CN120053755A

CN120053755A - Preparation method of facial filler

Info

Publication number: CN120053755A
Application number: CN202510525166.3A
Authority: CN
Inventors: 沈延臻; 张程皓; 张芳; 孙首禹; 田晓雷
Original assignee: Shandong Tongqi Medical Technology Co ltd
Current assignee: Shandong Tongqi Medical Technology Co ltd
Priority date: 2025-04-25
Filing date: 2025-04-25
Publication date: 2025-05-30

Abstract

本发明属于生物医疗技术领域，具体涉及面部填充剂的制备方法。包括以下步骤：（1）将琼脂糖与第一部分磷酸盐缓冲液混合，搅拌至琼脂糖完全溶解，降温得到琼脂糖凝胶；（2）将核苷酸类物质与第二部分磷酸盐缓冲液混合，得到核苷酸类物质溶液；（3）将盐酸利多卡因与第三部分磷酸盐缓冲液混合，得到盐酸利多卡因溶液；（4）将琼脂糖凝胶经筛网制粒后，与核苷酸类物质溶液和盐酸利多卡因溶液混合，经脱泡、灌装、辐照灭菌，得到面部填充剂。所述的制备方法，通过整合琼脂糖的逾渗阈值响应特性、核苷酸类物质的生物活性功能及盐酸利多卡因的麻醉效能，结合科学配比与工艺优化，形成了兼具安全性、功能性和临床实用性的创新型医美填充剂。The present invention belongs to the field of biomedical technology, and specifically relates to a method for preparing a facial filler. The method comprises the following steps: (1) mixing agarose with a first part of phosphate buffer, stirring until the agarose is completely dissolved, and cooling to obtain an agarose gel; (2) mixing a nucleotide substance with a second part of phosphate buffer to obtain a nucleotide substance solution; (3) mixing lidocaine hydrochloride with a third part of phosphate buffer to obtain a lidocaine hydrochloride solution; (4) granulating the agarose gel through a sieve, mixing it with a nucleotide substance solution and a lidocaine hydrochloride solution, degassing, filling, and irradiation sterilization to obtain a facial filler. The preparation method, by integrating the percolation threshold response characteristics of agarose, the biological activity function of nucleotide substances, and the anesthetic efficacy of lidocaine hydrochloride, combines scientific proportioning and process optimization, to form an innovative medical beauty filler with both safety, functionality, and clinical practicality.

Description

Preparation method of facial filler

Technical Field

The invention belongs to the technical field of biomedical treatment, and particularly relates to a preparation method of a facial filler.

Background

Reducing the risk of embolism is a critical safety concern in medical filler injection. Agarose fillers, because of their unique physicochemical properties, present certain advantages in reducing the risk of embolism. Agarose, a natural polysaccharide extracted from seaweed, has a high degree of biocompatibility and can be slowly degraded by the human body, usually for 6-24 months, which greatly reduces the risk of long-term foreign body reactions. The "percolation threshold response characteristic" of agarose gel is especially unique, and its physical state can show remarkable phase change behavior with the change of concentration. When the concentration exceeds a critical value (percolation threshold), the intermolecular crosslinking is enhanced to form a stable three-dimensional network structure, which shows the characteristics of solid gel, and when the concentration is lower than the threshold, the crosslinked network is dissociated, undergoes phase transition, and is converted into a free-flowing liquid state. Based on the dynamic reversible phase change mechanism, when the embolism complication is treated, the gel system is diluted by partially filling sufficient physiological saline, so that the environment concentration breaks through the percolation threshold critical point, thereby promoting the gel network of the embolism part to depolymerize and disperse, and realizing non-invasive embolism clearance.

