Polydeoxyribonucleotide solution for injection and preparation method thereof

Abstract

The invention discloses a polydeoxyribonucleotide solution for injection and a preparation method thereof, belongs to the field of passive implantation instruments of medical instruments, namely plastic and general surgery implants, and particularly relates to the polydeoxyribonucleotide solution for injection and the preparation method thereof. The poly-deoxyribonucleotide solution is prepared from sodium hyaluronate, poly-deoxyribonucleotide, mannitol, sodium dihydrogen phosphate, disodium hydrogen phosphate and water for injection, the sodium hyaluronate adopted by the invention is relatively large in molecular weight and can be used for filling superficial wrinkles of skin, and the poly-deoxyribonucleotide and the mannitol are supplemented, so that the skin wrinkles can be effectively removed, and the skin wrinkles can be effectively removed. On the basis of filling superficial wrinkles, the moisturizing, moisturizing and repairing effects are also achieved, and the skin state is improved. In addition, the preparation method of the polydeoxyribonucleotide solution is simple, large-scale production can be realized, and the application prospect is good.

Description

Technical Field

[0001] The present invention belongs to the field of passive implantable medical devices, namely plastic surgery and general surgery implants, and in particular relates to a polydeoxyribonucleotide solution for injection and a preparation method thereof. Background Art

[0002] Skin is the first protective layer of the human body. Moisturizing is the primary task in skin care, because the moisture of the skin is very easy to lose. Lack of sufficient water content in the skin will directly lead to abnormal skin metabolism and aging, wrinkles, spots and other skin problems. However, air conditioning, environmental pollution, temperature changes and pressure caused by seasonal changes, as well as skin aging, slow metabolism and other factors will cause skin moisture loss. Generally speaking, skin moisturizing refers to the skin's ability to hold water, which can slow down the rate of evaporation and loss of water in the skin.

[0003] Polydeoxyribonucleotide (PDRN for short) is a DNA fragment of specific specifications extracted from salmon germ cells. It is one of the complexes that generate DNA raw materials in cells. It has the function of stimulating the regeneration of collagen and elastic fibers, and has the effects of anti-inflammatory repair, hydration and moisturizing. It can be used to improve skin quality and revitalize skin. PDRN can be used as a fibroblast growth stimulator in wound repair. Furthermore, the nucleotides formed by DNA decomposition bind to adenosine A2A receptors, bidirectionally regulating related inflammatory and anti-inflammatory factors, and can exert a sustained anti-inflammatory effect. At the same time, it reduces the time required for healing and promotes human cell regeneration. Similar to injection beauty, it can activate the skin's self-healing ability, stimulate the continuous production of growth factors inside the skin, and promote fibroblast activity.

[0004] Therefore, PDRN directly acts on inflammation healing, injury repair, accelerates cell metabolism, improves skin repair and regeneration ability, and rebuilds healthy muscle base. The PDRN solution for injection is sterilized by sterilization filtration, which is easy to cause adverse reactions such as redness, swelling and pain, and has low safety. However, the terminal sterilization method will also affect the target molecular weight distribution of PDRN during the sterilization process. Therefore, it is urgent to develop a polydeoxyribonucleotide solution with good use effect, simple preparation, low cost, and terminal sterilization preparation process. Summary of the invention

[0005] Based on the above technical background, the purpose of the present invention is to provide a method for preparing a polydeoxyribonucleotide solution for injection in view of the defects and shortcomings of the prior art. The polydeoxyribonucleotide solution can not only fill wrinkles, but also promote the rapid recovery of the skin sebum membrane layer, and also has moisturizing, hydrating and repairing effects. In addition, it can reduce the risk of blood coagulation of sodium hyaluronate during clinical use, and can also reduce the incidence of adverse reactions such as redness, swelling and pain. The preparation method of the polydeoxyribonucleotide solution of the present invention is simple, can realize large-scale production, and has good application prospects.

[0006] In order to achieve the above object, the present invention adopts the following technical solutions:

[0007] The first aspect of the present invention is to provide a polydeoxyribonucleotide solution for injection, wherein the polydeoxyribonucleotide solution is prepared from polydeoxyribonucleotides, sodium hyaluronate, mannitol, sodium dihydrogen phosphate, sodium hydrogen phosphate and water for injection;

[0008] The mass ratio of the polydeoxyribonucleotide, sodium hyaluronate, mannitol, sodium dihydrogen phosphate, sodium hydrogen phosphate and water for injection is (0.01-8):(0.01-2):(0.01-2):(0.01-0.03):(0.02-0.05):100.

