Composite condensing gel based on photodynamic therapy and preparation method thereof

Abstract

The invention relates to the technical field of pharmaceutical preparations, in particular to composite condensing gel based on photodynamic therapy and a preparation method of the composite condensing gel. The preparation method of the composite condensate gel based on photodynamic therapy comprises the following steps: preparing a BSA (Bovine Serum Albumin) solution, uniformly mixing an inulin mother solution with the BSA solution, adjusting the pH value, heating in a water bath, cooling to room temperature, adding 5-aminolevulinic acid methyl ester hydrochloride into the condensate gel, adding an acid coagulant, diluting, and filling CO2 gas. According to the invention, CO2 is added into the composite condensation gel, so that the binding force of hemoglobin and O2 in tissues absorbing the composite condensation gel is reduced, and the concentration of free oxygen is improved, so that the composite condensation gel can transfer energy to surrounding oxygen and generate singlet oxygen with strong activity, thereby killing cancer cells and achieving the effect of improving the curative effect of photodynamic therapy.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of composite cold gel, in particular to a composite cold gel based on photodynamic therapy and a preparation method thereof. BACKGROUND

[0002] Photodynamic therapy is a new medical method for treating tumors, precancerous lesions, and proliferative skin diseases by using photosensitizing drugs and laser activation. By using specific wavelength irradiation of the lesion site, the photosensitizing drug selectively accumulated in the lesion tissue can be activated to initiate photochemical reactions to destroy the lesion. Clinically used photodynamic therapies include aminolevulinic acid photodynamic therapy, heimoporfin photodynamic therapy, and hematoporphytin monophotodynamic therapy. Compared with traditional therapies, photodynamic therapy has the advantages of precise and effective treatment with minimal side effects.

[0003] Aminolevulinic acid photodynamic therapy generally involves adding 5-aminolevulinic acid to a cold gel as a photosensitizer. Irradiation with specific wavelength laser excites the photosensitizer absorbed by the tissue, and the excited-state photosensitizer transfers energy to the surrounding oxygen to generate highly active singlet oxygen. The singlet oxygen and adjacent biological macromolecules undergo oxidation reactions, producing cytotoxic effects and killing cancer cells. However, the hypoxic environment of tumors can lead to low singlet oxygen concentration and insufficient oxidation reactions, affecting the efficacy of photodynamic therapy. Therefore, tumor hypoxia is a recognized technical challenge that limits the efficacy of photodynamic therapy. Traditional technical solutions focus only on developing new photosensitizers or oxygenation strategies, which do not fundamentally address the drawbacks of existing technologies, resulting in insignificant efficacy of photodynamic therapy. SUMMARY

[0004] In view of the above-mentioned deficiencies of the prior art, the present application aims to provide a composite cold gel based on photodynamic therapy and a preparation method thereof.

[0005] To achieve the above-mentioned purpose, the present application provides a composite cold gel based on photodynamic therapy, comprising 1-2 parts of bovine serum albumin solution, 2-3 parts of inulin mother liquor, 0.5-1.5 parts of 5-aminoacetylpropionic acid methyl ester hydrochloride solution, 2-3 parts of xanthan gum solution, 1-2 parts of acid coagulant, and an appropriate amount of CO2 gas. The acid coagulant is glucono-delta-lactone. The concentration of the bovine serum albumin solution is 7-10%, the concentration of the inulin mother liquor is 1-5%, and the solubility concentration of the acid coagulant is 1.5-2.5%.

[0006] Compared with the prior art, the application adds CO2 gas into the cold gel, because the inventor finds that by loading CO2 into the cold gel for local administration, the free oxygen concentration at the lesion site can be significantly increased through a unique physiological mechanism of reducing the affinity of hemoglobin to oxygen (Bohr effect), thereby reversing the inhibition of photodynamic therapy by the hypoxic microenvironment of the tumor. This technical solution is not conventional in the field of photodynamic therapy and cannot be deduced from commonly known techniques, and unexpected technical effects have been achieved.

