Injectable Gel Composition Containing Ablative Agent and Preparation Method Thereof

Abstract

The present application discloses an injectable gel composition carrying an ablating agent and a preparation method thereof. The injectable gel composition includes a gel and an ablating agent dispersed in the gel. The ablating agent is ethanol and the concentration is 0.1-14 wt%. After the injectable gel composition is implanted into the tumor site, it has good conformability and can slowly release the ablating agent, reducing the overflow of the ablating agent in the target area, thereby achieving effective ablation of tumor cells and protection of normal tissues around the tumor.

Description

Technical Field

[0001] The present application relates to the technical field of drug-loaded gel materials, and particularly to an injectable gel composition loaded with an ablative agent and a preparation method thereof. Background Art

[0002] The incidence of benign and malignant tumors is gradually increasing, seriously affecting human health and life. At present, the treatment options for tumors mainly include surgical methods, ablation methods, and radiotherapy.

[0003] The ablation method is mainly divided into physical ablation and chemical ablation. Among them, physical ablation refers to using high temperature or low temperature to damage tumor cells and cause the cells to lose their activity, such as physical technical means such as radiofrequency ablation, cryoablation, and high-intensity focused ultrasound ablation. Although physical ablation has a good therapeutic effect on many tumors, its conformability to certain special tumors is weak. For example, tumors with irregular shapes or at the renal hilum cannot completely ablate tumor cells, so the therapeutic effect is not good.

[0004] Chemical ablation refers to injecting an ablative agent into the tumor site to kill tumor cells. Its treatment mechanism is that the ablative agent, as a protein coagulant, can dehydrate and denature cells, resulting in the destruction of cell structure and coagulative necrosis. Compared with physical ablation, chemical ablation has good conformability. It can enter tumor cells along the intercellular space of tumor cell tissues when the tumor is large and has an uneven shape, inactivate tumor cells, and retain normal tissue cells, thereby protecting tissue function.

[0005] Ablative agents usually have sclerosis and chemical ablation effects. For example, ethanol has good application prospects in tumor treatment. However, it is very difficult to accurately control the dosage of directly injecting ethanol into the target site. And because ethanol has strong permeability, it will quickly spread to the surrounding tissue cells of the tumor, resulting in a decrease in the ethanol concentration in the target area and unable to achieve a satisfactory therapeutic effect. Ethanol in the non-target area will damage normal tissues and functions and cause pain to the patient. Summary of the Invention

[0006] In view of the problems existing in the prior art, the present application provides an injectable gel composition loaded with an ablative agent, which has good conformability after being implanted into the tumor site, and can slowly release the ablative agent, reduce the overflow of the ablative agent in the target area, so as to effectively ablate tumor cells and protect the normal tissues around the tumor.

[0007] The injectable gel composition loaded with an ablative agent provided by the present application includes a gel and an ablative agent dispersed in the gel. The ablative agent is ethanol, and the concentration is 0.1-14 wt%.

[0008] Optionally, the ethanol release rate of the injectable gel composition is 0.01-6 mg / h.

[0009] Optionally, the storage modulus of the injectable gel composition is 300 - 10000 Pa. Preferably it is 500 - 7000 Pa. More preferably it is 1000 - 3000 Pa.

[0010] Optionally, the concentration of the ethanol is 0.5 - 14 wt%.

[0011] Optionally, the concentration of the ethanol is 0.1 - 7 wt%.

[0012] Optionally, the gel has a three-dimensional network structure with a pore size of 10 - 100 μm.

[0013] Preferably, the pore size of the gel is 10 - 60 μm. More preferably it is 10 - 30 μm.

[0014] Optionally, the gel is formed by cross-linking a gel precursor with an alkaline earth metal salt.

[0015] Optionally, the gel precursor is at least one of alginate, oxidized alginate, alginate ester, silk fibroin, collagen, chitosan, carboxymethyl chitosan, cholate-modified chitosan, dextran. Preferably it is alginate.

[0016] Optionally, the alkaline earth metal salt is at least one of calcium nitrate, calcium carbonate, calcium chloride, magnesium chloride, barium chloride, barium sulfate, magnesium sulfate, calcium sulfate, calcium gluconate, calcium lactate, and calcium alginate.

