Composite whitening preparation containing liquid CGF and tranexamic acid

Abstract

The invention belongs to the technical field of biological medicine, and particularly relates to a composite whitening preparation containing liquid CGF and tranexamic acid and a preparation method of the composite whitening preparation. The preparation method of the composite whitening preparation containing the liquid CGF and the tranexamic acid comprises the following steps: performing differential centrifugation on blood to obtain a liquid CGF mixture, and mixing the liquid CGF mixture with a tranexamic acid injection according to a specific proportion to obtain the composite whitening preparation. According to the compound whitening preparation, the compound whitening preparation is injected to the chloasma in a subcutaneous injection mode, through the synergistic interaction effect of the liquid CGF and tranexamic acid, the compound whitening preparation can effectively relieve the chloasma and also can improve the transparency of the whole skin color and the darkness of local skin blocks, so that the skin has the whitening effect that the skin is white and red, and the whitening effect is good. The effect of the tranexamic acid injection is far better than that of conventional pure tranexamic acid injection or liquid CGF, and the tranexamic acid injection can be well applied to the freckle removing market and is safer and milder.

Description

Technical Field

[0001] The invention belongs to the technical field of biomedicine, and specifically relates to a composite whitening preparation containing liquid CGF and tranexamic acid and a preparation method thereof. Background Art

[0002] Melasma is a difficult-to-treat skin disease that often occurs on the face and is characterized by irregular pigmentation spots. Currently, the main treatment drugs include hydroquinone, retinoic acid, etc., but their side effects are significant.

[0003] Tranexamic acid, also known as tranexamic acid, has a chemical name of 1,5-diaminocaproic acid. It is mainly used for various bleedings caused by acute or chronic, localized or systemic primary hyperfibrinolysis, and secondary hyperfibrinolytic states caused by disseminated intravascular coagulation. It is a widely used hemostatic drug. In addition, tranexamic acid can effectively improve the rough base color of the skin, play a role in the enzyme layer inside the skin, improve resistance and metabolism, make the skin less rough and less prone to small pimples, and has an excellent effect on inhibiting the formation of melanin and preventing the appearance of dark spots and freckles. One of its possible mechanisms is that tranexamic acid is similar in structure to the tyrosine part that participates in melanin metabolism, and has a common carboxyl group, which can competitively inhibit tyrosinase and thus reduce the formation of melanin protein. Compared with general traditional whitening ingredients, tranexamic acid has good stability and is not easily affected by temperature environment and transmission system. It can directly hinder the activation of melanocytes and improve the active state of melanin. Its mild and non-irritating nature is particularly suitable for the effects of lightening spots, whitening and balancing the overall whiteness of the skin. In addition to fading spots, tranexamic acid can also improve the overall skin tone and transparency and the dullness of local skin areas, giving the skin a "white and rosy" whitening effect.

[0004] Tranexamic acid is currently mostly used for injection. The effect of low-dose long-term oral administration of tranexamic acid in treating chloasma is very certain, but there is no systematic report in China. At present, the general dosage for treating chloasma by oral administration is 0.5-1.5g / day. Studies have shown that after 1-2 months of oral administration of tranexamic acid, the symptoms are significantly alleviated. However, oral administration is prone to side effects of gastrointestinal discomfort and menstrual disorders.

[0005] CGF, or Concentrated Growth Factors, is a new generation of platelet concentrates obtained by extracting autologous venous blood and subjecting it to variable speed centrifugation. CGF is rich in growth factors, such as vascular endothelial growth factor (VEGF), epidermal growth factor (EGF), insulin-like growth factor 1 (IGF-1), and fibroblast growth factor (FGF), as well as CD34-positive cells (CD34+ stem cells) and fibrin. These substances have anti-inflammatory effects, promote tissue and skin regeneration, and improve local soft tissue nutrition, so they are also called "immortality factors" in the blood. Therefore, CGF is often used for anti-aging, increasing skin elasticity, repairing scalp hair follicles, improving scalp blood circulation, and promoting hair growth. Summary of the invention

[0006] In view of the deficiencies in the above-mentioned prior art, the present invention provides a composite whitening preparation containing liquid CGF and tranexamic acid and a preparation method thereof, which can reduce pigmentation and brighten the skin tone of patients with pigmented and pigment-dominant chloasma, and simultaneously improve the physiological function of the patient's skin. The effect is significantly better than that of conventional simple tranexamic acid injection or liquid CGF, and can be well applied to the freckle removal market, being safer and gentler.

