Zedoary turmeric oil / oxidized regenerated cellulose embolization microspheres, preparation method therefor, and use thereof

Abstract

The present application relates to the technical field of pharmaceutical formulations and provides zedoary turmeric oil / oxidized regenerated cellulose embolization microspheres, a preparation method therefor, and use thereof. In the present application, with zedoary turmeric oil as a core solution, a mixed solution of oxidized regenerated cellulose, sodium alginate, and gelatin as a shell layer solution, and a divalent or trivalent metal salt solution as a receiving solution, the zedoary turmeric oil / oxidized regenerated cellulose embolization microspheres are prepared by means of a coaxial electrostatic spraying method. The zedoary turmeric oil / oxidized regenerated cellulose embolization microspheres prepared in the present application have a marked core-shell structure, are uniform in size and good in sphericity, can effectively hide the pungent smell of zedoary turmeric oil, improves the stability of zedoary turmeric oil, and enables directed embolization in tumor blood vessels, thereby reducing toxic and side effects. In addition, the embolization microspheres prepared in the present application have good mechanical properties; therefore, the microspheres can effectively resist deformation under compression and prevent fragmentation within the body, enabling firm embolization of zedoary turmeric oil at the tumor vascular site.

Description

A zedoary turmeric oil / oxidized regenerated cellulose embolic microsphere and its preparation method and application

[0001] This application claims priority to the Chinese patent application filed with the Patent Office of China on March 6, 2024, with application number CN202410251122.1 and invention name “A kind of zedoary oil / oxidized regenerated cellulose embolic microspheres and their preparation method and application”, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The present application relates to the technical field of pharmaceutical preparations, and in particular to a zedoary oil / oxidized regenerated cellulose embolic microsphere, and a preparation method and application thereof. Background Art

[0003] Curcuma oil is an oily substance extracted from the plant and composed of a mixture of various compounds. Its chemical components include dozens of compounds, including curcumol, curdione, elemene, curdenol, isocurdenol, dehydrocurdione, zedoaryl, curdenone, bisabolol, curcumene, and others. Curcuma oil has a wide range of anti-tumor effects, including inhibiting tumor cell proliferation, inducing tumor cell apoptosis, directly destroying tumor cells, inhibiting tumor cell metastasis and invasion, inhibiting tumor angiogenesis, enhancing chemotherapy drug sensitivity, reversing drug resistance, and regulating immunity. It is effective against ovarian cancer, endometrial cancer, cervical cancer, breast cancer, lung cancer, gastric cancer, rectal cancer, and liver cancer.

[0004] Transarterial chemoembolization (TACE) is the treatment of choice for advanced liver cancer. This procedure involves inserting a catheter into the femoral artery, selectively inserting it into the artery feeding the liver cancer, and then embolizing and infusing chemotherapy drugs. Embolization blocks the main tumor vessels, blocking the blood supply and causing ischemic necrosis. Chemotherapy drugs are then infused to kill tumor cells. TACE is suitable for liver cancers with a rich blood supply. Its advantages include minimal trauma, mild side effects, the ability to repeat the treatment, improved quality of life, and prolonged life expectancy, making it an effective treatment for cancer.

[0005] Drug-loaded microspheres are a new type of embolic agent that can carry drugs, simultaneously embolizing the blood vessels that supply nutrients to tumors while also releasing chemical drugs that kill tumor cells, thereby achieving a dual anti-tumor effect. Currently, research on zedoary oil embolic microspheres has mostly focused on zedoary oil gelatin microspheres, which are typically prepared by emulsification and cross-linking. This complex preparation process results in uneven size and poor mechanical properties.

[0006] Summary of the Invention

[0007] In view of this, the present application provides a zedoary turmeric oil / oxidized regenerated cellulose embolic microsphere and its preparation method and application. The preparation method provided in the present application is simple to operate, and the obtained microspheres are uniform in size, have good mechanical properties, and have a high drug loading rate.

[0008] In order to achieve the above-mentioned invention objectives, this application provides the following technical solutions:

[0009] A method for preparing zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres comprises the following steps:

[0010] dissolving oxidized regenerated cellulose in a sodium hydroxide urea solution to obtain an oxidized regenerated cellulose solution; dissolving sodium alginate and gelatin in water to obtain a sodium alginate gelatin solution; and mixing the oxidized regenerated cellulose solution and the sodium alginate gelatin solution to obtain a shell solution;

[0011] Curcuma oil is used as the core solution; the shell solution and the core solution are sprayed into a receiving solution by a coaxial electrostatic spray method to obtain zedoary oil / oxidized regenerated cellulose embolic microspheres; the receiving solution is a metal salt solution; and the metal salt is a divalent metal salt or a trivalent metal salt.

[0012] Preferably, in the oxidized regenerated cellulose solution, the mass ratio of oxidized regenerated cellulose, sodium hydroxide, urea and water is (1-5):(1-5):(4-10):(30-60).

