Tofacitinib citrate gel as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of pharmaceutical preparations, and particularly relates to a tofacitinib citrate gel as well as a preparation method and application thereof. The invention provides a citric acid tofacitinib gel. The gel comprises the following components in percentage by mass: 0.1-5% of an active component citric acid tofacitinib, 0.5-15% of a gel matrix, 0.1-5% of a surfactant, 5-30% of a transdermal enhancer, 0.01-1% of a preservative and the balance of water, the tofacitinib citrate gel prepared by the preparation method disclosed by the invention is proper in viscosity, good in moisture retention, high in active ingredient content, relatively small in total impurity and single impurity and good in stability; the tofacitinib citrate gel has a remarkable effect of promoting hair follicle growth, has no irritation to skin, and can be used for treating alopecia related diseases.

Description

Technical Field

[0001] The present invention belongs to the technical field of pharmaceutical preparations, and in particular relates to a tofacitinib citrate gel, a preparation method and uses thereof. Background Art

[0002] Alopecia areata (AA) is a hair loss symptom with a high incidence rate worldwide. The clinical manifestations of this disease are sudden, unexplained localized or complete hair loss, and the formation of significant patchy hair loss, which can easily cause a heavy mental burden on patients. Alopecia areata symptoms often first occur in childhood, and people of any age, gender and race may suffer from alopecia areata. Public information shows that there are currently limited treatments for alopecia areata, and there are no approved drugs for specific treatment. Studies have shown that alopecia areata is an autoimmune disease caused by immune-mediated processes. Since many cytokines involved in the pathogenesis of alopecia areata are dependent on JAK-STAT signaling, the JAK-STAT signaling pathway is a cytokine-stimulated signal transduction pathway discovered in recent years. It is involved in many important biological processes such as cell proliferation, differentiation, apoptosis and immune regulation. Therefore, JAK-STAT is considered to be an attractive signaling pathway in the treatment of alopecia areata.

[0003] Tofacitinib, CAS No. 540737-29-9, is a selective JAK inhibitor that can selectively inhibit JAK1 and JAK3 and block the JAK-STAT pathway. Currently, it is used to treat adult patients with moderate to severe active rheumatoid arthritis (RA) who are inadequately treated with methotrexate or cannot tolerate it.

[0004] We have discovered a new use for tofacitinib citrate, which is to make tofacitinib citrate into an external preparation for hair growth, and found that it has a good hair growth effect. In clinical practice, the main way to administer tofacitinib citrate is oral administration. Guided by clinical needs, it is of great significance to develop an external preparation of tofacitinib citrate suitable for the treatment of alopecia areata. Summary of the invention

[0005] In view of the above technical problems, the object of the present invention is to provide a tofacitinib citrate gel and a preparation method and use thereof, which specifically include the following contents:

[0006] In a first aspect, the present invention provides a tofacitinib citrate gel, which comprises the following components by mass: 0.1-5% of tofacitinib citrate as an active ingredient, 0.5-15% of a gel matrix, 0.1-5% of a surfactant, 5-30% of a transdermal enhancer, 0.01-1% of a preservative, and the remainder of water;

[0007] The gel matrix is ​​selected from one or a mixture of carbomer 940, hypromellose, sodium carboxymethylcellulose;

[0008] The surfactant is selected from one or a mixture of Tween 80, Span 80, Poloxamer 188;

[0009] The transdermal enhancer is selected from one or a mixture of propylene glycol, glycerol, butylene glycol, laurocapram, and ethanol;

[0010] The preservative is selected from one of sodium bisulfite, sodium benzoate, ethyl p-hydroxybenzoate or a mixture of several of them.

[0011] Preferably, the gel matrix is ​​selected from hypromellose or sodium carboxymethylcellulose.

[0012] Preferably, the mass fraction of the gel matrix is ​​1-3%.

[0013] Preferably, the surfactant is selected from Tween 80 or Poloxamer 188.

[0014] Preferably, the mass fraction of the surfactant is 0.1-3%.

[0015] Preferably, the mass fraction of the active ingredient tofacitinib citrate is 1-3%.