[0007] The specific technical solutions are as follows:

[0008] The first object of the present invention is to provide a method for preparing a composite whitening preparation containing liquid CGF and tranexamic acid, comprising the following steps:

[0009] (1) Preparation of liquid CGF: The collected blood is injected into an EDTA-K2 blood collection tube, subjected to differential centrifugation, and then the liquid in the middle layer is taken to obtain liquid CGF;

[0010] (2) Mixing liquid CGF and tranexamic acid injection: Place the liquid CGF obtained in step (1), tranexamic acid injection and lidocaine in a sterile centrifuge tube, stir and shake evenly, and let stand to obtain the composite whitening preparation containing liquid CGF and tranexamic acid.

[0011] Furthermore, in step (2), the volume ratio of the liquid CGF, tranexamic acid injection and lidocaine is (3.0-4.8):(1.5-2.4):(0-1.2).

[0012] Furthermore, in step (2), the volume ratio of the liquid CGF, tranexamic acid injection and lidocaine is 4:2:1.

[0013] Furthermore, in step (2), the specification of the tranexamic acid injection is 5ml:0.25g or 10ml:1.0g.

[0014] Furthermore, in step (2), the specification of the tranexamic acid injection is 10ml:1.0g.

[0015] Furthermore, in step (1), the differential centrifugation procedure is as follows: accelerate for 25-35 seconds, centrifuge for 1.7-2.3 minutes after the speed reaches 2600-2800 rpm, centrifuge for 3.5-4.5 minutes after reducing the speed to 2300-2500 rpm, then accelerate to 2600-2800 rpm for 3.5-4.5 minutes, centrifuge for 2.5-3.5 minutes at 3000-3200 rpm, and finally decelerate for 25-35 seconds to stop, and stand for 2.5-3.5 minutes.

[0016] Furthermore, in step (1), the differential centrifugation procedure is as follows: accelerate for 30 seconds, centrifuge for 2 minutes after the speed reaches 2700 rpm, centrifuge for 4 minutes after reducing the speed to 2400 rpm, then accelerate to 2700 rpm for 4 minutes, centrifuge for 3 minutes at 3100 rpm, and finally decelerate for 30 seconds to stop and stand for 3 minutes.

[0017] Furthermore, in step (1), after the differential centrifugation, the blood in the EDTA-K2 blood collection tube is divided into three layers, the top layer is serum PPP, the middle layer is liquid CGF, and the bottom layer is red blood cells.

[0018] Furthermore, the liquid CGF obtained in step (1) includes substances between PPP (platelet-poor plasma) and αPRP (platelet-rich plasma), αPRP, CD34+, and substances between CD34+ and red blood cells.

[0019] The second object of the present invention is to provide a composite whitening preparation containing liquid CGF and tranexamic acid, which is prepared by the preparation method of the composite whitening preparation containing liquid CGF and tranexamic acid as described above.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] The main components of the composite whitening preparation provided by the present invention are liquid CGF and tranexamic acid. The present invention mixes the liquid CGF obtained by differential centrifugation with tranexamic acid injection in a specific ratio to obtain the composite whitening preparation; the composite whitening preparation of the present invention is injected into the chloasma by subcutaneous injection, and the synergistic effect of the liquid CGF and tranexamic acid is used to effectively alleviate the chloasma, improve the overall skin color transparency and the dullness of local skin patches, and make the skin have a "white and rosy" whitening effect, which is far superior to conventional simple tranexamic acid injection or liquid CGF, can be well applied to the freckle removal market, and is safer and milder. DETAILED DESCRIPTION

[0022] The principles and features of the present invention are described below in conjunction with examples. The examples are only used to explain the present invention and are not used to limit the scope of the present invention.