[0013] Preferably, in the sodium alginate gelatin solution, the concentration of sodium alginate is 0.5-5 wt %, and the concentration of gelatin is 0.1-3 wt %.

[0014] Preferably, sodium chloride is further added to the sodium alginate gelatin solution, and the concentration of sodium chloride in the sodium alginate gelatin solution is 0.1-2 wt %.

[0015] Preferably, the volume ratio of the oxidized regenerated cellulose solution to the sodium alginate gelatin solution is (1-5):(1-5).

[0016] Preferably, the divalent metal salt is one or more of calcium salt, barium salt, strontium salt, zinc salt and copper salt; the trivalent metal salt is one or more of trivalent iron salt and aluminum salt.

[0017] Preferably, the operating conditions of the coaxial electrostatic spray include: a shell solution injection speed of 0.1 to 1 mm / min, a core solution injection speed of 0.1 to 1 mm / min, a voltage of 10 to 20 V, and a receiving distance of 2 to 8 cm.

[0018] The present application also provides zedoary oil / oxidized regenerated cellulose embolic microspheres prepared by the preparation method described in the above scheme; the zedoary oil / oxidized regenerated cellulose embolic microspheres have a core-shell structure, the core is zedoary oil, and the shell components include gelatin, alginate and oxidized regenerated cellulose; the alginate is a divalent or trivalent metal salt of alginic acid.

[0019] The present application also provides the use of the zedoary oil / oxidized regenerated cellulose embolic microspheres described in the above scheme in the preparation of hepatic artery chemotherapy embolic agents.

[0020] The present application provides a method for preparing zedoary oil / oxidized regenerated cellulose embolic microspheres, comprising the following steps: dissolving oxidized regenerated cellulose in a sodium hydroxide / urea solution to obtain an oxidized regenerated cellulose solution; dissolving sodium alginate and gelatin in water to obtain a sodium alginate / gelatin solution; mixing the oxidized regenerated cellulose solution and the sodium alginate / gelatin solution to obtain a shell solution; using zedoary oil as a core solution; and spraying the shell solution and the core solution into a receiving liquid using a coaxial electrostatic spray method to obtain zedoary oil / oxidized regenerated cellulose embolic microspheres; the receiving liquid is a metal salt solution; and the metal salt is a divalent metal salt or a trivalent metal salt. The present application utilizes a coaxial electrostatic spray method to prepare zedoary oil / oxidized regenerated cellulose embolic microspheres, which can perfectly encapsulate the zedoary oil in the shell material, is simple to operate, and produces microspheres with uniform particle size.

[0021] In addition, the zedoary oil / oxidized regenerated cellulose embolic microspheres prepared in the present application have a distinct core-shell structure, are uniform in size, and have good sphericity. They can effectively mask the pungent odor of zedoary oil, improve the stability of zedoary oil, and achieve directional embolization in tumor blood vessels, thereby increasing the concentration of zedoary oil at the tumor site, reducing the concentration of zedoary oil in normal tissues, reducing toxic side effects, and achieving a targeted therapeutic effect. Moreover, the microspheres with a core-shell structure have a sustained-release effect and high bioavailability. The zedoary oil contains anti-tumor active ingredients such as curcumol, β-elemene, and curcumone, which have an effective therapeutic effect on tumors.

[0022] The present application also adds oxidized regenerated cellulose to the shell layer. The oxidized regenerated cellulose can form a cross-linked network in the microsphere shell layer, thereby improving the mechanical properties of the microspheres, effectively resisting the extrusion deformation of the microspheres, avoiding breakage in the body, and allowing the zedoary oil to firmly embolize the tumor blood vessels.

[0023] In summary, the preparation method provided in this application has simple steps and low cost, and the obtained microspheres have good mechanical properties and uniform particle size. They can protect zedoary oil, cover up the unpleasant odor of zedoary oil, and have a sustained-release effect. They have broad application prospects in the field of hepatic artery chemoembolization. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] FIG1 is a physical picture of the zedoary oil / oxidized regenerated cellulose embolic microspheres prepared using a 28G#20G coaxial needle in Example 1;

[0025] Figure 2 is an optical microscope photograph of zedoary oil / oxidized regenerated cellulose embolic microspheres prepared using different types of coaxial nozzles in Example 1, wherein A is 25G#18G, B is 28G#20G, C is 30G#21G, and D is 32G#22G;

[0026] FIG3 is a particle size frequency distribution diagram of the zedoary oil / oxidized regenerated cellulose embolic microspheres prepared using different types of coaxial nozzles in Example 1, wherein A is 25G#18G, B is 28G#20G, C is 30G#21G, and D is 32G#22G;

[0027] FIG4 is a scanning electron microscope image of zedoary oil / oxidized regenerated cellulose embolic microspheres prepared using a 28G#20G coaxial needle in Example 1;

[0028] FIG5 is an infrared spectrum of zedoary oil / oxidized regenerated cellulose embolic microspheres prepared using a 28G#20G coaxial needle in Example 1;

