Topical gel for jak inhibitor and use thereof

Abstract

The present application discloses a topical gel, containing 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile, or a crystal form and gel matrix thereof, and other pharmaceutical excipients.

Description

External-use gel for JAK inhibitor and use thereof Technical Field

[0001] The present application belongs to the field of pharmaceutical preparations and relates to a topical gel of a JAK inhibitor and its use. Background Art

[0002] 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile (Compound A):

[0003] It is a JAK class small molecule non-receptor tyrosine kinase inhibitor. Its JAK-STAT signaling pathway is closely related to inflammatory cytokines and tumors, and is widely involved in important biological processes such as cell proliferation, differentiation, metastasis, apoptosis, regulation of immune response and cell homeostasis in human health and disease.

[0004] Topically administered, locally effective preparations offer the advantage of being able to penetrate the skin and maintain prolonged retention. This, combined with minimal systemic exposure, is a key advantage. Furthermore, among topical preparations, gel formulations are often aqueous, making them non-greasy and easy to wash off after contact with clothing, resulting in excellent clinical compliance.

[0005] Summary of the Invention

[0006] The present application provides a prescription for a gel preparation for external use on the skin and a preparation method thereof. The gel preparation is non-irritating, uniform and fine, easy to apply, and quick-acting. It can quickly penetrate the skin and remain locally in the skin to exert its medicinal effect. It is an excellent topical drug delivery preparation.

[0007] According to one aspect, the present application provides a topical gel comprising

[0008] (a) 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile, a hydrate thereof, a pharmaceutically acceptable salt thereof, or a crystalline form thereof;

[0009] (b) gel matrix;

[0010] (c) 5% to 30% propylene glycol based on the total weight of the topical gel; and

[0011] (d) 2% to 20% polyethylene glycol based on the total weight of the topical gel.

[0012] According to some embodiments, the (a) in the topical gel of the present application is 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile hemihydrate.

[0013] According to some embodiments, the gel matrix in the topical gel of the present application may be carbomer, and the amount of carbomer is, for example, 0.8% to 1.5% of the total weight of the topical gel, and another example is 1%.

[0014] According to some embodiments, the average molecular weight of the polyethylene glycol in the topical gel of the present application may be 200-600, such as polyethylene glycol 200, polyethylene glycol 400, or polyethylene glycol 600. The amount of polyethylene glycol in the topical gel is, for example, 5% of the total weight of the topical gel.

[0015] According to some embodiments, the amount of propylene glycol in the topical gel of the present application is about 15% (w / w) of the total weight of the topical gel.

[0016] According to another aspect, the present application provides a topical gel comprising

[0017] (a) 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile hemihydrate crystalline form, the 2θ values ​​of the characteristic peaks in the powder X-ray diffraction pattern of the crystalline form are 7.93, 9.36, 11.29, 14.95, 20.96, 21.36, 21.77, 22.12, and 22.79, with a measurement error of ±0.2 degrees.

[0018] According to some embodiments, the topical gel of the present application further comprises:

[0019] (b) a gel matrix; and

[0020] (c) 5% to 30% propylene glycol based on the total weight of the topical gel.

[0021] According to some embodiments, the topical gel of the present application further comprises polyethylene glycol, wherein the average molecular weight of the polyethylene glycol in the topical gel of the present application may be 200 to 600, for example, polyethylene glycol 200, polyethylene glycol 400, or polyethylene glycol 600.

[0022] According to some embodiments, the amount of polyethylene glycol in the topical gel of the present application is 2% to 20% by weight of the total weight of the topical gel, for example, 5%.

[0023] According to some embodiments, the gel matrix in the topical gel of the present application may be carbomer. The amount of the gel matrix (such as carbomer) is, for example, 0.8% to 1.5% of the total weight of the topical gel, and another example is 1%.

[0024] According to some embodiments, the amount of propylene glycol in the topical gel of the present application is about 15% (w / w) of the total weight of the topical gel.