[0016] Preferably, the tofacitinib citrate gel comprises the following components by mass: 1.5% of active ingredient tofacitinib citrate, 2% of sodium carboxymethyl cellulose, 0.25% of poloxamer 188, 15% of propylene glycol, 5% of ethanol, 0.02% of ethyl parahydroxybenzoate, and the balance of water.

[0017] Preferably, the preparation process of the tofacitinib citrate gel comprises the following steps:

[0018] (1) dissolving a prescribed amount of ethyl p-hydroxybenzoate in ethanol to obtain an ethyl p-hydroxybenzoate solution;

[0019] (2) taking a prescribed amount of propylene glycol and mixing it with the ethyl hydroxybenzoate solution described in step (1) to obtain a mixed solution;

[0020] (3) taking a prescribed amount of sodium carboxymethyl cellulose, adding purified water to dissolve, and swelling overnight; adding the mixed solution obtained in step (2) to obtain a matrix;

[0021] (4) Take the prescribed amount of active ingredients tofacitinib citrate, poloxamer 188 and purified water equal to poloxamer 188, fully emulsify them, add the matrix described in step (3), and stir evenly to obtain tofacitinib citrate gel.

[0022] In a second aspect, the present invention provides use of the tofacitinib citrate gel described in the first aspect in the preparation of a medicament for preventing or treating hair loss.

[0023] Compared with the prior art, the beneficial effects of the present invention are as follows: the present invention provides a tofacitinib citrate gel, which comprises the following components by mass: 0.1-5% of tofacitinib citrate as an active ingredient, 0.5-15% of a gel matrix, 0.1-5% of a surfactant, 5-30% of a transdermal enhancer, 0.01-1% of a preservative, and a water balance; the tofacitinib citrate gel prepared by the present invention has suitable viscosity, good moisturizing properties, high content of active ingredients, small total impurities and single impurities, and good stability; and the tofacitinib citrate gel is non-irritating to the skin, has a significant effect of promoting hair follicle growth, and can be used to treat diseases related to hair loss. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 Vibrissa follicle length measurements. DETAILED DESCRIPTION

[0025] The technical solutions in the embodiments of the present invention will be clearly and completely described below in combination with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments.

[0026] Example 1 Preparation of Tofacitinib Citrate Gel 1

[0027] The formulation composition of the tofacitinib citrate gel described in this embodiment is shown in Table 1.

[0028] Table 1 Prescription composition of tofacitinib citrate gel

[0029] composition Content (w / w, %) Tofacitinib citrate 2 Hydroxypropyl methylcellulose 2 Twain 80 2 Propylene glycol 10 Butanediol 5 Sodium bisulfite 0.05 Purified water margin

[0030] Preparation method:

[0031] (1) dissolving a prescribed amount of sodium bisulfite in purified water to obtain a sodium bisulfite aqueous solution;

[0032] (2) taking a prescribed amount of propylene glycol and butylene glycol, mixing them, adding the sodium bisulfite aqueous solution described in step (1), mixing well, and obtaining a mixed solution;

[0033] (3) adding a prescribed amount of hypromellose to the mixed solution of step (2), mixing, and swelling overnight to obtain a matrix;

[0034] (4) Take the prescribed amount of active ingredient tofacitinib citrate, Tween 80 and purified water in an amount equal to that of Tween 80, fully emulsify them, add the matrix described in step (3), and stir evenly to obtain tofacitinib citrate gel 1.

[0035] Example 2 Preparation of Tofacitinib Citrate Gel 2

[0036] The formulation composition of the tofacitinib citrate gel described in this embodiment is shown in Table 2.

[0037] Table 2 Prescription composition of tofacitinib citrate gel

[0038]

[0039]

[0040] Preparation method:

[0041] (1) dissolving a prescribed amount of ethyl p-hydroxybenzoate in ethanol to obtain an ethanol solution of ethyl p-hydroxybenzoate;

[0042] (2) taking a prescribed amount of propylene glycol and mixing it with the ethyl hydroxybenzoate solution described in step (1) to obtain a mixed solution;

[0043] (3) Take a prescribed amount of sodium carboxymethyl cellulose, add purified water to dissolve, and swell overnight. Add the mixed solution obtained in step (2) to obtain a matrix;

[0044] (4) Take the prescribed amount of active ingredients tofacitinib citrate, poloxamer 188 and purified water equal to poloxamer 188, fully emulsify, add the matrix described in step (3), and stir evenly to obtain tofacitinib citrate gel 2.

