Crystal form of acetyl tetrapeptide-9 as well as preparation method and application of crystal form
By preparing a new crystal form of acetyl tetrapeptide-9, the problem of instability of acetyl tetrapeptide-9 in skin care products was solved, achieving the effects of low thermal stability and low hygroscopicity, making it suitable for use in cosmetics, especially lotions, day and night creams, and eye serums.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- SHENZHEN READLINE BIOTECH CO LTD
- Filing Date
- 2026-01-30
- Publication Date
- 2026-04-28
AI Technical Summary
The existing acetyl tetrapeptide-9 active ingredient has unstable performance in skin care products and cannot achieve the expected results.
A novel crystalline form of acetyl tetrapeptide-9 is provided by means of specific X-ray powder diffraction characteristic peaks (such as diffraction peaks at 2θ±0.2°) and preparation methods, including contact crystallization of acetyl tetrapeptide-9 aqueous solution with antisolvent, and control of temperature and stirring conditions, to prepare a crystalline form with good thermal stability and low hygroscopicity.
The acetyl tetrapeptide-9 crystal form exhibits good thermal stability and low hygroscopicity, making it suitable for large-scale industrial production and enhancing the stability and functionality of active ingredients in cosmetics.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of novel drug crystal forms, and more particularly to a crystal form of acetyl tetrapeptide-9, its preparation method, and its application. Background Technology
[0002] The polypeptide structure of acetyl tetrapeptide-9 is N-Ac-Gln-Asp-Val-His-OH, where acetyl modification enhances its stability and bioactivity. It improves skin firmness and elasticity by mimicking the mechanism of action of transforming growth factor β (TGFβ), binding to specific receptors on fibroblast membranes, and stimulating the synthesis of collagen, elastin, and extracellular matrix (ECM) components.
[0003] Acetyl tetrapeptide-9 is widely used in anti-wrinkle and anti-aging skincare products, such as lotions, day and night creams, and eye serums, to help improve skin quality and delay aging. However, existing acetyl tetrapeptide-9 active ingredients have performance instability issues in practical applications, failing to achieve the expected results. Summary of the Invention
[0004] In view of this, the technical problem to be solved by the present invention is to provide a crystal form of acetyl tetrapeptide-9, a preparation method and application thereof, wherein the prepared new crystal form has good thermal stability and low hygroscopicity.
[0005] To solve the above technical problems, the present invention provides a crystal form of acetyl tetrapeptide-9, which has diffraction peaks at 2θ±0.2° in its X-ray powder diffraction pattern, wherein 2θ includes: 9.444°, 10.812°, 12.717°, 17.672°, 19.143°, 19.606° and 23.613°;
[0006] Or include: 9.292°, 10.776°, 12.631°, 17.656°, 18.762°, 19.333° and 23.520°;
[0007] Or include: 9.340°, 10.806°, 12.721°, 17.723°, 18.778°, 19.374° and 23.549°.
[0008] Furthermore, the 2θ includes: 9.444°, 10.812°, 12.717°, 17.672°, 18.485°, 19.143°, 19.606°, 23.613°, 24.575°, 25.696°, 28.071°, and 37.392°.
[0009] Specifically, the X-ray powder diffraction pattern of the crystal form of acetyl tetrapeptide-9 described in this invention is as follows: Figure 1 ,2 Or as shown in Figure 3.
[0010] In this invention, the 2θ values of X-ray powder diffraction patterns may vary slightly between machines or samples, with differences of approximately 0.2 units or 0.1 units. Therefore, the values cited should not be interpreted as absolute values. Similarly, it should be understood that peak heights may also vary by approximately 5 units, 4 units, 3 units, 2 units, or 1 unit. Therefore, the XRPD trace intensities included in this invention are illustrative and not intended for absolute comparison.
[0011] This invention provides a method for preparing the above-mentioned acetyl tetrapeptide-9 crystal form, comprising the following steps:
[0012] An aqueous solution of acetyl tetrapeptide-9 was contacted with an antisolvent to induce crystallization and obtain the crystalline form of acetyl tetrapeptide-9.
[0013] The antisolvent includes one or more of acetone, dimethyl sulfoxide, and 1,4-dioxane.
[0014] The concentration of the aqueous solution of acetyl tetrapeptide-9 is preferably 10~20 g / L, and for example, it can be 10, 13, 15, 18, or 20 g / L.
