RNA m6A regulator composition as well as preparation method and application thereof

Abstract

The invention relates to an RNAm6A regulating agent composition as well as a preparation method and application thereof, and the composition comprises a compound as shown in a formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, an oil-phase substance 1, an oil-phase substance 2 and an oil-phase substance 3, and can also comprise a penetration enhancer and an antioxidant. In the RNAm6A regulator composition provided by the invention, the compound shown in the formula (I) has high exposure in skin, and can well prevent or treat skin diseases by inhibiting m6A methylation. Meanwhile, the composition disclosed by the invention has good stability and excellent appearance, viscosity, coating property, physical stability and pharmacokinetic properties.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical chemistry, and particularly relates to an RNA m6A regulator composition, a preparation method thereof, and an application thereof. Background Art

[0002] Hand-foot syndrome (HFS) and hand-foot skin reaction (HFSR) are erythematous skin lesions on the palms and soles, mainly caused by cytotoxic chemotherapy drugs and tumor-targeted drugs. In severe cases, patients may lose the ability to take care of themselves. The main pathological features of hand-foot syndrome and hand-foot skin reaction are vacuolar degeneration of basal keratinocytes, perivascular lymphocyte infiltration in the skin, keratinocyte apoptosis, and skin edema; inflammatory changes, vasodilation, edema, and leukocyte infiltration can be seen under the microscope.

[0003] Drugs that can cause hand-foot syndrome include chemotherapy drugs such as capecitabine, liposomal doxorubicin, cytarabine, docetaxel, vinorelbine, continuous infusion of doxorubicin, gemcitabine, etc.; drugs that can cause hand-foot skin reaction include targeted drugs such as sunitinib (Sutent), sorafenib (Nexavar), imatinib (Gleevec), erlotinib (Tarceva), etc. The World Health Organization (WHO) classifies HFS into 4 grades: Grade 1, numbness, paresthesia, or tingling sensation in the hands and feet; Grade 2, discomfort when holding objects and walking, painless swelling or erythema; Grade 3, painful erythema, edema of the palms and soles, perionychial erythema and swelling; Grade 4, peeling, ulceration, blistering, and severe pain. At present, when severe hand-foot syndrome and hand-foot skin reaction occur clinically, the symptoms are often only relieved by superficial skin care and even drug withdrawal may be required. The existing treatment methods only relieve the symptoms but do not address the root cause, which severely restricts the use of first-line oncology chemotherapy drugs. Therefore, there is a huge unmet medical need. There is a need to develop a drug with good efficacy, few side effects, and low cost for the prevention and treatment of hand-foot syndrome and hand-foot skin reaction to meet the growing global medical needs.

[0004] RNA m6A methylation modification is the most common RNA modification in mammals, which can regulate multiple signaling pathways and cellular processes (such as growth, development, and diseases, etc.) to play a key biological role (Frye M., et al. Science 2018, 361, 2073 - 2092; Yang C., et al. Cell Death & Disease 2020, 11, 960; Meyer K.D. & Jaffrey S.R. Nature Review Molecular Cell Biology 2014, 15, 313 - 326). The m6A methylation modification of mRNA, miRNA, circRNA, and lncRNA is a dynamic and reversible process. Methyltransferases (such as METTL3, METTL14, and METTL16, etc.) bind methyl groups to RNA, while demethylases (FTO and ALKBH5) can remove the methyl groups on RNA, thus constituting the regulatory basis of m6A. m6A affects processes such as RNA processing, translation, and degradation by recruiting specific binding proteins (such as YTHDF1, YTHDF2, YTHDC1, and IGF2BP, etc.), thereby leading to changes in downstream protein functions and cellular biological behaviors (Hsu P.J., et al. Journal of Biological Chemistry 2019, 294, 19889 - 19895; Yao Y., et al. FASEB Journal 2019, 33, 7529 - 7544).

[0005] There are few known direct associations between RNA m6A and skin-related diseases. Most of them are that m6A-related methylases or demethylases play roles in skin diseases (especially skin tumors). For example, the mRNA expression levels of METTL3 and ALKBH5 in the tumor tissues of patients with acral melanoma are significantly higher than those in the adjacent tissues, and the mRNA expression level of METTL3 in patients with advanced acral melanoma is significantly higher than that in early-stage patients (Le Zhanghui, Preliminary Study on the Pathogenesis of Acral Melanoma by LncRNA and RNA m6A Methylation, Dissertation, 2019). In the diagnosis of melanoma, the m6A-specific binding proteins YTHDF1 and HNRNPA2B1 can be used as novel biomarkers (Li T.D., et al. Cancer Cell 2020, 20, 239). In terms of keratinocytes, it has only been reported that long-term and low-level arsenic exposure can inhibit selective autophagy caused by m6A demethylase, thereby inducing the occurrence of skin tumors (Cui Y.H., et al. Nature Communications 2021, 12, 2183). In summary, there is currently no method for preventing and treating skin diseases by regulating RNA m6A methylation. Summary of the Invention

[0006] Problems to be Solved by the Invention

[0007] In order to solve the above technical problems, the object of the present invention is to provide an RNA m6A regulator and its preparation method and application to significantly improve skin diseases caused by RNA m6A methylation.

[0008] Solutions for Solving the Problems

[0009] The present invention provides an RNA m6A regulator composition, which comprises: a compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable cocrystal thereof, and an oil-phase substance.

[0010]

[0011] Preferably, the pharmaceutically acceptable salt includes hydrochloride, sulfate, phosphate, acetate, mesylate, benzenesulfonate, p-toluenesulfonate;

[0012] Preferably, the cocrystal-forming substances of the pharmaceutically acceptable cocrystal include nicotinamide, isonicotinamide, L-proline, glycolic acid;

[0013] Preferably, the oil-phase substance includes oil-phase substance 1, oil-phase substance 2 and oil-phase substance 3;

[0014] Preferably, the oil phase substance 1 is selected from one or more of lanolin, beeswax, white beeswax, and paraffin wax, and is preferably white beeswax;

[0015] Preferably, the oil phase substance 2 is selected from one or more of petrolatum and white petrolatum, and is preferably white petrolatum;

[0016] Preferably, the oil phase substance 3 is selected from one or more of light liquid paraffin and vegetable oil, and is preferably light liquid paraffin.

[0017] Preferably, by mass percentage, the composition comprises 0.1% - 20% of the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable cocrystal, 10% - 30% of oil phase substance 1, 30% - 70% of oil phase substance 2, and 15% - 40% of oil phase substance 3;

[0018] Preferably, by mass percentage, the composition comprises 0.2% - 15% of the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable cocrystal, 15% - 25% of oil phase substance 1, 35% - 64% of oil phase substance 2, and 20% - 35% of oil phase substance 3;

[0019] Preferably, by mass percentage, the composition comprises 0.3% - 10% of the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable cocrystal, 18% - 20% of oil phase substance 1, 40% - 58.7% of oil phase substance 2, and 22% - 30% of oil phase substance 3;

[0020] Preferably, by mass percentage, the composition comprises 0.3% of the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable cocrystal, 18% of oil phase substance 1, 58.7% of oil phase substance 2, and 23% of oil phase substance 3;

[0021] Preferably, by mass percentage, the composition comprises 1% of the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable cocrystal, 18% of oil phase substance 1, 58% of oil phase substance 2, and 23% of oil phase substance 3;

[0022] Preferably, by mass percentage, the composition comprises 3% of the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable cocrystal, 18% of oil phase substance 1, 57% of oil phase substance 2, and 22% of oil phase substance 3;

[0023] Preferably, by mass percentage, the composition comprises 10% of the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable cocrystal, 20% of oil phase substance 1, 40% of oil phase substance 2, and 30% of oil phase substance 3.

[0024] Preferably, the composition comprises the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable cocrystal, white beeswax, white petrolatum, and light liquid paraffin;

[0025] Preferably, by mass percentage, the composition comprises 0.1%-20% of the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 10%-30% of white beeswax, 30%-70% of white petrolatum, and 15%-40% of light liquid paraffin;

[0026] Preferably, by mass percentage, the composition comprises 0.2%-15% of the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 15%-25% of white beeswax, 35%-64% of white petrolatum, and 20%-35% of light liquid paraffin;

[0027] Preferably, by mass percentage, the composition comprises 0.3%-10% of the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 18%-20% of white beeswax, 40%-58.7% of white petrolatum, and 22%-30% of light liquid paraffin;

[0028] Preferably, by mass percentage, the composition comprises 0.3% of the compound represented by formula (I), 18% of white beeswax, 58.7% of white petrolatum, and 23% of light liquid paraffin;

[0029] Preferably, by mass percentage, the composition comprises 1% of the compound represented by formula (I), 18% of white beeswax, 58% of white petrolatum, and 23% of light liquid paraffin;

[0030] Preferably, by mass percentage, the composition comprises 3% of the compound represented by formula (I), 18% of white beeswax, 57% of white petrolatum, and 22% of light liquid paraffin;

[0031] Preferably, by mass percentage, the composition comprises 10% of the compound represented by formula (I), 20% of white beeswax, 40% of white petrolatum, and 30% of light liquid paraffin.

[0032] Preferably, the composition further comprises a penetration enhancer. More preferably, the penetration enhancer is selected from one or more of isopropyl myristate, isopropyl palmitate, propylene glycol dinonanoate, diethyl sebacate, azone, and propylene glycol. Even more preferably, the penetration enhancer is isopropyl myristate;

[0033] Preferably, the composition further comprises an antioxidant. More preferably, the antioxidant is selected from one or more of vitamin E, alkyl gallate, butylated hydroxyanisole (BHA), and dibutylhydroxytoluene (BHT). Even more preferably, the antioxidant is dibutylhydroxytoluene (BHT);

[0034] Preferably, the composition further comprises other pharmaceutically acceptable excipients.

[0035] Preferably, by mass percentage, the composition comprises 0.1%-20% of the compound shown by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 10%-30% of oil phase substance 1, 30%-70% of oil phase substance 2, 15%-30% of oil phase substance 3, 0.1%-20% of penetration enhancer, and 0%-2% of antioxidant;

[0036] Preferably, by mass percentage, the composition comprises 0.2%-15% of the compound shown by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 15%-25% of oil phase substance 1, 35%-63% of oil phase substance 2, 20%-25% of oil phase substance 3, 1%-15% of penetration enhancer, and 0%-1% of antioxidant;

[0037] Preferably, by mass percentage, the composition comprises 0.3%-10% of the compound shown by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 18%-20% of oil phase substance 1, 40%-58% of oil phase substance 2, 20%-21% of oil phase substance 3, 2%-10% of penetration enhancer, and 0%-0.2% of antioxidant.