[0029] FIG6 shows the drug loading and encapsulation efficiency test results of the zedoary oil / oxidized regenerated cellulose embolic microspheres prepared using different types of coaxial nozzles in Example 1;

[0030] FIG7 is an in vitro release curve of the zedoary oil / oxidized regenerated cellulose embolic microspheres prepared using different types of coaxial nozzles in Example 1;

[0031] FIG8 shows the force-compression curves of the zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres and the zedoary turmeric oil / sodium alginate gelatin embolic microspheres prepared in Example 1. DETAILED DESCRIPTION

[0032] The present application provides a method for preparing zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres, comprising the following steps:

[0033] dissolving oxidized regenerated cellulose in a sodium hydroxide urea solution to obtain an oxidized regenerated cellulose solution; dissolving sodium alginate and gelatin in water to obtain a sodium alginate gelatin solution; and mixing the oxidized regenerated cellulose solution and the sodium alginate gelatin solution to obtain a shell solution;

[0034] Use zedoary oil as core solution;

[0035] The shell solution and the core solution are sprayed into a receiving solution by a coaxial electrostatic spraying method to obtain zedoary oil / oxidized regenerated cellulose embolic microspheres; the receiving solution is a metal salt solution; and the metal salt is a divalent metal salt or a trivalent metal salt.

[0036] In the present application, oxidized regenerated cellulose is dissolved in a sodium hydroxide urea solution to obtain an oxidized regenerated cellulose solution. In the present application, the mass ratio of oxidized regenerated cellulose, sodium hydroxide, urea, and water in the oxidized regenerated cellulose solution is preferably (1-5):(1-5):(4-10):(30-60), and more preferably (2-3):(2-3):(5-8):(40-50).

[0037] In a specific embodiment of the present application, sodium hydroxide and urea are preferably first dissolved in water. The resulting sodium hydroxide-urea solution is placed in a -20°C refrigerator and frozen into an ice-water mixture. Oxidized regenerated cellulose is then added to the sodium hydroxide-urea solution and magnetically stirred for 10 minutes. After dissolution, the solution is stored at -20°C. Before use, the solution is preferably filtered three times through a 0.22 μm filter membrane to obtain a clear, transparent oxidized regenerated cellulose solution. In a specific embodiment of the present application, an oxidized cellulose membrane is preferably used as the raw material. The oxidized cellulose membrane is shredded and then added to the sodium hydroxide-urea solution for dissolution.

[0038] In the present application, sodium alginate and gelatin are dissolved in water to obtain a sodium alginate gelatin solution. In the present application, the concentration of sodium alginate in the sodium alginate gelatin solution is preferably 0.5-5wt%, more preferably 1-3wt%, and the concentration of gelatin is preferably 0.1-3wt%, more preferably 1-2wt%. The water used to dissolve the sodium alginate and gelatin is preferably double-distilled water. In a specific embodiment of the present application, sodium alginate and gelatin are preferably added to double-distilled water and magnetically stirred for more than 12 hours to obtain a transparent solution, namely the sodium alginate gelatin solution.

[0039] In the present application, sodium chloride is further added to the sodium alginate gelatin solution, and the concentration of sodium chloride in the sodium alginate gelatin solution is 0.1-2 wt %. When preparing the sodium alginate gelatin solution, the sodium chloride, sodium alginate and gelatin are added to double distilled water and stirred to dissolve.

[0040] After obtaining the oxidized regenerated cellulose solution and the sodium alginate gelatin solution, the present application mixes the oxidized regenerated cellulose solution and the sodium alginate gelatin solution to obtain a shell solution. In the present application, the volume ratio of the oxidized regenerated cellulose solution to the sodium alginate gelatin solution is preferably (1-5):(1-5), more preferably 1:1, 1:2, 1:3, 1:4, 1:5, 2:1, 3:1, 4:1 or 5:1.

[0041] After obtaining the shell solution, the present application uses zedoary turmeric oil as the core solution and uses a coaxial electrostatic spray method to spray the shell solution and core solution into a receiving solution to obtain zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres; the receiving solution is a metal salt solution. In the present application, the metal salt is a divalent metal salt or a trivalent metal salt; the divalent metal salt is preferably one or more of a calcium salt, a barium salt, a strontium salt, a zinc salt, and a copper salt; the trivalent metal salt is a trivalent iron salt or an aluminum salt; the divalent metal salt is preferably a divalent metal hydrochloride, and the trivalent metal salt is preferably a trivalent metal hydrochloride; in a specific embodiment of the present application, the metal salt is preferably calcium chloride, barium sulfate, zinc chloride, or copper sulfate; and the concentration of the metal salt solution is preferably 0.2 to 0.5 mol / L.