[0017] Preferably, the alkaline earth metal salt is at least one of calcium chloride, calcium gluconate, calcium lactate, and calcium alginate. More preferably it is at least one of calcium chloride and calcium alginate.

[0018] The present application also provides a preparation method of the injectable gel composition carrying an ablative agent, including:

[0019] Mixing an alkaline earth metal salt, an ablative agent, a gel precursor, and water, and reacting to obtain the injectable gel composition.

[0020] Optionally, the mixing method is:

[0021] Mixing an alkaline earth metal salt, an ablative agent, and water to obtain a mixed solution A;

[0022] Mixing a gel precursor and a part of water to obtain a mixed solution B;

[0023] Mixing the mixed solution A and the mixed solution B and reacting to obtain the injectable gel composition.

[0024] In this application, the above preparation process uses a cyclic extrusion method to rapidly prepare a gel, and continuously embeds ethanol during this process, improving the dispersion uniformity and stability of ethanol and enhancing the embedding effect.

[0025] Optionally, the specific operations of the preparation method include:

[0026] Provide a first container and a second container;

[0027] Place the mixture A and the mixture B in the first container and the second container respectively;

[0028] Connect the first container and the second container to mix and react the mixture A and the mixture B to obtain the injectable gel composition.

[0029] Optionally, the first container and the second container are syringes; by cyclically extruding the first container and the second container, the mixture A and the mixture B are mixed and reacted.

[0030] Optionally, the number of cyclic extrusion times is 20 - 60 times.

[0031] Optionally, the cyclic extrusion time is 10 - 60 s. Preferably, the cyclic extrusion time is 10 - 30 s. More preferably, it is 10 - 20 s.

[0032] Optionally, in the mixture A (approximate or without considering the ablative agent), the concentration of the alkaline earth metal salt aqueous solution is 0.1 - 10 wt%.

[0033] Optionally, in the mixture B, the concentration of the gel precursor is 0.5 - 10 wt%.

[0034] Optionally, the mass ratio of the alkaline earth metal salt solution to the gel precursor solution is 1:1 - 10.

[0035] Preferably, the mass ratio of the alkaline earth metal salt solution to the gel precursor solution is 1:1 - 5. More preferably, it is 1:1 - 3.

[0036] This application also provides a preparation method of an injectable gel composition carrying an ablative agent, which is characterized by including the following steps:

[0037] Mix the alkaline earth metal salt, the ablative agent and water to obtain a mixture A;

[0038] Mix the gel precursor and part of the water to obtain a mixture B;

[0039] Mix and react the mixture A and the mixture B to prepare the injectable gel composition;

[0040] In the mixture A (approximate or without considering the ablative agent), the concentration of the alkaline earth metal salt aqueous solution is 0.1 - 10 wt%;

[0041] In the mixture B, the concentration of the gel precursor is 0.5-10 wt%.

[0042] The mass ratio of the alkaline earth metal salt solution to the gel precursor solution is 1:1-10.

[0043] The present application also provides the application of the injectable gel composition loaded with the ablative agent in the treatment of tumors. For example, it has potential application prospects in the treatment of benign and malignant tumors, prostatic hypertrophy, myocardial hypertrophy, abnormal thickening of the endometrium and other diseases. The injectable gel composition can solve the problems of incomplete ablation or over-ablation in the current tissue ablation treatment methods.

[0044] Compared with the prior art, the present application uses a gel with a three-dimensional network structure to embed a low concentration of ablative agent (ethanol) to prepare an injectable gel composition, which has excellent conformability, can be injected according to the shape characteristics of the tissue to be ablated, and is not easy to overflow and spread after being implanted into the tumor area, avoiding the adverse effects on the proliferation and growth of normal cells and tissues; moreover, by slowly releasing ethanol from the gel, the purpose of long-term ablation of tumor cells is achieved. Description of the Drawings

[0045] Figure 1 It is a rheological mechanical property diagram of the injectable gel composition in Example 1;

[0046] Figure 2 It is a rheological mechanical property diagram of the injectable gel composition in Example 2;

[0047] Figure 3 It is a SEM diagram of the injectable gel composition in Example 2;

[0048] Figure 4 It is a rheological mechanical property diagram of the injectable gel composition in Example 3;

[0049] Figure 5 It is a rheological mechanical property diagram of the injectable gel composition in Example 4;

[0050] Figure 6 It is a rheological mechanical property diagram of the injectable gel composition in Example 5;

[0051] Figure 7 It is a diagram showing the relationship between the ethanol release amount and time in Example 6;

[0052] Figure 8 It is a diagram showing the relationship between the release rate of ethanol corresponding to different ethanol concentrations and the injectable gel composition in Example 7. Detailed Embodiments

[0053] The following further describes the technical solutions of the present application in combination with specific embodiments, but the present application is not limited thereto.

