0.2 mL / min, injection volume: 5 μL, column temperature: 35 °C. Mass spectrometry conditions were as follows: electrospray ionization source (ESI+) multiple reaction monitoring mode, nebulizer 50 psi, nebulizer gas temperature 550 °C, capillary voltage 5500 V, quantitative ion pairs of terazosin and internal standard prazosin were m / z 388.2→m / z 290.4 and m / z 384.2→m / z 247.3, respectively, declustering voltages were 122 V and 126 V, and collision voltages were 36 V and 39 V, respectively. Accurately measure 200 μL of plasma, add 10 μL of 200 ng / mL prazosin internal standard solution, mix well, add 0.4 mL of acetonitrile, vortex for 2 min, centrifuge at 12000 rpm for 10 min, and transfer the supernatant to a glass tube. Dry under nitrogen at 40℃, redissolve the residue with 200 μL of mobile phase, centrifuge at 12000 rpm for 10 min, and inject 5 μL of the supernatant for analysis. Calculate the plasma terazosin concentration-time curves for the two groups of samples using the plasma terazosin standard curve. Figure 16 ).

[0144] Depend on Figure 16 It was found that after direct injection of free terazosin [TZ], the plasma concentration of terazosin rose rapidly, reaching its peak at approximately 2 hours, at which point the plasma concentration was 14.66 ng / mL. It was completely metabolized in approximately 12 hours. 1 / 2 =5.48h; while after injection of liver-targeted terazosin sustained-release nanoparticles [CD44 IgG-nanoparticles + TZ], the plasma terazosin concentration rose slowly, reaching its peak at approximately 12 hours, at which point the plasma terazosin concentration was 5.73 ng / mL, and it was completely metabolized in approximately 48 hours. 1 / 2 =38.25h. Therefore, liver-targeted terazosin sustained-release nanoparticles can prolong the plasma half-life of terazosin.

[0145] Application Example 2: Application of liver-targeted terazosin sustained-release nanoparticles (liver-targeted sustained-release drugs) in the preparation of drugs for the prevention and treatment of liver fibrosis.

[0146] 1. Experimental Methods

[0147] Sixteen SPF-grade 8-week-old male C57BL / 6J mice, weighing 24.5-25.5g, were selected and divided into four groups of four mice each as biological replicates.

[0148] Following the preparation method described in Example 1, the concentration of terazosin was set to 0.5 mg / mL, and the concentration of CD44 IgG modified solution was set to 2 mg / mL, to prepare liver-targeted terazosin sustained-release nanoparticles with a particle size of approximately 150 nm. These nanoparticles were diluted with physiological saline to prepare a 0.375 mg / mL aqueous solution (containing 0.075 mg / mL terazosin and 0.3 mg / mL CD44 IgG nanosheets), named CD44 IgG nanosheets + TZ.

[0149] Using CD44 IgG nanosheets without terazosin loading as the control group, an aqueous solution of nanoparticles at a concentration of 0.3 mg / mL was prepared with physiological saline and named CD44 IgG nanosheets.

[0150] A mouse liver fibrosis model was induced by intraperitoneal injection of CCl4 with corn oil as the solvent, at a dose of 1 mL / kg body weight.

[0151] The specific treatment methods and dosages for each group are as follows:

[0152] [Corn oil] Solvent control group (ip corn oil 4 mL / kg, iv physiological saline 0.1 mL);

[0153] [CCl4] model group (ip 25% CCl4 solution 4mL / kg, iv 0.1mL physiological saline);

[0154] [CCl4 / CD44 IgG-nanoparticles] vector control group (ip 25% CCl4 solution 4 mL / kg, iv [CD44IgG-nanoparticles] 0.1 mL);

[0155] [CCl4 / CD44 IgG-nanoparticles + TZ] liver-targeted terazosin sustained-release nanoparticle treatment group (ip 25% CCl4 solution 4mL / kg, iv [CD44 IgG-nanoparticles + TZ] 0.1mL).

[0156] A one-week cycle was implemented, with intraperitoneal injections for model initiation on Mondays, Wednesdays, and Fridays, and intravenous drug administration on Tuesdays and Saturdays, for four consecutive weeks. Mice were fasted for 12 hours after the last administration and then sacrificed by cervical dislocation. The model initiation, drug administration, and sample collection time and procedures are as follows: Figure 17 As shown.

[0157] 2. Experimental Results

[0158] 2.1 Liver function index testing

[0159] Twenty-four hours after the last administration, blood was collected from the infraorbital venous plexus into anticoagulant tubes. After plasma separation, the ALT and AST levels in the plasma of the mice were measured using an ALT and AST assay kit (Changchun Huili, China). The results are as follows: Figure 18 A and Figure 18 As shown in Figure B, compared with the [CCl4] model group and the [CCl4 / CD44 IgG-nanoparticle] carrier control group, the [CCl4 / CD44 IgG-nanoparticle + TZ] liver-targeted terazosin sustained-release nanoparticles significantly reduced the levels of ALT and AST in mouse plasma, with therapeutic effects equivalent to low-dose, high-frequency injection of terazosin.