[0025] According to another aspect, the present application provides a topical gel comprising

[0026] (a) a crystalline form of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}, acetonitrile hemihydrate, wherein the 2θ values ​​of characteristic peaks in the powder X-ray diffraction pattern of the crystalline form are 7.93, 9.36, 11.29, 14.95, 20.96, 21.36, 21.77, 22.12, and 22.79, with a measurement error of ±0.2 degrees; and

[0027] (b) The gel matrix, which is, for example, carbomer, may contain 0.8% to 1.5%, for example, 1%, of the total weight of the topical gel.

[0028] According to some embodiments, the topical gel of the present application further comprises polyethylene glycol and propylene glycol.

[0029] According to some embodiments, the propylene glycol in the topical gel of the present application is 5% to 30% by weight of the total weight of the topical gel, such as 15%.

[0030] According to some embodiments, the polyethylene glycol in the topical gel of the present application is 2% to 20% by weight of the total weight of the topical gel, such as 5%.

[0031] According to some embodiments, the average molecular weight of the polyethylene glycol in the external gel of the present application may be 200 to 600, for example, polyethylene glycol 200, polyethylene glycol 400, or polyethylene glycol 600.

[0032] According to some embodiments, the topical gel of the present application further comprises one or more of the following pharmaceutical excipients: an antibacterial agent, an antioxidant, a chelating agent, and a pH regulator.

[0033] The gel matrix of the present application refers to a type of substance that can form a colloid and has a spatial network structure system.

[0034] In the topical gel of the present application, the gel matrix includes but is not limited to one or more of hydroxypropyl methylcellulose, methylcellulose, ethyl cellulose, and carbomer; preferably carbomer; more preferably one or more of carbomer homopolymer type A, carbomer homopolymer type B, and carbomer homopolymer type C.

[0035] In some embodiments, the polyethylene glycol can be selected from one or more of polyethylene glycol 200, polyethylene glycol 300, and polyethylene glycol 400; preferably polyethylene glycol 400.

[0036] In some embodiments, the chelating agent in the topical gel can be selected from one or more of edetate disodium and edetate sodium calcium; preferably edetate disodium.

[0037] In some embodiments, the antibacterial agent in the topical gel can be selected from one or more of phenoxyethanol, potassium sorbate, benzyl alcohol, methylparaben, propylparaben, sodium methylparaben, sodium propylparaben, chlorhexidine digluconate, chloroxylenol, chlorophenyl ether, dehydroacetic acid, diazolidinyl imidazole urea, DMDM ​​hydantoin, ethylparaben, iodopropynylbutylcarbamate, methylisothiazolinone, propylparaben, phenoxyethanol, phenoxyisopropanol, polyaminopropyl biguanide, benzoate, and salicylic acid; preferably phenoxyethanol, potassium sorbate, benzyl alcohol, methylparaben, propylparaben, sodium methylparaben, and sodium propylparaben; more preferably methylparaben, propylparaben, sodium methylparaben, and sodium propylparaben.

[0038] In some embodiments, the topical gel further comprises an antioxidant, which can be selected from one or more of butylated hydroxytoluene, butylated hydroxyanisole, propyl gallate, vitamin C, sodium bisulfite, sodium metabisulfite, and cysteine ​​hydrochloride; preferably butylated hydroxyanisole and propyl gallate.

[0039] In some embodiments, the topical gel further comprises a pH regulator, which can be selected from one or more of sodium hydroxide, potassium hydroxide, triethanolamine, and hydrochloric acid; preferably triethanolamine or sodium hydroxide.

[0040] {3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile (Compound A) exists in multiple crystalline forms, including Form I, Form IIa, Form IIb, and Form III. Form I and Form IIa are anhydrous forms, Form IIb is a hemihydrate, and Form III is an amorphous form. The X-ray powder diffraction data and spectra of the different crystalline forms are as follows:

[0041] Form IIa has a powder X-ray diffraction pattern with characteristic peaks at 2θ=7.61, 9.28, 12.44, 12.96, 14.28, 15.30, 17.56, 18.23, 21.05, 21.47, 22.28, 23.45, 24.87, and 26.03.