In addition, the nucleotide (PDRN/PN) as another key component contains deoxyribonucleotide fragments (extracted from salmon DNA, for example) that activate intracellular adenosine A2A receptors, stimulate proliferation of fibroblasts, and accelerate repair of damaged tissues. The process continuously promotes the synthesis of type I collagen, type III collagen and elastin, effectively improves the elasticity and compactness of the skin, and the effect is usually remarkably shown after 2-3 months and can be maintained for 6-12 months. Meanwhile, the three-dimensional support structure of the PDRN/PN also provides space for regeneration of extracellular matrix, and can temporarily play a role in improving wrinkles during volume maintenance.

Lidocaine hydrochloride, a local anesthetic, is widely used in medical procedures to reduce or eliminate pain. The action mechanism is mainly to reduce the conduction of nerve impulse by blocking the sodium channel on nerve fiber, thereby achieving the effect of local anesthesia. Clinically, lidocaine hydrochloride is commonly used for superficial anesthesia of skin, mucous membrane and nerve block anesthesia. In addition, because of its antiarrhythmic effect on the heart, it is also used to treat certain types of arrhythmias. Lidocaine hydrochloride is a safe and effective local anesthetic with well controlled doses.

However, although the existing medical fillers have achieved some success in improving skin condition, there are still some drawbacks in terms of safety, biocompatibility and risk control of embolism. Therefore, it is important to develop a medical filler that can significantly reduce the risk of embolism and improve biocompatibility and safety.

Disclosure of Invention

Aiming at the defects of the prior art, the invention aims to provide a preparation method of facial filler, which combines the percolation threshold response characteristic of agarose, the bioactivity function of nucleotide substances and the anesthetic efficacy of lidocaine hydrochloride, and combines scientific proportioning and process optimization to form an innovative medical filler with safety, functionality and clinical practicability.

The technical scheme adopted by the invention is as follows:

The facial filler comprises the following raw materials in parts by weight:

Agarose 1-5 parts;

0.2-1.5 parts of nucleotide substances;

0.1-0.5 part of lidocaine hydrochloride;

90-100 parts of phosphate buffer solution;

The nucleotide substance is Polydeoxyribonucleotide (PDRN) or Polynucleotide (PN);

The preparation method of the facial filler comprises the following steps:

(1) Mixing agarose with a first part of phosphate buffer solution, heating to 80-90 ℃, stirring until the agarose is completely dissolved, and then stopping stirring and reducing the system temperature to 0-35 ℃ to obtain agarose gel;

(2) Mixing the nucleotide substance with a second part of phosphate buffer solution, and stirring for dissolving to obtain a nucleotide substance solution;

(3) Mixing lidocaine hydrochloride with a third part of phosphate buffer solution, stirring and dissolving to obtain a lidocaine hydrochloride solution;

(4) Granulating agarose gel through a 60-mesh screen, mixing and stirring the agarose gel with nucleotide substance solution and lidocaine hydrochloride solution, filling the mixture into a prefilled syringe after vacuum defoamation treatment, and carrying out irradiation sterilization with the irradiation dose of 20-25 kGy to finally obtain the facial filler.

The phosphate buffer solution is an isotonic phosphate buffer solution with a pH value of 6.8-7.3.

In the step (1), the mass ratio of agarose to the first part of phosphate buffer solution is 1 (12-60).

In the step (2), the mass ratio of the nucleotide substance to the second part of phosphate buffer solution is 1 (20-150).

In the step (3), the mass ratio of lidocaine hydrochloride to the third part of phosphate buffer solution is 0.3 (3-18).

In the invention, the sepharose particles have stronger surface tension, and the irradiated sepharose particles are partially linked with the nucleotide substances through hydrogen bonds, so that the hydration capability is greatly improved, the friction force among the particles is reduced, and the pushing force is reduced. Under the condition of embolism, a large amount of physiological saline solution is injected, and water molecules in the physiological saline solution can quickly dilute and replace hydrogen bond links between nucleotide substances and agarose particles, so that the addition of the nucleotide substances can not only increase the comfort level during injection, but also ensure the embolism prevention effect of gel. And simultaneously, after PDRN irradiation sterilization, partial deoxynucleotides (such as deoxyadenylate, deoxyguanylate and the like) are decomposed into single-chain or short-chain deoxynucleotides (such as deoxyadenosine, deoxyguanylate and the like). These substances can be used as raw materials for cell repair and nucleic acid synthesis, and are involved in DNA damage repair or energy metabolism to provide instant nutrition for cell proliferation.