[0160] 2.2 Molecular immunoblotting detection of characteristic progression of liver fibrosis

[0161] Twenty-four hours after the last administration, mice were euthanized by cervical dislocation and their hearts were perfused. Liver tissue samples were then collected. Liver samples from each group were flash-frozen in liquid nitrogen and homogenized using a tissue homogenizer. The contents of Col1a1 and α-SMA were detected by Western blot. The results are as follows: Figure 19 As shown, compared with the [CCl4] model group and the [CCl4 / CD44 IgG-nanoparticle] carrier control group, the [CCl4 / CD44 IgG-nanoparticle + TZ] liver-targeted terazosin sustained-release nanoparticle treatment group reduced the protein content of Col1a1 and α-SMA in mouse liver tissue, which was equivalent to the treatment effect of low-dose high-frequency injection of terazosin.

[0162] 2.3 Detection of collagen fiber deposition in liver tissue

[0163] Furthermore, the inventors investigated collagen fiber deposition in mouse liver tissue using Masson's red and Sirius red staining. Twenty-four hours after the last administration, mice were euthanized by cervical dislocation and cardiac perfusion was performed, followed by liver tissue sampling. Liver samples from each group were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned. Staining and detection were performed using a Masson's red and Sirius red staining kit (Solepro, China). The results are as follows: Figure 20 As shown, compared with the [CCl4] model group and the [CCl4 / CD44 IgG-nanoparticle] carrier control group, the [CCl4 / CD44 IgG-nanoparticle + TZ] liver-targeted terazosin sustained-release nanoparticle treatment group reduced collagen fiber deposition in mouse liver tissue, and the treatment effect was equivalent to that of low-dose, high-frequency injection of terazosin.

[0164] Example 4: In vivo safety testing of terazosin, sheet-like nanoparticles, and liver-targeted terazosin sustained-release nanoparticles.

[0165] 1. Experimental Methods

[0166] Sixteen SPF-grade 8-week-old male C57BL / 6J mice, weighing 24.5-25.5g, were selected and divided into four groups of four mice each as biological replicates.

[0167] Terazosin hydrochloride dihydrate was dissolved in physiological saline to prepare an aqueous solution with a terazosin concentration of 0.075 mg / mL [TZ].

[0168] Following the preparation method described in Example 1, the concentration of terazosin was set to 0.5 mg / mL, and the concentration of the CD44 IgG modified solution was set to 2 mg / mL. Liver-targeted terazosin sustained-release nanoparticles with a particle size of approximately 150 nm were prepared. These nanoparticles were diluted with physiological saline to prepare a 0.375 mg / mL aqueous solution (containing 0.075 mg / mL terazosin and 0.3 mg / mL CD44 IgG nanosheets) [CD44 IgG nanosheets + TZ].

[0169] Using CD44 IgG nanosheets without terazosin loading as the control group, an aqueous solution of nanoparticles [CD44 IgG nanosheets] was prepared with physiological saline to a concentration of 0.3 mg / mL.

[0170] Mice that were intravenously injected with physiological saline served as the blank control group [blank].

[0171] The specific treatment methods and dosages for each group are as follows:

[0172] [Blank] Solvent control group (iv 0.1 mL physiological saline);

[0173] [TZ] Terazosin group (iv 0.075 mg / mL terazosin aqueous solution 0.1 mL);

[0174] [CD44 IgG-nanosheet] vector control group (iv [CD44 IgG-nanosheet] 0.1 mL);

[0175] [CD44 IgG-nanoparticles + TZ] liver-targeted terazosin sustained-release nanoparticle group (iv [CD44 IgG-nanoparticles + TZ] 0.1 mL).

[0176] Administer intravenously once a day for one week.

[0177] 2. Blood biochemical index testing

[0178] Twenty-four hours after the last injection, blood was collected from the infraorbital venous plexus into an anticoagulant tube. After plasma separation, the levels of lactate dehydrogenase, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, and blood urea nitrogen in the plasma of the mice were detected using a biochemical analysis kit (Toshiba, Japan).

[0179] The test results for lactate dehydrogenase, alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, and blood urea nitrogen are as follows: Figure 21 A, Figure 21 B, Figure 21 C, Figure 21 D, Figure 21 As shown in Figure E, the blood biochemical indicators of all drug injection groups were not different from those of the blank control group, indicating that terazosin, nanosheet carrier, and liver-targeted terazosin sustained-release nanoparticles have high in vivo safety.