[0042] Form IIb has characteristic peaks located at 2θ = 7.93, 9.36, 11.29, 14.95, 20.96, 21.36, 21.77, 22.12, and 22.79.

[0043] The crystal form IIb has a powder X-ray diffraction pattern with more characteristic peaks located at 2θ = 7.93, 9.36, 11.29, 14.69, 14.95, 15.13, 15.95, 19.93, 20.15, 20.96, 21.36, 21.77, 22.12, 22.79, 23.54, 24.14, 26.38, 28.74, and 29.15.

[0044] Form I has a powder X-ray diffraction pattern with characteristic peaks at 2θ = 6.48, 7.68, 9.90, 13.01, 14.81, 15.38, 16.43, 16.87, 19.61, 20.68, 23.47, 26.29 and 32.23.

[0045] Table 1 XRPD peak list of Form IIa

[0046] Table 2 XRPD peak list of Form IIb

[0047] Table 3 XRPD peak list of Form I

[0048] Form IIa converts to Form IIb within 1 hour at room temperature (58.6% RH). During the preparation of topical gels, Form I and Form IIb were used to prepare the gels. Surprisingly, after 3 days, the API was detected in the finished gel formulations as Form IIb (see Figures 5-6). BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 XRPD pattern of Form IIa

[0050] Figure 2 XRPD pattern of Form IIb

[0051] Figure 3 XRPD pattern of Form I

[0052] Figure 4 XRPD pattern of Form III

[0053] Figure 5 shows the XRPD results of the drug substance in the preparation after using Form I to prepare a gel (A); the transmission / reflection XRPD comparison of Form I (B); and the transmission / reflection XRPD comparison of Form IIb (C).

[0054] Figure 6 shows the XRPD results of the drug substance in the preparation after the gel was prepared using Form IIb.

[0055] Figure 7 Cumulative permeation of drugs in in vitro transdermal test

[0056] Figure 8 Drug retention in the epidermis during in vitro transdermal testing

[0057] Figure 9 Drug retention in the dermis during in vitro transdermal permeation testing

[0058] FIG10 shows the cumulative permeation amount of Form I (A) and Form IIb (B) in the rat transdermal test. DETAILED DESCRIPTION

[0059] The present application will be further described below in conjunction with specific embodiments so that those skilled in the art can better understand the present application and implement it, but the embodiments are not intended to limit the present application.

[0060] Preparation Example

[0061] The external-use gel was prepared according to the formulation in each example.

[0062] Example 1-Example 5:

[0063] Preparation process:

[0064] 1. Preparation of Phase I (Carbomer Phase)

[0065] Weigh an appropriate amount of purified water, slowly add carbomer, and continue stirring to disperse and swell completely.

[0066] 2. Phase II (active ingredient phase) preparation

[0067] Weigh propylene glycol and polyethylene glycol 400; add methylparaben and propylparaben and stir to dissolve completely; add compound A, continue stirring to disperse evenly, and then transfer to a homogenizer.

[0068] 3. Preparation of Phase III (Disodium Edetate Phase)

[0069] Weigh an appropriate amount of purified water, add disodium edetate, stir until completely dissolved, and then transfer to a homogenizer.

[0070] 4. Mixing and homogenization

[0071] Turn on the homogenizer to stir and homogenize the mixture.

[0072] 5. pH adjustment

[0073] Use sodium hydroxide solution to adjust the pH to 5.0-7.0.

[0074] 6. Vacuum stirring homogenization, vacuum stirring

[0075] After vacuum stirring and homogenization, stop the machine and discharge the material.

[0076] According to the preparation method of Examples 1-5, Examples 6-8 were prepared

[0077] Example 6

[0078] Example 7

[0079] Example 8

[0080] The gel samples of Example 3, Example 4, and Example 5 were subjected to evaluation of the transdermal behavior and skin retention of the active ingredients in the preparations using the ex vivo skin of one-month-old Bama miniature pigs in a Franz diffusion cell.

