Pharmaceutical composition comprising AAK1 inhibitor

Abstract

Provided are a pharmaceutical composition of a compound of formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof, and a pharmaceutical formulation thereof; and a preparation method for the pharmaceutical composition and the pharmaceutical formulation, and use of the pharmaceutical composition and the pharmaceutical formulation in the preparation of a medicament for treating and inhibiting or degrading AAK1-related diseases.

Description

[Article 26, 08.04.2025] A pharmaceutical composition containing an AAK1 inhibitor [Article 26, 08.04.2025] Technical field

[0001] [Article 26, 08.04.2025] The present invention relates to a pharmaceutical composition and pharmaceutical preparation of a compound of formula (I) or its stereoisomers, pharmaceutically acceptable salts, and its use in the preparation of drugs for treating diseases related to the inhibition of AAK1 inhibitors. [Article 26, 08.04.2025] Background technology

[0002] [Article 26 of the Rules, 08.04.2025] Neuropathic pain (NP) is a general term for a series of pains caused by damage and diseases of the somatic sensory nervous system, which is divided into peripheral neuropathic pain (peripherally-induced neuropathic pain, pNP) and central neuropathic pain. Clinically, pNP is more common and can be specifically divided into diabetic peripheral neuropathy, postherpetic neuralgia, trigeminal neuralgia, postoperative chronic neuralgia, etc. In clinical practice, pNP patients are often accompanied by symptoms such as spontaneous pain (pain without any external stimulation), allodynia (increased response to painful stimuli), hyperalgesia (feeling pain in response to stimuli that are normally painless) and paresthesia, which seriously affect the patient's quality of life.

[0003] [Article 26, 08.04.2025] AAK1 is a member of the Ark1 / Prk1 family of serine / threonine kinases and is widely expressed in the brain and spinal cord. Studies have shown that AAK1 knockout mice are endowed with responses to persistent pain or hypoalgesia, and that AAK1 knockout mice do not develop hyperalgesia in a spinal nerve ligation neuropathic pain model, confirming that AAK1 is a viable target for the treatment of pNP.

[0004] [Article 26, 08.04.2025] We provide an AAK1 inhibitor, but this compound has a low melting point and often sticks to the capsule punch or tablet punch when formulated into tablets or capsules. Based on this, we propose to provide a composition containing the AAK1 inhibitor that is well suited for the preparation of conventional solid dosage forms and exhibits good in vitro dissolution behavior and stability. [Article 26, 08.04.2025] Summary of the invention

[0005] [Article 26 of the Detailed Rules, 08.04.2025] The present invention provides a pharmaceutical composition and pharmaceutical preparation of a compound represented by formula (I) and its stereoisomers and pharmaceutically acceptable salts.

[0006] [Article 26 of the Detailed Rules, 08.04.2025] The pharmaceutical composition or pharmaceutical preparation of the present invention has good solubility, dissolution, bioavailability, oral performance, stable quality, good safety, low irritation, and meets the quality standards of pharmaceuticals.

[0007] [Article 26, 08.04.2025] In one aspect, the present invention provides a pharmaceutical composition comprising:

[0008] [Article 26, 08.04.2025] Active ingredient, the active ingredient is selected from the compound of formula (I) or its stereoisomers, pharmaceutically acceptable salts:

[0009] [Article 26, 08.04.2025] Among them,

[0010] [Rule 26, 08.04.2025] Z is selected from NH or O;

[0011] [Article 26, 08.04.2025]R 1 、R 2 Each independently selected from H, deuterium, halogen, amino, -COOH, cyano, sulfonyl, aminoacyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, wherein the alkyl group is optionally further substituted by 1-3 R A Substituent substitution;

[0012] [Article 26, 08.04.2025]R 41 、R 42 Each independently selected from H, deuterium, amino, C 1-6 Alkyl, halogen, cyano, hydroxyl, halo C 1-6 Alkyl, deuterated C 1-6 alkyl;

[0013] [Article 26, 08.04.2025]R 51 、R 52 Each is independently selected from H, deuterium, amino, halogen;

[0014] [Article 26, 08.04.2025]R 61 、R 62 、R 63 Each independently selected from H, deuterium, halogen, amino, cyano, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 alkyl;

[0015] [Article 26, 08.04.2025] or, R 51 and R 61 , or R 61 and R 62 Together with the carbon atoms to which they are attached, they form double bonds;