[0042] In the present application, the operating conditions of the coaxial electrostatic spray preferably include: the shell solution injection speed is 0.1-1 mm / min, preferably 0.3-0.5 mm / min; the core solution injection speed is 0.1-1 mm / min, preferably 0.1-0.5 mm / min; the voltage is 10-25 V, preferably 15-20 V, and the receiving distance is 2-10 cm, preferably 5-9 cm; the coaxial needle model used in the coaxial electrostatic spray is preferably 32G#22G, 30G#21G, 28G#20G or 25G#18G, wherein, taking 32G#22G as an example, 32G represents The model of the needle sleeved inside the coaxial needle, 22G represents the model of the outer needle; G represents the needle tube size, specifically, 18G inner diameter is 0.9mm, outer diameter is 1.3mm, 20G inner diameter is 0.6mm, outer diameter is 0.9mm, 21G inner diameter is 0.5mm, outer diameter is 0.8mm, 22G inner diameter is 0.4mm, outer diameter is 0.7mm, 25G inner diameter is 0.2mm, outer diameter is 0.5mm, 28G inner diameter is 0.15mm, outer diameter is 0.35mm, 30G inner diameter is 0.12mm, outer diameter is 0.32mm, 32G inner diameter is 0.06mm, outer diameter is 0.24mm. In a specific embodiment of the present application, the shell solution is injected into the shell flow chamber of the coaxial nozzle, the core solution is injected into the core flow chamber of the coaxial nozzle, and then coaxial electrostatic spraying is performed under the above conditions. This application has no special requirements for the device used for coaxial electrostatic spraying, and the coaxial electrostatic spraying can be performed using an electrostatic spinning apparatus well known to those skilled in the art.

[0043] In the present application, after the shell solution and the core solution are sprayed into the receiving liquid by coaxial electrostatic spraying, sodium alginate and metal ions react to form a gel, gelatin and alginate gel form a shell layer, and at the same time, the oxidized regenerated cellulose is doped in the shell layer to form a skeleton structure, and the shell layer coats the zedoary oil to form core-shell microspheres; after obtaining the core-shell microspheres, the present application preferably collects the microspheres and rinses them with double distilled water to remove the receiving liquid on the surface, and then stores them in double distilled water.

[0044] The present application also provides zedoary oil / oxidized regenerated cellulose embolic microspheres (also known as zedoary oil core-shell microspheres) prepared by the preparation method described in the above scheme; the zedoary oil / oxidized regenerated cellulose embolic microspheres have a core-shell structure, the core is zedoary oil, and the shell components include gelatin, alginate and oxidized regenerated cellulose; the alginate is a divalent or trivalent metal salt of alginic acid, preferably calcium alginate; the drug loading amount of zedoary oil in the zedoary oil / oxidized regenerated cellulose embolic microspheres is preferably 28.62-43.20%.

[0045] This application also provides the use of the zedoary oil / oxidized regenerated cellulose embolic microspheres described in the above-mentioned scheme in the preparation of a hepatic artery chemoembolic agent. The zedoary oil / oxidized regenerated cellulose embolic microspheres provided herein exhibit sustained-release properties, high bioavailability, and excellent mechanical properties. They effectively resist extrusion and deformation of the microspheres, preventing them from breaking in the body. This allows the zedoary oil to securely embolize tumor vessels, and thus has broad application prospects.

[0046] The following will be combined with the embodiments of this application to clearly and completely describe the technical solutions in this application. Obviously, the embodiments described are only part of the embodiments of this application, not all of them. Based on the embodiments of this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0047] Example 1

[0048] 1. Preparation of Reagents

[0049] 1) Preparation of Sodium Alginate-Gelatin Solution: Weigh 0.6 g of sodium alginate, 0.2 g of gelatin, and 0.18 g of sodium chloride. Add double-distilled water to a volume of 20 mL. Stir overnight on a magnetic stirrer until a clear solution forms.

[0050] To prepare the oxidized regenerated cellulose solution: Dissolve 2.8g of NaOH and 4.8g of urea in 40mL of water. Freeze the resulting solution in a -20°C freezer until it becomes an ice-water mixture. Cut 2g of oxidized regenerated cellulose fiber membrane into pieces and dissolve them in a chilled alkaline-urea solution. Stir magnetically for 10 minutes to form a clear solution. Store in a -20°C freezer. Filter the solution twice through a 0.22μm filter to prevent needle clogging.

[0051] 6 mL of sodium alginate gelatin solution and 2 mL of oxidized regenerated cellulose solution were mixed and magnetically stirred to obtain a shell solution.

[0052] 2) Take 5 mL of zedoary turmeric oil as the core solution.

[0053] 2. Synthesis of Curcuma Oil / Oxidized Regenerated Cellulose Embolic Microspheres

[0054] Oil / water (O / W) core-shell microspheres were prepared using zedoary turmeric oil as the oil phase and oxidized regenerated cellulose sodium alginate gelatin solution as the aqueous phase. Preparation was performed using a coaxial electrospray method using an electrospinning apparatus. The steps were as follows: 5 mL of shell solution and 5 mL of core solution were drawn into a syringe and placed into the injection section of the electrospinning apparatus. The injection speeds for the shell solution and the core solution were set at 0.3 mm / min and 0.1 mm / min, respectively, with a voltage of 20 V. The ambient conditions required stability. 100 mL of 0.2 mol / L calcium chloride solution was used as the receiving solution, and the receiving distance was 8 cm. The coaxial needles used were 32G / 22G, 30G / 21G, 28G / 20G, and 25G / 18G, respectively. Core-shell zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres were obtained. The microspheres were rinsed multiple times with double-distilled water to remove excess surface solution, and then stored in double-distilled water at 4°C.