[0081] The results of the in vitro permeation test are as follows:

[0082] In vitro cumulative permeation results

[0083] In vitro release intradermal retention results

[0084] The results of the in vitro release cumulative penetration test can be seen in FIG7 , the results of the epidermal retention test can be seen in FIG8 , and the results of the dermal retention test can also be seen in FIG9 .

[0085] An unexpected discovery was made through in vitro permeation tests. Compared with the use of propylene glycol or polyethylene glycol alone, the combined use of propylene glycol and polyethylene glycol increased the drug skin permeation by 4 to 11 times, and the intradermal retention in the epidermis and dermis increased by nearly 1 time. The in vitro permeation results showed that the combined use of propylene glycol and polyethylene glycol can significantly improve the skin permeation ability of the active ingredient 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile.

[0086] Example 9 Preparation of Polymorphs of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile

[0087] (A) Preparation of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile Form I

[0088] In a 250 mL flask, 20.0 g of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile and 100 mL of N,N-dimethylformamide were added, stirred to dissolve, filtered, and the insoluble matter was removed. 250 mL of purified water was added to the filtrate, stirred for crystallization, and filtered. The resulting solid was dried and transferred to a 250 mL flask. 200 mL of anhydrous ethanol was further added, the temperature was raised to reflux for 3 h, cooled, and filtered. The resulting solid was dried at 55-60° C. to constant weight to give 18.8 g of an off-white solid with a yield of 94.0% and a chemical purity of 99.62%.

[0089] (B) Preparation of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile Form IIa

[0090] In a 100 mL flask, 5.0 g of type I crystals of compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile were added, and 100 mL of ethanol (95%) was added. The mixture was heated, stirred, and refluxed for 5 h. The temperature was lowered and the mixture was filtered to obtain a solid. The obtained solid was dried at 80°C under vacuum (0.1 MPa) for 12 h to obtain 4.8 g of a solid with a yield of 96.0% and a purity of 99.50% as determined by HPLC.

[0091] (C) Preparation of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile Form IIb

[0092] In a 100 mL flask, 6.2 g of the compound 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile and 25 mL of N,N-dimethylformamide were added. The mixture was stirred to dissolve and filtered to remove insoluble matter. 50 mL of purified water was added to the filtrate, and the mixture was stirred for crystallization and filtered. The resulting solid was dried at 55-60°C to constant weight to obtain 5.6 g of solid with a yield of 88.9%.

[0093] (D) Preparation of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile Form III

[0094] 15 mg of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile was added to 3 mL of dioxane, and the temperature was raised to 40°C to dissolve and clarify. After filtration, the solution was evaporated at 40°C to crystallize to obtain Form III. This form has poor crystallinity and stability. After stability is allowed to stand, nuclear magnetic resonance shows that about 10% of impurities are generated.

[0095] Investigation of the solubility of each crystal form

[0096] Stability investigation of each crystal form

[0097] In accordance with the stability study guidelines issued by the State Food and Drug Administration, the stability of Forms I, IIa, and IIb was investigated. The samples were packaged in pharmaceutical low-density polyethylene bags. The long-term stability was investigated for 24 months. The results are as follows:

[0098] Long-term 24-month stability of Form I

[0099] Long-term 24-month stability of Form IIa

[0100] Long-term 24-month stability of Form IIb

[0101] The stability studies of the various crystalline forms above demonstrate that Forms I and IIb of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile exhibit excellent chemical stability. Form IIa gradually absorbs water over time, transforming into Form IIb. This water absorption increases the sample's weight, leading to a decrease in sample content.

[0102] Transdermal test of crystal form

[0103] Crystal forms I and IIb were selected and prepared into topical gels according to the formulation of Example 1. Transdermal tests were then immediately performed to determine the transdermal penetration rates of the respective gels. The test results are shown in Figures 10A and 10B, which indicate that crystal form I has better transdermal performance.