[0054] The purpose of the present application is to provide an injectable gel composition carrying an ablative agent with good conformability to effectively ablate tumors with irregular shapes or at the renal hilum. To achieve this purpose, the solution adopted in the present application is as follows:

[0055] An injectable gel composition carrying an ablative agent is provided, including a gel and an ablative agent dispersed in the gel, wherein the ablative agent is ethanol and the concentration is 0.1-14 wt%.

[0056] In the scope of the present application, the term "conformability" refers to the ability of the injected substance to form a gel property suitable for the shape of the ablation site at the injection site according to the treatment requirements, mainly involving two aspects: one is that the injected substance has a certain fluidity at the initial injection stage in the body and can smoothly diffuse into the tumor tissue interstices. Therefore, it is applicable to tumors growing in certain parts such as irregularly shaped tumors in the liver, kidney and other parts; the other is that after entering the tumor tissue interstices, it can in-situ form a three-dimensional network structure with anti-deformation ability, so as to resist the in-situ pressure in the tissue, realize the controlled slow release of ethanol, and reduce the risk of ethanol overflowing to surrounding normal cells, thereby effectively ablating tumors with specific shapes.

[0057] The storage modulus is one of the indicators to measure the rheological mechanics of the injectable gel composition, and can reflect the fluidity and the magnitude of the anti-deformation ability of the gel composition. To ensure the conformability of the injectable gel composition, its storage modulus is required to be 300-10000 Pa. On the one hand, ethanol is embedded in the gel with this storage modulus and is not easily released quickly, achieving the effect of slow release; on the other hand, since the storage modulus of tumor tissue is positively correlated with the deterioration of tumor tissue, the storage modulus of the injectable gel composition should be lower than the modulus of tumor cells while ensuring a certain anti-deformation ability, so as to avoid serious deformation (or overgrowth) of tumor tissue and cause deterioration.

[0058] Generally, the storage modulus of most tumor cells is usually 3000-8000 Pa, and the storage modulus of tumor tissue is 58000-120000 Pa. Therefore, the storage modulus of the injectable gel composition is preferably 500-7000 Pa, more preferably 1000-3000 Pa.

[0059] In some embodiments, the release rate of the ablative agent in the injectable gel composition is 0.01-6 mg / h. In this application, the release rate is obtained from in vitro pharmacokinetic experiments, which are carried out in physiological saline to simulate the human physiological environment. Therefore, the addition amount of ethanol in the injectable gel composition and the release rate of ethanol in vitro have certain guiding significance for in vivo experiments. For example, the injectable gel composition with the best in vitro experimental effect can be directly applied to animal experiments or clinical experiments to save experimental costs.

[0060] For tumors of different sizes, the addition amount of ethanol in the injectable gel composition will be different to achieve a better ablation effect.

[0061] For example, for tumors with a volume of not less than 700 mm 3 ³, the concentration of ethanol is 7-14 wt%, and the ethanol sustained-release rate is 1-6 mg / h; for tumors with a volume less than 700 mm 3 ³, the concentration of ethanol is 0.1-7 wt%, and the ethanol sustained-release rate is 0.05-1 mg / h.

[0062] The gel used in this application has a three-dimensional network structure with a pore size of 10-100 μm. On the one hand, it can effectively entrap ethanol, and on the other hand, it can slowly release ethanol at the tumor site. In some preferred embodiments, the pore size of the gel is 10-60 μm.

[0063] In addition, the diameter of most tumor cells is 12-25 μm. To effectively inhibit the growth and proliferation of tumor cells, the pore size of the gel is more preferably 10-30 μm.