[0055] Example 2

[0056] 1. Preparation of Reagents

[0057] 1) Preparation of Sodium Alginate-Gelatin Solution: Weigh 0.4 g of sodium alginate, 0.2 g of gelatin, and 0.18 g of sodium chloride. Add double-distilled water to 20 mL and stir overnight on a magnetic stirrer until a clear solution forms.

[0058] To prepare the oxidized regenerated cellulose solution: Dissolve 2.8g of NaOH and 4.8g of urea in 40mL of water. Freeze the resulting solution in a -20°C freezer until it becomes an ice-water mixture. Cut 2g of oxidized regenerated cellulose fiber membrane into pieces and dissolve them in a chilled alkaline-urea solution. Stir magnetically for 10 minutes to form a clear solution. Store in a -20°C freezer. Filter the solution twice through a 0.22μm filter to prevent needle clogging.

[0059] 6 mL of sodium alginate gelatin solution and 2 mL of oxidized regenerated cellulose solution were mixed and magnetically stirred to obtain a shell solution.

[0060] 2) Take 5 mL of zedoary turmeric oil as the core solution.

[0061] 2. Synthesis of Curcuma Oil / Oxidized Regenerated Cellulose Embolic Microspheres

[0062] Oil / water (O / W) core-shell microspheres were prepared using zedoary turmeric oil as the oil phase and oxidized regenerated cellulose sodium alginate gelatin solution as the aqueous phase. Preparation was performed using a coaxial electrospray method using an electrospinning apparatus. The steps were as follows: 5 mL of the shell solution and 5 mL of the core solution were drawn into a syringe and placed into the electrospinning apparatus's injection section. The injection speeds were set at 0.3 mm / min for the shell solution and 0.1 mm / min for the core solution, respectively, at a voltage of 20 V. The ambient conditions required stability. 100 mL of 0.4 mol / L barium sulfate solution was used as the receiving solution, with a receiving distance of 8 cm. Coaxial needles of 32G / 22G, 30G / 21G, 28G / 20G, and 25G / 18G were used, respectively. Core-shell zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres were obtained. The microspheres were rinsed multiple times with double-distilled water to remove excess surface solution, and then stored in double-distilled water at 4°C.

[0063] Example 3

[0064] 1. Preparation of Reagents

[0065] 1) Preparation of Sodium Alginate-Gelatin Solution: Weigh 0.5 g of sodium alginate, 0.2 g of gelatin, and 0.18 g of sodium chloride. Add double-distilled water to 20 mL and stir overnight on a magnetic stirrer until a clear solution forms.

[0066] To prepare the oxidized regenerated cellulose solution: Dissolve 2.8g of NaOH and 4.8g of urea in 40mL of water. Freeze the resulting solution in a -20°C freezer until it becomes an ice-water mixture. Cut 2g of oxidized regenerated cellulose fiber membrane into pieces and dissolve them in a chilled alkaline-urea solution. Stir magnetically for 10 minutes to form a clear solution. Store in a -20°C freezer. Filter the solution twice through a 0.22μm filter to prevent needle clogging.

[0067] 2 mL of sodium alginate gelatin solution and 2 mL of oxidized regenerated cellulose solution were mixed and magnetically stirred to obtain a shell solution.

[0068] 2) Take 5 mL of zedoary turmeric oil as the core solution.

[0069] 2. Synthesis of Curcuma Oil / Oxidized Regenerated Cellulose Embolic Microspheres

[0070] Oil / water (O / W) core-shell microspheres were prepared using zedoary turmeric oil as the oil phase and oxidized regenerated cellulose sodium alginate gelatin solution as the aqueous phase. Preparation was performed using a coaxial electrospray method using an electrospinning apparatus. The steps were as follows: 5 mL of shell solution and 5 mL of core solution were drawn into a syringe and placed into the injection section of the electrospinning apparatus. The injection speeds for the shell solution and the core solution were set at 0.5 mm / min and 0.1 mm / min, respectively, with a voltage of 12 V. The ambient conditions required stability. 100 mL of 0.2 mol / L zinc chloride solution was used as the receiving solution, with a receiving distance of 8 cm. Coaxial needles of 32G / 22G, 30G / 21G, 28G / 20G, and 25G / 18G were used, respectively. Core-shell zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres were obtained. The microspheres were rinsed multiple times with double-distilled water to remove excess surface solution, and then stored in double-distilled water at 4°C.