[0016] [Article 26, 08.04.2025]R A Selected from deuterium, halogen, amino, cyano, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, deuterated C 1-6 Alkoxy or hydroxy C 1-6 alkyl;

[0017] [Rule 26, 08.04.2025] Provided that, when Z is selected from O, The following structure is not formed:

[0018] [Article 26, 08.04.2025] Inactive ingredients;

[0019] [Article 26, 08.04.2025] wherein the content of the active ingredient in the pharmaceutical composition is 5% to 90% w / w, preferably 5% to 80% w / w, more preferably 10% to 80% w / w;

[0020] [Article 26, 08.04.2025] In some embodiments, the content of the active ingredient (calculated as free base) in the pharmaceutical composition is 5% to 85% w / w, preferably 8% to 80% w / w, more preferably 10% to 80% w / w;

[0021] [Article 26, 08.04.2025] In some embodiments, the content of the active ingredient (calculated as free base) in the pharmaceutical composition is 8% to 85% w / w, preferably 10% to 85% w / w, more preferably 10% to 80% w / w;

[0022] [Article 26 of the Detailed Rules, 08.04.2025] In some embodiments, the content of the active ingredient (calculated as free base) in the pharmaceutical composition is 5-80% w / w, 5%-70% w / w, 5%-60% w / w, 5%-50% w / w, 5%-40% w / w, 5%-30% w / w, 5%-20% w / w, 10%-80% w / w, 10%-70% w / w, 10%-60% w / w, 10%-50% w / w, 10%-40% w / w, 10%-30% w / w, 10%-20% w / w.

[0023] [Rule 26, 08.04.2025] In some embodiments, in the compound of formula (I), Z is O;

[0024] [Article 26, 08.04.2025]R 1 、R 2 Selected from halogenated C 1-2 alkyl;

[0025] [Article 26, 08.04.2025]R 51 、R 52 Each independently selected from H, deuterium;

[0026] [Article 26, 08.04.2025]R 41 、R 42 Each independently selected from amino, C 1-2 alkyl;

[0027] [Article 26, 08.04.2025]R 61 、R 62 、R 63 Each independently selected from H, deuterium, C 1-2 Alkyl, halogenated C 1-2 alkyl;

[0028] [Article 26, 08.04.2025] or, R 51 and R 61 , or R 61 and R 62 Together with the carbon atoms to which they are attached, they form double bonds;

[0029] [Rule 26, 08.04.2025] In some embodiments, in the compound of formula (I),

[0030] [Article 26, 08.04.2025]R 1 、R 2 Each is independently selected from -CH2F, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, -CHFCH2F, -CHFCHF2, -CHFCF3, -CF2CH2F, -CF2CHF2, -CF2CF3, -CH2Cl, -CHCl2, -CCl3, -CH2CH2Cl, -CH2CHCl2, -CH2CCl3, -CHClCH2Cl, -CHClCHCl2, -CHClCCl3, -CCl2CH2Cl, -CCl2CHCl2, -CCl2CCl3;

[0031] [Article 26, 08.04.2025]R51 、R 52 Each independently selected from H, deuterium;

[0032] [Article 26, 08.04.2025]R 41 、R 42 Each independently selected from amino, -CH3, -CH2CH3;

[0033] [Article 26, 08.04.2025]R 61 、R 62 、R 63 each independently selected from H, deuterium, -CH3, -CH2CH3, -CH2F, -CHF2, -CF3, -CH2CH2F, -CH2CHF2, -CH2CF3, -CHFCH2F, -CHFCHF2, -CHFCF3, -CF2CH2F, -CF2CHF2, -CF2CF3, -CH2Cl, -CHCl2, -CCl3, -CH2CH2Cl, -CH2CHCl2, -CH2CCl3, -CHClCH2Cl, -CHClCHCl2, -CHClCCl3, -CCl2CH2Cl, -CCl2CHCl2, -CCl2CCl3;

[0034] [Article 26, 08.04.2025] or, R 51 and R 61 , or R 61 and R 62 Together with the carbon atoms to which they are attached, they form double bonds;

[0035] [Rule 26, 08.04.2025] In some embodiments, in the compound of formula (I),

[0036] [Article 26, 08.04.2025] Z is O;

[0037] [Article 26, 08.04.2025]R 1 、R 2 Each independently selected from -CH2F, -CHF2, -CF3;

[0038] [Article 26, 08.04.2025]R 51 、R 52 Each independently selected from H, deuterium;

[0039] [Article 26, 08.04.2025]R 41 、R 42 Each is independently selected from amino, -CH3;