[0064] Based on the application scenario of the injectable gel composition in the living body, the gel used generally adopts a material with excellent biocompatibility. For example, in one embodiment, the gel is formed by cross-linking a gel precursor with an alkaline earth metal salt. The gel precursor can be at least one of alginate, oxidized alginate, alginate ester, silk fibroin, collagen, chitosan, carboxymethyl chitosan, cholate-modified chitosan, and dextran. The alkaline earth metal salt can be at least one of calcium nitrate, calcium carbonate, calcium chloride, magnesium chloride, barium chloride, barium sulfate, magnesium sulfate, calcium sulfate, calcium gluconate, calcium lactate, and calcium alginate; preferably at least one of calcium chloride, calcium gluconate, calcium lactate, and calcium alginate; more preferably calcium chloride, which has better water solubility, good dispersion, is easy to prepare and store, and is beneficial to preparing a gel with uniform texture.

[0065] In the field of tissue engineering applications, it is often required that tissue fillers do not affect normal biological functions, that is, it is usually necessary to maintain the generation and conduction of cell electrical signals. Therefore, the tissue filler (the gel of the present application) needs to have "biological inertness", and the "biological inertness" includes that the tissue filler lacks specific recognition sites for mammalian cells, has less protein adsorption, and has relatively weak biological activity and biological induction ability, etc.

[0066] The gel precursor is one of the key raw materials for preparing the gel and affects the "biological inertness" of the gel. For example, in one embodiment, the gel precursor is alginate, and the prepared gel will not disrupt cell electrical signal conduction in vivo and can avoid many adverse reactions and clinical adverse events.

[0067] The present application also provides a preparation method of an injectable gel composition loaded with an ablating agent, including: mixing an alkaline earth metal salt, an ablating agent, a gel precursor and water, and reacting to obtain the injectable gel composition.

[0068] In the above preparation method, the mixing method is as follows:

[0069] Mix the alkaline earth metal salt, the ablating agent and water to obtain a mixed solution A;

[0070] Mix the gel precursor and a part of water to obtain a mixed solution B;

[0071] Mix and react the mixed solution A and the mixed solution B to obtain the injectable gel composition.

[0072] In the present application, the above preparation process uses a cyclic extrusion method to quickly prepare the gel, and continuously embeds ethanol during this process to improve the dispersion uniformity and stability of ethanol and enhance the embedding effect.

[0073] For example, in one embodiment, the specific operations of the preparation method include:

[0074] Provide a first container and a second container;

[0075] Place the mixed solution A and the mixed solution B in the first container and the second container respectively;

[0076] Connect the first container and the second container to make the mixed solution A and the mixed solution B mix and react to obtain an injectable gel composition.

[0077] Among them, the first container and the second container are syringes; by cyclically extruding the first container and the second container, the mixed solution A and the mixed solution B are mixed and reacted.

[0078] The effect of the mixing reaction is related to the number of cyclic extrusion times. In this application, the number of cyclic extrusion times is 20 - 60 times, which can make the embedding rate of ethanol close to 100%. In addition, gel is continuously generated during the extrusion mixing process. Only by controlling the gel generation rate can a gel composition with good texture uniformity be obtained. Therefore, the cyclic extrusion time should not be too long or too short. If the time is too long, the uniformity of the prepared gel composition is poor; if the time is too short, the mixing reaction is insufficient. In this application, the cyclic extrusion time is 10 - 60 s; preferably 10 - 30 s; more preferably 10 - 20 s.

[0079] Regarding the effects of the gel precursor and metal salt on the gel pore size and storage modulus, it mainly includes the following two points:

[0080] 1. The gel precursor and metal salt form a three-dimensional network pore-like spatial structure. The larger the mass ratio of the two and the higher the metal salt concentration, the more compact the structure, the smaller the pore size, and the higher the storage modulus.

[0081] 2. By optimizing the mass ratio of the two, the storage modulus of the prepared gel is within the preferred range, close to the mechanical properties of the target tissue cells, with better conformability, a better ethanol sustained-release effect, and ultimately a better ablation effect of the hydrogel within this preferred range on tumor tissue.

[0082] To prepare the expected gel, in mixture A (approximate or without considering ethanol), the concentration of the alkaline earth metal salt aqueous solution is 0.1 - 10 wt%. In mixture B, the concentration of the gel precursor is 0.5 - 10 wt%.