[0038] Preferably, the composition comprises the compound shown by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, white beeswax, white petrolatum, light liquid paraffin, and isopropyl myristate;

[0039] Preferably, by mass percentage, the composition comprises 0.1%-20% of the compound shown by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 10%-30% of white beeswax, 30%-70% of white petrolatum, 15%-30% of light liquid paraffin, and 0.1%-20% of isopropyl myristate;

[0040] Preferably, by mass percentage, the composition comprises 0.2%-15% of the compound shown by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 15%-25% of white beeswax, 35%-63% of white petrolatum, 20%-25% of light liquid paraffin, and 1%-15% of isopropyl myristate;

[0041] Preferably, by mass percentage, the composition comprises 0.3%-10% of the compound shown by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 18%-20% of white beeswax, 40%-58% of white petrolatum, 20%-21% of light liquid paraffin, and 2%-10% of isopropyl myristate;

[0042] Preferably, by mass percentage, the composition comprises 1% of the compound shown by formula (I), 18% of white beeswax, 58% of white petrolatum, 21% of light liquid paraffin, and 2% of isopropyl myristate;

[0043] Preferably, by mass percentage, the composition comprises 10% of the compound represented by formula (I), 20% of white beeswax, 40% of white petrolatum, 20% of light liquid paraffin, and 10% of isopropyl myristate.

[0044] Preferably, the composition comprises the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, white beeswax, white petrolatum, light liquid paraffin, isopropyl myristate, and butylated hydroxytoluene (BHT);

[0045] Preferably, by mass percentage, the composition comprises 0.1%-20% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 10%-30% of white beeswax, 30%-70% of white petrolatum, 15%-30% of light liquid paraffin, 0.1%-20% of isopropyl myristate, and 0%-2% of butylated hydroxytoluene (BHT);

[0046] Preferably, by mass percentage, the composition comprises 0.2%-15% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 15%-25% of white beeswax, 35%-63% of white petrolatum, 20%-25% of light liquid paraffin, 1%-15% of isopropyl myristate, and 0%-1% of butylated hydroxytoluene (BHT);

[0047] Preferably, by mass percentage, the composition comprises 0.3%-10% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 18%-20% of white beeswax, 40%-58% of white petrolatum, 20%-21% of light liquid paraffin, 2%-10% of isopropyl myristate, and 0%-0.2% of butylated hydroxytoluene (BHT);

[0048] More preferably, by mass percentage, the composition comprises 1% of the compound represented by formula (I), 18% of white beeswax, 58% of white petrolatum, 20.9% of light liquid paraffin, 2% of isopropyl myristate, and 0.1% of butylated hydroxytoluene (BHT).

[0049] The present invention also provides a method for preparing the composition as described above, characterized in that

[0050] when the composition does not contain a penetration enhancer and / or an antioxidant, the method comprises the following steps:

[0051] (1) Weigh oil phase substances 1 and 2, heat and melt them in a water bath, and obtain a homogeneous mixture A after homogenization;

[0052] (2) Weigh oil phase substance 3, add the weighed compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, heat and stir in a water bath to obtain a homogeneous mixture B;

[0053] (3) After cooling the mixture A, add the mixture B thereto, stir and then homogenize, and obtain the composition after cooling;

[0054] When the composition contains a penetration enhancer and / or an antioxidant, the method comprises the following steps:

[0055] (1) Weigh the oil phase substance 1, oil phase substance 2, penetration enhancer and / or antioxidant, heat and melt in a water bath, and obtain a uniform mixture A after homogenization;

[0056] (2) Weigh the oil phase substance 3, add the compound shown in formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal thereto, heat and stir in a water bath to obtain a uniform mixture B;

[0057] (3) After cooling the mixture A, add the mixture B thereto, stir and then homogenize, and obtain the composition after cooling.

[0058] The present invention also provides an application of the RNA m6A regulator composition as described above, or the method as described above in the preparation of a preparation for preventing or treating skin diseases caused by RNA m6A methylation;

[0059] Preferably, the skin diseases include acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction and dermatitis;

[0060] More preferably, the skin diseases include hand-foot syndrome and hand-foot skin reaction.

[0061] Preferably, the preparation includes ointment, cream, gel, frost, lotion, lotion, solution, paste, film, oil preparation or patch;

[0062] Preferably, the preparation is an ointment;

[0063] More preferably, the preparation is a suspension ointment.

[0064] Effects of the invention

[0065] In the RNA m6A regulator composition provided by the present invention, the exposure amount of the compound shown in formula (I) in the skin is high, and it can well prevent or treat skin diseases by inhibiting RNA m6A methylation. The composition of the present invention has good stability, and its appearance, viscosity, spreadability, physical stability and pharmacokinetic properties are all excellent. Description of the drawings

[0066] Figure 1Exemplary results of the mRNA expression levels of the glial cell differentiation markers KRT1, KRT10, Loricrin, and Involucrin measured by RT-PCR 24 hours after administering human recombinant HBEGF protein and the compound represented by formula (I) to keratinocytes HaCaT.

[0067] Figure 2 Exemplary results of the measurement of the stratum corneum thickness of the plantar skin of the rat paw in Example 4.

[0068] Figure 3 Exemplary histopathological results of the epithelial vesicles, inflammatory cell infiltration, and skin tissue congestion in the plantar skin of the rat paw after tissue staining in Example 4.

[0069] Figure 4 Exemplary results of the histopathological scoring of the epithelial vesicles, inflammatory cell infiltration, and skin tissue congestion in the plantar skin of the rat paw after tissue staining in Example 4.

[0070] Figure 5 XRPD pattern of the pharmaceutical cocrystal formed by the compound represented by formula (I) and nicotinamide.

[0071] Figure 6 XRPD pattern of the pharmaceutical cocrystal formed by the compound represented by formula (I) and isonicotinamide.

[0072] Figure 7 XRPD pattern of the pharmaceutical cocrystal formed by the compound represented by formula (I) and L-proline.

[0073] Figure 8 XRPD pattern of the pharmaceutical cocrystal formed by the compound represented by formula (I) and glycolic acid. Detailed Description of the Invention

[0074] To make the technical solutions and beneficial effects of the present invention more obvious and understandable, the following provides a detailed description by way of enumerating specific examples. Among them, the drawings are not necessarily drawn to scale, and local features can be enlarged or reduced to more clearly show the details of the local features; unless otherwise defined, the technical and scientific terms used herein have the same meanings as those in the technical and scientific fields to which this application belongs.

[0075] The present invention provides an RNA m6A regulator composition, the composition comprising: a compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable cocrystal thereof, and an oil phase substance.

[0076]

[0077]

[0078] In certain embodiments, the pharmaceutically acceptable salts include hydrochloride, sulfate, phosphate, acetate, mesylate, benzenesulfonate, and p-toluenesulfonate.

[0079] In certain embodiments, the eutectic forming substances of the pharmaceutically acceptable eutectics include nicotinamide, isonicotinamide, L-proline, and glycolic acid.

[0080] In certain embodiments, the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable eutectic exists in the form of solid particles.

[0081] In certain embodiments, the D90 of the solid particles is 0.01 - 50 μm.

[0082] In certain embodiments, the D90 of the solid particles is 0.01 - 20 μm.

[0083] In certain embodiments, the D90 of the solid particles is 0.01 - 10 μm.

[0084] In certain embodiments, the oil phase substances include oil phase substance 1, oil phase substance 2, and oil phase substance 3;

[0085] In certain embodiments, the oil phase substance 1 is selected from one or more of lanolin, beeswax, white beeswax, and paraffin wax.

[0086] In certain embodiments, the oil phase substance 1 is white beeswax.

[0087] In certain embodiments, the oil phase substance 2 is selected from one or more of petrolatum and white petrolatum.

[0088] In certain embodiments, the oil phase substance 2 is white petrolatum.

[0089] In certain embodiments, the oil phase substance 3 is selected from one or more of light liquid paraffin and vegetable oil.

[0090] In certain embodiments, the oil phase substance 3 is light liquid paraffin.

[0091] In certain embodiments, by mass percentage, the composition includes 0.1% - 20% of the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable eutectic, 10% - 30% of oil phase substance 1, 30% - 70% of oil phase substance 2, and 15% - 40% of oil phase substance 3.

[0092] In certain embodiments, the composition comprises, by mass percentage, 0.1%, or 0.2%, or 0.3%, or 0.4%, or 0.5%, or 0.6%, or 0.7%, or 0.8%, or 0.9%, or 1%, or 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or 11%, or 12%, or 13%, or 14%, or 15%, or 16%, or 17%, or 18%, or 19%, or 20% of the compound of formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable co-crystal thereof.

[0093] In certain embodiments, the composition comprises, by mass percentage, 10%, or 11%, or 12%, or 13%, or 14%, or 15%, or 16%, or 17%, or 18%, or 19%, or 20%, or 21%, or 22%, or 23%, or 24%, or 25%, or 26%, or 27%, or 28%, or 29%, or 30% of oil phase substance 1.

[0094] In certain embodiments, the composition comprises, by mass percentage, 30%, or 31%, or 32%, or 33%, or 34%, or 35%, or 36%, or 37%, or 38%, or 39%, or 40%, or 41%, or 42%, or 43%, or 44%, or 45%, or 46%, or 47%, or 48%, or 49%, or 50%, or 51%, or 52%, or 53%, or 54%, or 55%, or 56%, or 57%, or 58%, or 59%, or 60%, or 61%, or 62%, or 63%, or 64%, or 65%, or 66%, or 67%, or 68%, or 69%, or 70% of oil phase substance 2.

[0095] In certain embodiments, the composition comprises, by mass percentage, 15%, or 16%, or 17%, or 18%, or 19%, or 20%, or 21%, or 22%, or 23%, or 24%, or 25%, or 26%, or 27%, or 28%, or 29%, or 30%, or 31%, or 32%, or 33%, or 34%, or 35%, or 36%, or 37%, or 38%, or 39%, or 40% of oil phase substance 3.

[0096] In certain embodiments, the composition comprises 0.2% - 15% of the compound of formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable co-crystal thereof, 15% - 25% of oil phase substance 1, 35% - 64% of oil phase substance 2, and 20% - 35% of oil phase substance 3.

[0097] In certain embodiments, the composition comprises 0.3% - 10% of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable co-crystal thereof, 18% - 20% of oil phase substance 1, 40% - 58.7% of oil phase substance 2, and 22% - 30% of oil phase substance 3.

[0098] In certain embodiments, the composition comprises 0.3% of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable co-crystal thereof, 18% of oil phase substance 1, 58.7% of oil phase substance 2, and 23% of oil phase substance 3.

[0099] In certain embodiments, the composition comprises 1% of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable co-crystal thereof, 18% of oil phase substance 1, 58% of oil phase substance 2, and 23% of oil phase substance 3.

[0100] In certain embodiments, the composition comprises 3% of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable co-crystal thereof, 18% of oil phase substance 1, 57% of oil phase substance 2, and 22% of oil phase substance 3.

[0101] In certain embodiments, the composition comprises 10% of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable co-crystal thereof, 20% of oil phase substance 1, 40% of oil phase substance 2, and 30% of oil phase substance 3.

[0102] In certain embodiments, the composition comprises the compound represented by formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable co-crystal thereof, white beeswax, white petrolatum, and light liquid paraffin.

[0103] In certain embodiments, by mass percentage, the composition comprises 0.1% - 20% of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable co-crystal thereof, 10% - 30% of white beeswax, 30% - 70% of white petrolatum, and 15% - 40% of light liquid paraffin.

[0104] In certain embodiments, by mass percentage, the composition comprises 0.1%, or 0.2%, or 0.3%, or 0.4%, or 0.5%, or 0.6%, or 0.7%, or 0.8%, or 0.9%, or 1%, or 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or 11%, or 12%, or 13%, or 14%, or 15%, or 16%, or 17%, or 18%, or 19%, or 20% of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable co-crystal thereof.