[0040] [Article 26, 08.04.2025]R61 、R 62 、R 63 Each independently selected from H, deuterium, -CH3, CF3;

[0041] [Rule 26, 08.04.2025] In some embodiments, in the compound of formula (I),

[0042] [Article 26, 08.04.2025] Z is O;

[0043] [Article 26, 08.04.2025]R 1 、R 2 Each independently selected from -CH2F, -CHF2, -CF3;

[0044] [Article 26, 08.04.2025]R 51 、R 52 Together with the carbon atoms to which they are attached, they form double bonds;

[0045] [Article 26, 08.04.2025]R 41 、R 42 Each is independently selected from amino, -CH3;

[0046] [Article 26, 08.04.2025]R 61 、R 62 、R 63 Each independently selected from H, deuterium, -CH3, CF3;

[0047] [Rule 26, 08.04.2025] In some embodiments, in the compound of formula (I),

[0048] [Article 26, 08.04.2025] Z is O;

[0049] [Article 26, 08.04.2025]R 1 、R 2 Each independently selected from -CH2F, -CHF2, -CF3;

[0050] [Article 26, 08.04.2025]R 51 、R 52 Each independently selected from H, deuterium;

[0051] [Article 26, 08.04.2025]R 41 、R 42 Each is independently selected from amino, -CH3;

[0052] [Article 26, 08.04.2025]R 63Each independently selected from H, deuterium, -CH3, CF3;

[0053] [Article 26, 08.04.2025]R 61 and R 62 Together with the carbon atoms to which they are attached, they form double bonds;

[0054] [Rule 26, 08.04.2025] In some embodiments, the compound of formula (I) is selected from one of the following structures:

[0055] [Article 26, 08.04.2025] or

[0056] [Rule 26, 08.04.2025] In some embodiments, the compound of formula (I) is selected from one of the following structures:

[0057] [Article 26, 08.04.2025] or

[0058] [Rule 26, 08.04.2025] In some embodiments, the inactive ingredient comprises a wetting agent;

[0059] [Rule 26, 08.04.2025] In some embodiments, the inactive ingredient comprises a disintegrant;

[0060] [Rule 26, 08.04.2025] In some embodiments, the inactive ingredient comprises a diluent;

[0061] [Rule 26, 08.04.2025] In some embodiments, the inactive ingredient comprises a lubricant;

[0062] [Article 26, 08.04.2025] In some embodiments, the wetting agent is selected from one or more silicates; preferably one or more of silicon dioxide, magnesium silicate, magnesium trisilicate, magnesium aluminum silicate, and talc; more preferably one or more of fumed silica, precipitated silica, sol-gel silica, magnesium silicate, magnesium trisilicate, magnesium aluminum silicate, and talc;

[0063] [Article 26, 08.04.2025] In some embodiments, the disintegrant is selected from one or more of cross-linked sodium carboxymethyl cellulose, cross-linked polyvinylpyrrolidone, starch and its derivatives, low-substituted hydroxypropyl cellulose, low-substituted sodium hydroxymethyl cellulose, surfactants, alginic acid and sodium alginate, and clays; preferably, it is one or more of cross-linked sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, low-substituted sodium hydroxymethyl cellulose, cross-linked polyvinylpyrrolidone, starch, sodium carboxymethyl starch, hydroxypropyl starch, polysorbate 80, sodium lauryl sulfate, bentonite, and colloidal magnesium aluminum silicate; more preferably, it is one or more of cross-linked sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, low-substituted sodium hydroxymethyl cellulose, sodium carboxymethyl starch, and cross-linked polyvinylpyrrolidone;

[0064] [Article 26, 08.04.2025] In some embodiments, the diluent is selected from one or more of starch, pregelatinized starch, dextrin, lactose monohydrate, anhydrous lactose, sucrose, microcrystalline cellulose, inorganic salts, and sugar alcohols; preferably, it is one or more of pregelatinized starch, dextrin, lactose, sucrose, microcrystalline cellulose, calcium sulfate, dibasic calcium phosphate dihydrate, anhydrous dibasic calcium phosphate, calcium phosphate, calcium carbonate, calcium stearate, magnesium oxide, aluminum hydroxide, mannitol, xylitol, and sorbitol; more preferably, it is one or more of pregelatinized starch, lactose, sucrose, microcrystalline cellulose, dibasic calcium phosphate dihydrate, anhydrous dibasic calcium phosphate, calcium phosphate, mannitol, and dextrin;