[0045] Example 3 Preparation of Tofacitinib Citrate Gel

[0046] The formulation composition of the tofacitinib citrate gel described in this embodiment is shown in Table 3.

[0047] Table 3 Prescription composition of tofacitinib citrate gel

[0048] composition Content (w / w, %) Tofacitinib citrate 1 Carbomer 940 0.5 Span 80 1.5 Laurocapram 2 Glycerol 5 Sodium Benzoate 0.02 Purified water margin

[0049] Preparation method:

[0050] (1) dissolving a prescribed amount of sodium benzoate in purified water to obtain a sodium benzoate aqueous solution;

[0051] (2) taking a prescribed amount of laurocapram and glycerol, mixing them, adding the prepared sodium benzoate aqueous solution thereto, and mixing well to obtain a mixed solution;

[0052] (3) taking the prescribed amount of Carbomer 940 and adding it to purified water, stirring rapidly until uniform and fully swollen, and mixing evenly with the mixed solution described in step (2) above to obtain a matrix;

[0053] (4) Take the prescribed amount of tofacitinib citrate, Span 80 and purified water in an amount equal to Span 80, fully emulsify them, add the matrix described in step (3), and stir evenly to obtain tofacitinib citrate gel 3.

[0054] Example 4 Preparation of Tofacitinib Citrate Gel

[0055] The formulation composition of the tofacitinib citrate gel described in this embodiment is shown in Table 4.

[0056] Table 4 Tofacitinib citrate gel formulation composition

[0057] composition Content (w / w, %) Tofacitinib citrate 1 Sodium Carboxymethyl Cellulose 4 Twain 80 3 Glycerol 8 Butanediol 10 Ethanol 8 Ethyl paraben 0.03 Purified water margin

[0058] Preparation method:

[0059] (1) dissolving a prescribed amount of ethyl p-hydroxybenzoate in ethanol to obtain an ethanol solution of ethyl p-hydroxybenzoate;

[0060] (2) taking a prescribed amount of propylene glycol and butylene glycol, mixing them, adding the prepared ethanol solution of ethyl p-hydroxybenzoate thereto, and mixing well to obtain a mixed solution;

[0061] (3) slowly adding hypromellose to the mixed solution of step (2), stirring while adding, stirring thoroughly until the temperature reaches room temperature, and swelling overnight to obtain a matrix;

[0062] (4) Take the prescribed amount of tofacitinib citrate, Tween 80 and purified water in an amount equal to that of Tween 80, fully emulsify them, add the matrix, and stir evenly to obtain tofacitinib citrate gel 4.

[0063] Example 5 Evaluation of Tofacitinib Citrate Gel

[0064] 1. Appearance inspection

[0065] The scoring criteria for the three indicators of appearance, viscosity and moisturizing performance are divided into 5 levels based on sensory scores. The details are shown in the table below. The corresponding scoring results in Table 5 are the scores of multiple people, and the average value is taken.

[0066] Table 5 Appearance, viscosity, and moisturizing performance scoring criteria

[0067]

[0068]

[0069] The appearance test scoring results of the tofacitinib citrate gel prepared in Examples 1-4 are shown in Table 6. The appearance test results show that compared with Example 3, the tofacitinib citrate gel prepared in Examples 1, 2, and 4 described in the present application has suitable viscosity and good moisturizing properties.

[0070] Table 6 Appearance, viscosity, and moisturizing performance ratings

[0071] sample Appearance Viscosity Moisturizing properties Example 1 Colorless transparent gel Moderate viscosity Moist, comfortable to apply Example 2 Colorless transparent gel Moderate viscosity Moist, comfortable to apply Example 3 Colorless transparent gel Low viscosity Poor humidity and poor comfort Example 4 Colorless transparent gel High viscosity Moist, comfortable to apply

[0072] 2. Quality inspection

[0073] The content and related substances of the tofacitinib citrate gel samples prepared in Examples 1-4 were quality tested.