Compared with the prior art, the invention has the following beneficial effects:

(1) The gel system constructed based on agarose concentration-dependent phase change characteristics breaks through the limitation of difficult post-treatment of the traditional filler embolism. By precisely controlling the percolation threshold critical point, if vascular embolism occurs after injection, reversible depolymerization of the gel network can be realized by local normal saline perfusion, and the embolism can be removed without invasive surgical intervention. This feature not only greatly improves the fault tolerance of the clinical procedure, but also reduces the risk of tissue necrosis due to embolism. Meanwhile, the biodegradation period (6-24 months) of agarose is matched with the human tissue repair rhythm, so that long-term foreign body reaction is avoided, and the safety foundation of the material is further enhanced;

(2) The invention forms a double construction system with complementary functions through polydeoxyribonucleotide or polynucleotide and agarose, wherein a three-dimensional network of agarose provides physical support for regeneration of extracellular matrixes, and the polydeoxyribonucleotide or polynucleotide continuously stimulates collagen synthesis through activating an adenosine A2A receptor, and the polydeoxyribonucleotide or polynucleotide realizes the double effects of 'instant filling and long-acting repair' through the synergistic effect of the polydeoxyribonucleotide or the polynucleotide. The bioactive components of the polydeoxyribonucleotide or the polynucleotide are gradually released within 2-3 months, so that the continuous regeneration of type I collagen, type III collagen and elastin is promoted, the filling effect is transited from pure mechanical support to self-tissue reconstruction, the maintenance period of the cosmetic effect is prolonged to 6-12 months, and the cosmetic effect is obviously superior to the single-phase action mode of the traditional filling agent;

(3) The lidocaine hydrochloride integrated in the formula provided by the invention avoids the risk of cardiovascular side effects while realizing effective local anesthesia through accurate dosage control. The standardized application of the phosphate buffer system ensures the pH stability and osmotic pressure balance of the gel, and combines irradiation sterilization and vacuum defoaming treatment, so that the final product has excellent biocompatibility and clinical stability. The direct filling design of the prefilled syringe further reduces the possibility of secondary pollution, and improves the convenience and safety of clinical use.

Detailed Description

The invention is further illustrated below with reference to examples, which are not intended to limit the practice of the invention.

The raw materials used in examples and comparative examples are conventional commercial raw materials unless otherwise specified, and the process methods used in examples and comparative examples are conventional in the art unless otherwise specified.

Some of the raw materials used in the examples and comparative examples are described below:

polydeoxyribonucleotides, purchased from the biotechnology company, regimine (shandong);

polynucleotides, available from the biotechnology company, regimine (shandong);

the phosphate buffer solution is isotonic phosphate buffer solution with the pH value of 7+/-0.2, and the preparation method comprises the steps of accurately weighing 7.5g of sodium dihydrogen phosphate (NaH ₂PO₄), 33g of disodium hydrogen phosphate (Na ₂HPO₄) and 1350g of sodium chloride (NaCl), adding into a clean liquid preparation tank, adding a proper amount of deionized water for preliminary dissolution, continuing adding deionized water until the total volume of the solution is 150L, and finely adjusting the solution to the target pH range of 7+/-0.2 by dropwise adding a dilute hydrochloric acid (HCl) or sodium hydroxide (NaOH) solution.