[0007] The second aspect of the application provides a preparation method of a composite cold gel based on photodynamic therapy, specifically comprising the following steps: S1: mix inositol mother liquor and BSA solution and adjust ph 2-3 parts of inositol mother liquor are added to the BSA solution, and slowly stirred for 10-30 min to mix the inositol mother liquor and the BSA solution, and at this time, the sensor senses that the gravity exceeds the threshold, and the valve of the NaOH solution storage box is opened; The NaOH solution storage box starts to drop NaOH solution into the stirring box, and after 0.8-1.5 parts of NaOH solution are added to the stirrer, the ph is adjusted to 6-7; When the acid-base detector detects that the ph in the stirrer is between 6-7, the valve is closed, and the stirrer is closed, to obtain a BSA-inositol composite solution; S2: heat in a water bath and cool to room temperature The BSA-inositol solution is sealed in a 20-30ml white cap bottle, heated in a 80-90℃ water bath for 15-25min, and cooled with ice water to room temperature to obtain a BSA-inositol composite gel; S3: add 5-amino levulinic acid methyl ester hydrochloride to the composite gel 0.5-1.5 parts of 5-amino levulinic acid methyl ester hydrochloride solution are weighed and added to a centrifugal stirrer, and then the BSA-inositol composite gel is added to the centrifugal stirrer, and the 5-amino levulinic acid methyl ester hydrochloride solution and the BSA-inositol composite gel are stirred for 5-15 min by the centrifugal stirrer to dissolve, to obtain a mixed solution; The xanthan gum solution is swelled by adding a diluent to the xanthan gum solution, and the concentration of the xanthan gum solution is adjusted to 1.5-2%; The xanthan gum solution and the mixed solution are added to the centrifugal stirrer in a ratio of 1:2-3, and stirred for 2-5 min to mix; S4: add an acid coagulant and dilute An acid coagulant with a dissolved concentration of 1.5-2.5% and 0.2-0.4 parts of a diluent are added to the centrifugal stirrer, and stirred for 4h to obtain a cold gel; S5: fill in CO2 gas The cold gel is placed in an air-cooled cooling environment box at 0-10 DEG C, and the gas in the environment box is extracted to a low pressure environment, and the cold gel is solidified into a solid state under low temperature and low pressure environment, and then CO2 gas is introduced into the cold gel for 20-30 min, and then the environment box is placed at room temperature to obtain the BSA-inulin composite cold gel co-loaded with 5-amino levulinic acid methyl ester and CO2.

[0008] Further, the diluent is stored in a diluent storage box, and the diluent is added into the storage container storing the xanthan gum solution by adjusting the two-way valve at the discharge port of the diluent storage box, and the xanthan gum is swelled, and then in the step S4, the diluent is injected into the centrifugal stirrer through the pipeline by reversely adjusting the two-way valve of the storage container, and the solution in the centrifugal stirrer is diluted, and then the centrifugal stirring is performed.

[0009] Compared with the prior art, the application reduces the binding force of hemoglobin in the tissue absorbing the composite cold gel to O2, increases the concentration of free oxygen, enables the composite cold gel to transfer energy to the surrounding oxygen to generate singlet oxygen with strong activity, and thus kills cancer cells, thereby improving the curative effect of photodynamic therapy.

[0010] Meanwhile, the application adds the inulin solution into the cold gel when preparing the composite cold gel, so that the state of the cold gel is more stable, and the cold gel is convenient to freeze and add CO2, thereby improving the curative effect of photodynamic therapy. BRIEF DESCRIPTION OF DRAWINGS

[0011] Figure 1 The preparation method flow chart of the composite cold gel based on photodynamic therapy used in the embodiments of the application. DETAILED DESCRIPTION

[0012] The technical solutions in the embodiments of the application will be clearly and completely described below with reference to the drawings in the embodiments of the application. Obviously, the described embodiments are only part of the embodiments of the application, rather than all the embodiments of the application. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative work fall within the protection scope of the application. Embodiment 1

[0013] A composite cold gel based on photodynamic therapy comprises 1 part of a bovine serum albumin solution, 2 parts of an inulin mother liquor, 1 part of a 5-amino levulinic acid methyl ester hydrochloride solution, 3 parts of a xanthan gum solution, 1 part of an acid coagulant, and appropriate CO2 gas, wherein the acid coagulant is glucono-delta-lactone, and the mass fraction of the bovine serum albumin solution is 8%, and the concentration of the inulin mother liquor is 2%.

