Somatic cell injection for acupoint injection and preparation method thereof

Abstract

The invention provides a somatic cell injection for acupoint injection and a preparation method, and belongs to the technical field of drugs, the somatic cell injection for acupoint injection is composed of bone marrow mesenchymal stem cells, an injection carrier for diluting and stabilizing the bone marrow mesenchymal stem cells, and auxiliary materials for improving the stability and safety of the injection; by mixing the auxiliary materials in the injection, the safety and stability of the injection can be improved.

Description

Technical Field

[0001] The present invention relates to the field of drugs for injection, and in particular to a somatic cell injection solution for acupoint injection and a preparation method thereof. Background Art

[0002] Acupuncture injection therapy is a unique technique in Traditional Chinese Medicine (TCM). It involves injecting small doses of medication into specific acupuncture points, leveraging the combined effects of acupuncture, acupuncture, and medication to achieve the desired therapeutic effect. This therapy has been widely used in clinical practice, demonstrating its effectiveness and wide indications.

[0003] Although acupuncture injection therapy has shown significant advantages in clinical applications, the currently used acupuncture injection solutions still face a series of challenges in their preparation and use: such as the stability of the injection solution: after preparation, some acupuncture injection solutions are difficult to maintain their stability for a long time due to factors such as complex ingredients and limited storage conditions. This may lead to reduced drug activity and increased impurities, thereby affecting the therapeutic effect and safety.

[0004] Invention application content In view of this, the present invention provides a somatic cell injection solution for acupoint injection and a preparation method thereof. The present invention can improve the safety and stability of the injection solution by mixing excipients into the injection solution.

[0005] In order to solve the above technical problems, the present invention provides a somatic cell injection solution for acupoint injection and a preparation method. The somatic cell injection solution for acupoint injection is composed of bone marrow mesenchymal stem cells, an injection carrier for diluting and stabilizing the bone marrow mesenchymal stem cells, and excipients for improving the stability and safety of the injection solution.

[0006] Excipients include pH regulators, osmotic pressure regulators, and antioxidants.

[0007] Each dose of bone marrow mesenchymal stem cell injection includes 1E6~4E7 living mesenchymal stem cells, 48.5ml~97ml injection vehicle, 0.1~1ml antioxidant, 0.1~1ml osmotic pressure regulator, and 0.1~1ml pH regulator.

[0008] The injection carrier is made of physiological saline (0.9% sodium chloride solution) as the main material and supplemented with a trace amount of glucose solution and cell culture medium.

[0009] The antioxidant is vitamin C or vitamin E.

[0010] The osmotic pressure regulator was mannitol or glucose.

[0011] pH adjuster is sodium hydroxide or hydrochloric acid A method for preparing a somatic cell injection for acupoint injection: S1, collecting mesenchymal stem cells and separating them; S2. Cultivating and expanding the collected cells; S3. Performing quality inspection on the expanded mesenchymal stem cells; S4, mixing the mesenchymal stem cells with the injection carrier to obtain a mixed solution; S5. Add appropriate amount of excipients to the mixed solution and mix to obtain an injection solution; S6. Sterile treatment of the injection solution; S7. Conduct quality inspection on the injection solution; S8. Packaging.

[0012] In summary, compared with the prior art, this application has at least one of the following beneficial technical effects: 1. By adding excipients to the injection solution, the safety and stability of the injection solution can be improved. Antioxidants such as vitamin C and vitamin E can scavenge free radicals in the body, reduce cell damage, and prolong cell survival. The addition of pH and osmotic pressure regulators ensures that the pH and osmotic pressure of the injection solution are similar to those of human tissue fluid, reducing cellular stress responses in the body and improving therapeutic efficacy and safety.

[0013] 2. The preparation method of the present invention maximizes the preservation of the original quality and activity of mesenchymal stem cells through meticulous cell extraction, culture expansion, and purification steps. These steps not only remove impurities and unwanted cell types but also ensure the genetic stability and functional integrity of the cells during the expansion process. Highly active mesenchymal stem cells can function more effectively after injection, promoting tissue repair and regeneration.

[0014] 3. Compared with traditional intravenous injection, the present invention has a stronger local effect when injected into the human body through acupuncture points: acupuncture point injection can directly deliver mesenchymal stem cells to specific acupuncture points and local tissues, thereby producing a stronger therapeutic effect locally; Reduce systemic side effects: Since mesenchymal stem cells act locally, they may reduce systemic side effects such as fever and allergic reactions.