[0071] Example 4

[0072] 1. Preparation of Reagents

[0073] 1) Preparation of Sodium Alginate-Gelatin Solution: Weigh 0.5 g of sodium alginate, 0.2 g of gelatin, and 0.18 g of sodium chloride. Add double-distilled water to a volume of 20 mL. Stir overnight on a magnetic stirrer until a clear solution forms.

[0074] To prepare the oxidized regenerated cellulose solution: Dissolve 2.8g of NaOH and 4.8g of urea in 40mL of water. Freeze the resulting solution in a -20°C freezer until it becomes an ice-water mixture. Cut 2g of oxidized regenerated cellulose fiber membrane into pieces and dissolve them in a chilled alkaline-urea solution. Stir magnetically for 10 minutes to form a clear solution. Store in a -20°C freezer. Filter the solution twice through a 0.22μm filter to prevent needle clogging.

[0075] 4 mL of sodium alginate gelatin solution and 1 mL of oxidized regenerated cellulose solution were mixed and magnetically stirred to obtain a shell solution.

[0076] 2) Take 5 mL of zedoary turmeric oil as the core solution.

[0077] 2. Synthesis of Curcuma Oil / Oxidized Regenerated Cellulose Embolic Microspheres

[0078] Oil / water (O / W) core-shell microspheres were prepared using zedoary turmeric oil as the oil phase and oxidized regenerated cellulose sodium alginate gelatin solution as the aqueous phase. Preparation was performed using a coaxial electrospray method using an electrospinning apparatus. The steps were as follows: 5 mL of the shell solution and 5 mL of the core solution were drawn into a syringe and placed into the electrospinning apparatus's injection section. The injection speeds were set at 0.5 mm / min for the shell solution and 0.1 mm / min for the core solution, respectively, at a voltage of 15 V. The ambient conditions required stability. 100 mL of 0.2 mol / L copper sulfate solution was used as the receiving solution, with a receiving distance of 8 cm. Coaxial needles of 32G / 22G, 30G / 21G, 28G / 20G, and 25G / 18G were used, respectively. Core-shell zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres were obtained. The microspheres were rinsed multiple times with double-distilled water to remove excess surface solution, and then stored in double-distilled water at 4°C.

[0079] Example 5

[0080] All other conditions were the same as in Example 1, except that the sodium alginate gelatin solution was prepared as follows: 0.1 g of sodium alginate, 0.02 g of gelatin, and 0.02 g of sodium chloride were weighed and double-distilled water was added to 20 mL. The mixture was stirred overnight on a magnetic stirrer until a transparent solution formed. Using a 28G / 20G coaxial needle, the conditions of Example 1 were followed to produce core-shell zedoary oil / oxidized regenerated cellulose embolic microspheres.

[0081] Example 6

[0082] All other conditions were the same as in Example 1, except that the sodium alginate gelatin solution was prepared as follows: 1 g of sodium alginate, 0.6 g of gelatin, and 0.4 g of sodium chloride were weighed and double-distilled water was added to 20 mL. The mixture was stirred overnight on a magnetic stirrer until a transparent solution formed. Using a 28G / 20G coaxial needle, the conditions of Example 1 were followed to produce core-shell zedoary oil / oxidized regenerated cellulose embolic microspheres.

[0083] Example 7

[0084] All other conditions were the same as in Example 1, except that sodium chloride was omitted when preparing the sodium alginate gelatin solution. Furthermore, when preparing the shell solution, 1 mL of the sodium alginate gelatin solution and 3 mL of the oxidized regenerated cellulose solution were mixed. Using a 28G / 20G coaxial needle, the preparation was performed according to the conditions of Example 1 to obtain core-shell zedoary oil / oxidized regenerated cellulose embolic microspheres.

[0085] Example 8

[0086] Other conditions were the same as those in Example 1, except that when preparing the oxidized regenerated cellulose solution, the amount of NaOH was changed to 1 g, the amount of urea was changed to 4 g, the amount of water was changed to 30 mL, and the amount of oxidized regenerated cellulose was changed to 1 g.

[0087] Furthermore, when preparing the shell solution, 5 mL of sodium alginate gelatin solution and 1 mL of oxidized regenerated cellulose solution were mixed. Using a 28G / 20G coaxial needle, the preparation was carried out according to the conditions of Example 1 to obtain core-shell structured zedoary oil / oxidized regenerated cellulose embolic microspheres.

[0088] Example 9

[0089] Other conditions were the same as those in Example 1, except that when preparing the oxidized regenerated cellulose solution, the amount of NaOH was changed to 5 g, the amount of urea was changed to 10 g, the amount of water was changed to 60 mL, and the amount of oxidized regenerated cellulose was changed to 5 g.

[0090] Furthermore, when preparing the shell solution, 1 mL of sodium alginate gelatin solution and 5 mL of oxidized regenerated cellulose solution were mixed. Using a 28G / 20G coaxial needle, the preparation was carried out according to the conditions of Example 1 to obtain core-shell structured zedoary oil / oxidized regenerated cellulose embolic microspheres.