[0023] Example 1

[0024] A method for preparing a composite whitening preparation containing liquid CGF and tranexamic acid comprises the following steps:

[0025] (1) Preparation of liquid CGF: Before surgery, venous blood was drawn from the patient's arm, and 8 ml of the collected venous blood was injected into an EDTA-K2 blood collection tube for differential centrifugation. The differential centrifugation procedure was as follows: accelerate for 30 seconds, centrifuge for 2 minutes after the speed reaches 2700 rpm, centrifuge for 4 minutes after the speed is reduced to 2400 rpm, then accelerate to 2700 rpm for 4 minutes, centrifuge for 3 minutes at 3100 rpm, and finally decelerate for 30 seconds to stop, let it stand for 3 minutes, then take the liquid in the middle layer to obtain liquid CGF, and take 4 ml;

[0026] (2) Mixing of liquid CGF and tranexamic acid injection: Place 4 ml of the liquid CGF obtained in step (1), 2 ml of tranexamic acid injection (specification: 10 ml: 1.0 g), and 1 ml of lidocaine in a 15 ml sterile centrifuge tube, stir and shake evenly, and let stand to obtain the composite whitening preparation containing liquid CGF and tranexamic acid.

[0027] Example 2

[0028] A method for preparing a composite whitening preparation containing liquid CGF and tranexamic acid comprises the following steps:

[0029] (1) Preparation of liquid CGF: Before surgery, venous blood was drawn from the patient's arm, and 8 ml of the collected venous blood was injected into an EDTA-K2 blood collection tube for differential centrifugation. The differential centrifugation procedure was as follows: accelerate for 35 seconds, centrifuge for 1.7 minutes after the speed reaches 2800 rpm, centrifuge for 3.5 minutes after the speed is reduced to 2500 rpm, then accelerate to 2800 rpm for 3.5 minutes, centrifuge for 2.5 minutes at 3200 rpm, and finally decelerate for 35 seconds to stop, let stand for 3.5 minutes, then take the liquid in the middle layer to obtain liquid CGF, and take 4.8 ml;

[0030] (2) Mixing of liquid CGF and tranexamic acid injection: Place 4.8 ml of the liquid CGF obtained in step (1), 1.5 ml of tranexamic acid injection (specification: 10 ml: 1.0 g), and 1.2 ml of lidocaine in a 15 ml sterile centrifuge tube, stir and shake evenly, and let stand to obtain the composite whitening preparation containing liquid CGF and tranexamic acid.

[0031] Example 3

[0032] A method for preparing a composite whitening preparation containing liquid CGF and tranexamic acid comprises the following steps:

[0033] (1) Preparation of liquid CGF: Before surgery, venous blood was drawn from the patient's arm, and 8 ml of the collected venous blood was injected into an EDTA-K2 blood collection tube for differential centrifugation. The differential centrifugation procedure was as follows: accelerate for 25 seconds, centrifuge for 2.3 minutes after the speed reaches 2600 rpm, centrifuge for 4.5 minutes after the speed is reduced to 2300 rpm, then accelerate to 2600 rpm for 4.5 minutes, centrifuge for 3.5 minutes at 3000 rpm, and finally decelerate for 25 seconds to stop, let stand for 2.5 minutes, then take the liquid in the middle layer to obtain liquid CGF, and take 3 ml;

[0034] (2) Mixing of liquid CGF and tranexamic acid injection: Place 3 ml of the liquid CGF obtained in step (1) and 2.4 ml of tranexamic acid injection (specification: 10 ml: 1.0 g) in a 15 ml sterile centrifuge tube, stir and shake evenly, and let stand to obtain the composite whitening preparation containing liquid CGF and tranexamic acid.

[0035] Example 4

[0036] A method for preparing a composite whitening preparation containing liquid CGF and tranexamic acid, which is different from Example 1 in that the specification of the tranexamic acid injection is 5ml:0.25g.

[0037] Comparative Example 1

[0038] A method for preparing a whitening preparation containing tranexamic acid comprises the following steps:

[0039] Place 4 ml of normal saline, 2 ml of tranexamic acid injection (specification: 10 ml: 1.0 g) and 1 ml of lidocaine in a 15 ml sterile centrifuge tube, stir and shake evenly, and let stand for later use.

[0040] Comparative Example 2

[0041] A method for preparing a whitening preparation containing liquid CGF comprises the following steps:

[0042] (1) Preparation of liquid CGF: refer to Example 1;

[0043] (2) Place 4 ml of the liquid CGF obtained in step (1), 2 ml of normal saline and 1 ml of lidocaine in a 15 ml sterile centrifuge tube, stir and shake evenly, and let stand for later use.

[0044] test

[0045] Subjects:

[0046] A total of 180 patients with melasma were selected as subjects and randomly divided into six groups, with 30 patients in each group.