[0015] 4. The advantages of this acupuncture point injection method over traditional intravenous injection are as follows: Supported by Traditional Chinese Medicine theory: Acupuncture point injection combines traditional Chinese medicine meridian theory with modern medicine stem cell technology, providing a new treatment method for traditional Chinese medicine. By stimulating specific acupuncture points, it can regulate the body's physiological functions and achieve the purpose of treating diseases.

[0016] Personalized treatment: Traditional Chinese medicine focuses on individualized treatment. Acupoint injection can select appropriate acupoints for injection based on the patient's specific symptoms and constitution, thereby achieving a more personalized treatment plan.

[0017] Reduced pain and discomfort: For some patients, intravenous injections may cause pain or discomfort. Acupuncture injections, on the other hand, are usually performed in more comfortable body positions and acupuncture points, which can improve patient acceptance and comfort.

[0018] Psychological comfort effect: As a traditional treatment, TCM acupoint injection may have a certain psychological comfort effect on some patients, helping to relieve anxiety and fear. DETAILED DESCRIPTION

[0019] To make the purpose, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with embodiments 1 to 3 of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the described embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field fall within the scope of protection of the present invention.

[0020] Embodiment 1; The somatic cell injection for acupuncture injection consists of bone marrow mesenchymal stem cells, an injection carrier for diluting and stabilizing the bone marrow mesenchymal stem cells, and excipients for improving the stability and safety of the injection; The excipients include pH regulators, osmotic pressure regulators, and antioxidants; pH regulators can be used to adjust the pH value of the injection solution, that is, sodium hydroxide or hydrochloric acid can be used to maintain the pH value of the injection solution within the range of 6.0-8.0; Precise pH adjustment can ensure the stability and effectiveness of the injection. The appropriate pH value helps maintain the activity and function of stem cells, thereby achieving better results during the treatment process; An inappropriate pH value may cause adverse reactions in patients after receiving the injection, such as fever and allergies. By adjusting the pH value to an appropriate range, the occurrence of these adverse reactions can be reduced, improving treatment safety and patient comfort.

[0021] Appropriate osmotic pressure helps maintain the stability of the injection solution and prevents adverse changes such as deformation, rupture or aggregation of cells during storage and transportation. In this embodiment, the osmotic pressure regulator can be glucose, mannitol or sodium chloride. That is, by precisely adjusting the osmotic pressure, the normal physiological function of cells can be maintained, the cell survival rate can be improved, the stability of the injection solution can be ensured, and the occurrence of adverse reactions can be reduced.

[0022] Antioxidants can be vitamin C, vitamin E, tea polyphenols, ascorbic acid, and its derivatives. The addition of antioxidants can improve the stability of the injection solution and prevent oxidative deterioration during storage and transportation. This helps maintain the active ingredients and therapeutic effects of the injection solution, extending its shelf life. They can also enhance the antioxidant capacity of mesenchymal stem cells by regulating the antioxidant enzyme system within the cells. This helps the cells better cope with oxidative stress challenges in the external environment and maintain their normal physiological functions.

[0023] The injected vector can provide a good microenvironment for stem cells, helping to protect them from damage from the external environment, thereby increasing their survival rate; The injection carrier is made of physiological saline (0.9% sodium chloride solution) as the main material and supplemented with a small amount of glucose solution and cell culture medium; As an injection carrier, physiological saline can provide a good microenvironment for stem cells, helping to protect them from damage by the external environment, thereby increasing their survival rate. Physiological saline has good biocompatibility and safety, and generally does not cause adverse reactions during use. Compared with other carriers, physiological saline may have a lower risk of allergies and fewer complications. Glucose solution has a wide range of applications in medical treatment as an injection carrier. Its use is simple, safe and effective, and can provide patients with multiple therapeutic effects such as energy supplementation, drug delivery, improved osmotic pressure, and blood sugar level regulation. The amount of glucose solution to be added should be determined based on actual conditions during use. Cell culture medium enables the injected cells to continue to proliferate and differentiate in a suitable microenvironment; this helps to repair damaged tissues, replace diseased cells, and thus restore the normal structure and function of the tissues.