[0014] A preparation method of a composite cold gel based on photodynamic therapy, as shown in Figure 1 Specifically comprises the following steps: S1: Mix inositol mother liquor and BSA solution, adjust ph Add 2 parts of inositol mother liquor to the BSA solution, slowly stir by the blender at 200 r / min for 20 min, mix the inositol mother liquor and the BSA solution, at this time the solution is acidic, the sensor senses that the gravity exceeds the threshold, the valve of the NaOH solution storage box is opened, the NaOH solution is used to adjust the pH of the solution, and the sodium ion is a common ion in human tissues, the introduction of sodium ion will not affect the effect of the composite cold gel; The NaOH solution storage box starts to add NaOH solution into the stirring tank, after adding 1 part of NaOH solution into the blender, the ph is adjusted to 7; When the acid-base detector detects that the ph in the blender is between 6 and 7, the valve is closed, and the stirrer is closed, to obtain a BSA-inositol composite solution, which can accurately adjust the ph of the solution, making it easier to obtain a cold gel in the subsequent steps.

[0015] S2: After water bath heating, cool to room temperature Seal the BSA-inositol solution in a 20ml white cap bottle. Heat in a 90℃ water bath for 20 min, cool to room temperature with ice water, and obtain a BSA-inositol composite gel.

[0016] S3: Add 5-amino levulinic acid methyl ester hydrochloride to the composite gel Weigh 1 part of 5-amino levulinic acid methyl ester hydrochloride solution into a centrifugal stirrer, then add BSA-inositol composite gel to the centrifugal stirrer, stir the 5-amino levulinic acid methyl ester hydrochloride solution and BSA-inositol composite gel for 10 min to dissolve, and obtain a mixed solution; Swelling of xanthan gum solution by adding NaCl solution to xanthan gum solution, adjusting the concentration of xanthan gum solution to 1.5%; Add the xanthan gum solution and the mixed solution to the centrifugal stirrer in a 1:3 ratio, and mix well by centrifugal stirring for 3 min.

[0017] S4: Add acid coagulant and dilute To the centrifugal stirrer, 1 part of glucose acid-delta-lactone solution with a dissolution concentration of 2% and 0.2 part of NaCl solution are added, centrifugal stirring is performed for 4 h to obtain the cold gel, and the NaCl solution is stored in the diluent storage box. In step 4.2, the NaCl solution is added to the storage container storing the xanthan gum solution by adjusting the two-way valve at the discharge port of the diluent storage box to swell the xanthan gum, and then in step S5, the NaCl solution is injected into the centrifugal stirrer through the pipeline by reversely adjusting the two-way valve of the storage container to dilute the solution in the centrifugal stirrer, followed by centrifugal stirring for 4 h to obtain the cold gel.

[0018] S5: CO2 gas is filled Specifically, the cold gel is placed in an air-cooled cooling environment box to 5℃, and then the gas in the environment box is extracted to a low-pressure environment. Under the low-temperature and low-pressure environment, the cold gel is solidified into a solid state, at this time, the cold gel has many voids. Then CO2 gas is introduced into the cold gel for 30 min. The CO2 gas is filled into the voids of the cold gel. Then the environment box is placed at room temperature to obtain the BSA-inulin composite cold gel co-loaded with 5-amino levulinic acid methyl ester and CO2. Example 2

[0019] A composite cold gel based on photodynamic therapy, comprising 2 parts of bovine serum albumin solution, 3 parts of inulin mother liquor, 0.5 parts of 5-amino levulinic acid methyl ester hydrochloride solution, 2 parts of xanthan gum solution, 1 part of acid coagulant and appropriate amount of CO2 gas; wherein the acid coagulant is glucose acid-delta-lactone, the mass fraction of the bovine serum albumin solution is 8%, and the concentration of the inulin mother liquor is 2%.