[0065] [Rule 26, 08.04.2025] In some embodiments, the lubricant is selected from one or more of talc, stearic acid, metal stearate, stearic acid ester, glyceryl behenate, sodium lauryl sulfate, or colloidal silicon dioxide; in some embodiments, the lubricant is selected from one or more of talc, stearic acid, metal stearate, stearic acid ester, glyceryl behenate, sodium lauryl sulfate; preferably, one or more of talc, calcium stearate, magnesium stearate, zinc stearate, polyoxyethylene stearate, glyceryl monostearate, glyceryl palmitostearate; more preferably, one or more of talc, calcium stearate, magnesium stearate, glyceryl monostearate; more preferably, one or more of magnesium stearate, talc, or colloidal silicon dioxide;

[0066] [Article 26, 08.04.2025] In some embodiments, the weight ratio of the active ingredient to the wetting agent is 1:0.05 to 1:5, preferably 1:0.08 to 1:3, further preferably 1:0.12 to 1:2, more preferably 1:0.125, 1:0.2, 1:0.25, 1:0.375, 1:0.5, 1:0.5, 1:1, 1:1.5, 1:2;

[0067] [Article 26, 08.04.2025] In some embodiments, the weight ratio of the active ingredient to the diluent is 1:0.05 to 1:10, preferably 1:0.1 to 1:8.5, further preferably 1:0.18 to 1:8.5, more preferably 1:0.18125, 1:0.1875, 1:0.9875, 1:1, 1:1.1125, 1:1.125, 1:1.2375, 1:1.25, 1:1.3625, 1:1.375, 1:3.725, 1:3.75, 1:6.45, 1:6.95, 1:7.45, 1:7.95, 1:8.25, 1:8.45;

[0068] [Article 26, 08.04.2025] In some embodiments, the weight ratio of the active ingredient to the disintegrant is 1:0.01 to 1:3, preferably 1:0.03 to 1:1.5, further preferably 1:0.03 to 1:0.6, more preferably 1:0.0375, 1:0.0625, 1:0.075, 1:0.125, 1:0.15, 1:0.25, 1:0.5;

[0069] [Article 26, 08.04.2025] In some embodiments, the weight ratio of the active ingredient to the lubricant is 1:0.001 to 1:2, preferably 1:0.006 to 1:0.1, more preferably 1:0.00625, 1:0.0125, 1:0.025, 1:0.05, 1:0.1. The present invention provides a pharmaceutical composition comprising an active ingredient and lactose, microcrystalline cellulose, low-substituted-hydroxypropyl cellulose, and magnesium stearate.

[0070] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and silicon dioxide, lactose, microcrystalline cellulose, low-substituted-hydroxypropyl cellulose and magnesium stearate.

[0071] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and silicon dioxide, talc, lactose, microcrystalline cellulose, low-substituted-hydroxypropyl cellulose and magnesium stearate.

[0072] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and microcrystalline cellulose, mannitol, crospovidone and magnesium stearate.

[0073] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and silicon dioxide, magnesium silicate, microcrystalline cellulose, mannitol, crospovidone and magnesium stearate.

[0074] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and pregelatinized starch, lactose, sodium carboxymethyl starch and talc.

[0075] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and silicon dioxide, pregelatinized starch, lactose, sodium carboxymethyl starch and talc.

[0076] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and microcrystalline cellulose, lactose, croscarmellose sodium and magnesium stearate.

[0077] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and silicon dioxide, microcrystalline cellulose, lactose, croscarmellose sodium and magnesium stearate.

[0078] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and lactose, microcrystalline cellulose, croscarmellose sodium and magnesium stearate.

[0079] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and lactose, cross-linked sodium carboxymethyl cellulose and magnesium stearate.

[0080] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and microcrystalline cellulose, croscarmellose sodium and magnesium stearate.

[0081] [Article 26 of the Detailed Rules, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient, mannitol, low-substituted carboxymethyl cellulose sodium and talc.

[0082] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and microcrystalline cellulose, low-substituted carboxymethyl cellulose sodium and talc.

[0083] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and mannitol, microcrystalline cellulose, low-substituted carboxymethyl cellulose sodium and talc.

[0084] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and pregelatinized starch, sodium carboxymethyl starch and magnesium stearate.

[0085] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and dextrin, sodium carboxymethyl starch and magnesium stearate.

[0086] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and pregelatinized starch, dextrin, sodium carboxymethyl starch and magnesium stearate.