[0083] In one embodiment, the mass ratio of the alkaline earth metal salt solution to the gel precursor solution is 1:1 - 10; preferably 1:1 - 5; more preferably 1:1 - 3.

[0084] Example 1

[0085] (1) Prepare an aqueous solution of CaCl with a concentration of 0.1 wt%, weigh 0.1 g of ethanol and mix it with 0.413 g of the CaCl aqueous solution to obtain mixture A, and transfer mixture A to a 3 mL syringe, denoted as syringe 1; 2 aqueous solution, weigh 0.1 g of ethanol and mix it with 0.413 g of the CaCl 2 aqueous solution to obtain mixture A, and transfer mixture A to a 3 mL syringe, denoted as syringe 1;

[0086] (2) Prepare a medical-grade sodium alginate with a concentration of 10 wt% to obtain solution B, and take 0.910 g of the sodium alginate solution and transfer it to a 3 mL syringe, denoted as syringe 2;

[0087] (3) Connect syringe 1 and syringe 2 using a three-way valve, and mix and crosslink mixture A and solution B by cyclic injection 60 times within 30 seconds to obtain an injectable gel composition.

[0088] The rheological mechanical properties of the injectable gel composition were measured using an MCR-302 rheometer. The test parameters were: 20 °C, a fixed frequency of 1 Hz, and a strain sweep of 0.01 - 100%.

[0089] The stress-strain sweep of the injectable gel composition prepared in this example is as Figure 1 shown. The storage modulus of the injectable gel composition in the linear viscoelastic region is 1800 Pa.

[0090] Example 2

[0091] (1) Prepare an aqueous solution of CaCl with a concentration of 0.1 wt%, weigh 0.08 g of ethanol and mix it with 0.413 g of the aqueous solution of CaCl 2 to obtain a mixed solution A. Transfer the mixed solution A to a 3 mL syringe and label it as syringe 1; 2

[0092] (2) Prepare a medical-grade sodium alginate solution with a concentration of 8 wt% to obtain solution B. Weigh 0.910 g of the medical-grade sodium alginate solution and transfer it to a 3 mL syringe, label it as syringe 2;

[0093] (3) Connect syringe 1 and syringe 2 using a three-way valve, and mix and crosslink the mixed solution A and solution B by cyclic injection 60 times within 30 seconds to obtain an injectable gel composition.

[0094] Refer to the method of Example 1 to measure the rheological mechanical properties of the injectable gel composition.

[0095] The stress-strain sweep of the injectable gel composition prepared in this example is as Figure 2 shown. The storage modulus of the injectable gel composition in the linear viscoelastic region is 2800 Pa.

[0096] The gel composition was observed using a scanning electron microscope. As Figure 3 shown, the prepared injectable gel composition has a three-dimensional network structure, the size distribution of the network pore diameters is uniform, and the pore diameter is 25 μm.

[0097] Example 3

[0098] (1) Prepare an aqueous solution of Ca(NO 3 ) 2 with a concentration of 0.5 wt%, weigh 0.04 g of ethanol and mix it with 0.413 g of the aqueous solution of Ca(NO 3 ) 2 to obtain a mixed solution A. Transfer it to a 3 mL syringe and label it as syringe 1;

[0099] (2) Weigh an appropriate amount of medical-grade sodium alginate and prepare a sodium alginate solution with a concentration of 1.0 wt%, obtaining Solution B. Take 0.910 g of Solution B and transfer it to a 3 mL syringe, denoted as Syringe 2;

[0100] (3) Connect Syringe 1 and Syringe 2 using a three-way valve, and mix and crosslink Mixture A and Solution B by performing 20 cycles of injection within 30 seconds to obtain an injectable gel composition.

[0101] Measure the rheological and mechanical properties of the injectable gel composition with reference to the method of Example 1.

[0102] The stress-strain scan of the injectable gel composition prepared in this example is as Figure 4 shown. The storage modulus of the injectable gel composition within the linear viscoelastic region is 982 Pa.