[0024] The injection solution was dosed according to the above description. Each 100 ml of bone marrow mesenchymal stem cell injection solution included 4E7 living mesenchymal stem cells, 1 ml of antioxidant, 1 ml of osmotic pressure regulator, 1 ml of pH regulator, and 97 ml of injection vehicle.

[0025] Embodiment 2; The somatic cell injection for acupuncture injection consists of bone marrow mesenchymal stem cells, an injection carrier for diluting and stabilizing the bone marrow mesenchymal stem cells, and excipients for improving the stability and safety of the injection; The excipients include pH regulators, osmotic pressure regulators, and antioxidants; pH regulators can be used to adjust the pH value of the injection solution, that is, sodium hydroxide or hydrochloric acid can be used to maintain the pH value of the injection solution within the range of 6.0-8.0; Precise pH adjustment can ensure the stability and effectiveness of the injection. The appropriate pH value helps maintain the activity and function of stem cells, thereby achieving better results during the treatment process; An inappropriate pH value may cause adverse reactions in patients after receiving the injection, such as fever and allergies. By adjusting the pH value to an appropriate range, the occurrence of these adverse reactions can be reduced, improving treatment safety and patient comfort.

[0026] Appropriate osmotic pressure helps maintain the stability of the injection solution and prevents adverse changes such as deformation, rupture or aggregation of cells during storage and transportation. In this embodiment, the osmotic pressure regulator can be glucose, mannitol or sodium chloride. That is, by precisely adjusting the osmotic pressure, the normal physiological function of cells can be maintained, the cell survival rate can be improved, the stability of the injection solution can be ensured, and the occurrence of adverse reactions can be reduced.

[0027] Antioxidants can be vitamin C, vitamin E, tea polyphenols, ascorbic acid, and its derivatives. The addition of antioxidants can improve the stability of the injection solution and prevent oxidative deterioration during storage and transportation. This helps maintain the active ingredients and therapeutic effects of the injection solution, extending its shelf life. They can also enhance the antioxidant capacity of mesenchymal stem cells by regulating the antioxidant enzyme system within the cells. This helps the cells better cope with oxidative stress challenges in the external environment and maintain their normal physiological functions.

[0028] The injected vector can provide a good microenvironment for stem cells, helping to protect them from damage from the external environment, thereby increasing their survival rate; The injection carrier is made of physiological saline (0.9% sodium chloride solution) as the main material and supplemented with a small amount of glucose solution and cell culture medium; As an injection carrier, physiological saline can provide a good microenvironment for stem cells, helping to protect them from damage by the external environment, thereby increasing their survival rate. Physiological saline has good biocompatibility and safety, and generally does not cause adverse reactions during use. Compared with other carriers, physiological saline may have a lower risk of allergies and fewer complications. Glucose solution has a wide range of applications in medical treatment as an injection carrier. Its use is simple, safe and effective, and can provide patients with multiple therapeutic effects such as energy supplementation, drug delivery, improved osmotic pressure, and blood sugar level regulation. The amount of glucose solution to be added should be determined based on actual conditions during use. Cell culture medium enables the injected cells to continue to proliferate and differentiate in a suitable microenvironment; this helps to repair damaged tissues, replace diseased cells, and thus restore the normal structure and function of the tissues.

[0029] The injection solution was dosed according to the above description. Each 50 ml of bone marrow mesenchymal stem cell injection solution included 1E6 living mesenchymal stem cells, 0.5 ml of antioxidant, 0.5 ml of osmotic pressure regulator, 0.5 ml of pH regulator, and 48.5 ml of injection vehicle.

[0030] Embodiment 3; The somatic cell injection for acupuncture injection consists of bone marrow mesenchymal stem cells, an injection carrier for diluting and stabilizing the bone marrow mesenchymal stem cells, and excipients for improving the stability and safety of the injection; The excipients include pH regulators, osmotic pressure regulators, and antioxidants; pH regulators can be used to adjust the pH value of the injection solution, that is, sodium hydroxide or hydrochloric acid can be used to maintain the pH value of the injection solution within the range of 6.0-8.0; Precise pH adjustment can ensure the stability and effectiveness of the injection. The appropriate pH value helps maintain the activity and function of stem cells, thereby achieving better results during the treatment process; An inappropriate pH value may cause adverse reactions in patients after receiving the injection, such as fever and allergies. By adjusting the pH value to an appropriate range, the occurrence of these adverse reactions can be reduced, improving treatment safety and patient comfort.