[0087] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and calcium phosphate, cross-linked polyvinylpyrrolidone and talc.

[0088] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and sucrose, cross-linked polyvinylpyrrolidone and talc.

[0089] [Rule 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising an active ingredient and calcium phosphate, sucrose, crospovidone, and talc. The present invention provides a pharmaceutical composition comprising 10-80 w / w%, 10-60 w / w%, 10-45 w / w%, 10-40 w / w%, 5-35 w / w%, 5-20 w / w%, 10%, 20%, 40%, or 80% of the active ingredient.

[0090] [Rule 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 10-85w / w%, 10-80w / w%, 10-70w / w%, 10-60w / w%, 10-50w / w%, 10-40w / w%, 10-30w / w%, 14.5%, 15%, 39.5%, 40%, 44.5%, 45%, 49.5%, 50%, 54.5%, 55%, 64.5%, 69.5%, 74.5%, 75%, 79.5%, 80%, 82.5%, 84.5% of a diluent.

[0091] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 1-8w / w%, 1-5w / w%, 1-3w / w%, 3-5w / w%, 5-8w / w%, 3w / w%, 5w / w%, or 8w / w% of a disintegrant.

[0092] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 0.1-2w / w%, 0.1-1.5w / w%, 0.1-1w / w%, 0.1-0.5w / w%, 0.5-2w / w%, 0.5-1w / w%, 0.5w / w%, 1w / w%, 2w / w% of a lubricant.

[0093] [Article 26 of the Detailed Rules, 08.04.2025] The present invention provides a pharmaceutical composition comprising 2-35w / w%, 2-30w / w%, 2-25w / w%, 2-20w / w%, 2-15w / w%, 2-10w / w%, 2-5w / w%, 5-35w / w%, 5-30w / w%, 5-25w / w%, 5-20w / w%, 5-15w / w%, 5-10w / w%, 2w / w%, 5w / w%, 10w / w%, 15w / w%, 20w / w% of a wetting agent.

[0094] [Rule 26, 08.04.2025] The pharmaceutical composition according to any one of the present invention comprises the active ingredient and the inactive ingredient according to any one of the aforementioned embodiments, including:

[0095] [Article 26, 08.04.2025] active ingredient;

[0096] [Rule 26, 08.04.2025] a diluent, the diluent being selected from one or more of pregelatinized starch, lactose, microcrystalline cellulose, mannitol, dextrin, calcium phosphate or sucrose;

[0097] [Article 26, 08.04.2025] disintegrant, the disintegrant being selected from one or more of croscarmellose sodium, sodium carboxymethyl starch, cross-linked povidone or low-substituted hydroxypropyl cellulose;

[0098] [Rule 26, 08.04.2025] Lubricant, the lubricant being selected from one or more of magnesium stearate, talc or colloidal silicon dioxide;

[0099] [Article 26, 08.04.2025] Optionally, the pharmaceutical composition further comprises a wetting agent selected from silicon dioxide, talc, magnesium silicate

[0100] [Rule 26, 08.04.2025] one or more.

[0101] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 5-35w / w% active ingredient, 60-85w / w% diluent, 1-8w / w% disintegrant, and 0.1-2w / w% lubricant.

[0102] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 5-20 w / w% active ingredient, 70-85 w / w% diluent, 1-8 w / w% disintegrant, and 0.1-2 w / w% lubricant.

[0103] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 10-80 w / w% active ingredient, 2-35 w / w% wetting agent, 15-85 w / w% diluent, 1-10 w / w% disintegrant, and 0.1-5 w / w% lubricant.

[0104] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 10-60 w / w% active ingredient, 2-35 w / w% wetting agent, 25-80 w / w% diluent, 1-10 w / w% disintegrant, and 0.1-5 w / w% lubricant.

[0105] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 10-45 w / w% active ingredient, 2-25 w / w% wetting agent, 30-70 w / w% diluent, 1-10 w / w% disintegrant, and 0.1-5 w / w% lubricant.

[0106] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 10-40 w / w% active ingredient, 5-20 w / w% wetting agent, 35-60 w / w% diluent, 1-10 w / w% disintegrant, and 0.1-5 w / w% lubricant. The present invention provides a pharmaceutical composition comprising 10 w / w% active ingredient, 24.5 w / w% lactose, 60 w / w% microcrystalline cellulose, 5 w / w% low-substituted hydroxypropyl cellulose, and 0.5 w / w% magnesium stearate.