[0015] The volume ratio of the aqueous solution of acetyl tetrapeptide-9 to the antisolvent is 1:(1~3), more preferably 1:(1.5~2), and for example, it can be 1:1.5, 1:1.6, 1:1.7, 1:1.8, 1:1.9, or 1:2.
[0016] The contact between the aqueous solution of acetyl tetrapeptide-9 and the antisolvent is specifically achieved by adding the antisolvent dropwise into the aqueous solution of acetyl tetrapeptide-9.
[0017] The preferred dripping rate is 300-500 mL / h, and for example, it can be 300, 350, 400, 450, or 500 mL / h.
[0018] Preferably, the dripping is carried out under stirring conditions; the stirring speed is preferably 50~250 rpm, more preferably 100~200 rpm, and for example, it can be 100, 150, or 200 rpm.
[0019] Preferably, after the addition is completed, the system temperature is maintained and stirring is continued for 1 to 48 hours to grow crystals. More preferably, stirring is continued for 4 to 10 hours. For example, it can be 4, 5, 6, 7, 8, 9, or 10 hours.
[0020] The preferred temperature for contacting the aqueous solution of acetyl tetrapeptide-9 with the antisolvent is 15~45℃, more preferably 20~35℃, and for example, it can be 20, 25, 30, or 35℃.
[0021] The crystallization temperature is preferably 4~14℃, more preferably 5~10℃, and for example, it can be 5, 6, 7, 8, 9, or 10℃.
[0022] The crystallization time is preferably 8 to 24 hours, and for example, it can be 8, 10, 14, 20, or 24 hours.
[0023] Preferably, the crystallization is specifically performed by cooling the temperature to 4-14°C at a rate of 3-10°C / h, and the cooling rate is more preferably 4-7°C / h. For example, it can be 4, 5, 6, or 7°C / h.
[0024] After crystallization, filter and dry the crystals.
[0025] In some specific embodiments, the method for preparing the acetyl tetrapeptide-9 crystal form includes the following steps:
[0026] Prepare an aqueous solution of acetyl tetrapeptide-9 with a concentration of 10-20 g / L and pH 4.5 at 15-45℃. Stir at a low speed of 50-250 rpm and add 1-3 times the volume of antisolvent dropwise at a rate of 400-500 mL / h. After the addition is complete, maintain the temperature and continue stirring for 6 hours to grow crystals. Then, reduce the temperature to 10℃ at a rate of 5℃ per hour and continue crystallizing for 8-24 hours. Filter the solution, wash the powder with an appropriate amount of the corresponding antisolvent, and then vacuum dry at 45℃ until the moisture content is ≤8%.
[0027] The present invention also provides the application of the crystal form of the above-mentioned acetyl tetrapeptide-9 or the crystal form of acetyl tetrapeptide-9 prepared by the above preparation method in the preparation of cosmetics.
[0028] The cosmetics mentioned can be any type of cosmetics known to those skilled in the art, including but not limited to serums, face creams, lotions, eye creams, cleansers, face masks, sunscreens, foundations, cushion compacts, etc.
[0029] Based on this, the present invention also provides a cosmetic composition comprising the crystal form of acetyl tetrapeptide-9 described above or the crystal form of acetyl tetrapeptide-9 prepared by the above preparation method.
[0030] Depending on the cosmetic dosage form, the cosmetic composition may also include suitable excipients, which may be added as needed by those skilled in the art, without any special limitations.
[0031] The cosmetic may also include other functional ingredients, including but not limited to whitening, moisturizing, and antioxidant ingredients. Those skilled in the art can add them as needed, and the present invention does not impose any special limitations on this.
[0032] Compared with the prior art, the present invention provides a crystal form of acetyl tetrapeptide-9, the X-ray powder diffraction pattern of which has diffraction peaks at 2θ±0.2°, wherein 2θ includes: 9.444°, 10.812°, 12.717°, 17.672°, 19.143°, 19.606° and 23.613°; or includes: 9.292°, 10.776°, 12.631°, 17.656°, 18.762°, 19.333° and 23.520°; or includes: 9.340°, 10.806°, 12.721°, 17.723°, 18.778°, 19.374° and 23.549°.