[0104] Blood samples were collected at 0.25, 0.5, 1, 2, 4, 6, 8, and 24 hours after transdermal administration. After isoflurane anesthesia using a small animal anesthesia machine, 0.3 mL of whole blood was collected from the retinal venous plexus. The blood was placed in a heparinized tube and centrifuged at 4000 rpm for 5 minutes at 4°C. The plasma was transferred to a centrifuge tube and stored at -80°C until analysis. The plasma samples were extracted using protein precipitation, and the extracts were analyzed by LC / MS / MS. After washing the epidermal drug, the rats' skin at the administration site was peeled and homogenized, followed by LC / MS / MS analysis of the drug content in the skin.

[0105] Table 4. Drug concentrations in the skin 24 hours after transdermal administration

[0106] Form I and Form IIb are characterized by high crystallinity and good stability. Form I has higher solubility than Form IIb. Although Form IIb has inferior transdermal absorption to Form I, the drug concentration retained in the skin of its topical preparations is significantly higher than that of Form I, by several times. This unexpected feature makes it more suitable for topical gel preparations with less systemic penetration and higher drug retention concentration in the skin.

Claims

1. A gel for external use, comprising (a) 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile, a hydrate thereof, a pharmaceutically acceptable salt thereof or a crystalline form thereof; (b) gel matrix; (c) 5% to 30% propylene glycol based on the total weight of the topical gel; and (d) 2% to 20% polyethylene glycol based on the total weight of the topical gel.

2. The topical gel according to claim 1, wherein the gel base is selected from carbomer.

3. The topical gel according to claim 2, wherein the amount of the carbomer is 0.8% to 1.5% of the total weight of the topical gel.

4. A gel for external use, comprising (a) 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile hemihydrate crystalline form, the 2θ values ​​of characteristic peaks in the powder X-ray diffraction pattern of the crystalline form are 7.93, 9.36, 11.29, 14.95, 20.96, 21.36, 21.77, 22.12, and 22.79, and the measurement error is ±0.20 degrees.

5. The external use gel according to claim 4, wherein The external gel also comprises (b) a gel matrix; and (c) 5% to 30% propylene glycol based on the total weight of the topical gel.

6. The external gel according to claim 4 or 5, wherein the external gel further comprises polyethylene glycol.

7. The topical gel according to claim 6, wherein the amount of the polyethylene glycol is 2% to 20% of the total weight of the topical gel.

8. The topical gel according to any one of claims 4 to 7, wherein the gel matrix is ​​selected from carbomer.

9. The topical gel according to claim 8, wherein the amount of the carbomer is 0.8% to 1.5% of the total weight of the topical gel.

10. A gel for external use, comprising (a) a crystalline form of 2-{3-[3-amino-4-(7H-pyrrolo[2,3-d]pyrimidin-4-yl)-1H-pyrazol-1-yl]-1-(isopropylsulfonyl)azetidin-3-yl}acetonitrile hemihydrate, wherein the 2θ values ​​of characteristic peaks in the powder X-ray diffraction pattern of the crystalline form are 7.93, 9.36, 11.29, 14.95, 20.96, 21.36, 21.77, 22.12, and 22.79, and the measurement error is ±0.2 degrees; and (b) A gel base which is 0.8% to 1.5% carbomer based on the total weight of the topical gel.

11. The topical gel of claim 10, wherein the topical gel further comprises polyethylene glycol and propylene glycol.

12. The topical gel according to claim 11, wherein the propylene glycol is 5% to 30% of the total weight of the topical gel; and the polyethylene glycol is 2% to 20% of the total weight of the topical gel.

13. The external gel according to any one of claims 1 to 12, wherein the average molecular weight of the polyethylene glycol is 200 to 600.

14. The topical gel according to any one of claims 1 to 13, wherein the topical gel further comprises one or more of the following pharmaceutical excipients: an antibacterial agent, an antioxidant, and a pH regulator.

15. Use of the external gel according to any one of claims 1 to 14 in the preparation of a medicament for inhibiting JAK.

Images