[0180] The above efficacy verification experiments show that terazosin can significantly inhibit the progression of liver fibrosis, and the liver-targeted sustained-release drug prepared by this invention can also better exert the application effect of terazosin or its pharmaceutically acceptable salt in the prevention and / or treatment of liver fibrosis.

[0181] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. Use of terazosin or a pharmaceutically acceptable salt thereof in the preparation of medicaments for the prevention and / or treatment of liver fibrosis.

2. The use of terazosin or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of a medicament for the prevention and / or treatment of liver fibrosis, characterized in that, The terazosin is used to achieve any one or more of the following objectives (a)-(d): (a) Improves liver function; (b) Reduce the content of α-SMA, a marker of activated hepatic stellate cells, in liver tissue; (c) Reduce the content of type I collagen in liver tissue; (d) Reduces collagen deposition in liver tissue.

3. The use of terazosin or a pharmaceutically acceptable salt thereof according to claim 1 in the preparation of a medicament for the prevention and / or treatment of liver fibrosis, characterized in that, The drug is prepared in unit dose form, each unit dose containing 0.000001 mg to 20 mg of terazosin or a pharmaceutically acceptable salt thereof; And / or, the dosage form of the drug is tablets, capsules, granules, pills, powders, ointments, oral liquids, infusions, or injections.

4. A liver-targeted sustained-release drug, characterized in that, This includes therapeutically effective amounts of terazosin or its pharmaceutically acceptable salts, as well as pharmaceutically acceptable carriers or excipients; The liver-targeted sustained-release drug comprises PEG- b - PLLA self-assembled sheet-like nanoparticles and an outer surface modifier that targets and binds to liver cells, wherein the outer surface modifier is attached to the outer surface of the sheet-like nanoparticles and the core of the sheet-like nanoparticles is loaded with the terazosin or a pharmaceutically acceptable salt thereof; The PEG- b -PLLA has a hydrophilic end with exposed maleimide; The outer surface modification is CD44 IgG; The preparation method of the liver-targeted sustained-release drug includes the following steps: (1) Dissolve the PEG-b-PLLA and the terazosin in an organic solvent to obtain an organic phase; (2) The organic phase is dispersed in water, and the organic solvent is removed after cooling to obtain an aqueous solution of drug-loaded sheet-like nanoparticles; (3) Add a freeze-drying protectant to the aqueous solution of the drug-loaded sheet nanoparticles to obtain a freeze-dried powder of the drug-loaded sheet nanoparticles; (4) Add the N-succinimide-S-acetylthioacetate solution to the modification solution containing the outer surface modifier, mix and incubate, then add hydroxylamine solution, mix and incubate to remove excess small molecules, and obtain a mixed aqueous solution; (5) Add the mixed aqueous solution to the lyophilized powder of the drug-loaded nanoparticles, incubate, and then perform solid-liquid separation.

5. The liver-targeted sustained-release drug according to claim 4, characterized in that, The number-average molecular weight ratio of PEG to PLLA is 0.02-1, and the number-average molecular weight of PEG is 1000-5000.

6. The liver-targeted sustained-release drug according to claim 4 or 5, characterized in that, The sheet-like nanoparticles exhibit an encapsulation efficiency of 56.22% and a loading rate of 19.08% for the terazosin. And / or, the average particle size of the sheet-like nanoparticles is 100-200 nm and the average thickness is 8-30 nm.

7. The liver-targeted sustained-release drug according to claim 6, characterized in that, The loading rate of the sheet-like nanoparticles for the CD44 IgG was 38.71%.

8. The method for preparing the liver-targeted sustained-release drug according to any one of claims 4-7, characterized in that, Includes the following steps: (1) Dissolve the PEG-b-PLLA and the terazosin in an organic solvent to obtain an organic phase; (2) The organic phase is dispersed in water, and the organic solvent is removed after cooling to obtain an aqueous solution of drug-loaded sheet-like nanoparticles; (3) Add a freeze-drying protectant to the aqueous solution of the drug-loaded sheet nanoparticles to obtain a freeze-dried powder of the drug-loaded sheet nanoparticles; (4) Add the N-succinimide-S-acetylthioacetate solution to the modification solution containing the outer surface modifier, mix and incubate, then add hydroxylamine solution, mix and incubate to remove excess small molecules, and obtain a mixed aqueous solution; (5) Add the mixed aqueous solution to the lyophilized powder of the drug-loaded nanoparticles, incubate, and then perform solid-liquid separation.

9. The method for preparing the liver-targeted sustained-release drug according to claim 8, characterized in that, In the organic phase, the concentration of PEG-b-PLLA is 0.5 mg / mL-5 mg / mL, and the concentration of terazosin is 0.001 mg / mL-1 mg / mL; And / or, the freeze-drying protectant includes at least one of sucrose, trehalose, mannitol, glycerol, and arginine, and its addition amount is 2%-20%.

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