[0033] Crystal form differences affect the physicochemical properties of drugs (active ingredients), manifesting in melting point, solubility, and stability. These properties are particularly important in cosmetic applications. For example, solubility determines the dispersibility and absorption of the active ingredient in the formulation, while stability affects its performance during storage, preventing quality problems caused by crystal form changes. The functionality of active ingredients (such as anti-aging and moisturizing) also depends on their physicochemical properties, which are in turn affected by crystal form. Furthermore, the compatibility characteristics of active ingredients with excipients (such as compatibility, stability, and release properties) directly determine their application effect in actual products.
[0034] The novel crystalline form of acetyl tetrapeptide-9 provided by this invention has good thermal stability and low hygroscopicity, giving it the advantage of large-scale industrial production. Attached Figure Description
[0035] Figure 1 Powder diffraction pattern of the acetyl tetrapeptide-9 crystal form prepared in Example 1;
[0036] Figure 2 Powder diffraction pattern of the acetyl tetrapeptide-9 crystal form prepared in Example 2;
[0037] Figure 3 Powder diffraction pattern of the acetyl tetrapeptide-9 crystal form prepared in Example 3;
[0038] Figure 4 Powder diffraction pattern of the acetyl tetrapeptide-9 crystal form prepared in Example 1. Detailed Implementation
[0039] To further illustrate the present invention, the crystal form and preparation method of acetyl tetrapeptide-9 provided by the present invention are described in detail below with reference to embodiments.
[0040] Example 1
[0041] At 25℃, 1000 mL of a 15 g / L acetyl tetrapeptide-9 aqueous solution was prepared. The mixture was stirred continuously at 200 rpm, and acetone was added dropwise at a rate of 400 mL / h, twice the volume of the solution. After the addition was complete, the temperature was maintained and stirred for 6 h to allow crystal growth. The temperature was then lowered to 10℃ at a rate of 6℃ per hour, and crystallization continued for 8 h. The mixture was filtered, and the powder was washed with an appropriate amount of acetone. It was then vacuum dried at 45℃ until the moisture content met the requirements. The yield was 95%, and the purity was 99.7%. Figure 1 The powder X-ray diffraction pattern shown has characteristic peaks at 2θ = 9.444°, 10.812°, 12.717°, 17.672°, 19.143°, 19.606° and 23.613°.
[0042] The diffraction results of the prepared acetyl tetrapeptide-9- powder are as follows:
[0043] Table 1
[0044]
[0045] Example 2
[0046] At 20℃, 1000 mL of a 10 g / L acetyl tetrapeptide-9 aqueous solution was prepared and stirred continuously at 150 rpm. 1,4-dioxane was added dropwise at a rate of 350 mL / h, with 1.5 times its volume of acetyl tetrapeptide-9 added. After the addition was complete, the temperature was maintained and stirred for 6 h to allow crystal growth. The temperature was then lowered to 10℃ at a rate of 5℃ per hour, and crystallization continued for 14 h. The mixture was filtered, and the powder was washed with an appropriate amount of 1,4-dioxane. The powder was then vacuum dried at 45℃ until the moisture content met the requirements. The yield was 90%, and the purity was 99.2%. Figure 2 The powder X-ray diffraction pattern shown, measured with Cu Kα rays, has characteristic peaks at 2θ = 9.292°, 10.776°, 12.631°, 17.656°, 18.762°, 19.333° and 23.520°, indicating that it is the same crystal form as that prepared in Example 1.
[0047] Example 3
[0048] At 35℃, 1000 mL of a 18 g / L acetyl tetrapeptide-9 aqueous solution was prepared and stirred continuously at 150 rpm. 1.6 times the volume of dimethyl sulfoxide was added dropwise at a rate of 450 mL / h. After the addition was complete, the temperature was maintained and stirred for 6 h to allow crystal growth. The temperature was then lowered to 10℃ at a rate of 5℃ per hour, and crystallization continued for 20 h. The mixture was filtered, and the powder was washed with an appropriate amount of dimethyl sulfoxide. The powder was then vacuum dried at 45℃ until the moisture content met the requirements. The yield was 93%, and the purity was 99.1%. Figure 3The powder X-ray diffraction pattern shown, measured with Cu Kα rays, has characteristic peaks at 2θ = 9.340°, 10.806°, 12.721°, 17.723°, 18.778°, 19.374°, and 23.549°, indicating that it is the same crystal form as that prepared in Example 1.