[0009] Preferably, the mass ratio of the polydeoxyribonucleotide, sodium hyaluronate, mannitol, sodium dihydrogen phosphate, sodium hydrogen phosphate and water for injection is (0.5-1.0):(0.2-0.4):(0.5-1):(0.02-0.03):(0.03-0.04):100.

[0010] More preferably, the mass ratio of the polydeoxyribonucleotide, sodium hyaluronate, mannitol, sodium dihydrogen phosphate, sodium hydrogen phosphate and water for injection is 0.8:0.3:1.0:0.02:0.03:100.

[0011] Preferably, the molecular weight of the sodium hyaluronate is 300K to 1500KDa;

[0012] Preferably, the polydeoxynucleotide is a salmon testis extract, and its molecular weight is 5KD to 1000KD.

[0013] The second aspect of the present invention is to provide a method for preparing the polydeoxyribonucleotide solution for injection according to the first aspect of the present invention, the method comprising the following steps:

[0014] Step 1, adding mannitol, sodium dihydrogen phosphate and disodium hydrogen phosphate to water for injection, stirring evenly, filtering and obtaining a mixed solution;

[0015] Step 2: Add polydeoxyribonucleotides and sodium hyaluronate to the mixed solution in sequence. After each raw material is added and stirred evenly, the next raw material is added. After sodium hyaluronate is added, stir evenly, sterilize, and obtain a polydeoxyribonucleotide solution for injection.

[0016] Preferably, in step 2, the sterilization temperature is 121° C., and the sterilization time is 15 to 25 minutes.

[0017] Compared with the prior art, the beneficial effects of the present invention are as follows: the present invention provides a method for preparing a polydeoxyribonucleotide solution for injection, which can reduce the risk of blood coagulation of sodium hyaluronate during clinical use, and can also reduce the incidence of adverse reactions such as redness, swelling and pain caused by PDRN; it can be prepared by a mechanized method, and the product process and quality have good stability and high reproducibility. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The molecular weight test graph of the polydeoxyribonucleotide solution prepared in Example 5 at different sterilization times is shown;

[0019] Figure 2 A comparison diagram of the red area image taken by VISIA-CR of the polydeoxyribonucleotide solution prepared in Example 5 is shown; DETAILED DESCRIPTION

[0020] This specific implementation method (Example 1) adopts the following technical solution: The operation steps of this embodiment are as follows:

[0021] The first aspect of the present invention is to provide a polydeoxyribonucleotide solution for injection, wherein the polydeoxyribonucleotide solution is prepared from polydeoxyribonucleotides, sodium hyaluronate, mannitol, sodium dihydrogen phosphate, sodium hydrogen phosphate and water for injection;

[0022] The mass ratio of the polydeoxyribonucleotide, sodium hyaluronate, mannitol, sodium dihydrogen phosphate, sodium hydrogen phosphate and water for injection is (0.01-8):(0.01-2):(0.01-2):(0.01-0.03):(0.02-0.05):100.

[0023] Preferably, the mass ratio of the polydeoxyribonucleotide, sodium hyaluronate, mannitol, sodium dihydrogen phosphate, sodium hydrogen phosphate and water for injection is (0.5-1.0):(0.2-0.4):(0.5-1):(0.02-0.03):(0.03-0.04):100.

[0024] More preferably, the mass ratio of the polydeoxyribonucleotide, sodium hyaluronate, mannitol, sodium dihydrogen phosphate, sodium hydrogen phosphate and water for injection is 0.8:0.3:1.0:0.02:0.03:100.

[0025] Preferably, the molecular weight of the sodium hyaluronate is 300K to 1500KDa;

[0026] Preferably, the polydeoxynucleotide is a salmon testis extract, and its molecular weight is 5KD to 1000KD.

[0027] Mannitol maintains cell structure and can prevent cells from being damaged due to water loss, pressure changes, etc. during the sterilization process, thereby maintaining the cell's morphology and structural integrity. It has a protective effect on biological macromolecules such as proteins and nucleic acids, preventing them from denaturing and inactivating due to physical or chemical factors during the sterilization process.

[0028] The second aspect of the present invention is to provide a method for preparing the polydeoxyribonucleotide solution for injection according to the first aspect of the present invention, the method comprising the following steps:

[0029] Step 1, adding mannitol, sodium dihydrogen phosphate and disodium hydrogen phosphate to water for injection, stirring evenly to obtain a mixed solution;

[0030] Step 2: Add polydeoxyribonucleotides and sodium hyaluronate to the mixed solution in sequence. After each raw material is added and stirred evenly, the next raw material is added. After sodium hyaluronate is added, stir evenly, sterilize, and obtain a polydeoxyribonucleotide solution for injection.