[0107] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 10 w / w% active ingredient, 2 w / w% silicon dioxide, 24.5 w / w% lactose, 58 w / w% microcrystalline cellulose, 5 w / w% low-substituted hydroxypropyl cellulose, and 0.5 w / w% magnesium stearate.

[0108] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 10 w / w% active ingredient, 5 w / w% silicon dioxide, 24.5 w / w% lactose, 55 w / w% microcrystalline cellulose, 5 w / w% low-substituted hydroxypropyl cellulose, and 0.5 w / w% magnesium stearate.

[0109] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 10 w / w% active ingredient, 10 w / w% silicon dioxide, 24.5 w / w% lactose, 50 w / w% microcrystalline cellulose, 5 w / w% low-substituted hydroxypropyl cellulose, and 0.5 w / w% magnesium stearate.

[0110] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 10 w / w% active ingredient, 10 w / w% silicon dioxide, 5 w / w% talc, 24.5 w / w% lactose, 45 w / w% microcrystalline cellulose, 5 w / w% low-substituted hydroxypropyl cellulose, and 0.5 w / w% magnesium stearate.

[0111] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 10 w / w% active ingredient, 10 w / w% silicon dioxide, 10 w / w% talc, 24.5 w / w% lactose, 40 w / w% microcrystalline cellulose, 5 w / w% low-substituted hydroxypropyl cellulose, and 0.5 w / w% magnesium stearate.

[0112] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 20 w / w% active ingredient, 29.5 w / w% mannitol, 45 w / w% microcrystalline cellulose, 5 w / w% cross-linked polyvinylpyrrolidone, and 0.5 w / w% magnesium stearate.

[0113] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 40 w / w% active ingredient, 19.5 w / w% mannitol, 35 w / w% microcrystalline cellulose, 5 w / w% cross-linked polyvinylpyrrolidone, and 0.5 w / w% magnesium stearate.

[0114] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 80 w / w% active ingredient, 4.5 w / w% mannitol, 10 w / w% microcrystalline cellulose, 5 w / w% cross-linked polyvinylpyrrolidone, and 0.5 w / w% magnesium stearate.

[0115] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 40 w / w% active ingredient, 3 w / w% silicon dioxide, 2 w / w% magnesium silicate, 19.5 w / w% mannitol, 30 w / w% microcrystalline cellulose, 5 w / w% cross-linked polyvinylpyrrolidone, and 0.5 w / w% magnesium stearate.

[0116] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 40 w / w% active ingredient, 6 w / w% silicon dioxide, 4 w / w% magnesium silicate, 19.5 w / w% mannitol, 25 w / w% microcrystalline cellulose, 5 w / w% cross-linked polyvinylpyrrolidone, and 0.5 w / w% magnesium stearate.

[0117] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 40w / w% active ingredient, 10w / w% silicon dioxide, 5w / w% magnesium silicate, 19.5w / w% mannitol, 20w / w% microcrystalline cellulose, 5w / w% cross-linked polyvinylpyrrolidone, and 0.5w / w% magnesium stearate.

[0118] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 20 w / w% active ingredient, 40 w / w% pregelatinized starch, 35 w / w% lactose, 3 w / w% sodium carboxymethyl starch, and 2 w / w% talc.

[0119] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 40 w / w% active ingredient, 30 w / w% pregelatinized starch, 25 w / w% lactose, 3 w / w% sodium carboxymethyl starch, and 2 w / w% talc.

[0120] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 80 w / w% active ingredient, 10 w / w% pregelatinized starch, 5 w / w% lactose, 3 w / w% sodium carboxymethyl starch, and 2 w / w% talc.

[0121] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 40 w / w% active ingredient, 5 w / w% silicon dioxide, 30 w / w% pregelatinized starch, 20 w / w% lactose, 3 w / w% sodium carboxymethyl starch, and 2 w / w% talc.

[0122] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 40 w / w% active ingredient, 10 w / w% silicon dioxide, 25 w / w% pregelatinized starch, 20 w / w% lactose, 3 w / w% sodium carboxymethyl starch, and 2 w / w% talc.

[0123] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 40 w / w% active ingredient, 15 w / w% silicon dioxide, 20 w / w% pregelatinized starch, 20 w / w% lactose, 3 w / w% sodium carboxymethyl starch, and 2 w / w% talc.

[0124] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 20 w / w% active ingredient, 45 w / w% microcrystalline cellulose, 29.5 w / w% lactose, 5 w / w% croscarmellose sodium, and 0.5 w / w% magnesium stearate.