[0105] In certain embodiments, the composition comprises, by mass percentage, 10%, or 11%, or 12%, or 13%, or 14%, or 15%, or 16%, or 17%, or 18%, or 19%, or 20%, or 21%, or 22%, or 23%, or 24%, or 25%, or 26%, or 27%, or 28%, or 29%, or 30% white beeswax.

[0106] In certain embodiments, the composition comprises, by mass percentage, 30%, or 31%, or 32%, or 33%, or 34%, or 35%, or 36%, or 37%, or 38%, or 39%, or 40%, or 41%, or 42%, or 43%, or 44%, or 45%, or 46%, or 47%, or 48%, or 49%, or 50%, or 51%, or 52%, or 53%, or 54%, or 55%, or 56%, or 57%, or 58%, or 59%, or 60%, or 61%, or 62%, or 63%, or 64%, or 65%, or 66%, or 67%, or 68%, or 69%, or 70% white petrolatum.

[0107] In certain embodiments, the composition comprises, by mass percentage, 15%, or 16%, or 17%, or 18%, or 19%, or 20%, or 21%, or 22%, or 23%, or 24%, or 25%, or 26%, or 27%, or 28%, or 29%, or 30% light liquid paraffin.

[0108] In certain embodiments, the composition comprises, by mass percentage, 0.2% - 15% of the compound of formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable co-crystal thereof, 15% - 25% white beeswax, 35% - 64% white petrolatum, 20% - 35% light liquid paraffin.

[0109] In certain embodiments, the composition comprises, by mass percentage, 0.3% - 10% of the compound of formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable co-crystal thereof, 18% - 20% white beeswax, 40% - 58.7% white petrolatum, 22% - 30% light liquid paraffin.

[0110] In certain embodiments, the composition comprises the compound of formula (I), white beeswax, white petrolatum, and light liquid paraffin.

[0111] In certain embodiments, the composition comprises, by mass percentage, 0.1% - 20% of the compound of formula (I), 10% - 30% white beeswax, 30% - 70% white petrolatum, 15% - 40% light liquid paraffin.

[0112] In certain embodiments, by mass percentage, the composition comprises 0.2% - 15% of the compound represented by formula (I), 15% - 25% white beeswax, 35% - 64% white petrolatum, and 20% - 35% light liquid paraffin.

[0113] In certain embodiments, by mass percentage, the composition comprises 0.3% - 10% of the compound represented by formula (I), 18% - 20% white beeswax, 40% - 58.7% white petrolatum, and 22% - 30% light liquid paraffin.

[0114] In certain embodiments, by mass percentage, the composition comprises 0.3% of the compound represented by formula (I), 18% white beeswax, 58.7% white petrolatum, and 23% light liquid paraffin.

[0115] In certain embodiments, by mass percentage, the composition comprises 1% of the compound represented by formula (I), 18% white beeswax, 58% white petrolatum, and 23% light liquid paraffin.

[0116] In certain embodiments, by mass percentage, the composition comprises 3% of the compound represented by formula (I), 18% white beeswax, 57% white petrolatum, and 22% light liquid paraffin.

[0117] In certain embodiments, by mass percentage, the composition comprises 10% of the compound represented by formula (I), 20% white beeswax, 40% white petrolatum, and 30% light liquid paraffin.

[0118] In certain embodiments, the composition consists of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, white beeswax, white petrolatum, and light liquid paraffin.

[0119] In certain embodiments, by mass percentage, the composition consists of 0.1% - 20% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 10% - 30% white beeswax, 30% - 70% white petrolatum, and 15% - 40% light liquid paraffin.

[0120] In certain embodiments, by mass percentage, the composition consists of 0.2% - 15% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 15% - 25% white beeswax, 35% - 64% white petrolatum, and 20% - 35% light liquid paraffin.

[0121] In certain embodiments, by mass percentage, the composition consists of 0.3% - 10% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 18% - 20% white beeswax, 40% - 58.7% white petrolatum, and 22% - 30% light liquid paraffin.

[0122] In certain embodiments, the composition consists of the compound represented by formula (I), white beeswax, white petrolatum, and light liquid paraffin.

[0123] In certain embodiments, by mass percentage, the composition consists of 0.1% - 20% of the compound represented by formula (I), 10% - 30% white beeswax, 30% - 70% white petrolatum, and 15% - 40% light liquid paraffin.

[0124] In certain embodiments, by mass percentage, the composition consists of 0.2% - 15% of the compound represented by formula (I), 15% - 25% white beeswax, 35% - 64% white petrolatum, and 20% - 35% light liquid paraffin.

[0125] In certain embodiments, by mass percentage, the composition consists of 0.3% - 10% of the compound represented by formula (I), 18% - 20% white beeswax, 40% - 58.7% white petrolatum, and 22% - 30% light liquid paraffin.

[0126] In certain embodiments, by mass percentage, the composition consists of 0.3% of the compound represented by formula (I), 18% white beeswax, 58.7% white petrolatum, and 23% light liquid paraffin.

[0127] In certain embodiments, by mass percentage, the composition consists of 1% of the compound represented by formula (I), 18% white beeswax, 58% white petrolatum, and 23% light liquid paraffin.

[0128] In certain embodiments, by mass percentage, the composition consists of 3% of the compound represented by formula (I), 18% white beeswax, 57% white petrolatum, and 22% light liquid paraffin.

[0129] In certain embodiments, by mass percentage, the composition consists of 10% of the compound represented by formula (I), 20% white beeswax, 40% white petrolatum, and 30% light liquid paraffin.

[0130] In certain embodiments, the composition further includes a penetration enhancer.

[0131] In certain embodiments, the penetration enhancer is selected from one or more of isopropyl myristate, isopropyl palmitate, propylene glycol dinonanoate, diethyl sebacate, azone, and propylene glycol.

[0132] In certain embodiments, the penetration enhancer is isopropyl myristate.

[0133] In certain embodiments, the composition further includes an antioxidant.

[0134] In some embodiments, the antioxidant is selected from one or more of vitamin E, alkyl gallate, butylated hydroxyanisole (BHA), and dibutylhydroxytoluene (BHT).

[0135] In some embodiments, the antioxidant is dibutylhydroxytoluene (BHT).

[0136] In some embodiments, the composition further comprises other pharmaceutically acceptable excipients.

[0137] In some embodiments, by mass percentage, the composition comprises 0.1% - 20% of the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable cocrystal, 10% - 30% of oil phase substance 1, 30% - 70% of oil phase substance 2, 15% - 30% of oil phase substance 3, 0.1% - 20% of penetration enhancer, and 0% - 2% of antioxidant.

[0138] In some embodiments, by mass percentage, the composition comprises 0.1%, or 0.2%, or 0.3%, or 0.4%, or 0.5%, or 0.6%, or 0.7%, or 0.8%, or 0.9%, or 1%, or 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or 11%, or 12%, or 13%, or 14%, or 15%, or 16%, or 17%, or 18%, or 19%, or 20% of the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable cocrystal.

[0139] In some embodiments, by mass percentage, the composition comprises 10%, or 11%, or 12%, or 13%, or 14%, or 15%, or 16%, or 17%, or 18%, or 19%, or 20%, or 21%, or 22%, or 23%, or 24%, or 25%, or 26%, or 27%, or 28%, or 29%, or 30% of oil phase substance 1.

[0140] In some embodiments, by mass percentage, the composition comprises 30%, or 31%, or 32%, or 33%, or 34%, or 35%, or 36%, or 37%, or 38%, or 39%, or 40%, or 41%, or 42%, or 43%, or 44%, or 45%, or 46%, or 47%, or 48%, or 49%, or 50%, or 51%, or 52%, or 53%, or 54%, or 55%, or 56%, or 57%, or 58%, or 59%, or 60%, or 61%, or 62%, or 63%, or 64%, or 65%, or 66%, or 67%, or 68%, or 69%, or 70% of oil phase substance 2.

[0141] In certain embodiments, the composition comprises, by mass percentage, 15%, or 16%, or 17%, or 18%, or 19%, or 20%, or 21%, or 22%, or 23%, or 24%, or 25%, or 26%, or 27%, or 28%, or 29%, or 30% of oil phase substance 3.

[0142] In certain embodiments, the composition comprises, by mass percentage, 0.1%, or 0.2%, or 0.3%, or 0.4%, or 0.5%, or 0.6%, or 0.7%, or 0.8%, or 0.9%, or 1%, or 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or 11%, or 12%, or 13%, or 14%, or 15%, or 16%, or 17%, or 18%, or 19%, or 20% of penetration enhancer.

[0143] In certain embodiments, the composition comprises, by mass percentage, 0%, or 0.1%, or 0.2%, or 0.3%, or 0.4%, or 0.5%, or 0.6%, or 0.7%, or 0.8%, or 0.9%, or 1%, or 1.1%, or 1.2%, or 1.3%, or 1.4%, or 1.5%, or 1.6%, or 1.7%, or 1.8%, or 1.9%, or 2% of antioxidant.

[0144] In certain embodiments, the composition comprises, by mass percentage, 0.2% - 15% of the compound of formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 15% - 25% of oil phase substance 1, 35% - 63% of oil phase substance 2, 20% - 25% of oil phase substance 3, 1% - 15% of penetration enhancer, 0% - 1% of antioxidant.

[0145] In certain embodiments, the composition comprises, by mass percentage, 0.3% - 10% of the compound of formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 18% - 20% of oil phase substance 1, 40% - 58% of oil phase substance 2, 20% - 21% of oil phase substance 3, 2% - 10% of penetration enhancer, 0% - 0.2% of antioxidant.

[0146] In certain embodiments, the composition comprises, by mass percentage, 0.1% - 20% of the compound of formula (I), 10% - 30% of oil phase substance 1, 30% - 70% of oil phase substance 2, 15% - 30% of oil phase substance 3, 0.1% - 20% of penetration enhancer, 0% - 2% of antioxidant.

[0147] In certain embodiments, by mass percentage, the composition comprises 0.2% - 15% of the compound represented by formula (I), 15% - 25% of oil phase substance 1, 35% - 63% of oil phase substance 2, 20% - 25% of oil phase substance 3, 1% - 15% of penetration enhancer, and 0% - 1% of antioxidant.

[0148] In certain embodiments, by mass percentage, the composition comprises 0.3% - 10% of the compound represented by formula (I), 18% - 20% of oil phase substance 1, 40% - 58% of oil phase substance 2, 20% - 21% of oil phase substance 3, 2% - 10% of penetration enhancer, and 0% - 0.2% of antioxidant.

[0149] In certain embodiments, the composition comprises the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, white beeswax, white petrolatum, light liquid paraffin, and isopropyl myristate.

[0150] In certain embodiments, by mass percentage, the composition comprises 0.1% - 20% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 10% - 30% of white beeswax, 30% - 70% of white petrolatum, 15% - 30% of light liquid paraffin, and 0.1% - 20% of isopropyl myristate.

[0151] In certain embodiments, by mass percentage, the composition comprises 0.1%, or 0.2%, or 0.3%, or 0.4%, or 0.5%, or 0.6%, or 0.7%, or 0.8%, or 0.9%, or 1%, or 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or 11%, or 12%, or 13%, or 14%, or 15%, or 16%, or 17%, or 18%, or 19%, or 20% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof.