[0031] Preferably, in step 2, the sterilization temperature is 121° C., and the sterilization time is 15 to 25 minutes.

[0032] Example 1

[0033] Add mannitol, sodium dihydrogen phosphate and disodium hydrogen phosphate to water for injection, stir thoroughly and evenly, filter to obtain a mixed solution, in which the mass fractions of mannitol, disodium hydrogen phosphate and sodium dihydrogen phosphate are 0.1%, 0.03% and 0.02% respectively.

[0034] Add polydeoxyribonucleotide to the buffer solution in an amount of 0.5% of the total weight of the water for injection, stir evenly, then add sodium hyaluronate with a molecular weight of 300K to 1500KDa to the mixed solution in an amount of 1.0% of the total weight of the water for injection, stir evenly to obtain a polydeoxyribonucleotide solution for injection.

[0035] Example 2

[0036] Add mannitol, sodium dihydrogen phosphate and disodium hydrogen phosphate to water for injection, stir thoroughly and evenly, filter to obtain a mixed solution, in which the mass fractions of mannitol, disodium hydrogen phosphate and sodium dihydrogen phosphate are 0.1%, 0.03% and 0.02% respectively.

[0037] Add polydeoxyribonucleotide to the buffer solution in an amount of 0.7% of the total weight of the water for injection, stir evenly, then add sodium hyaluronate with a molecular weight of 300K to 1500KDa to the mixed solution in an amount of 0.3% of the total weight of the water for injection, stir evenly to obtain a polydeoxyribonucleotide solution for injection.

[0038] Example 3

[0039] Add mannitol, sodium dihydrogen phosphate and disodium hydrogen phosphate to water for injection, stir thoroughly and evenly, filter to obtain a mixed solution, in which the mass fractions of mannitol, disodium hydrogen phosphate and sodium dihydrogen phosphate are 0.1%, 0.03% and 0.02% respectively.

[0040] Add polydeoxyribonucleotide to the buffer solution in an amount of 0.6% of the total weight of the water for injection, stir evenly, then add sodium hyaluronate with a molecular weight of 300K to 1500KDa to the mixed solution in an amount of 0.4% of the total weight of the water for injection, stir evenly to obtain a polydeoxyribonucleotide solution for injection.

[0041] Example 4

[0042] Add mannitol, sodium dihydrogen phosphate and disodium hydrogen phosphate to water for injection, stir thoroughly and evenly, filter to obtain a mixed solution, in which the mass fractions of mannitol, disodium hydrogen phosphate and sodium dihydrogen phosphate are 0.1%, 0.03% and 0.02% respectively.

[0043] Add polydeoxyribonucleotide to the buffer solution in an amount of 0.8% of the total weight of the water for injection, stir evenly, then add sodium hyaluronate with a molecular weight of 300K to 1500KDa to the mixed solution in an amount of 0.4% of the total weight of the water for injection, stir evenly to obtain a polydeoxyribonucleotide solution for injection.

[0044] Example 5

[0045] Add mannitol, sodium dihydrogen phosphate and disodium hydrogen phosphate to water for injection, stir thoroughly and evenly, filter to obtain a mixed solution, in which the mass fractions of mannitol, disodium hydrogen phosphate and sodium dihydrogen phosphate are 0.1%, 0.03% and 0.02% respectively.

[0046] Add polydeoxyribonucleotide to the buffer solution in an amount of 0.8% of the total weight of the water for injection, stir evenly, then add sodium hyaluronate with a molecular weight of 300K to 1500KDa to the mixed solution in an amount of 0.3% of the total weight of the water for injection, stir evenly to obtain a polydeoxyribonucleotide solution for injection.

[0047] The hyperchromic effect data of the prepared polydeoxyribonucleotide solution are compared with those of Examples 1, 2, 3, and 4. Hyperchromic effect refers to the phenomenon that the molar absorption coefficient ε increases due to the change of the polymer structure, also known as the hyperchromic effect. In biological research, the hyperchromic effect usually refers to the increase in light absorption caused by DNA denaturation, that is, the effect of enhanced ultraviolet absorption of the DNA solution after denaturation. DNA molecules have the characteristic of absorbing ultraviolet light with a wavelength of 250 to 280 nm, and their absorption peak is at 260 nm. The interaction of electrons between bases in DNA molecules is the structural basis of ultraviolet absorption, but the orderly stacking of bases in the double helix structure "constrains" this effect.