Example 1

The preparation method of the facial filler comprises the following steps:

(1) Mixing 3 parts by weight of agarose with 60 parts by weight of a first part of phosphate buffer solution, heating to 90 ℃, stirring for 30min at a rotating speed of 500rpm until the agarose is completely dissolved, and then stopping stirring and reducing the system temperature to 20 ℃ to obtain agarose gel;

(2) Mixing 1 part by weight of Polydeoxyribonucleotide (PDRN) with 30 parts by weight of a second part of phosphate buffer solution, and stirring for 60min at a rotation speed of 500rpm to obtain a nucleotide solution;

(3) Mixing 0.3 part by weight of lidocaine hydrochloride with 6 parts by weight of phosphate buffer solution, and stirring for 30min at a rotating speed of 500rpm to obtain a lidocaine hydrochloride solution;

(4) Granulating agarose gel through a 60-mesh screen, mixing and stirring the agarose gel with nucleotide substance solution and lidocaine hydrochloride solution, filling the mixture into a prefilled syringe after vacuum defoamation treatment, and carrying out irradiation sterilization after filling, wherein the irradiation dose is 20kGy, thus finally obtaining the facial filler.

Example 2

The preparation method of the facial filler comprises the following steps:

(1) Mixing 1 part by weight of agarose with 60 parts by weight of a first part of phosphate buffer solution, heating to 90 ℃, stirring for 30min at a rotating speed of 500rpm until the agarose is completely dissolved, and then stopping stirring and reducing the system temperature to 20 ℃ to obtain agarose gel;

(2) Mixing 0.2 parts by weight of Polydeoxyribonucleotide (PDRN) with 30 parts by weight of a second part of phosphate buffer solution, and stirring for 60min at a rotation speed of 500rpm to obtain a nucleotide solution;

(3) Mixing 0.1 part by weight of lidocaine hydrochloride with 6 parts by weight of phosphate buffer solution, and stirring for 30min at a rotating speed of 500rpm to obtain a lidocaine hydrochloride solution;

Example 3

The preparation method of the facial filler comprises the following steps:

(1) Mixing 5 parts by weight of agarose with 60 parts by weight of a first part of phosphate buffer solution, heating to 90 ℃, stirring for 30min at a rotating speed of 500rpm until the agarose is completely dissolved, and then stopping stirring and reducing the system temperature to 20 ℃ to obtain agarose gel;

(2) Mixing 1.5 parts by weight of Polydeoxyribonucleotide (PDRN) with 30 parts by weight of a second part of phosphate buffer, and stirring for 60min at a rotation speed of 500rpm to obtain a nucleotide solution;

(3) Mixing 0.5 part by weight of lidocaine hydrochloride with 6 parts by weight of phosphate buffer solution, and stirring for 30min at a rotating speed of 500rpm to obtain a lidocaine hydrochloride solution;

Example 4

The preparation method of the facial filler comprises the following steps:

(2) Mixing 1 part by weight of Polynucleotide (PN) with 30 parts by weight of a second part of phosphate buffer solution, and stirring for 60 minutes at a rotating speed of 500rpm to obtain a nucleotide solution;

Example 5

The preparation method of the facial filler comprises the following steps:

(1) Mixing 3 parts by weight of agarose with 60 parts by weight of a first part of phosphate buffer solution, heating to 80 ℃, stirring for 30min at a rotating speed of 500rpm until the agarose is completely dissolved, and then stopping stirring and reducing the system temperature to 0 ℃ to obtain agarose gel;

Example 6

The preparation method of the facial filler comprises the following steps:

(1) Mixing 3 parts by weight of agarose with 60 parts by weight of a first part of phosphate buffer, heating to 90 ℃, stirring for 30min at a rotating speed of 500rpm until the agarose is completely dissolved, and then stopping stirring and reducing the system temperature to 35 ℃ to obtain agarose gel;

Example 7

The preparation method of the facial filler comprises the following steps:

(4) Granulating agarose gel through a 60-mesh screen, mixing and stirring the agarose gel with nucleotide substance solution and lidocaine hydrochloride solution, filling the mixture into a prefilled syringe after vacuum defoamation treatment, and carrying out irradiation sterilization after filling, wherein the irradiation dose is 25kGy, thus finally obtaining the facial filler.

Comparative example 1

The difference from example 1 is that agarose is 0.5 parts by weight, otherwise the same as in example 1.