[0091] Performance testing:

[0092] 1. Topography testing and standards

[0093] Figure 1 shows a photo of the zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres prepared using a 28G / 20G coaxial needle in Example 1. Figure 1 demonstrates that the zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres prepared in this application exhibit uniform particle size and good dispersibility. Furthermore, observations indicate that Examples 2 through 9 all produced zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres with uniform particle size and good dispersibility.

[0094] Figure 2 shows optical microscopic images of zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres prepared using different coaxial nozzle types in Example 1. A shows 25G on 18G, B shows 28G on 20G, C shows 30G on 21G, and D shows 32G on 22G. The scale is 200 μm. Figure 2 demonstrates that using different coaxial nozzle types, zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres of varying sizes can be produced, and the resulting microspheres all exhibit uniform particle size.

[0095] Figure 3 shows the particle size frequency distribution of the zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres prepared using different coaxial nozzle types in Example 1, with A being 25G and 18G, B being 28G and 20G, C being 30G and 21G, and D being 32G and 22G. Figure 3 also shows a particle size distribution histogram calculated using ImageJ software for 100 microspheres. Figure 3 shows that the zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres prepared in this application have a narrow particle size distribution.

[0096] The zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres prepared using a 28G / 20G coaxial needle in Example 1 were freeze-dried and then subjected to scanning electron microscopy. The results are shown in Figure 4. A clear core-shell structure can be observed in Figure 4, and the zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres exhibit surface shrinkage after freeze-drying.

[0097] 2. Infrared spectrum test

[0098] FIG5 is an infrared spectrum of the zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres prepared using a 28G#20G coaxial needle in Example 1. As can be seen from FIG5 , the 3436 cm -1 The peaks near 1606cm correspond to the stretching vibration of -OH. -1 and 1415cm -1 The peaks at 3436 cm-1 belong to the stretching vibration of C=O and CO, respectively, and belong to the -COO- group of sodium alginate. -1 The peak at 1630 cm is assigned to -OH or -NH groups. -1 and 1518cm -1 The peaks at 1722 cm-1 correspond to C=O and CN groups respectively. -1 The peak is -COOH group. The peak of zedoary oil is 1697cm -1 A characteristic peak appears, which is caused by the stretching vibration of the carbonyl group (C=O) in zedoary oil, 2926cm -1 There is a peak near it, which is the stretching vibration peak of hydroxyl group, at 1444cm -1 and 1365cm -1 The double peaks near the 2926cm-1 peak are caused by the bending vibration of the saturated CH bond plane. -1 The characteristic peaks indicate that the zedoary oil is stably contained in the microspheres and has stable properties. -1 and 1452cm -1The characteristic peak at α-C was significantly enhanced compared to that of sodium alginate gelatin. The possible explanation is that the carboxyl groups of oxidized regenerated cellulose formed a cross-linking network with the adjacent alginate gelatin molecules. These results indicate that there is ionic interaction and molecular compatibility between sodium alginate gelatin and oxidized regenerated cellulose.

[0099] 3. Drug loading and encapsulation efficiency test

[0100] Figure 6 shows the drug loading and encapsulation efficiency test results of the zedoary oil / oxidized regenerated cellulose embolic microspheres prepared using different types of coaxial nozzles in Example 1. Specific test data are shown in Table 1.

[0101] Table 1 Drug loading and encapsulation efficiency of zedoary oil core-shell microspheres (mean ± SD, n = 3, %)

[0102] According to FIG6 and Table 1, the drug loading of the zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres prepared in the present application is 28.62-43.20%, and the encapsulation efficiency is 92.84-99.72%.

[0103] 4. In vitro release experiment

[0104] The in vitro drug release characteristics of the zedoary oil / oxidized regenerated cellulose embolic microspheres prepared in Example 1 were determined using a dialysis bag method. The specific steps were as follows: 10 mL of wet zedoary oil / oxidized regenerated cellulose embolic microspheres prepared using four different coaxial nozzles were precisely pipetted, divided equally into three portions, and placed into pretreated dialysis bags (molecular weight cutoff 3500). Each of the dialysis bags was placed into an Erlenmeyer flask containing 40 mL of release medium (PBS, pH 7.4, containing 0.5% Tween-80) and placed in a constant temperature water bath shaker at (37±0.5)°C and 100 rpm. 2 mL of dialysate was withdrawn from the Erlenmeyer flask at 0, 0.25, 0.5, 0.75, 1, 2, 4, 6, 8, 12, 24, 48, 72, and 96 h, and an equal amount of fresh release medium was simultaneously replenished. Repeat three times. Dispense 0.2 mL of the above solution into a 2 mL EP tube, dilute to 2 mL with vanillin-sulfuric acid solution, and shake thoroughly. Protect from light and allow to develop for 1 hour. Measure the absorbance at 508 nm and calculate the cumulative release rate to investigate the release behavior. The same experiment was also performed using free zedoary turmeric oil for comparison.

[0105] Figure 7 shows the in vitro release curves of the zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres prepared using different coaxial nozzle types in Example 1, where ZTO represents free zedoary turmeric oil. Figure 7 clearly shows that the zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres prepared in this application exhibit superior sustained-release properties.