[0049] Comparative Example 1
[0050] At 25℃, 1000 mL of a 15 g / L acetyl tetrapeptide-9 aqueous solution was prepared, rapidly frozen at -80℃ under a vacuum of 50 Pa and an ambient temperature of 20℃, and freeze-dried for 24 h to obtain an amorphous powder with a yield of 92% and a purity of 99.5%. Its XRD pattern is shown below. Figure 4 .
[0051] Application Example 1
[0052] The hygroscopicity test method was as follows: Nine new 20 mL glass bottles were selected, dried in an oven at 105℃ until constant weight, and numbered. After the products prepared in Example 1, Example 2 and Comparative Example 1 were dried to constant weight, 1 g of powder was filled into each glass bottle and placed in a constant temperature and humidity chamber (room temperature, 95% humidity). The samples were weighed every 24 h until the samples reached constant weight. The results are shown in Table 2. The results show that the crystal form prepared by this invention is more stable in a high humidity environment.
[0053] Table 2 Hygroscopicity Results
[0054]
[0055] The temperature stability test method is as follows: 4-5g of the products prepared in Example 1, Example 2 and Comparative Example 1, dried to constant weight, are packed into a clean glass bottle. The glass bottle is placed in an incubator at 45℃ for 30 days. The powder composition is then tested. The results are shown in Table 3. The results show that the crystal form prepared in this invention has better temperature stability.
[0056] Table 3 Temperature stability test results
[0057]
[0058] The results of hygroscopicity and temperature stability tests show that the new crystal form of acetyl tetrapeptide-9 prepared in this invention has higher thermal stability and lower hygroscopicity.
[0059] The above description of the embodiments is only for the purpose of helping to understand the method and core ideas of the present invention. It should be noted that those skilled in the art can make several improvements and modifications to the present invention without departing from the principles of the present invention, and these improvements and modifications also fall within the protection scope of the claims of the present invention.
Claims
1. A crystal form of acetyl tetrapeptide-9, characterized in that, The X-ray powder diffraction pattern shows diffraction peaks at 2θ ± 0.2°, where 2θ includes: 9.444°, 10.812°, 12.717°, 17.672°, 19.143°, 19.606°, and 23.613°. Or include: 9.292°, 10.776°, 12.631°, 17.656°, 18.762°, 19.333° and 23.520°; Or include: 9.340°, 10.806°, 12.721°, 17.723°, 18.778°, 19.374° and 23.549°.
2. The crystal form of acetyl tetrapeptide-9 according to claim 1, characterized in that, The 2θ includes: 9.444°, 10.812°, 12.717°, 17.672°, 18.485°, 19.143°, 19.606°, 23.613°, 24.575°, 25.696°, 28.071°, and 37.392°.
3. The crystal form of acetyl tetrapeptide-9 according to claim 1, characterized in that, The X-ray powder diffraction pattern is shown in Figure 1, 2, or 3.
4. A method for preparing the acetyl tetrapeptide-9 crystal form, comprising the following steps: An aqueous solution of acetyl tetrapeptide-9 was contacted with an antisolvent to induce crystallization and obtain the crystalline form of acetyl tetrapeptide-9. The antisolvent includes one or more of acetone, dimethyl sulfoxide, and 1,4-dioxane.
5. The preparation method according to claim 4, characterized in that, The concentration of the aqueous solution of acetyl tetrapeptide-9 is 10~20 g / L.
6. The preparation method according to claim 4, characterized in that, The volume ratio of the aqueous solution of acetyl tetrapeptide-9 to the antisolvent is 1:(1~3).
7. The preparation method according to claim 4, characterized in that, The contact between the aqueous solution of acetyl tetrapeptide-9 and the antisolvent is specifically as follows: The antisolvent was added dropwise to an aqueous solution of acetyl tetrapeptide-9; The dripping rate is 300~500 mL / h; The addition is carried out under stirring conditions; The stirring speed is 50~250 rpm.
8. The preparation method according to claim 4, characterized in that, The aqueous solution of acetyl tetrapeptide-9 is in contact with the antisolvent at a temperature of 15~45℃; The crystallization temperature is 4~14℃; The crystallization time is 8~24h.
9. The preparation method according to claim 4, characterized in that, The crystallization process specifically involves cooling the temperature to 4~14℃ at a rate of 5℃ / h to induce crystallization.
10. The use of the crystal form of acetyl tetrapeptide-9 according to any one of claims 1 to 3 or the crystal form of acetyl tetrapeptide-9 prepared by the preparation method according to any one of claims 4 to 9 in the preparation of cosmetics.