[0020] A preparation method of a composite cold gel based on photodynamic therapy, as shown in Figure 1 specifically comprising the following steps: S1: mix inulin mother liquor and BSA solution and adjust ph 3 parts of inulin mother liquor are added to the BSA solution, and the inulin mother liquor and the BSA solution are mixed uniformly by slowly stirring with a stirrer at 200 r / min for 20 min. At this time, the solution is acidic. When the sensor senses that the gravity exceeds the threshold value, the valve of the NaOH solution storage box is opened. The NaOH solution is used to adjust the pH of the solution, and the introduction of sodium ions will not affect the effect of the composite cold gel because sodium ions are common ions in human tissues. The NaOH solution storage box starts to add NaOH solution into the stirring tank. After 1 part of NaOH solution is added into the stirrer, the ph is adjusted to 7. When the acid-base detector detects that the ph in the stirrer is between 6 and 7, the valve is closed, and the stirrer is closed to obtain the BSA-inulin composite solution. In this way, the ph value of the solution can be accurately adjusted, so that the solution is more easily obtained in the subsequent steps.

[0021] S2: cooling to room temperature after water bath heating The BSA-inulin solution was sealed in a 20ml white cap bottle. After heating in a 90℃ water bath for 20min, the solution was cooled to room temperature with ice water to obtain a BSA-inulin complex gel.

[0022] S3: adding 5-amino levulinic acid methyl ester hydrochloride to the complex gel 0.5 parts of 5-amino levulinic acid methyl ester hydrochloride solution was weighed into a centrifugal stirrer, and then the BSA-inulin complex gel was added into the centrifugal stirrer. The 5-amino levulinic acid methyl ester hydrochloride solution and the BSA-inulin complex gel were stirred for 10min by the centrifugal stirrer to dissolve, to obtain a mixed solution. The xanthan gum solution was swelled by adding the NaCl solution to the xanthan gum solution, and the concentration of the xanthan gum solution was adjusted to 1.5%. The xanthan gum solution and the mixed solution were added into the centrifugal stirrer at a ratio of 1:2, and then mixed uniformly by centrifugal stirring for 3min.

[0023] S4: adding an acid coagulant and diluting 1 part of a 2% dissolved concentration of glucono-delta-lactone and 0.2 parts of a NaCl solution were added into the centrifugal stirrer, and then centrifugal stirring was performed for 4h to obtain a cold gel. The NaCl solution was stored in a diluent storage box. In step 4.2, the NaCl solution was added into a storage container storing the xanthan gum solution by adjusting a two-way valve at the discharge port of the diluent storage box to swell the xanthan gum. Then, in step S5, the NaCl solution was injected into the centrifugal stirrer through a pipeline by reversely adjusting the two-way valve of the storage container to dilute the solution in the centrifugal stirrer, and then centrifugal stirring was performed for 4h to obtain a cold gel.

[0024] S5: filling with CO2 gas Specifically, the cold gel was placed in an environmental box cooled to 5℃ by air cooling, and then the gas in the environmental box was extracted to a low-pressure environment. Under the low-temperature and low-pressure environment, the cold gel was coagulated into a solid state, and at this time, there were many voids in the cold gel. Then, CO2 gas was introduced into the cold gel for 30min, and the CO2 gas was filled into the voids of the cold gel. Then, the environmental box was placed at room temperature to obtain a BSA-inulin complex cold gel co-loaded with 5-amino levulinic acid methyl ester and CO2.