[0047] Treatment options:

[0048] Inject the whitening preparations of Examples 1-4 and Comparative Examples 1-2 into the chloasma area of ​​the patient respectively;

[0049] Inject 0.05 ml of whitening preparation solution subcutaneously every 1 cm at the patient's melasma site once a week for 12 weeks, and continuously record the results.

[0050] Observation indicators:

[0051] ①Clinical efficacy

[0052] The efficacy was judged by referring to the following standards: if the area of ​​facial pigmentation of the patient decreased by more than 90% after the course of treatment, the patient was cured; if the area of ​​facial pigmentation of the patient decreased by 60% to 90% after the course of treatment, and the color became obviously lighter, the patient was markedly effective; if the area of ​​facial pigmentation of the patient decreased by 30% to 59% after the course of treatment, and the color faded a little, the patient was effective; if the area of ​​facial pigmentation of the patient decreased by less than 30% after the course of treatment, and there was no obvious change in the color of the pigmentation, the patient was ineffective; the total of cured, markedly effective and effective was the total effective.

[0053] ②Melasma area and severity index (MASI) scores before and after treatment

[0054] The MASI score ranges from 0 to 48, and the higher the score, the more severe the melasma symptoms.

[0055] ③Adverse reactions and recurrence

[0056] Adverse reactions include redness, swelling, pain, pigmentation, coagulation abnormalities, etc.

[0057] Statistical methods:

[0058] SPSS 27.0 software was used for data analysis. Measurement data were expressed as mean ± standard deviation (x ± s), and one-way analysis of variance and paired sample t test were used; count data were expressed as rate, and chi-square test was used. P < 0.05 was considered statistically different, and P < 0.01 was considered statistically significant.

[0059] result:

[0060] 1. Comparison of clinical efficacy

[0061] The clinical efficacy of each embodiment and comparative example is compared, and the results are shown in Table 1, where n represents the number of people.

[0062] Table 1 Comparison of clinical efficacy of each group [n (%)]

[0063] Group n get well Significant effect efficient invalid Total effective Example 1 30 19(63.33) 8(26.67) 2(6.67) 1(3.33) 29(96.67) Example 2 30 18(60.00) 9(30.00) 2(6.67) 1(3.33) 29(96.67) Example 3 30 17(56.67) 8(26.67) 3(10.00) 2(6.67) 28(93.33) Example 4 30 17(56.67) 8(26.67) 2(6.67) 3(10.00) 27(90.00) Comparative Example 1 30 11(36.67) 6(20.00) 4(13.33) 9(30.00) 21(70.00) Comparative Example 2 30 13(43.33) 4(13.33) 5(16.67) 8(26.67) 22(73.33)

[0064] The total effective rate of treatment in Example 1 was 96.67%, the total effective rate of treatment in Example 2 was 96.67%, the total effective rate of treatment in Example 3 was 93.33%, and the total effective rate of treatment in Example 4 was 90.00%, which were higher than 70.00% and 73.33% of Comparative Example 1 and Comparative Example 2 respectively (P < 0.05, the difference was statistically significant).

[0065] 2. Comparison of MASI scores before and after treatment

[0066] The MASI scores of the embodiments and comparative examples are compared, and the results are shown in Table 2, where n represents the number of people.

[0067] Table 2 Comparison of MASI scores before and after treatment in each group (x±s, points)

[0068] Group n Before treatment After treatment Example 1 30 10.03±1.96 3.9±1.75 Example 2 30 10.05±2.09 3.8±2.04 Example 3 30 10.03±1.75 3.9±1.69 Example 4 30 10.04±2.04 4.0±1.96 Comparative Example 1 30 10.06±2.09 7.1±2.17 Comparative Example 2 30 10.01±2.17 6.17±2.04

[0069] Comparison of MASI scores before and after treatment in Examples 1, 2, 3, and 4 showed significant differences (P < 0.01);

[0070] Meanwhile, the MASI scores after treatment in Examples 1, 2, 3, 4 were compared with those in Comparative Examples 1 and 2, and the differences were significant (P < 0.01).

[0071] Comparison of adverse reactions and recurrence

[0072] During the treatment, no obvious adverse reactions occurred in the three groups.