[0103] Example 4

[0104] (1) Prepare an aqueous solution of CaSO with a concentration of 3.0 wt%, weigh 0.2 g of ethanol and mix it with 0.413 g of the aqueous solution of CaSO to obtain Mixture A, and transfer Solution A to a 3 mL syringe, denoted as Syringe 1; 4 aqueous solution, weigh 0.2 g of ethanol and mix it with 0.413 g of the aqueous solution of CaSO 4 aqueous solution to obtain Mixture A, and transfer Solution A to a 3 mL syringe, denoted as Syringe 1;

[0105] (2) Weigh an appropriate amount of medical-grade sodium alginate and dissolve it in distilled water to prepare a solution with a concentration of 5.0 wt%. Take 0.910 g of the solution and transfer it to a 3 mL syringe, denoted as Syringe 2;

[0106] (3) Connect Syringe 1 and Syringe 2 using a three-way valve, and mix and crosslink Mixture A and Solution B by performing 60 cycles of injection within 30 seconds to obtain a gel composition.

[0107] Measure the rheological and mechanical properties of the injectable gel composition with reference to the method of Example 1.

[0108] The stress-strain scan of the injectable gel composition prepared in this example is as Figure 5 shown. The storage modulus of the injectable gel composition within the linear viscoelastic region is 2100 Pa.

[0109] Example 5

[0110] (1) Prepare an aqueous solution of CaCO with a concentration of 8.0 wt%, weigh 0.08 g of ethanol and mix it with 0.413 g of the aqueous solution of CaCO to obtain Mixture A, and transfer the mixture to a 3 mL syringe, denoted as Syringe 1; 3 aqueous solution, weigh 0.08 g of ethanol and mix it with 0.413 g of the aqueous solution of CaCO 3 aqueous solution to obtain Mixture A, and transfer the mixture to a 3 mL syringe, denoted as Syringe 1;

[0111] (2) Weigh an appropriate amount of medical-grade sodium alginate and dissolve it in distilled water to prepare Solution B with a concentration of 5.0 wt%. Take 0.910 g of Solution B and transfer it to a 3 mL syringe, denoted as Syringe 2;

[0112] (3) Connect Syringe 1 and Syringe 2 using a three-way valve, and mix and crosslink the mixture A and Solution B by cyclic injection 40 times within 30 seconds to obtain a gel composition.

[0113] Refer to the method of Example 1 to measure the rheological and mechanical properties of the injectable gel composition.

[0114] The stress-strain scan of the injectable gel composition prepared in this example is as Figure 5 shown. The storage modulus of the injectable gel composition within the linear viscoelastic region is 2800 Pa.

[0115] Comparative Example 1

[0116] The components of the mixture are ethanol, calcium chloride solution, and sodium alginate gel solution, with the same amounts as in Example 1. The mixing method of the gel composition is magnetic stirring, with a stirring speed of 1500 rpm and a temperature of room temperature.

[0117] The following method is used to measure the encapsulation efficiency of ethanol in Examples 1 to 5 and Comparative Example 1. The specific steps are as follows:

[0118] First, place the prepared gel composition in a 15 mL centrifuge tube containing physiological saline. Part of the ethanol will be released into the physiological saline, and after 5 days, samples are taken to detect the ethanol content in the physiological saline.

[0119] Analyze using an Aligent6890N gas chromatograph, detect using an FID detector, and measure the ethanol content using the external standard method.

[0120] Calculation method or formula for encapsulation efficiency:

[0121] The encapsulation efficiency refers to the percentage of the encapsulated substance (such as a certain drug) in the suspension relative to the total amount of the drug. It is commonly expressed as the percentage encapsulation efficiency (Encapsulation percentage, EN%), and EN% = (1 - Cf / Ct) × 100%. In the formula, Cf is the amount of free drug; Ct is the total amount of drug in the suspension.

[0122] The injectable gel compositions prepared by the in-situ embedding method in Examples 1 to 5 of this application can completely encapsulate ethanol in the gel composition. Among them, the encapsulation efficiency of ethanol in Example 1 is 100%, while the encapsulation efficiency of ethanol in Comparative Example 1 is 53%. There is a difference in encapsulation efficiency. See Table 1.