[0025] The above examples are only illustrative of the principles and effects of the present application, and are not intended to limit the present application. Any person skilled in the art can modify or change the above examples without departing from the spirit and scope of the present application. Therefore, all equivalent modifications or changes made by those skilled in the art without departing from the spirit and technical thought disclosed by the present application should be covered by the claims of the present application.

Claims

1. A composite cold gel based on photodynamic therapy, characterized in that, The BSA solution, 2-3 parts of inulin mother liquor, 0.5-1.5 parts of 5-aminolevulinic acid methyl ester hydrochloride solution, 2-3 parts of xanthan gum solution, 1-2 parts of acid coagulant and a proper amount of CO2 gas; wherein the acid coagulant is glucono-delta-lactone; The concentration of the BSA solution is 7-10%, the concentration of the inulin mother liquor is 1-5%, and the dissolved concentration of the acid coagulant is 1.5-2.5%.

2. A method for preparing a composite cold gel based on photodynamic therapy, characterized by, Specifically comprising the following steps: S1: Mix the inulin mother liquor and the BSA solution, and adjust the ph 2-3 parts of inulin mother liquor are added to the BSA solution, and the inulin mother liquor and the BSA solution are mixed uniformly at 150-300 r / min for 10-30 min. At this time, the inductor senses that the gravity exceeds the threshold, and the valve of the NaOH solution storage box is opened. The NaOH solution storage box starts to add NaOH solution into the stirring box. After 0.8-1.5 parts of NaOH solution are added into the stirrer, the ph is adjusted to 6-7. When the acid-base detector detects that the ph in the stirrer is between 6-7, the valve is closed, and the stirrer is turned off, obtaining a BSA-inulin composite solution. S2: After water bath heating, cool to room temperature The BSA-inulin solution is sealed in a 20-30 ml white cap bottle, heated in a 80-90℃ water bath for 15-25 min, and cooled with ice water to room temperature, obtaining a BSA-inulin composite gel. S3: Add 5-aminolevulinic acid methyl ester hydrochloride to the composite gel 0.5-1.5 parts of 5-aminolevulinic acid methyl ester hydrochloride solution is weighed and added to the centrifugal stirrer, and then the BSA-inulin composite gel is added to the centrifugal stirrer. The 5-aminolevulinic acid methyl ester hydrochloride solution and the BSA-inulin composite gel are stirred for 5-15 min by the centrifugal stirrer to dissolve, obtaining a mixed solution. The xanthan gum solution is swelled by adding a diluent to the xanthan gum solution, and the concentration of the xanthan gum solution is adjusted to 1.5-2%. The xanthan gum solution and the mixed solution are added to the centrifugal stirrer in a ratio of 1:2-3, and are mixed uniformly by centrifugal stirring for 2-5 min. S4: Add acid coagulant and diluent An acid coagulant with a dissolved concentration of 1.5-2.5% and 0.2-0.4 parts of diluent are added to the centrifugal stirrer, and are centrifugally stirred for 4 h, obtaining a cold gel. S5: Fill CO2 gas The cold gel is placed in an environmental box cooled by air cooling to 0-10℃, and then the gas in the environmental box is extracted. The gas is extracted to a low pressure environment. Under the condition of low temperature and low pressure, the cold gel is solidified into a solid state. Then CO2 gas is introduced into the cold gel for 20-30 min. Subsequently, the environmental box is placed at room temperature, obtaining a BSA-inulin composite cold gel co-loaded with 5-aminolevulinic acid methyl ester and CO2.

3. The method for preparing a composite cryogel based on photodynamic therapy according to claim 2, characterized in that, The diluent is stored in a diluent storage box. By adjusting the two-way valve at the discharge port of the diluent storage box, the diluent is added to the storage container storing the xanthan gum solution to swell the xanthan gum. Then in step S4, by reversely adjusting the two-way valve of the storage container, the diluent is injected into the centrifugal stirrer through the pipeline to dilute the solution in the centrifugal stirrer, and then the centrifugal stirring is performed.

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