Comparative example 2

The difference from example 1 is that the agarose is 8 parts by weight, and the other is the same as in example 1.

Comparative example 3

The difference from example 1 is that in the step (4), the wet heat sterilization is performed after the filling, and the condition of the wet heat sterilization is that the temperature is 121 ℃ for 8min, and the same as in example 1 is adopted.

Comparative example 4

The difference from example 1 is that step (2) is not included, and the other steps are the same as those in example 1.

Comparative example 5

The difference from example 1 is that agarose in step (1) is replaced with an equal weight part of crosslinked sodium hyaluronate gel, in step (4), the filling is followed by a wet heat sterilization (121 ℃ C., 8 min) treatment;

The preparation method of the crosslinked sodium hyaluronate gel comprises the steps of weighing 2g of sodium hyaluronate, placing the sodium hyaluronate in 10mL of sodium hydroxide solution containing 0.1mol of sodium hydroxide, adding 0.2g of 1, 4-butanediol diglycidyl ether after the sodium hyaluronate is fully dissolved, reacting for 24 hours at 30 ℃, taking out the formed gel after the reaction is finished, cutting the formed gel into blocks with the mass of 1g, and then placing the cut gel blocks into phosphate buffer solution to fully swell the gel blocks until the mass reaches 120g. And finally, granulating the swelled gel through a 60-mesh screen to obtain the granular crosslinked sodium hyaluronate gel.

Comparative example 6

The difference from example 1 is that in the above-mentioned step (1), the temperature of stirring was 70℃and the same as in example 1 was followed.

Comparative example 7

The difference from example 1 is that in the above-mentioned step (1), stirring was stopped and the system temperature was lowered to-10℃with the other matters of example 1.

The final facial fillers of examples 1 to 7 and comparative examples 1 to 7 were tested for performance by the following test methods:

Respectively taking 20 parts of the facial fillers prepared in examples 1-7 and comparative examples 1-7, placing the facial fillers into a stability test box, standing for 5 months at the temperature of 45 ℃ with the humidity of 75%, observing whether layering exists in the samples, and counting the number of the layered samples;

Sensitization is tested by referring to GB/T16886.10-2017;

The pushing force (N) is tested by a universal tester, the temperature is 25 ℃, the pushing speed is 30mm/min, and the injector is a disposable sterile injector with the volume of 2 mL;

elastic modulus G' (Pa) rheological measurements were made using a rheometer at 25 ℃;

The anti-embolism effect is that 1mL of the facial filler prepared in examples 1-7 and comparative examples 1-7 is injected into an external arterial vessel model of the last branch, the embolism occurrence condition is simulated, and then 20mL of physiological saline is injected into the same part, and the dredging condition of the vessel model is observed.

The test results are shown in Table 1.

TABLE 1 Performance test results

As can be seen from the test data in Table 1, the facial fillers prepared in examples 1 to 3 showed a tendency to gradually increase in both the pushing force and the elastic modulus G' with the gradual increase in the agarose content. This phenomenon is mainly due to the excellent mechanical properties of agarose itself. In addition, the experimental results show that the content change of lidocaine hydrochloride has no significant effect on the product performance of the facial filler.

In example 4, using Polynucleotide (PN) as a starting material, it was found that it did not significantly affect the product properties of the facial filler.

In examples 5-6, the preparation was performed by changing to gel temperature conditions, and experimental data showed that the change in gel temperature also had no significant effect on the product properties of the facial fillers.

In example 7, the irradiation dose was increased to 25kGy, and the detection result showed that the pushing force of the product was reduced and the elastic modulus G' was also reduced. This is because irradiation treatment breaks chemical bonds between agarose molecules, which in turn leads to a decrease in the mechanical strength of the gel.

In comparative example 1, since the agarose content is at a low level, the gel forming property of the gel is inferior, thereby resulting in insufficient stability of the gel and significant decrease in mechanical properties.