[0152] In certain embodiments, by mass percentage, the composition comprises 10%, or 11%, or 12%, or 13%, or 14%, or 15%, or 16%, or 17%, or 18%, or 19%, or 20%, or 21%, or 22%, or 23%, or 24%, or 25%, or 26%, or 27%, or 28%, or 29%, or 30% of white beeswax.

[0153] In certain embodiments, by mass percentage, the composition comprises 30%, or 31%, or 32%, or 33%, or 34%, or 35%, or 36%, or 37%, or 38%, or 39%, or 40%, or 41%, or 42%, or 43%, or 44%, or 45%, or 46%, or 47%, or 48%, or 49%, or 50%, or 51%, or 52%, or 53%, or 54%, or 55%, or 56%, or 57%, or 58%, or 59%, or 60%, or 61%, or 62%, or 63%, or 64%, or 65%, or 66%, or 67%, or 68%, or 69%, or 70% white petrolatum.

[0154] In certain embodiments, by mass percentage, the composition comprises 15%, or 16%, or 17%, or 18%, or 19%, or 20%, or 21%, or 22%, or 23%, or 24%, or 25%, or 26%, or 27%, or 28%, or 29%, or 30% light liquid paraffin.

[0155] In certain embodiments, by mass percentage, the composition comprises 0.1%, or 0.2%, or 0.3%, or 0.4%, or 0.5%, or 0.6%, or 0.7%, or 0.8%, or 0.9%, or 1%, or 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or 11%, or 12%, or 13%, or 14%, or 15%, or 16%, or 17%, or 18%, or 19%, or 20% isopropyl myristate.

[0156] In certain embodiments, by mass percentage, the composition comprises 0.2% - 15% of the compound of formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 15% - 25% white beeswax, 35% - 63% white petrolatum, 20% - 25% light liquid paraffin, 1% - 15% isopropyl myristate.

[0157] In certain embodiments, by mass percentage, the composition comprises 0.3% - 10% of the compound of formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 18% - 20% white beeswax, 40% - 58% white petrolatum, 20% - 21% light liquid paraffin, 2% - 10% isopropyl myristate.

[0158] In certain embodiments, by mass percentage, the composition comprises 0.1% - 20% of the compound of formula (I), 10% - 30% white beeswax, 30% - 70% white petrolatum, 15% - 30% light liquid paraffin, 0.1% - 20% isopropyl myristate.

[0159] In certain embodiments, by mass percentage, the composition comprises 0.2%-15% of the compound represented by formula (I), 15%-25% white beeswax, 35%-63% white petrolatum, 20%-25% light liquid paraffin, and 1%-15% isopropyl myristate.

[0160] In certain embodiments, by mass percentage, the composition comprises 0.3%-10% of the compound represented by formula (I), 18%-20% white beeswax, 40%-58% white petrolatum, 20%-21% light liquid paraffin, and 2%-10% isopropyl myristate.

[0161] In certain embodiments, by mass percentage, the composition comprises 1% of the compound represented by formula (I), 18% white beeswax, 58% white petrolatum, 21% light liquid paraffin, and 2% isopropyl myristate.

[0162] In certain embodiments, by mass percentage, the composition comprises 10% of the compound represented by formula (I), 20% white beeswax, 40% white petrolatum, 20% light liquid paraffin, and 10% isopropyl myristate.

[0163] In certain embodiments, the composition consists of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, white beeswax, white petrolatum, light liquid paraffin, and isopropyl myristate.

[0164] In certain embodiments, by mass percentage, the composition consists of 0.1%-20% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 10%-30% white beeswax, 30%-70% white petrolatum, 15%-30% light liquid paraffin, and 0.1%-20% isopropyl myristate.

[0165] In certain embodiments, by mass percentage, the composition consists of 0.2%-15% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 15%-25% white beeswax, 35%-63% white petrolatum, 20%-25% light liquid paraffin, and 1%-15% isopropyl myristate.

[0166] In certain embodiments, by mass percentage, the composition consists of 0.3%-10% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 18%-20% white beeswax, 40%-58% white petrolatum, 20%-21% light liquid paraffin, and 2%-10% isopropyl myristate.

[0167] In certain embodiments, by mass percentage, the composition consists of 0.1%-20% of the compound represented by formula (I), 10%-30% white beeswax, 30%-70% white petrolatum, 15%-30% light liquid paraffin, and 0.1%-20% isopropyl myristate.

[0168] In certain embodiments, by mass percentage, the composition consists of 0.2%-15% of the compound represented by formula (I), 15%-25% white beeswax, 35%-63% white petrolatum, 20%-25% light liquid paraffin, and 1%-15% isopropyl myristate.

[0169] In certain embodiments, by mass percentage, the composition consists of 0.3%-10% of the compound represented by formula (I), 18%-20% white beeswax, 40%-58% white petrolatum, 20%-21% light liquid paraffin, and 2%-10% isopropyl myristate.

[0170] In certain embodiments, by mass percentage, the composition consists of 1% of the compound represented by formula (I), 18% white beeswax, 58% white petrolatum, 21% light liquid paraffin, and 2% isopropyl myristate.

[0171] In certain embodiments, by mass percentage, the composition consists of 10% of the compound represented by formula (I), 20% white beeswax, 40% white petrolatum, 20% light liquid paraffin, and 10% isopropyl myristate.

[0172] In certain embodiments, the composition includes the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, white beeswax, white petrolatum, light liquid paraffin, isopropyl myristate, and butylated hydroxytoluene (BHT).

[0173] In certain embodiments, by mass percentage, the composition includes 0.1%-20% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 10%-30% white beeswax, 30%-70% white petrolatum, 15%-30% light liquid paraffin, 0.1%-20% isopropyl myristate, and 0%-2% butylated hydroxytoluene (BHT).

[0174] In certain embodiments, by mass percentage, the composition comprises 0.1%, or 0.2%, or 0.3%, or 0.4%, or 0.5%, or 0.6%, or 0.7%, or 0.8%, or 0.9%, or 1%, or 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or 11%, or 12%, or 13%, or 14%, or 15%, or 16%, or 17%, or 18%, or 19%, or 20% of the compound represented by formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutically acceptable co-crystal thereof.

[0175] In certain embodiments, by mass percentage, the composition comprises 10%, or 11%, or 12%, or 13%, or 14%, or 15%, or 16%, or 17%, or 18%, or 19%, or 20%, or 21%, or 22%, or 23%, or 24%, or 25%, or 26%, or 27%, or 28%, or 29%, or 30% white beeswax.

[0176] In certain embodiments, by mass percentage, the composition comprises 30%, or 31%, or 32%, or 33%, or 34%, or 35%, or 36%, or 37%, or 38%, or 39%, or 40%, or 41%, or 42%, or 43%, or 44%, or 45%, or 46%, or 47%, or 48%, or 49%, or 50%, or 51%, or 52%, or 53%, or 54%, or 55%, or 56%, or 57%, or 58%, or 59%, or 60%, or 61%, or 62%, or 63%, or 64%, or 65%, or 66%, or 67%, or 68%, or 69%, or 70% white petrolatum.

[0177] In certain embodiments, by mass percentage, the composition comprises 15%, or 16%, or 17%, or 18%, or 19%, or 20%, or 21%, or 22%, or 23%, or 24%, or 25%, or 26%, or 27%, or 28%, or 29%, or 30% light liquid paraffin.

[0178] In certain embodiments, by mass percentage, the composition comprises 0.1%, or 0.2%, or 0.3%, or 0.4%, or 0.5%, or 0.6%, or 0.7%, or 0.8%, or 0.9%, or 1%, or 2%, or 3%, or 4%, or 5%, or 6%, or 7%, or 8%, or 9%, or 10%, or 11%, or 12%, or 13%, or 14%, or 15%, or 16%, or 17%, or 18%, or 19%, or 20% isopropyl myristate.

[0179] In certain embodiments, the composition comprises, by mass percentage, 0%, or 0.1%, or 0.2%, or 0.3%, or 0.4%, or 0.5%, or 0.6%, or 0.7%, or 0.8%, or 0.9%, or 1%, or 1.1%, or 1.2%, or 1.3%, or 1.4%, or 1.5%, or 1.6%, or 1.7%, or 1.8%, or 1.9%, or 2% of dibutylhydroxytoluene (BHT).

[0180] In certain embodiments, the composition comprises, by mass percentage, 0.2% - 15% of the compound of formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 15% - 25% of white beeswax, 35% - 63% of white petrolatum, 20% - 25% of light liquid paraffin, 1% - 15% of isopropyl myristate, and 0% - 1% of dibutylhydroxytoluene (BHT).

[0181] In certain embodiments, the composition comprises, by mass percentage, 0.3% - 10% of the compound of formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 18% - 20% of white beeswax, 40% - 58% of white petrolatum, 20% - 21% of light liquid paraffin, 2% - 10% of isopropyl myristate, and 0% - 0.2% of dibutylhydroxytoluene (BHT).

[0182] In certain embodiments, the composition comprises, by mass percentage, 0.1% - 20% of the compound of formula (I), 10% - 30% of white beeswax, 30% - 70% of white petrolatum, 15% - 30% of light liquid paraffin, 0.1% - 20% of isopropyl myristate, and 0% - 2% of dibutylhydroxytoluene (BHT).

[0183] In certain embodiments, the composition comprises, by mass percentage, 0.2% - 15% of the compound of formula (I), 15% - 25% of white beeswax, 35% - 63% of white petrolatum, 20% - 25% of light liquid paraffin, 1% - 15% of isopropyl myristate, and 0% - 1% of dibutylhydroxytoluene (BHT).

[0184] In certain embodiments, the composition comprises, by mass percentage, 0.3% - 10% of the compound of formula (I), 18% - 20% of white beeswax, 40% - 58% of white petrolatum, 20% - 21% of light liquid paraffin, 2% - 10% of isopropyl myristate, and 0% - 0.2% of dibutylhydroxytoluene (BHT).

[0185] In certain embodiments, the composition comprises, by mass percentage, 1% of the compound of formula (I), 18% of white beeswax, 58% of white petrolatum, 20.9% of light liquid paraffin, 2% of isopropyl myristate, and 0.1% of dibutylhydroxytoluene (BHT).

[0186] In certain embodiments, the composition consists of a compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, white beeswax, white petrolatum, light liquid paraffin, isopropyl myristate, and butylated hydroxytoluene (BHT).

[0187] In certain embodiments, by mass percentage, the composition consists of 0.1% - 20% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 10% - 30% white beeswax, 30% - 70% white petrolatum, 15% - 30% light liquid paraffin, 0.1% - 20% isopropyl myristate, and 0% - 2% butylated hydroxytoluene (BHT).

[0188] In certain embodiments, by mass percentage, the composition consists of 0.2% - 15% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 15% - 25% white beeswax, 35% - 63% white petrolatum, 20% - 25% light liquid paraffin, 1% - 15% isopropyl myristate, and 0% - 1% butylated hydroxytoluene (BHT).

[0189] In certain embodiments, by mass percentage, the composition consists of 0.3% - 10% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 18% - 20% white beeswax, 40% - 58% white petrolatum, 20% - 21% light liquid paraffin, 2% - 10% isopropyl myristate, and 0% - 0.2% butylated hydroxytoluene (BHT).