[0125] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 40 w / w% active ingredient, 35 w / w% microcrystalline cellulose, 19.5 w / w% lactose, 5 w / w% cross-linked sodium carboxymethyl cellulose, and 0.5 w / w% magnesium stearate.

[0126] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 80 w / w% active ingredient, 10 w / w% microcrystalline cellulose, 4.5 w / w% lactose, 5 w / w% cross-linked sodium carboxymethyl cellulose, and 0.5 w / w% magnesium stearate.

[0127] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 40 w / w% active ingredient, 5 w / w% silicon dioxide, 30 w / w% microcrystalline cellulose, 19.5 w / w% lactose, 5 w / w% cross-linked sodium carboxymethyl cellulose, and 0.5 w / w% magnesium stearate.

[0128] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 40 w / w% active ingredient, 10 w / w% silicon dioxide, 25 w / w% microcrystalline cellulose, 19.5 w / w% lactose, 5 w / w% croscarmellose sodium, and 0.5 w / w% magnesium stearate.

[0129] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 40 w / w% active ingredient, 15 w / w% silicon dioxide, 20 w / w% microcrystalline cellulose, 19.5 w / w% lactose, 5 w / w% cross-linked sodium carboxymethyl cellulose, and 0.5 w / w% magnesium stearate.

[0130] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 10 w / w% active ingredient, 22 w / w% lactose, 62 w / w% microcrystalline cellulose, 5 w / w% croscarmellose sodium, and 1 w / w% magnesium stearate.

[0131] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 25w / w% active ingredient, 30w / w% mannitol, 40w / w% microcrystalline cellulose, 3w / w% low-substituted carboxymethyl cellulose sodium, and 2w / w% talc.

[0132] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 25w / w% active ingredient, 37.5w / w% pregelatinized starch, 35w / w% dextrin, 1.5w / w% sodium carboxymethyl starch, and 1w / w% magnesium stearate.

[0133] [Article 26, 08.04.2025] The present invention provides a pharmaceutical composition comprising 33 w / w% active ingredient, 27 w / w% calcium phosphate, 38.33 w / w% sucrose, 1 w / w% crospovidone, and 0.67 w / w% talc. In any of the above pharmaceutical compositions, the active ingredient is selected from a compound of formula (I) or a stereoisomer or a pharmaceutically acceptable salt thereof.

[0134] [Rule 26, 08.04.2025] In one aspect, the present invention provides a pharmaceutical preparation comprising any one of the pharmaceutical compositions described above;

[0135] [Rule 26, 08.04.2025] In some embodiments, the amount of the active ingredient in a unit preparation of the pharmaceutical preparation is 1 mg to 100 mg; in some embodiments, the amount of the active ingredient in a unit preparation of the pharmaceutical preparation is 5 mg, 10 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, or 100 mg.

[0136] [Article 26 of the Detailed Rules, 08.04.2025] In some embodiments, the dosage form of the pharmaceutical preparation is selected from tablets, granules, capsules, dry suspensions, oral solutions, soft capsules, and emulsions; in some embodiments, the dosage form of the pharmaceutical preparation is selected from tablets, granules, capsules, and soft capsules.

[0137] [Article 26 of the Detailed Rules, 08.04.2025] In one aspect, the present invention provides the use of any of the above-mentioned pharmaceutical compositions or any of the above-mentioned pharmaceutical preparations in the preparation of a medicament for treating a disease associated with the inhibition or degradation of AAK1, wherein the disease is pain, preferably inflammatory pain, postoperative pain, trigeminal neuralgia, acute postherpetic neuralgia and postherpetic neuralgia, diabetic peripheral neuropathy, causalgia, occipital neuralgia, fibromyalgia, phantom limb pain, burn pain and other forms of neuralgia, neuropathy and spontaneous pain syndrome.

[0138] [Article 26 of the Detailed Rules, 08.04.2025] The preparation process of the pharmaceutical composition or pharmaceutical preparation of the present invention is one or more of direct mixing, wet granulation, dry granulation, fluidized bed granulation, spray drying, freeze drying, hot melt extrusion, and pellet coating, preferably direct mixing, wet granulation, fluidized bed granulation, dry granulation, and spray drying, and more preferably direct mixing and dry granulation.

[0139] [Article 26, 08.04.2025] In some embodiments, the preparation process is one or more of direct mixing, wet granulation, dry granulation, fluidized bed granulation, spray drying, freeze drying, hot melt extrusion, pellet coating, and extrusion spheronization, preferably spray drying and hot melt extrusion;

[0140] [Article 26, 08.04.2025] In some embodiments, the preparation process is dry granulation tableting, direct powder tableting and direct powder capsule filling.