[0074] The test results are shown in Table 7. The quality test results show that compared with Examples 3-4, the tofacitinib citrate gel prepared in Examples 1-2 described in the present application has a higher content, and the total impurities and single impurities are smaller. Among them, the tofacitinib citrate gel prepared in Example 2 has the highest content, and the total impurities and single impurities are the smallest.

[0075] Table 7 Content and related substances determination of different samples

[0076] sample content(%) Single impurity (%) Total impurities (%) Example 1 98.8 0.02 0.16 Example 2 99.3 0.015 0.13 Example 3 97.6 0.03 0.18 Example 4 96.4 0.05 0.19

[0077] 3. Stability test

[0078] According to the general rule 9001 of the 2020 edition of the Chinese Pharmacopoeia, the stability test of the tofacitinib citrate gel samples prepared in Examples 1-4 was carried out. The accelerated test conditions were 30°C and RH 65%; the long-term test conditions were 25°C and RH 60%.

[0079] The results are shown in Table 8. Compared with Examples 3-4, the tofacitinib citrate gel prepared in Examples 1-2 described in the present application has stable appearance and stable content, and the related substances (single impurities and total impurities) have a small change trend with the factors of temperature, humidity and time. Among them, the tofacitinib citrate gel prepared in Example 2 has a stable appearance and stable content, and the related substances (single impurities and total impurities) have the smallest change trend with the factors of temperature, humidity and time.

[0080] Table 8 Stability test results

[0081]

[0082] Example 6 Effect Evaluation of Tofacitinib Citrate Gel

[0083] 1. Skin irritation test

[0084] Eight white guinea pigs weighing 300-400 g were divided into two groups: a single-dose group and a multiple-dose group. 24 hours before administration, the hair on both sides of the spine of the guinea pigs' backs was removed, with a hair removal area of ​​20 cm on each side. 2. Skin irritation test for single administration: The left and right sides of the same subject are compared, the left hair removal area is coated with the test sample, and the right hair removal area is coated with the matrix as a blank control. The test sample (tofacitinib citrate gel prepared in Examples 1 and 2) and the blank control are fixed with gauze and adhesive tape. After 24 hours of application of the test sample (tofacitinib citrate gel prepared in Examples 1 and 2) and the blank control, the residual test sample and the blank control are removed with warm water. After removing the test sample and the blank control, the naked eye is observed for 1, 24, 48, and 72 hours, and the presence of erythema and edema at the application site is recorded. The irritation intensity is judged according to the scoring standard and the intensity standard. (2) Skin irritation test for multiple administration: The test method and observation are still carried out according to the single administration test, but the drug is administered once a day for 1 week. After removing the test sample and the blank control for 1, 24, 48, and 72 hours, the naked eye is observed, and the average reaction score of each group of the test substance and the matrix is ​​recorded. The irritation intensity is judged according to the scoring standard and the intensity standard. (3) Scoring criteria for irritation symptoms: 1 point for barely visible erythema, 2 points for moderate erythema, 3 points for severe erythema, 4 points for purple-red spots with eschar formation, 1 point for barely visible edema, 2 points for clear skin bulges, and 4 points for edema bulges of about 1-2 mm and expanding in range. (4) Irritation intensity evaluation criteria: A score of 6.0 is strong irritation.

[0085] The results are shown in Table 9. The tofacitinib citrate gels prepared in Examples 1 and 2 of the present application are non-irritating, and Example 1 is slightly better than Example 2, meeting clinical requirements.

[0086] Table 9 Skin irritation test results

[0087]

[0088] 2. Mouse in vitro hair follicle test

[0089] The hair follicles of C57BL / 6 mice were isolated and cultured, and samples were added on the next day of culture. The culture medium containing the blank control (matrix), the positive control (minoxidil liniment was prepared into a concentration of 10 μM using culture medium), and the test sample (tofacitinib citrate gel prepared in Examples 1 and 2) was added to the corresponding wells, and the medium was changed every 3 days. The length of the vibrissa follicles was photographed on the 1st day and the 14th day, and the length of the vibrissa follicles was morphologically observed and measured.