[0031] Appropriate osmotic pressure helps maintain the stability of the injection solution and prevents adverse changes such as deformation, rupture or aggregation of cells during storage and transportation. In this embodiment, the osmotic pressure regulator can be glucose, mannitol or sodium chloride. That is, by precisely adjusting the osmotic pressure, the normal physiological function of cells can be maintained, the cell survival rate can be improved, the stability of the injection solution can be ensured, and the occurrence of adverse reactions can be reduced.

[0032] Antioxidants can be vitamin C, vitamin E, tea polyphenols, ascorbic acid, and its derivatives. The addition of antioxidants can improve the stability of the injection solution and prevent oxidative deterioration during storage and transportation. This helps maintain the active ingredients and therapeutic effects of the injection solution, extending its shelf life. They can also enhance the antioxidant capacity of mesenchymal stem cells by regulating the antioxidant enzyme system within the cells. This helps the cells better cope with oxidative stress challenges in the external environment and maintain their normal physiological functions.

[0033] The injected vector can provide a good microenvironment for stem cells, helping to protect them from damage from the external environment, thereby increasing their survival rate; The injection carrier is made of physiological saline (0.9% sodium chloride solution) as the main material and supplemented with a small amount of glucose solution and cell culture medium; As an injection carrier, physiological saline can provide a good microenvironment for stem cells, helping to protect them from damage by the external environment, thereby increasing their survival rate. Physiological saline has good biocompatibility and safety, and generally does not cause adverse reactions during use. Compared with other carriers, physiological saline may have a lower risk of allergies and fewer complications. Glucose solution has a wide range of applications in medical treatment as an injection carrier. Its use is simple, safe and effective, and can provide patients with multiple therapeutic effects such as energy supplementation, drug delivery, improved osmotic pressure, and blood sugar level regulation. The amount of glucose solution to be added should be determined based on actual conditions during use. Cell culture medium enables the injected cells to continue to proliferate and differentiate in a suitable microenvironment; this helps to repair damaged tissues, replace diseased cells, and thus restore the normal structure and function of the tissues.

[0034] The injection solution was dosed according to the above description. Each 75 ml of bone marrow mesenchymal stem cell injection solution included 2.05E7 living mesenchymal stem cells, 0.75 ml of antioxidant, 0.75 ml of osmotic pressure regulator, 0.75 ml of pH regulator, and 72.5 ml of injection vehicle. A method for preparing a somatic cell injection for acupoint injection: S1, collecting mesenchymal stem cells and separating them; That is, healthy donors are selected and a comprehensive physical examination and serological tests are performed to rule out infectious diseases and other related diseases.

[0035] Evaluate the donor's age, gender, weight, and other factors to determine the optimal cell collection quantity and quality.

[0036] Cell collection; Depending on the specific conditions of the donor, bone marrow, adipose tissue, umbilical cord blood or placenta are selected as the cell source.

[0037] Cells were collected using minimally invasive techniques or standard surgical methods, ensuring aseptic operation during the collection process.

[0038] Cell separation; The collected tissue samples are initially processed, such as mincing and enzymatic hydrolysis, to release cells.

[0039] Density gradient centrifugation, red blood cell lysis, magnetic bead sorting and other methods are used to remove impurities, red blood cells and other non-mesenchymal stem cell components.

[0040] Cell purification; Mesenchymal stem cells are further purified through adherent culture, flow cytometer sorting and other methods to improve cell purity and activity.

[0041] Count and detect the activity of purified cells to ensure that the cell quantity and quality meet the requirements S2. Cultivating and expanding the collected cells; Prepare appropriate culture medium according to cell type and growth requirements, including basal culture medium, growth factors, cytokines, etc.

[0042] Ensure the sterility and suitability of the culture medium to avoid contamination and cell damage.

[0043] Cell seeding; The purified mesenchymal stem cells are seeded into a culture flask containing culture medium, and the seeding density is controlled so that the cells can be evenly distributed and grow.

[0044] cell culture; The culture flask was placed in an incubator at 37°C and 5% CO2 for culture, and the cell growth was observed regularly.