[0123] Table 1 Encapsulation rates of ethanol in Example 1 and Comparative Example 1

[0124]

[0125] Example 6 Relationship between Ethanol Release Amount and Time

[0126] (1) Prepare an aqueous solution of CaCl with a concentration of 2 wt%, weigh 0.2 g of ethanol and mix it with 0.413 g of the CaCl 2 aqueous solution to obtain a mixed solution A. Transfer the mixed solution A to a 3 mL syringe, denoted as syringe 1; 2

[0127] (2) Weigh an appropriate amount of medical-grade sodium alginate and dissolve it in distilled water to prepare a solution B with a concentration of 0.5 wt%. Take 0.910 g of solution B and transfer it to a 3 mL syringe, denoted as syringe 2;

[0128] (3) Connect syringe 1 and syringe 2 using a three-way valve, and mix and crosslink the mixed solution A and solution B by 30 cycles of injection to obtain a gel composition.

[0129] Inject the gel composition into a centrifuge tube containing 20 mL of normal saline. Take samples every 2 h within 24 h and measure the ethanol content, and draw a relationship diagram of ethanol content vs. time, see Figure 7 .

[0130] By simulating the release of ethanol from the injectable gel composition under physiological conditions in vitro, it can be seen from Figure 7 the results that the injectable gel composition rapidly releases ethanol within the first 5 h, and then the ethanol release amount gradually reaches a dynamic equilibrium and maintains at a certain release level.

[0131] Example 7 Ethanol Mass Concentration and Ethanol Release Rate

[0132] The preparation method of Example 7 is the same as that of Example 1. Measure the sustained release rate corresponding to different mass concentrations of ethanol in each group, and the results are shown in Table 2.

[0133] Table 2 Ethanol Mass Concentration and Corresponding Ethanol Release Rate

[0134] Mass fraction of ethanol (%) Ethanol sustained-release rate (mg / h) 0.75019 0.05 1.4892 0.1125 2.9347 0.225 4.33839 0.2875 5.70207 0.5625 7.03 4.075 13.13198 1.2

[0135] Figure 8 shows the influence of in vitro simulated ethanol concentration on the release of ethanol from the injectable gel composition under physiological conditions. It can be seen from Figure 8 the results that as the concentration of ethanol in the injectable gel composition increases, the release rate of ethanol from the composition first increases and then decreases.

[0136] Test Example 1 In Vitro Tumor Cell Experiment

[0137] ​In this experiment, the human hepatoma cell line (HepG2) was selected. A negative control group (with normal saline as the negative control) and a drug group (containing ethanol) were set up. The CCK8 colorimetric method was used to measure the inhibitory activity of the drug against tumor cells. The absorbance value (A490) of each well was measured at a wavelength of 490 nm on an enzyme-linked immunosorbent assay detector. The inhibition rate of the drug on the growth of tumor cells was calculated according to the following formula, and the results of the experiment are listed in Table 3.

[0138] Inhibition rate of tumor cell growth (IR) = (1 - A490 of drug group / A490 of control group) × 100%.

[0139] Table 3 Inhibition rate of tumor cells

[0140]

[0141] The experimental results showed that the 24-hour and 48-hour tumor cell inhibition rates of the ethanol-alginate gel composition were higher than those of the ethanol group, indicating that ethanol in the concentration range of 0.1 - 14 wt% could more effectively inhibit the growth of tumor cells by slow release in vivo through gel embedding.

[0142] Test Example 2 In vivo tumor inhibition test

[0143] Five-week-old severe combined immunodeficiency nude mice were used in the experiment. After the nude mice were purchased, they were raised in the laboratory for 1 - 2 weeks to allow their bodies to adapt to the environment and reduce uncontrollable errors. Each nude mouse was inoculated with 2 × 10 6 MCF-7 cells (human breast cancer cells). When the volume of the tumor grew to 700 mm 3 it was recorded as the first day. The formula for calculating the tumor volume was V = d 2 × D / 2, where d represents the shortest diameter of the tumor and D represents the longest diameter of the tumor, with the unit of mm. The qualified tumor-bearing nude mice were divided into 3 groups, with 5 mice in each group (n = 5). The 3 groups were the normal saline group as the control group, the pure ethanol injection group (0.1 g), and the injectable gel composition injection group (ethanol concentration of 7.03 wt%). The injection times were 0, 3, 6, 9, 12, 15, 18, and 20 days. Each time, the body weight (mg), D, and d of the nude mice were recorded as the basis for the anti-tumor effect of the drug. Finally, the tumors were removed and photographed.