[0106] 5. Mechanical properties test

[0107] The mechanical properties of the zedoary oil / oxidized regenerated cellulose embolic microspheres prepared in Example 1 were tested. Simultaneously, zedoary oil / sodium alginate gelatin embolic microspheres were prepared and tested for mechanical properties, and compared with the mechanical properties of the zedoary oil / oxidized regenerated cellulose embolic microspheres. The zedoary oil / sodium alginate gelatin embolic microspheres were prepared using the same conditions as in Example 1, except that the oxidized regenerated cellulose in the shell layer was omitted. Specifically, coaxial electrospraying was performed using a sodium alginate gelatin solution as the shell solution and zedoary oil as the core solution, according to the conditions of Example 1, to produce the zedoary oil / sodium alginate gelatin embolic microspheres.

[0108] Figure 8 shows the force-compression curves for the zedoary oil / oxidized regenerated cellulose embolic microspheres and the zedoary oil / sodium alginate gelatin embolic microspheres prepared in Example 1. As shown in Figure 8 , as the elastic deformation gradually increases from 10% to 50%, the zedoary oil / oxidized regenerated cellulose embolic microspheres require a gradually greater pressure than the zedoary oil / sodium alginate gelatin embolic microspheres under the same deformation conditions, exhibiting stronger compression resistance and better elastic properties. This demonstrates that the zedoary oil / oxidized regenerated cellulose embolic microspheres prepared in this application have excellent mechanical properties, effectively resisting compression deformation of the microspheres and preventing them from breaking in vivo, allowing the zedoary oil to securely embolize within the tumor vasculature.

[0109] The above is only a preferred embodiment of the present application. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present application. These improvements and modifications should also be regarded as the scope of protection of the present application.

Claims

1. A method for preparing zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres, characterized in that: The following steps are involved: dissolving oxidized regenerated cellulose in a sodium hydroxide urea solution to obtain an oxidized regenerated cellulose solution; dissolving sodium alginate and gelatin in water to obtain a sodium alginate gelatin solution; and mixing the oxidized regenerated cellulose solution and the sodium alginate gelatin solution to obtain a shell solution; Use zedoary oil as core solution; The shell solution and the core solution are sprayed into a receiving solution by a coaxial electrostatic spraying method to obtain zedoary oil / oxidized regenerated cellulose embolic microspheres; the receiving solution is a metal salt solution; and the metal salt is a divalent metal salt or a trivalent metal salt.

2. The preparation method according to claim 1, characterized in that In the oxidized regenerated cellulose solution, the mass ratio of the oxidized regenerated cellulose, sodium hydroxide, urea and water is (1-5): (1-5): (4-10): (30-60).

3. The preparation method according to claim 1, characterized in that In the sodium alginate gelatin solution, the concentration of sodium alginate is 0.5-5 wt %, and the concentration of gelatin is 0.1-3 wt %.

4. The preparation method according to claim 1, characterized in that Sodium chloride is further added to the sodium alginate gelatin solution, and the concentration of sodium chloride in the sodium alginate gelatin solution is 0.1-2 wt %.

5. The preparation method according to claim 1, characterized in that The volume ratio of the oxidized regenerated cellulose solution to the sodium alginate gelatin solution is (1-5):(1-5).

6. The preparation method according to claim 1, characterized in that The divalent metal salt is one or more of calcium salt, barium salt, strontium salt, zinc salt and copper salt; the trivalent metal salt is one or more of trivalent iron salt and aluminum salt.

7. The preparation method according to claim 6, characterized in that The metal salt is calcium chloride, barium sulfate, zinc chloride or copper sulfate.

8. The preparation method according to claim 1, characterized in that The concentration of the metal salt solution is 0.2-0.5 mol / L.

9. The preparation method according to claim 1, characterized in that The operating conditions of the coaxial electrostatic spray include: a shell solution injection speed of 0.1 to 1 mm / min, a core solution injection speed of 0.1 to 1 mm / min, a voltage of 10 to 20 V, and a receiving distance of 2 to 8 cm.

10. The preparation method according to claim 9, characterized in that The coaxial needle used in the coaxial electrostatic sprayer has a model of 32G#22G, 30G#21G, 28G#20G or 25G#18G.

11. The zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres prepared by the method according to any one of claims 1 to 10, characterized in that: The zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres have a core-shell structure, wherein the core is zedoary turmeric oil and the shell components include gelatin, alginate and oxidized regenerated cellulose; the alginate is a divalent or trivalent metal salt of alginic acid.

12. The zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres according to claim 11, characterized in that: The drug loading amount of the zedoary turmeric oil in the zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres is 28.62-43.20 wt %, and the encapsulation efficiency is 92.84-99.72%.

13. Use of the zedoary turmeric oil / oxidized regenerated cellulose embolic microspheres according to claim 11 or 12 in the preparation of a hepatic artery chemoembolization agent.

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