In comparative example 2, the agarose content was too high, which directly resulted in too high a pushing force, greatly reducing the comfort during injection. In addition, too high an agarose content can adversely affect the "percolation threshold response" of the agarose gel, thereby impairing its anti-embolic efficacy.

In comparative example 3, the gel was melted again in the syringe by the wet heat sterilization and was coagulated into a block, so that it could not be smoothly pushed out for use.

Comparative example 4, in contrast to example 1, was not added Polydeoxyribonucleotide (PDRN). In this case, strong surface tension is exhibited between individual agarose particles, which directly results in a significant increase in the pushing force of the product. In the embodiment 1, the irradiated agarose particles and the nucleotide substances form partial links through hydrogen bonds, so that the hydration capacity is greatly improved, and the friction force and the pushing force among the particles are effectively reduced. In addition, in the case of possible embolism, when physiological saline solution is injected, water molecules thereof can be rapidly diluted and replace hydrogen bond links between nucleotide substances and agarose particles. Thus, comparative example 4 was not only reduced in comfort upon injection due to the lack of addition of nucleotide substances, but also reduced in anti-embolic effect of the gel accordingly.

In comparative example 5, compared with example 1, since the crosslinked sodium hyaluronate gel was used, it was difficult to effectively dredge the gel by using normal saline infusion washing when embolism occurred, and since the characteristic of the crosslinked sodium hyaluronate gel, only a wet heat sterilization mode was used, which only partially changed the molecular weight of PDRN, and the double helix structure could not be broken. In contrast, example 1 employs irradiation sterilization to partially decompose PDRN into single-stranded or short-chain deoxynucleotides (e.g., deoxyadenylate, deoxyguanylate, etc.) and deoxynucleosides (e.g., deoxyadenosine, deoxyguanylate, etc.), which can be used as raw materials for cell repair and nucleic acid synthesis, and are involved in DNA damage repair or energy metabolism to provide immediate nutrition for cell proliferation.

In comparative example 6, the gel forming temperature was low, resulting in gel failure.

In comparative example 7, when the temperature is lowered below the freezing point, a part of gel is precipitated, thereby adversely affecting the stability of the product.

Claims

1. A method for preparing a facial filler, characterized in that the facial filler comprises the following raw materials in parts by weight:

Agarose: 1~5 parts;

Nucleotide substances: 0.2~1.5 parts;

Lidocaine hydrochloride: 0.1~0.5 parts;

Phosphate buffer: 90~100 parts;

The preparation method of the facial filler comprises the following steps:

(1) Mixing agarose with the first part of phosphate buffer, heating to 80-90°C, stirring until the agarose is completely dissolved, then stopping stirring and lowering the system temperature to 0-35°C to obtain agarose gel;

(2) mixing the nucleotide substance with the second portion of phosphate buffer, stirring to dissolve, and obtaining a nucleotide substance solution;

(3) mixing lidocaine hydrochloride with the third portion of phosphate buffer, stirring to dissolve, and obtaining a lidocaine hydrochloride solution;

(4) After the agarose gel is granulated through a sieve, it is mixed with a nucleotide solution and a lidocaine hydrochloride solution, and then degassed, filled, and irradiated for sterilization at a dose of 20 to 25 kGy to finally obtain a facial filler.

2. The method for preparing a facial filler according to claim 1, wherein the nucleotide substance is polydeoxyribonucleotide or polynucleotide.

3. The method for preparing a facial filler according to claim 1, wherein the pH of the phosphate buffer is 6.8-7.3.

4. The method for preparing a facial filler according to claim 1, characterized in that in the step (1), the mass ratio of agarose to the first part of phosphate buffer is 1:(12-60).

5. The method for preparing a facial filler according to claim 1, characterized in that in the step (2), the mass ratio of the nucleotide substance to the second part of the phosphate buffer is 1:(20-150).

6. The method for preparing a facial filler according to claim 1, characterized in that in the step (3), the mass ratio of lidocaine hydrochloride to the third part of phosphate buffer is 0.3:(3-18).