[0144] The test results showed that the order of the tumor volume size was: control group > pure ethanol injection group > injectable gel composition injection group.

[0145] The above description of the embodiments is to enable those of ordinary skill in the art to understand and use the present application. Those skilled in the art to which the present application pertains may also make appropriate changes and modifications to the above-described embodiments. Therefore, the present application is not limited to the specific embodiments disclosed and described above, and some modifications and changes to the present application should also fall within the scope of protection of the claims of the present application.

Claims

1. Injectable gel composition containing an ablating agent, characterized in that, it comprises a gel and an ablating agent dispersed in the gel, the ablating agent being ethanol and having a concentration of 0.1-14 wt%; the gel has a three-dimensional network structure with a pore size of 10-100 μm; The preparation method of the injectable gel composition containing an ablating agent comprises the following steps: Mix an alkaline earth metal salt, an ablating agent, a gel precursor and water, and react to obtain the injectable gel composition; The mixing method is: Mix the alkaline earth metal salt, the ablating agent and water to obtain a mixed solution A; Mix the gel precursor and part of the water to obtain a mixed solution B; Mix and react the mixed solution A and the mixed solution B to obtain the injectable gel composition; The gel precursor is sodium alginate.

2. The injectable gel composition containing an ablating agent according to claim 1, characterized in that, the storage modulus of the injectable gel composition is 300-10000 Pa.

3. The injectable gel composition containing an ablating agent according to claim 2, characterized in that, the storage modulus of the injectable gel composition is 500-7000 Pa.

4. The injectable gel composition containing an ablating agent according to claim 3, characterized in that, the storage modulus of the injectable gel composition is 1000-3000 Pa.

5. The injectable gel composition containing an ablating agent according to claim 1, characterized in that, the release rate of the ablating agent is 0.01-6 mg / h.

6. The injectable gel composition containing an ablating agent according to claim 1, characterized in that, the pore size of the gel is 10-60 μm.

7. The injectable gel composition containing an ablating agent according to claim 6, characterized in that, the pore size of the gel is 10-30 μm.

8. The injectable gel composition containing an ablating agent according to claim 1, characterized in that, the gel is formed by cross-linking a gel precursor and an alkaline earth metal salt; the alkaline earth metal salt is at least one of calcium nitrate, calcium carbonate, calcium chloride, magnesium chloride, barium chloride, barium sulfate, magnesium sulfate, calcium sulfate, calcium gluconate, calcium lactate and calcium alginate.

9. The injectable gel composition containing an ablating agent according to claim 1, characterized in that, the specific operation of the preparation method is: Provide a first container and a second container; Place the mixed solution A and the mixed solution B in the first container and the second container respectively; Connect the first container and the second container, and mix and react the mixed solution A and the mixed solution B to obtain the injectable gel composition.

10. The injectable gel composition containing an ablating agent according to claim 9, characterized in that, the first container and the second container are syringes, and by cyclically pushing the first container and the second container, the mixed solution A and the mixed solution B are mixed and reacted.

11. The injectable gel composition containing an ablating agent according to claim 1, characterized in that, in the mixed solution A, the concentration of the alkaline earth metal salt aqueous solution is 0.1-10 wt%; In the mixture B, the concentration of the gel precursor is 0.5 to 10 wt%.

12. The injectable gel composition carrying an ablative agent according to claim 1, wherein, the mass ratio of the alkaline earth metal salt solution to the gel precursor solution is 1:1 to 10.

13. A method for preparing an injectable gel composition carrying an ablative agent, wherein, it includes the following steps: Mix an alkaline earth metal salt, an ablative agent and water to obtain a mixture A; Mix the gel precursor and a part of water to obtain a mixture B; Mix and react the mixture A and the mixture B to prepare the injectable gel composition; In the mixture A, the concentration of the alkaline earth metal salt aqueous solution is 0.1-10 wt%; In the mixture B, the concentration of the gel precursor is 0.5 to 10 wt%; the mass ratio of the alkaline earth metal salt solution to the gel precursor solution is 1:1 to 10; the ablative agent is ethanol and the concentration is 0.1 to 14 wt%; the gel has a three-dimensional network structure and its pore size is 10-100 μm.

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