[0190] In certain embodiments, by mass percentage, the composition consists of 0.1% - 20% of the compound represented by formula (I), 10% - 30% white beeswax, 30% - 70% white petrolatum, 15% - 30% light liquid paraffin, 0.1% - 20% isopropyl myristate, and 0% - 2% butylated hydroxytoluene (BHT).

[0191] In certain embodiments, by mass percentage, the composition consists of 0.2% - 15% of the compound represented by formula (I), 15% - 25% white beeswax, 35% - 63% white petrolatum, 20% - 25% light liquid paraffin, 1% - 15% isopropyl myristate, and 0% - 1% butylated hydroxytoluene (BHT).

[0192] In certain embodiments, by mass percentage, the composition consists of 0.3% - 10% of the compound represented by formula (I), 18% - 20% white beeswax, 40% - 58% white petrolatum, 20% - 21% light liquid paraffin, 2% - 10% isopropyl myristate, and 0% - 0.2% butylated hydroxytoluene (BHT).

[0193] In some embodiments, by mass percentage, the composition consists of 1% of the compound represented by formula (I), 18% white beeswax, 58% white petrolatum, 20.9% light liquid paraffin, 2% isopropyl myristate, and 0.1% butylated hydroxytoluene (BHT).

[0194] The present invention also provides a method for preparing the composition as described above, characterized in that

[0195] When the composition does not contain a penetration enhancer and / or an antioxidant, the method comprises the following steps:

[0196] (1) Weigh oil phase substance 1 and oil phase substance 2, melt them by heating in a water bath, and obtain a homogeneous mixture A after homogenization;

[0197] (2) Weigh oil phase substance 3, add the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal that has been weighed, heat and stir in a water bath to obtain a homogeneous mixture B;

[0198] (3) After cooling the mixture A, add the mixture B thereto, stir and then homogenize, and obtain the composition after cooling;

[0199] When the composition contains a penetration enhancer and / or an antioxidant, the method comprises the following steps:

[0200] (1) Weigh oil phase substance 1, oil phase substance 2, the penetration enhancer and / or the antioxidant, melt them by heating in a water bath, and obtain a homogeneous mixture A after homogenization;

[0201] (2) Weigh oil phase substance 3, add the compound represented by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal that has been weighed, heat and stir in a water bath to obtain a homogeneous mixture B;

[0202] (3) After cooling the mixture A, add the mixture B thereto, stir and then homogenize, and obtain the composition after cooling.

[0203] In some embodiments, the heating in the water bath in step (1) is heating to 65 - 70 °C.

[0204] In some embodiments, the homogenization in step (1) is homogenization at 350 rpm for 15 - 20 minutes.

[0205] In some embodiments, the heating in the water bath in step (2) is heating to 40 °C.

[0206] In some embodiments, the stirring in step (2) is stirring at 300 - 400 rpm for 10 minutes.

[0207] In some embodiments, the cooling in step (3) is cooling to 40 °C.

[0208] In some embodiments, the speed of the homogenizer in step (3) is 10,000 rpm and the homogenization time is 2 minutes.

[0209] In some embodiments, the cooling in step (3) is cooling to room temperature.

[0210] The present invention also provides an application of the RNA m6A regulator composition as described above, or the method as described above, in the preparation of a preparation for preventing or treating skin diseases caused by RNA m6A methylation.

[0211] In some embodiments, the skin diseases include acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction, and dermatitis.

[0212] In some embodiments, the skin diseases include hand-foot syndrome and hand-foot skin reaction.

[0213] In some embodiments, the preparation includes ointment, cream, gel, frost, lotion, lotion, solution, paste, film, oil preparation or patch.

[0214] In some embodiments, the preparation is an ointment.

[0215] In some embodiments, the preparation is a suspension ointment.

[0216] Example 1: Preparation of the compound shown in formula (I)

[0217]

[0218] Ethyl 3-bromo-2-oxocyclohexane-1-carboxylate (20 mg, 0.08 mmol) and 4-chloroaniline (25 mg, 0.2 mmol) were mixed and then heated to 150 °C. After reacting for 3 hours, the reaction solution was cooled to room temperature, diluted with 100 mL of dichloromethane, washed 3 times with 100 mL of 1N HCl and once with 100 mL of saturated NaHCO3. The organic layer was dried over anhydrous Na2SO4 and concentrated in vacuo. Purification by silica gel column chromatography gave ethyl 6-chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylate (16 mg). 10 mg of ethyl 6-chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxylate (0.036 mmol) was dissolved in 10 mL of ethanol, 2 mL of 2M LiOH solution was added, and the mixture was stirred at room temperature for 1 hour. After rotary evaporation, 20 mL of water was added for dilution, the pH was adjusted to 2, and then extracted 3 times with 50 mL of dichloromethane. The combined organic phases were dried and concentrated, ammonia water was added, and the reaction was carried out at 60 °C for 24 hours. Purification by silica gel column chromatography gave a white solid, 6-chloro-2,3,4,9-tetrahydro-1H-carbazole-1-carboxamide (Compound 1, 7.6 mg). The (S)-6-chloro-2,3,4,9-tetrahydro-1H-carbazolecarboxamide (the compound shown in Formula (I)) was obtained by separation using a Chiralpak AD chiral column. The compound shown in Formula (I) LCMS [M+H] + : 249. 1 1H-NMR (400 MHz, DMSO-d6): δ 10.79 (s, 1H), 7.37 - 7.39 (m, 2H), 7.28 (d, J = 8.0 Hz, 1H), 7.08 (s, 1H), 6.98 - 7.01 (m, 1H), 3.64 - 3.67 (m, 1H), 2.58 - 2.61 (m, 2H), 1.66 - 2.04 (m, 4H).

[0219] Example 2: Effect of the compound shown in Formula (I) on the mRNA methylation level of THP-1 cells

[0220] 8×10 6THP-1 cells were placed in a culture dish and added with DMEM medium, an equal volume of DMSO was added to the control group, 2 μM UZH1a (Shanghai Haoyuan Pharmaceutical Co., Ltd.) was added to the positive reference group 2, and 5 nM, 100 nM and 2 μM of the compound represented by formula (I) were added to sample groups A, B and C, respectively, and placed in an incubator containing 5% carbon dioxide at 37°C for 24 hours, washed with PBS, centrifuged at high speed, and RLT lysis buffer (QIAGEN) was added after the supernatant was discarded. After adding 400 μL of 70% ethanol to dilute the sample, 700 μL of the sample was taken out and total RNA was extracted with Trizol reagent (Sigma Aldrich), and the RNA sample was purified with Oligo (dT) magnetic beads. Take 200ng of purified mRNA sample, add 20μL of nuclease P1 mixture (1unit nuclease P1, 0.4μL of 2M NaCl, 2μL of 0.1M ZnCl2, 17.1μL of PCR grade water) and react at 37℃ for 2 hours, add 2μL of NH4HCO3 solution and 1unit of alkaline phosphatase and react at 37℃ for 2 hours, add 1.2M HCl to neutralize and centrifuge, collect 20μL of supernatant and detect ion peak area (150.000549 / 150.000549) by LC-MS. The inhibition rate of m6A is calculated as follows, and the results are shown in Table 1:

[0221] m6A inhibition rate = (peak area (control group) – peak area (sample group)) / peak area (control group) * 100%

[0222] Table 1: m6A inhibition rate of control group and sample group AC

[0223] Group Compound Compound Concentration m6A Inhibition Rate Control Group None None 0 Sample Group A Compound Shown in Formula (I) 5nM 18.2％ Sample Group B Compound Shown in Formula (I) 100nM 29.7％ Sample Group C Compound Shown in Formula (I) 2μM 56.8％ Positive Reference Group UZH1a 2μM 41.5％

[0224] UZH1a is a nanomolar inhibitor of the selective RNA m6A methylase METTL3 (Moroz-Omori EV et al. ChemMedChem 2021, 2021, 16(19): 3035-3043). In THP-1 cells, the inhibition rate of UZH1a for RNA m6A is about 41.5%. The compound represented by formula (I) inhibits m6A in a dose-dependent manner, and the inhibition rate of RNA m6A is higher (56.8%) at the same concentration as UZH1a.

[0225] Example 3: Inhibitory effect of the compound represented by formula (I) on differentiation of keratinocytes HaCaT

[0226] Human keratinocytes HaCaT were cultured in DMEM medium containing 10% fetal bovine serum (Gibco, 10099141), 100 U / mL penicillin, and 100 μg / mL streptomycin. Cells were seeded in 96-well plates at a density of 2*10 6 cells per square centimeter and cultured in an incubator with 5% carbon dioxide at 37 °C for 24 hours. Equal volume of DMSO was added to control well 1, and 2.5 ng / mL human recombinant HBEGF protein (Abcam, ab205523) was added to control well 2. Compounds of formula (I) at corresponding concentrations and 2.5 ng / mL human recombinant HBEGF protein were added to sample wells. After continuous culture for 24 hours, cells were collected by high-speed centrifugation after washing with PBS. Total RNA was extracted using Trizol reagent (SigmaAldrich), and 1 μg of RNA was reverse transcribed into cDNA using a cDNA reverse transcription kit (Transgene Biotech, AT311-03). A 20 μL reaction solution was prepared by adding 1.25 μL of primers (primer sequences are shown in Table 2), 10 μL of iTag Universal SYBR Green supermix (Bio-Rad, 172-5125), and appropriate amount of DEPC ultrapure water for RT-PCR reaction. After the reaction, the reaction solution was subjected to agarose gel electrophoresis to determine the mRNA expression levels of keratinocyte differentiation markers KRT1, KRT10, Loricrin, and Involucrin.

[0227] Table 2: Information on partial primer sequences

[0228]

[0229] As Figure 1 shown, after induction with human recombinant HBEGF protein, the mRNA expression levels of differentiation markers KRT1, KRT10, Loricrin, and Involucrin in HaCaT increased significantly. However, after adding the compounds of formula (I), the mRNA expression levels of differentiation markers KRT1, KRT10, Loricrin, and Involucrin in HaCaT were significantly inhibited.

[0230] Example 4: Inhibitory effect of the compound of formula (I) on keratinocyte differentiation in a rat hand-foot skin reaction model

[0231] After feeding and acclimating SD rats (weighing approximately 200 g) for one week, they were evenly grouped by body weight, with 12 rats in each group. The modeling drugs (sorafenib, erlotinib, afatinib, and osimertinib) were separately dissolved in a solution containing 5% DMSO, 45% PEG400, and 50% H2O, and the modeling drugs were made up to the required concentration. They were administered by gavage once a day at the doses shown in Table 3. One hour after gavage administration, 0.05 g of ointment containing the compound shown in formula (I) with different mass ratios was evenly applied to the left paw of the rats, while the right paw was used as a self-control and applied with a blank matrix. The rats in the blank control group were not medicated. After applying the medicine, the four limbs were fixed for another 2 hours, then the residual medicine was wiped off with water, the fixation was released, and the rats were allowed to resume free movement. The modeling drugs and the test compound ointment were both administered once a day. After continuous administration for 30 days, the rats were euthanized, and the skin tissue of the paw sole was taken, fixed in 10% neutral formaldehyde, cut into 5-μm sections, dehydrated, and embedded in paraffin.