[0045] According to the cell growth rate and metabolic needs, the culture medium should be replaced in a timely manner and growth factors and cytokines should be supplemented.

[0046] cell expansion; When the cells grow to about 80% of the bottom of the bottle, subculture is performed to expand the cell number.

[0047] During the subculture process, the subculture ratio and cell density should be controlled to avoid excessive cell growth and aging.

[0048] S3. Performing quality inspection on the expanded mesenchymal stem cells; The expanded mesenchymal stem cells were counted and tested for activity to ensure that the cell quantity and quality met the requirements.

[0049] Trypan blue staining, flow cytometry, and other methods were used to detect cell activity to ensure that the proportion of live cells was no less than 95%.

[0050] S4, mixing the mesenchymal stem cells with the injection carrier to obtain a mixed solution; Mix MSCs with saline according to the desired injection volume and cell concentration.

[0051] S5. Add appropriate amount of excipients to the mixed solution and mix to obtain an injection solution; Add appropriate amounts of antioxidants (such as vitamin C), pH regulators (such as sodium hydroxide or hydrochloric acid) and osmotic pressure regulators (such as mannitol) to maintain cell stability and activity.

[0052] S6. Sterile treatment of the injection solution; Use a sterile filter to filter and sterilize the injection solution to ensure the sterility of the injection solution.

[0053] The pore size of the filter should be less than 0.22 μm to remove bacteria, fungi and other microbial contamination.

[0054] S7. Conduct quality inspection on the injection solution; The prepared injection solution is subjected to quality testing, including indicators such as cell activity, purity, pH value, and osmotic pressure.

[0055] Ensure that the quality of the injection meets relevant standards and requirements S8, packaging; The qualified injection solution should be sterilely packaged and marked with product name, batch number, expiration date and other information.

[0056] The packaging materials should have good sealing and sterility to avoid contamination and deterioration.

[0057] Store the prepared injection at 2-8°C, away from direct sunlight and high temperature.

[0058] Refrigerated conditions should be maintained during transportation to ensure the stability and activity of the injection.

[0059] The above is a preferred embodiment of the present invention. 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 described in the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A somatic cell injection solution for acupoint injection, characterized in that: It is composed of bone marrow mesenchymal stem cells, an injection carrier for diluting and stabilizing the bone marrow mesenchymal stem cells, and excipients for improving the stability and safety of the injection solution.

2. The somatic cell injection solution for acupoint injection according to claim 1, characterized in that: The auxiliary materials include pH regulator, osmotic pressure regulator and antioxidant.

3. The somatic cell injection solution for acupoint injection according to claim 2, characterized in that: Each bone marrow mesenchymal stem cell injection includes 1E6-4E7 living mesenchymal stem cells, 48.5ml-97ml injection carrier, 0.1-1ml antioxidant, 0.1-1ml osmotic pressure regulator, and 0.1-1ml pH regulator.

4. The somatic cell injection solution for acupoint injection according to claim 3, characterized in that: The injection carrier is made of physiological saline (0.9% sodium chloride solution) as the main material and is supplemented with a trace amount of glucose solution and cell culture medium.

5. The somatic cell injection solution for acupoint injection according to claim 3, characterized in that: The antioxidant is vitamin C or vitamin E.

6. The somatic cell injection solution for acupoint injection according to claim 3, characterized in that: The osmotic pressure regulator is mannitol or glucose.

7. The somatic cell injection solution for acupoint injection according to claim 3, characterized in that: The pH adjuster is sodium hydroxide or hydrochloric acid.

8. A method for preparing a somatic cell injection solution for acupoint injection, characterized in that: S1. Collecting mesenchymal stem cells and separating them; S2, culturing and amplifying the collected cells; S3, conducting quality inspection on the expanded mesenchymal stem cells; S4, mixing the mesenchymal stem cells with the injection carrier to obtain a mixed solution; S5. Add appropriate amount of auxiliary materials into the mixed solution and mix to obtain an injection solution; S6. Treat the injection solution aseptically; S7. Conduct quality inspection on the injection solution; S8. Packaging.

9. The method for preparing a somatic cell injection solution for acupoint injection according to claim 8, characterized in that: The prepared injection solution should be kept at a temperature of 2 to 8° for transportation or storage.