[0073] During the follow-up period, 1 case of relapse occurred in Example 1, 2 cases of relapse occurred in Example 2, 1 case of relapse occurred in Example 3, 3 cases of relapse occurred in Example 4, 9 cases of relapse occurred in Comparative Example 1, and 8 cases of relapse occurred in Comparative Example 2. The relapse rate of Example 1 was 3.3%, the relapse rate of Example 2 was 6.6%, the relapse rate of Example 3 was 3.3%, and the relapse rate of Example 4 was 10%, which were all lower than 30.0% of Comparative Example 1 and 26.7% of Comparative Example 1 (P<0.05, the difference was statistically significant).

[0074] It can be seen from the above test results that the composite whitening preparation containing liquid CGF and tranexamic acid in the embodiment of the present invention can effectively alleviate chloasma through the synergistic effect of liquid CGF and tranexamic acid, and can also improve the overall skin transparency and the dullness of local skin patches, so that the skin has a "white and rosy" whitening effect, and the effect is far better than the conventional simple tranexamic acid injection (Comparative Example 1) or liquid CGF (Comparative Example 2), and can be well applied to the freckle removal market, which is safer and milder.

[0075] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a composite whitening preparation containing liquid CGF and tranexamic acid, characterized in that, The steps include: (1) Preparation of liquid CGF: The collected blood is injected into an EDTA-K2 blood collection tube, subjected to differential centrifugation, and then the liquid in the middle layer is taken to obtain liquid CGF; (2) Mixing liquid CGF and tranexamic acid injection: Place the liquid CGF obtained in step (1), tranexamic acid injection and lidocaine in a sterile centrifuge tube, stir and shake evenly, and let stand to obtain the composite whitening preparation containing liquid CGF and tranexamic acid.

2. The preparation method of the composite whitening preparation containing liquid CGF and tranexamic acid according to claim 1, characterized in that, In step (2), the volume ratio of the liquid CGF, tranexamic acid injection and lidocaine is (3.0-4.8):(1.5-2.4):(0-1.2).

3. The preparation method of the composite whitening preparation containing liquid CGF and tranexamic acid according to claim 2, characterized in that, In step (2), the volume ratio of the liquid CGF, tranexamic acid injection and lidocaine is 4:2:

1.

4. The method for preparing a composite whitening preparation containing liquid CGF and tranexamic acid according to any one of claims 1 to 3, characterized in that: In step (2), the specification of the tranexamic acid injection is 5ml: 0.25g or 10ml: 1.0g.

5. The preparation method of the composite whitening preparation containing liquid CGF and tranexamic acid according to claim 4, characterized in that, In step (2), the specification of the tranexamic acid injection is 10ml:1.0g.

6. The preparation method of the composite whitening preparation containing liquid CGF and tranexamic acid according to claim 1, characterized in that, In step (1), the differential centrifugation procedure is as follows: accelerate for 25-35 seconds, centrifuge for 1.7-2.3 minutes after the speed reaches 2600-2800 rpm, centrifuge for 3.5-4.5 minutes after reducing the speed to 2300-2500 rpm, then accelerate to 2600-2800 rpm for 3.5-4.5 minutes, centrifuge for 2.5-3.5 minutes at 3000-3200 rpm, and finally decelerate for 25-35 seconds to stop, and stand for 2.5-3.5 minutes.

7. The preparation method of the composite whitening preparation containing liquid CGF and tranexamic acid according to claim 6, characterized in that, In step (1), the differential centrifugation procedure is as follows: accelerate for 30 seconds, centrifuge for 2 minutes after the speed reaches 2700 rpm, centrifuge for 4 minutes after the speed decreases to 2400 rpm, then accelerate to 2700 rpm for 4 minutes, centrifuge for 3 minutes at 3100 rpm, and finally decelerate for 30 seconds to stop and stand for 3 minutes.

8. The preparation method of the composite whitening preparation containing liquid CGF and tranexamic acid according to claim 7, characterized in that, In step (1), after the differential centrifugation, the blood in the EDTA-K2 blood collection tube is divided into three layers, the top layer is serum PPP, the middle layer is liquid CGF, and the bottom layer is red blood cells.

9. The method for preparing a composite whitening preparation containing liquid CGF and tranexamic acid according to claim 8, wherein: The liquid CGF obtained in step (1) includes substances between PPP and αPRP, αPRP, CD34+, and substances between CD34+ and red blood cells.

10. A composite whitening preparation containing liquid CGF and tranexamic acid, characterized in that, The whitening preparation is prepared by the method for preparing the composite whitening preparation containing liquid CGF and tranexamic acid as described in any one of claims 1 to 9.