[0232] Tissue staining: After dewaxing and rehydrating the skin tissue sections of the rat paw soles described above, they were stained in a hematoxylin solution for several minutes, washed, and then soaked in 1% acidic alcohol until the sections faded to light blue-red. After washing with running water for 5 minutes, they were stained with eosin for 2-3 minutes, washed to remove excess dye, dehydrated, then made transparent with xylene for several minutes, and sealed with neutral resin. The stratum corneum was observed under an optical microscope, and the thickness of the epidermal stratum corneum was measured using Dmetrix software.

[0233] Immunohistochemical staining: After dewaxing and hydrating the skin tissue sections of the rat paw soles described above, they were incubated with 3% H2O2 at room temperature for 30 minutes. After antigen retrieval, they were blocked with 10% goat serum for 30 minutes. Anti-KRT1 antibody (Abcam, ab93652) and anti-Loricrin antibody (Abcam, ab183646) were added dropwise and incubated overnight at 4°C. HRP secondary antibody (ZSGB-BIO, PV-6001) and DAB kit (ZSG-BIO, ZLI9017) were added for color development, counterstained with hematoxylin, washed, and sealed with neutral resin. The expression levels of KRT1 and Loricrin were observed under an optical microscope.

[0234] The criteria for determining the successful establishment of the rat model described above are as follows: (i) symptoms such as erythema, swelling, desquamation, ulceration, or blisters appear in the paw area; and / or (ii) the thickness of the stratum corneum in tissue staining is significantly higher than that of normal rats; and / or (iii) the markers such as KRT1, KRT5, and Loricrin are significantly elevated. The incidence rate is calculated as the proportion of animals in each group that meet the criteria for successful model establishment, that is, incidence rate = (number of rats with successful modeling / total number of rats in the group) * 100%.

[0235] As described above, the pathological scoring criteria for tissue staining of rats are as follows: no blisters, score 0; the number of blisters is 1 - 3, score 1; the number of blisters is 4 - 6, score 2; the number of blisters is 7 - 9, score 3; the number of blisters is more than 10, score 4. The inflammatory area accounts for less than 10% of the total area of the section, score 0; the inflammatory area accounts for 10 - 25% of the total area of the section, score 1; the inflammatory area accounts for 25 - 50% of the total area of the section, score 2; the inflammatory area accounts for 50 - 75% of the total area of the section, score 3; the inflammatory area accounts for more than 75% of the total area of the section, score 4. The hyperemic area accounts for less than 10% of the total area of the section, score 0; the hyperemic area accounts for 10 - 25% of the total area of the section, score 1; the hyperemic area accounts for 25 - 50% of the total area of the section, score 2; the hyperemic area accounts for 50 - 75% of the total area of the section, score 3; the hyperemic area accounts for more than 75% of the total area of the section, score 4. The scoring adopts a double - blind scoring system. After the statistics of each group of data are completed, it is presented in the form of Mean + SEM (N = 12).

[0236] Table 3: Drug dosage and experimental results in the rat model

[0237]

[0238]

[0239] As shown in Table 3, sorafenib, erlotinib, afatinib, and osimertinib were administered by gavage once a day to SD rats at doses of 100 mg / kg, 70 mg / kg, 50 mg / kg, and 60 mg / kg respectively, and the success rates of establishing the hand - foot skin reaction model were 75%, 83.3%, 66.7%, and 75% respectively. The compound shown in formula (I) can significantly reduce the incidence of hand - foot skin reactions at the drug - applied site at low doses (for example, the mass content of the compound in the ointment is 1%), and it can significantly reduce the incidence of hand - foot skin reactions in all paw pads of rats at high mass contents (for example, the mass content of the compound in the ointment is 3% and 10%).

[0240] As Figure 2 shown in the measurement results of the stratum corneum thickness, in the rats successfully modeled with sorafenib, erlotinib, afatinib, and osimertinib, the stratum corneum thickness of the plantar skin of the paw is significantly higher than that of normal rats. The compound shown in formula (I) can effectively reduce the stratum corneum thickness of the plantar skin of the paw in the modeled rats.

[0241] As Figure 3As shown in the tissue staining results, the skin toxicity caused by the modeling drug can form blisters under the subcutaneous and epidermal layers of the rat paw plantar, with inflammatory cell infiltration in the dermis layer and hyperemia symptoms in the skin tissue. The compound shown in formula (I) can effectively improve the formation of blisters under the subcutaneous and epidermal layers caused by the above modeling drug, inhibit the inflammatory cell infiltration in the dermis layer, and significantly improve the skin hyperemia phenomenon.

[0242] As Figure 4 shown in the pathological scoring results, the modeling drug can cause blisters to form under the subcutaneous and epidermal layers of the rat paw plantar, with inflammatory cell infiltration in the dermis layer and hyperemia symptoms in the skin tissue. The compound shown in formula (I) can effectively improve the formation of blisters under the subcutaneous and epidermal layers caused by the above modeling drug, inhibit the inflammatory cell infiltration in the dermis layer, and significantly improve the skin hyperemia phenomenon.

[0243] The immunohistochemical staining results also show that in rats successfully modeled with sorafenib, erlotinib, afatinib, and osimertinib, KRT1 and Loricrin are significantly increased in the tissue. However, with the addition of the compound shown in formula (I), KRT1 and Loricrin are significantly decreased in the tissue. In summary, the compound shown in formula (I) can effectively inhibit the differentiation of keratinocytes in vitro and in vivo and has the potential to treat hyperkeratosis-related diseases.

[0244] Example 5: Preparation of a drug cocrystal formed by the compound shown in formula (I) and nicotinamide

[0245] Weigh approximately 20 mg of the compound shown in formula (I) and place it in a 2 mL glass bottle with 10.3 mg of nicotinamide, then add acetonitrile (0.25 mL) to obtain a suspension. The obtained sample is stirred under suspension at 5 °C for 3 days. The obtained suspension is centrifuged at 14,000 rpm through a 0.45 μm nylon filter membrane, and the obtained solid is vacuum dried at 25 °C for 4 hours to obtain the product. After X-ray powder diffraction detection, this product is a drug cocrystal formed by the compound shown in formula (I) and nicotinamide, and the XRPD spectrum is as Figure 5 shown, and its characteristic peak positions are shown in Table 4. The DSC spectrum shows that the melting T onset @156.75 °C.

[0246] Table 4: XRPD diffraction peak data of the drug cocrystal formed by the compound shown in formula (I) and nicotinamide

[0247]

[0248]

[0249] Example 6: Preparation of a drug cocrystal formed by the compound shown in formula (I) and isonicotinamide

[0250] Weigh approximately 20 mg of the compound shown in formula (I), place it in a 2 mL glass bottle together with 10.2 mg of isonicotinamide, add ethanol (0.2 mL) to obtain a suspension. The obtained sample is stirred in suspension at 5 °C for 3 days. The obtained suspension is centrifuged at 14,000 rpm through a 0.45 μm nylon filter membrane, and the obtained solid is dried in vacuo at 25 °C for 4 hours to obtain the product. After detection by X-ray powder diffraction, this product is a drug cocrystal formed by the compound shown in formula (I) and isonicotinamide, and the XRPD pattern is as shown in Figure 6 shown, and the characteristic peak positions are shown in Table 5. The DSC pattern shows that the melting T onset is 160.45 °C.

[0251] Table 5: XRPD diffraction peak data of the drug cocrystal formed by the compound shown in formula (I) and isonicotinamide

[0252]

[0253] Example 7: Preparation of a drug cocrystal formed by the compound shown in formula (I) and L-proline

[0254] Weigh approximately 20 mg of the compound shown in formula (I), place it in a 2 mL glass bottle together with 9.7 mg of L-proline, add ethanol (0.25 mL) to obtain a suspension. The obtained sample is stirred in suspension at 5 °C for 3 days. The obtained suspension is centrifuged at 14,000 rpm through a 0.45 μm nylon filter membrane, and the obtained solid is dried in vacuo at 25 °C for 4 hours to obtain the product. After detection by X-ray powder diffraction, this product is a drug cocrystal formed by the compound shown in formula (I) and L-proline, and the XRPD pattern is as shown in Figure 7 shown, and the characteristic peak positions are shown in Table 6. The DSC pattern shows that the melting T onset is @183.55 °C.

[0255] Table 6: XRPD diffraction peak data of the drug cocrystal formed by the compound shown in formula (I) and L-proline

[0256]

[0257]

[0258] Example 8: Preparation of a drug cocrystal formed by the compound shown in formula (I) and glycolic acid

[0259] Weigh approximately 20 mg of the compound shown in formula (I), place it together with 6.3 mg of glycolic acid in a 2 mL glass bottle, add isopropyl acetate (0.2 mL) to obtain a suspension. The obtained sample is stirred in suspension at 5 °C for 3 days. The obtained suspension is centrifuged at 14,000 rpm through a 0.45 μm nylon filter membrane, and the obtained solid is dried in vacuo at 25 °C for 4 hours to obtain the product. After detection by X-ray powder diffraction, this product is a drug cocrystal formed by the compound shown in formula (I) and glycolic acid, and the XRPD spectrum is as Figure 8 shown, and the characteristic peak positions are shown in Table 7. The DSC spectrum shows that the melting T onset @ 135.68 °C.

[0260] Table 7: XRPD diffraction peak data of the drug cocrystal formed by the compound shown in formula (I) and glycolic acid

[0261]

[0262]

[0263] Example 9: Suspension-type ointment preparation (1%) containing the compound shown in formula (I)

[0264] Prepare a suspension-type ointment preparation containing the compound shown in formula (I), and the specific prescription is shown in Table 8:

[0265] Table 8: Prescription information of the suspension-type ointment preparation (1%) containing the compound shown in formula (I)

[0266] Ingredient Function Dosage % (w / w) Compound Shown in Formula (I) Active Ingredient 1 White Beeswax Ointment Base 18 White Petrolatum Ointment Base 58 Light Liquid Paraffin Ointment Base 23

[0267] Preparation method:

[0268] (1) Weigh white beeswax and white petrolatum, heat them in a water bath to 65 - 70 °C until melted, and homogenize at a stirring speed of 350 rpm for 20 minutes to obtain a uniform mixture A;

[0269] (2) Weigh light liquid paraffin, add the weighed compound shown in formula (I) thereto, heat it in a water bath to about 40 °C and stir for 10 minutes until the active ingredient is completely dispersed, then stir at a speed of 400 rmp to obtain a uniform mixture B;

[0270] (3) Cool mixture A to 40 °C, add mixture B thereto, continuously stir, and then homogenize at a speed of 10,000 rmp with a homogenizer for 2 minutes, and then cool to room temperature to obtain a suspension-type ointment preparation containing the compound shown in formula (I).

[0271] Example 10: Suspension-type ointment preparation (3%) containing the compound shown in formula (I)

[0272] Prepare a suspension ointment preparation containing the compound shown in formula (I). The preparation method is the same as that in Example 9, and the specific prescription is shown in Table 9.

[0273] Table 9: Prescription Information of Suspension Ointment Preparation (3%) Containing the Compound Shown in Formula (I)

[0274] Ingredient Function Dosage % (w / w) Compound Shown in Formula (I) Active Ingredient 3 White Beeswax Ointment Base 18 White Petrolatum Ointment Base 57 Light Liquid Paraffin Ointment Base 22

[0275] Example 11: Suspension Ointment Preparation (10%) Containing the Compound Shown in Formula (I)

[0276] Prepare a suspension ointment preparation containing the compound shown in formula (I). The preparation method is the same as that in Example 9, and the specific prescription is shown in Table 10.