[0090] The results are shown in Table 10 and Figure 1 As shown, the tofacitinib citrate gel prepared in Examples 1 and 2 of the present application is better than the positive control group in promoting the growth of in vitro hair follicles, and the hair follicle growth effect of Example 1 is significant compared with the blank control group.

[0091] Table 10 Hair follicle growth in different groups

[0092] Group Growth length (μm) SD p Blank control group 51.33 2.12 / Positive control group 169.33 35.10 0.028* Example 1 246.67 35.00 0.011* Example 2 283.33 37.81 0.009**

[0093] Note: When using the t-test method for statistical analysis, the significance of the sample group and PC group compared with the BC group is indicated by *, and the p-value

[0094] p-value < 0.05 is indicated by *, p-value < 0.01 is indicated by **.

[0095] After statistical analysis, compared with the control group, the hair follicle growth length in the positive group was statistically different (p<0.05); compared with the control group, the hair follicle growth length under the action of Example 1 was statistically different (p<0.05), and the hair follicle growth length under the action of Example 1 was statistically significantly different (p<0.01).

[0096] Conclusion: Under the experimental conditions, the samples of Example 1 and Example 2 have corresponding effects on promoting hair growth, and the effect of Example 2 is better.

Claims

1. A tofacitinib citrate gel, characterized in that: The tofacitinib citrate gel comprises the following components by mass: 0.1-5% of tofacitinib citrate as an active ingredient, 0.5-15% of a gel matrix, 0.1-5% of a surfactant, 5-30% of a transdermal enhancer, 0.01-1% of a preservative, and the remainder of water; The gel matrix is ​​selected from one or a mixture of carbomer 940, hypromellose, sodium carboxymethylcellulose; The surfactant is selected from one or a mixture of Tween 80, Span 80, Poloxamer 188; The transdermal enhancer is selected from one or a mixture of propylene glycol, glycerol, butylene glycol, laurocapram, and ethanol; The preservative is selected from one of sodium bisulfite, sodium benzoate, ethyl p-hydroxybenzoate or a mixture of several of them.

2. The tofacitinib citrate gel according to claim 1, characterized in that The gel matrix is ​​selected from hypromellose or sodium carboxymethylcellulose.

3. The tofacitinib citrate gel according to claim 2, characterized in that: The mass fraction of the gel matrix is ​​1-3%.

4. The tofacitinib citrate gel according to claim 1, characterized in that: The surfactant is selected from Tween 80 or Poloxamer 188.

5. The tofacitinib citrate gel according to claim 4, characterized in that: The mass fraction of the surfactant is 0.1-3%.

6. The tofacitinib citrate gel according to claim 1, characterized in that: The mass fraction of the active ingredient tofacitinib citrate is 1-3%.

7. The tofacitinib citrate gel according to claims 1-7, characterized in that: The components of the tofacitinib citrate gel by weight are: 1.5% of tofacitinib citrate as an active ingredient, 2% of sodium carboxymethyl cellulose, 0.25% of poloxamer 188, 15% of propylene glycol, 5% of ethanol, 0.02% of ethyl p-hydroxybenzoate, and the balance of water.

8. The preparation process of tofacitinib citrate gel according to claim 7, comprising the following steps: (1) dissolving a prescribed amount of ethyl p-hydroxybenzoate in ethanol to obtain an ethyl p-hydroxybenzoate solution; (2) taking a prescribed amount of propylene glycol and mixing it with the ethyl hydroxybenzoate solution described in step (1) to obtain a mixed solution; (3) taking a prescribed amount of sodium carboxymethyl cellulose, adding purified water to dissolve, and swelling overnight; adding the mixed solution obtained in step (2) to obtain a matrix; (4) Take the prescribed amount of active ingredients tofacitinib citrate, poloxamer 188 and purified water equal to poloxamer 188, fully emulsify them, add the matrix described in step (3), and stir evenly to obtain tofacitinib citrate gel.

9. Use of the tofacitinib citrate gel according to any one of claims 1 to 7 in the preparation of a medicament for preventing or treating hair loss.

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