[0048] Table 1 Comparison of color enhancement effect data of Examples 1, 2, 3, 4, and 5

[0049]

[0050] Test results: Example 5 The color enhancement effect of the polydeoxyribonucleotide solution of the present invention is better than that of the formulations with other proportions, further confirming that Example 5 is the preferred formulation.

[0051] The prepared polydeoxyribonucleotide for injection is analyzed according to the red area of ​​the skin. The red area images taken by VISIA-CR are analyzed by the color spot analysis in the skin comprehensive analysis module of Image Pro Plus software to measure the changes in the proportion of the red area of ​​the skin in the AOI selection area (radius 3.5cm) at different time points. By comparing the changes in the proportion of the red area of ​​the skin before and after use, the repair effect of the product can be reflected.

[0052] The more the red area ratio at different revisit time points decreases compared with the baseline value before use (DO), the smaller the skin redness area is, indicating that the product has a better effect in alleviating redness.

[0053] Red area percentage reduction rate = (red area percentage before using the product - red area percentage after using the product) / red area percentage before using the product × 100%

[0054] Table 2 Descriptive statistics of skin red area ratio (%, n = 33)

[0055]

[0056]

[0057] Table 3 Analysis of the difference in skin red area ratio (%, x±s, n=33)

[0058]

[0059] Test results: Compared with the baseline value (DO) before use, the red area ratio in the specific area of ​​the face showed a downward trend after using the product, and decreased by 26.75% after 28 days of use (D28), which was significantly different from the baseline value (p<0.01). The red area ratio in the specific area of ​​the face after using the product was lower than the baseline value. This shows that the polydeoxyribonucleotide repair effect of the present invention is significant, better than existing similar products, and can promote the rapid repair of the skin sebum membrane layer.

[0060] For those skilled in the art, they can modify the technical solutions described in the aforementioned embodiments and make equivalent substitutions for some of the technical features. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A polydeoxyribonucleotide solution, characterized in that The polydeoxyribonucleotide solution is prepared from polydeoxyribonucleotide, sodium hyaluronate, mannitol, sodium dihydrogen phosphate, disodium hydrogen phosphate and water for injection. The mass ratio of the polydeoxyribonucleotide, sodium hyaluronate, mannitol, sodium dihydrogen phosphate, sodium hydrogen phosphate and water for injection is (0.01-8):(0.01-2):(0.01-2):(0.01-0.03):(0.02-0.05):

100.

2. The polydeoxyribonucleotide solution for injection according to claim 1, characterized in that: The mass ratio of the polydeoxyribonucleotide, sodium hyaluronate, mannitol, sodium dihydrogen phosphate, sodium hydrogen phosphate and water for injection is (0.5-1.0):(0.2-0.4):(0.5-1):(0.02-0.03):(0.03-0.04):

100.

3. The sodium hyaluronate composite solution for injection according to claim 2, characterized in that: The mass ratio of the polydeoxyribonucleotide, sodium hyaluronate, mannitol, sodium dihydrogen phosphate, sodium hydrogen phosphate and water for injection is 0.8:0.3:1.0:0.02:0.03:

100.

4. The polydeoxyribonucleotide solution for injection according to claim 1, characterized in that: The sodium hyaluronate comprises cross-linked sodium hyaluronate, sodium hyaluronate and zinc hyaluronate.

5. The polydeoxyribonucleotide solution for injection according to claim 2, characterized in that: The molecular weight of the sodium hyaluronate is 300K to 1500KDa.

6. A polydeoxyribonucleotide solution according to claim 2, characterized in that: The polydeoxyribonucleotide is a salmon testis extract, and its molecular weight is 5KD-1000KD.

7. A method for preparing the polydeoxyribonucleotide solution for injection according to any one of claims 1 to 6, characterized in that: The method comprises the following steps: Step 1, adding mannitol, sodium dihydrogen phosphate and disodium hydrogen phosphate to water for injection, stirring evenly, filtering and obtaining a mixed solution; Step 2: Add polydeoxyribonucleotides and sodium hyaluronate to the mixed solution in sequence. After each raw material is added and stirred evenly, the next raw material is added. After sodium hyaluronate is added, stir evenly, sterilize, and obtain a polydeoxyribonucleotide solution for injection.

8. The method according to claim 7, characterized in that In step 2, the sterilization temperature is 121° C., and the sterilization time is 15 to 25 minutes.

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