[0277] Table 10: Prescription Information of Suspension Ointment Preparation (10%) Containing the Compound Shown in Formula (I)

[0278] Ingredient Function Dosage % (w / w) Compound Shown in Formula (I) Active Ingredient 10 White Beeswax Ointment Base 20 White Petrolatum Ointment Base 40 Light Liquid Paraffin Ointment Base 30

[0279] Example 12: Suspension Ointment Preparation (1%) Containing the Compound Shown in Formula (I) and a Penetration Enhancer

[0280] Prepare a suspension ointment preparation containing the compound shown in formula (I) and a penetration enhancer. The specific prescription is shown in Table 11:

[0281] Table 11: Prescription Information of Suspension Ointment Preparation (1%) Containing the Compound Shown in Formula (I) and a Penetration Enhancer

[0282] Ingredient Function Dosage % (w / w) Compound Shown in Formula (I) Active Ingredient 1 White Beeswax Ointment Base 18 White Petrolatum Ointment Base 58 Light Liquid Paraffin Ointment Base 21 Isopropyl Myristate Penetration Enhancer 2

[0283] Preparation Method:

[0284] (1) Weigh white beeswax, white petrolatum and isopropyl myristate, heat them in a water bath to 65 - 70 °C until melted, homogenize at a stirring speed of 350 rpm for 15 minutes to obtain a uniform mixture A;

[0285] (2) Weigh light liquid paraffin, add the weighed compound shown in formula (I) thereto, heat it in a water bath to about 40 °C and stir for 10 minutes until the active ingredient is completely dispersed, then stir at a speed of 300 rmp to obtain a uniform mixture B;

[0286] (3) Cool mixture A to 40 °C, add mixture B thereto, continue stirring, then homogenize at a speed of 10000 rmp for 2 minutes with a homogenizer, and then cool to room temperature to obtain a suspension ointment preparation containing the compound shown in formula (I) and a penetration enhancer.

[0287] Example 13: Suspension Ointment Preparation (10%) Containing the Compound Shown in Formula (I) and a Penetration Enhancer

[0288] Prepare a suspension-type ointment preparation containing the compound shown in formula (I) and a penetration enhancer. The preparation method is the same as that of Example 12, and the specific prescription is shown in Table 12.

[0289] Table 12: Prescription information of the suspension-type ointment preparation (10%) containing the compound shown in formula (I) and a penetration enhancer

[0290] Ingredient Function Dosage % (w / w) Compound Shown in Formula (I) Active Ingredient 10 White Beeswax Ointment Base 20 White Petrolatum Ointment Base 40 Light Liquid Paraffin Ointment Base 20 Isopropyl Myristate Penetration Enhancer 10

[0291] Example 14: Suspension-type ointment preparation (1%) containing the compound shown in formula (I), a penetration enhancer and an antioxidant

[0292] Prepare a suspension-type ointment preparation containing the compound shown in formula (I), a penetration enhancer and an antioxidant. The specific prescription is shown in Table 13:

[0293] Table 13: Prescription information of the suspension-type ointment preparation (1%) containing the compound shown in formula (I), a penetration enhancer and an antioxidant

[0294] Ingredient Function Dosage % (w / w) Compound Shown in Formula (I) Active Ingredient 1 White Beeswax Ointment Base 18 White Petrolatum Ointment Base 58 Light Liquid Paraffin Ointment Base 20.9 Isopropyl Myristate Penetration Enhancer 2 Butylated Hydroxytoluene (BHT) Antioxidant 0.1

[0295] Preparation method:

[0296] (1) Weigh white beeswax, white petrolatum, isopropyl myristate and butylated hydroxytoluene (BHT), heat them in a water bath to 65 - 70 °C until melted, homogenize at a stirring speed of 350 rpm for 15 minutes to obtain a uniform mixture A;

[0297] (2) Weigh light liquid paraffin, add the weighed compound shown in formula (I) thereto, heat in a water bath to about 40 °C and stir for 10 minutes until the active ingredient is completely dispersed, then stir at a speed of 300 rmp to obtain a uniform mixture B;

[0298] (3) Cool the mixture A to 40 °C, add the mixture B thereto, continuously stir, then homogenize at a speed of 10000 rmp for 2 minutes with a homogenizer, and then cool to room temperature to obtain a suspension-type ointment preparation containing the compound shown in formula (I), a penetration enhancer and an antioxidant.

[0299] Example 15: Suspension-type ointment preparation (0.3%) containing the compound shown in formula (I)

[0300] Prepare a suspension-type ointment preparation containing the compound shown in formula (I). The preparation method is the same as that of Example 9, and the specific prescription is shown in Table 14.

[0301] Table 14: Prescription information of the suspension-type ointment preparation (0.3%) containing the compound shown in formula (I)

[0302] Ingredient Function Dosage % (w / w) Compound Shown in Formula (I) Active Ingredient 0.3 White Beeswax Ointment Base 18 White Petrolatum Ointment Base 58.7 Light Liquid Paraffin Ointment Base 23

[0303] Example 16: Pharmacokinetic Study after Single Skin Application of the Ointment Containing the Compound Shown in Formula (I) to CD-1 Mice

[0304] Six CD-1 mice were divided into two groups of three each, and each group received 1 mg / cm 2 The ointment containing the compound shown in formula (I) (3% suspension ointment) was applied once to the back, and the application area was 1 cm 2 . The plasma concentration and skin tissue homogenate concentration of the compound shown in formula (I) were determined by LC-MS / MS method.

[0305] The blood drug concentration of the compound shown in formula (I) in the first group of three CD-1 mice at 4 h was 6.15 ng·h / mL. At the same time, the skin tissue homogenate of the back of CD-1 mice at 4 h was taken, and the average drug concentration of the compound shown in formula (I) in the skin taken was 33033 ng / g.

[0306] The area under the blood drug concentration-time curve (AUC 0-last ) of the compound shown in formula (I) in the second group of three CD-1 mice from time 0 to the last quantifiable concentration time (24 h) was 40.7 ng·h / mL on average. The C max average value of the compound shown in formula (I) in plasma was 14.0 ng / mL, while the peak time (T max ) appeared at 1.67 h after administration, and the elimination half-life (T 1 / 2 ) was 5.15 h on average. The skin tissue homogenate of the back of these three CD-1 mice was sampled at the end point of 24 h. The average drug concentration of the compound shown in formula (I) in the skin taken was 7907 ng / g, and the drug concentration of the compound shown in formula (I) in the plasma sample of CD-1 mice at 24 h was below the lower limit of detection.

[0307] The concentration of the compound shown in formula (I) in the skin tissue was much higher than the drug concentration in plasma.

[0308] At this dose, all animals were tolerable. No abnormal manifestations were observed during the whole study.

[0309] The summary of the average PK parameters of the compound shown in formula (I) after single back skin application to CD-1 mice is shown in Table 15.

[0310] Table 15: Pharmacokinetic Parameters of the Compound Shown in Formula (I) after Single Transdermal Administration to CD-1 Mice

[0311]

[0312]

[0313] Example 17: Pharmacokinetic Study after Single Skin Application of Ointment Containing the Compound Shown in Formula (I) to Bama Miniature Pigs

[0314] Two Bama miniature pigs were each administered 20 mg / cm 2 The ointment containing the compound shown in formula (I) (3% suspension ointment) was applied once to the back and abdominal skin. The total application area was 6.8% of the body surface area, and the ratio of the application area on the back to that on the abdomen was approximately 1:1. The plasma concentration and skin tissue homogenate concentration of the compound shown in formula (I) were determined by LC-MS / MS method.

[0315] The mean area under the plasma concentration-time curve (AUC 0-last ) of the compound shown in formula (I) in two Bama miniature pigs from time 0 to the last quantifiable concentration time (24 h) was 304 ng·h / mL. The mean C max of the compound shown in formula (I) in plasma was 21.6 ng / mL, and the time to peak concentration (T max ) occurred at 16 h after dosing. The mean plasma drug concentration at the end point of 24 h was 17.7 ng / mL.

[0316] The sampling end point of skin tissue homogenate in two Bama miniature pigs was 24 h. The mean drug concentration of the compound shown in formula (I) in the dorsal dermis skin tissue sampled at 24 h was 88336 ng / g, the mean drug concentration of the compound shown in formula (I) in the dorsal epidermis skin tissue sample taken at 24 h was 1109245 ng / g, and the mean drug concentration of the compound shown in formula (I) in the abdominal skin tissue sample taken at 24 h was 110952 ng / g. The concentration of the compound shown in formula (I) in each skin tissue was much higher than the drug concentration in plasma.

[0317] All animals were tolerable at this dose. No abnormal manifestations were observed during the whole study.

[0318] The summary of the average PK parameters of the compound shown in formula (I) after single dorsal skin application to Bama miniature pigs is shown in Table 16.

[0319] Table 16: Pharmacokinetic Parameters of the Compound Shown in Formula (I) after Single Transdermal Administration to Bama Miniature Pigs

[0320]

[0321]

[0322] Example 18: Pharmacokinetic Study after Single Skin Application of Ointment Containing the Compound Shown in Formula (I) to SD Rats Six SD rats were divided into two groups of three each. One group received 20 mg / cm 2The ointment containing the compound shown in formula (I) (3% suspension ointment) was administered once by topical application to the back skin, with an application area of 20 cm 2 , and another group received 20 mg / cm 2 The ointment containing the compound shown in formula (I) (10% suspension ointment) was administered once by topical application to the back skin, with an application area of 20 cm 2 . The plasma concentration and the concentration in the homogenate of the back skin tissue of the compound shown in formula (I) were determined by LC-MS / MS method.

[0323] The mean plasma drug concentration at the end point of 24 h of 3 SD rats receiving the 3% suspension ointment was 3.21 ng / mL, and the drug concentration in the homogenate of the back skin tissue at the end point of 24 h was 161167 ng / g. The mean plasma drug concentration at the end point of 24 h of 3 SD rats receiving the 10% suspension ointment was 17.3 ng / mL, and the drug concentration in the homogenate of the back skin tissue at the end point of 24 h was 799500 ng / g. The concentration of the compound shown in formula (I) in the skin tissue was much higher than the drug concentration in the plasma.

[0324] At this dose, all animals could be tolerated. No abnormal manifestations were observed during the whole study.

[0325] The summary of the average PK parameters of the compound shown in formula (I) after single topical application to the back skin of SD rats is shown in Table 17.

[0326] Table 17: Pharmacokinetic parameters of the compound shown in formula (I) after single transdermal administration in SD rats

[0327]

[0328] It can be seen from the above PK results that after topical application of the ointment containing the compound shown in formula (I) in mice, rats and pigs, the exposure in plasma was much lower than that in the skin. The drug ointment has low systemic exposure and high safety in the treatment of skin-related diseases.

[0329] Example 19: Stability study of the prescription of suspension ointment

[0330] The 1% suspension ointment preparation, 3% suspension ointment preparation and 10% suspension ointment preparation prepared according to the methods of Example 9, Example 10 and Example 11 were respectively placed open in containers and stored for 4 weeks under three different conditions of 25°C / 60% RH, 30°C / 65% RH and 40°C / 75% RH. The change in the content of related substances was detected by HPLC to observe the stability of the prescription. The results are shown in Table 18. After storage for 4 weeks under the three conditions, the change in related substances of the preparation was small, and the prescription was generally stable.

[0331] Table 18: Stability study of suspension ointment preparation

[0332]

[0333] Example 20: Stability Study of Suspension Ointment Prescription

[0334] The 10% penetration enhancer-containing suspension ointment preparation prepared according to the method of Example 13 was placed open in a container and stored at 40°C / 75% RH for 4 weeks. The change in the content of related substances was detected by HPLC to observe the stability of the prescription. The results are shown in Table 19. After being stored at 40°C / 75% RH under accelerated conditions for 4 weeks, the change in related substances of the preparation was small, and the prescription was generally stable.

[0335] Table 19: Stability Study of Suspension Ointment Preparation

[0336]

[0337] It should be understood that the above embodiments are all exemplary and do not cover all possible implementation manners included in the claims. Without departing from the scope of the present disclosure, various modifications and changes can be made based on the above embodiments. Similarly, any combination of the technical features of the above embodiments can be made to form additional embodiments of the present invention that may not be explicitly described. Therefore, the above embodiments only represent several implementation manners of the present invention and do not limit the protection scope of the present invention patent.

Claims

1. An RNA m6A regulator composition, characterized in that, The composition comprises: a compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable co-crystal thereof, and an oil phase substance, 2. The composition according to claim 1, characterized in that, The pharmaceutically acceptable salts include hydrochloride, sulfate, phosphate, acetate, mesylate, benzenesulfonate, p-toluenesulfonate; Preferably, the co-crystal formers of the pharmaceutically acceptable co-crystals include nicotinamide, isonicotinamide, L-proline, glycolic acid; Preferably, the oil phase substance includes oil phase substance 1, oil phase substance 2 and oil phase substance 3; Preferably, the oil phase substance 1 is selected from one or more of lanolin, beeswax, white beeswax, paraffin wax, and preferably white beeswax; Preferably, the oil phase substance 2 is selected from one or more of petrolatum, white petrolatum, and preferably white petrolatum; Preferably, the oil phase substance 3 is selected from one or more of light liquid paraffin, vegetable oil, and preferably light liquid paraffin.

3. The composition according to claim 2, characterized in that, By mass percentage, the composition comprises 0.1% - 20% of the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable co-crystal thereof, 10% - 30% of oil phase substance 1, 30% - 70% of oil phase substance 2, 15% - 40% of oil phase substance 3; Preferably, by mass percentage, the composition comprises 0.2% - 15% of the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable co-crystal thereof, 15% - 25% of oil phase substance 1, 35% - 64% of oil phase substance 2, 20% - 35% of oil phase substance 3; Preferably, by mass percentage, the composition comprises 0.3% - 10% of the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable co-crystal thereof, 18% - 20% of oil phase substance 1, 40% - 58.7% of oil phase substance 2, 22% - 30% of oil phase substance 3; Preferably, by mass percentage, the composition comprises 0.3% of the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable co-crystal thereof, 18% of oil phase substance 1, 58.7% of oil phase substance 2, 23% of oil phase substance 3; Preferably, by mass percentage, the composition comprises 1% of the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable co-crystal thereof, 18% of oil phase substance 1, 58% of oil phase substance 2, 23% of oil phase substance 3; Preferably, by mass percentage, the composition comprises 3% of the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable co-crystal thereof, 18% of oil phase substance 1, 57% of oil phase substance 2, 22% of oil phase substance 3; Preferably, by mass percentage, the composition comprises 10% of the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable co-crystal thereof, 20% of oil phase substance 1, 40% of oil phase substance 2, 30% of oil phase substance 3.

4. The composition according to claim 3, characterized in that, The composition comprises the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable co-crystal thereof, white beeswax, white petrolatum, light liquid paraffin; Preferably, by mass percentage, the composition comprises 0.1% - 20% of the compound represented by formula (I), or a pharmaceutically acceptable salt thereof, or a pharmaceutically acceptable co-crystal thereof, 10% - 30% of white beeswax, 30% - 70% of white petrolatum, 15% - 40% of light liquid paraffin; Preferably, by mass percentage, the composition comprises 0.2%-15% of the compound shown by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 15%-25% of white beeswax, 35%-64% of white petrolatum, and 20%-35% of light liquid paraffin; Preferably, by mass percentage, the composition comprises 0.3%-10% of the compound shown by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 18%-20% of white beeswax, 40%-58.7% of white petrolatum, and 22%-30% of light liquid paraffin; Preferably, by mass percentage, the composition comprises 0.3% of the compound shown by formula (I), 18% of white beeswax, 58.7% of white petrolatum, and 23% of light liquid paraffin; Preferably, by mass percentage, the composition comprises 1% of the compound shown by formula (I), 18% of white beeswax, 58% of white petrolatum, and 23% of light liquid paraffin; Preferably, by mass percentage, the composition comprises 3% of the compound shown by formula (I), 18% of white beeswax, 57% of white petrolatum, and 22% of light liquid paraffin; Preferably, by mass percentage, the composition comprises 10% of the compound shown by formula (I), 20% of white beeswax, 40% of white petrolatum, and 30% of light liquid paraffin.

5. The composition according to any one of claims 1-4, wherein Preferably, the composition further comprises a penetration enhancer. More preferably, the penetration enhancer is selected from one or more of isopropyl myristate, isopropyl palmitate, propylene glycol dinonanoate, diethyl sebacate, azone, and propylene glycol. Further preferably, the penetration enhancer is isopropyl myristate; Preferably, the composition further comprises an antioxidant. More preferably, the antioxidant is selected from one or more of vitamin E, alkyl gallate, butylated hydroxyanisole (BHA), and dibutylhydroxytoluene (BHT). Further preferably, the antioxidant is dibutylhydroxytoluene (BHT); Preferably, the composition further comprises other pharmaceutically acceptable excipients.

6. The composition according to claim 5, characterized in that, By mass percentage, the composition comprises 0.1%-20% of the compound shown by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 10%-30% of oil phase substance 1, 30%-70% of oil phase substance 2, 15%-30% of oil phase substance 3, 0.1%-20% of penetration enhancer, and 0%-2% of antioxidant; Preferably, by mass percentage, the composition comprises 0.2%-15% of the compound shown by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 15%-25% of oil phase substance 1, 35%-63% of oil phase substance 2, 20%-25% of oil phase substance 3, 1%-15% of penetration enhancer, and 0%-1% of antioxidant; Preferably, by mass percentage, the composition comprises 0.3%-10% of the compound shown by formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal, 18%-20% of oil phase substance 1, 40%-58% of oil phase substance 2, 20%-21% of oil phase substance 3, 2%-10% of penetration enhancer, and 0%-0.2% of antioxidant.

7. The composition according to claim 6, characterized in that, The composition comprises a compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, white beeswax, white petrolatum, light liquid paraffin, and isopropyl myristate; Preferably, by mass percentage, the composition comprises 0.1%-20% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 10%-30% of white beeswax, 30%-70% of white petrolatum, 15%-30% of light liquid paraffin, and 0.1%-20% of isopropyl myristate; Preferably, by mass percentage, the composition comprises 0.2%-15% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 15%-25% of white beeswax, 35%-63% of white petrolatum, 20%-25% of light liquid paraffin, and 1%-15% of isopropyl myristate; Preferably, by mass percentage, the composition comprises 0.3%-10% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 18%-20% of white beeswax, 40%-58% of white petrolatum, 20%-21% of light liquid paraffin, and 2%-10% of isopropyl myristate; Preferably, by mass percentage, the composition comprises 1% of the compound represented by formula (I), 18% of white beeswax, 58% of white petrolatum, 21% of light liquid paraffin, and 2% of isopropyl myristate; Preferably, by mass percentage, the composition comprises 10% of the compound represented by formula (I), 20% of white beeswax, 40% of white petrolatum, 20% of light liquid paraffin, and 10% of isopropyl myristate; Preferably, the composition comprises a compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, white beeswax, white petrolatum, light liquid paraffin, isopropyl myristate, and butylated hydroxytoluene (BHT); Preferably, by mass percentage, the composition comprises 0.1%-20% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 10%-30% of white beeswax, 30%-70% of white petrolatum, 15%-30% of light liquid paraffin, 0.1%-20% of isopropyl myristate, and 0%-2% of butylated hydroxytoluene (BHT); Preferably, by mass percentage, the composition comprises 0.2%-15% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 15%-25% of white beeswax, 35%-63% of white petrolatum, 20%-25% of light liquid paraffin, 1%-15% of isopropyl myristate, and 0%-1% of butylated hydroxytoluene (BHT); Preferably, by mass percentage, the composition comprises 0.3%-10% of the compound represented by formula (I) or a pharmaceutically acceptable salt or a pharmaceutically acceptable co-crystal thereof, 18%-20% of white beeswax, 40%-58% of white petrolatum, 20%-21% of light liquid paraffin, 2%-10% of isopropyl myristate, and 0%-0.2% of butylated hydroxytoluene (BHT); Preferably, by mass percentage, the composition comprises 1% of the compound shown in formula (I), 18% white beeswax, 58% white petrolatum, 20.9% light liquid paraffin, 2% isopropyl myristate, and 0.1% butylated hydroxytoluene (BHT).

8. A method for preparing the composition according to any one of claims 1-7, characterized in that when the composition does not contain a penetration enhancer and / or an antioxidant, the method comprises the following steps: (1) Weigh oil phase substance 1 and oil phase substance 2, melt them by water bath heating, and obtain a homogeneous mixture A after homogenization; (2) Weigh oil phase substance 3, add the compound shown in formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal that has been weighed, heat it by water bath and stir to obtain a homogeneous mixture B; (3) After cooling the mixture A, add the mixture B thereto, stir and then homogenize, and obtain the composition after cooling; when the composition contains a penetration enhancer and / or an antioxidant, the method comprises the following steps: (1) Weigh oil phase substance 1, oil phase substance 2, a penetration enhancer and / or an antioxidant, melt them by water bath heating, and obtain a homogeneous mixture A after homogenization; (2) Weigh oil phase substance 3, add the compound shown in formula (I) or its pharmaceutically acceptable salt or its pharmaceutically acceptable co-crystal that has been weighed, heat it by water bath and stir to obtain a homogeneous mixture B; (3) After cooling the mixture A, add the mixture B thereto, stir and then homogenize, and obtain the composition after cooling.

9. Use of an RNA m6A regulator composition according to any one of claims 1-7, or a method according to claim 8 in the preparation of a preparation for preventing or treating skin diseases caused by RNA m6A methylation; Preferably, the skin diseases include acanthosis, eczema, ichthyosis, psoriasis, keratosis, systemic lupus erythematosus, hand-foot syndrome, hand-foot skin reaction and dermatitis; More preferably, the skin diseases include hand-foot syndrome and hand-foot skin reaction.

10. The application according to claim 9, wherein The preparation includes ointment, cream, gel, frost, lotion, lotion, solution, paste, film, oil preparation or patch; Preferably, the preparation is an ointment; More preferably, the preparation is a suspension-type ointment.

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