[0141] [Article 26, 08.04.2025] In some embodiments, the preparation process of the pharmaceutical composition further comprises pre-treating the active ingredient, wherein the pre-treatment method is selected from one or more of pulverization, solid dispersion, and nano-grinding.

[0142] [Rule 26, 08.04.2025] Unless otherwise stated, the terms used in the description and claims shall have the following meanings:

[0143] [Article 26, 08.04.2025] “Pharmaceutically acceptable salts” means salts that are safe, non-toxic and neither biologically nor otherwise undesirable, and include salts thereof that are pharmaceutically acceptable for veterinary use as well as for human pharmaceutical use and that possess the desired pharmacological activity.

[0144] [Article 26 of the Regulations, 08.04.2025] “Stereoisomers” refer to isomers resulting from different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers and conformational isomers.

[0145] [Rule 26, 08.04.2025] “Optional” or “optionally” or “selectively” or “optionally” means that the subsequently described event or circumstance may but need not occur, and that the description includes instances where the event or circumstance occurs as well as instances where it does not. For example, “a heterocyclyl optionally substituted with an alkyl group” means that the alkyl group may but need not be present, and that the description includes instances where the heterocyclyl group is substituted with an alkyl group as well as instances where the heterocyclyl group is not substituted with an alkyl group. [Article 26, 08.04.2025] Illustrations

[0146] [Article 26, 08.04.2025] Figure 1 is the time-MPT curve of the mechanical pain threshold (MPT) of mice in Example 5.

[0147] [Article 26 of the Regulations, 08.04.2025] Figure 2 is a dissolution curve of the tablet of Example 7.

[0148] [Article 26, 08.04.2025] Figure 3 is a dissolution curve of the capsule of Example 7. [Article 26, 08.04.2025] Specific implementation methods

[0149] [Article 26 of the Detailed Rules, 08.04.2025] The following examples illustrate the technical problems, technical solutions, and beneficial effects to be solved by the present invention. It should be understood that the specific embodiments described herein are only used to illustrate the present invention and are not intended to limit the present invention.

[0150] [Article 26, 08.04.2025] The compound of formula (I) was prepared using the following synthesis scheme:

[0151] [Article 26, 08.04.2025] Example 1: Preparation of active ingredient

[0152] [Article 26, 08.04.2025] Intermediate 1:

[0153] [Article 26, 08.04.2025] Step 1:

[0154] [Article 26 of the Detailed Rules, 08.04.2025] The raw material 1A (10 g, 49 mmol) was dissolved in 200 mL of dichloromethane, cooled to -20°C, and DAST (11.7 mL, 88 mmol) was added. The temperature was slowly raised to room temperature and the reaction was allowed to react for 5 h. After the disappearance of the raw material by plate monitoring, the reaction was quenched with saturated aqueous sodium bicarbonate solution and extracted with ethyl acetate. The organic phase was dried and passed through a silica gel column (petroleum ether:ethyl acetate = 20:1) to obtain the target compound intermediate 1 (9.8 g, 89%).

[0155] [Article 26, 08.04.2025] 1 H NMR (400MHz, CDCl3) δ7.65-7.58(m,1H),7.46-7.40(m,1H),6.85-6.56(m,1H).

[0156] [Article 26, 08.04.2025] Intermediate 2:

[0157] [Article 26, 08.04.2025] Step 1:

[0158] [Article 26, 08.04.2025] 2A (5 g, 24 mmol), Xphos PdG2 (189 mg, 0.24 mmol, CAS: 1310584-14-5), Xphos (229 mg, 0.48 mmol, CAS 564483-18-7), biboronic acid pinacol ester (9.14 g, 36 mmol), and KOAc (7.07 g, 72 mmol) were added to a flask. After nitrogen displacement, 200 mL of ethanol was added and the reaction was heated to 80°C for 5 h. After the disappearance of the starting material, water was added to quench the reaction. The ethanol in the system was evaporated and extracted with ethyl acetate. The organic phase was dried to give intermediate 2 (5.1 g).

[0159] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 174.1 [M+H] + .

[0160] [Article 26, 08.04.2025] Intermediate 3:

[0161] [Article 26, 08.04.2025] Step 1:

[0162] [Article 26, 08.04.2025] Under a nitrogen atmosphere, add 62 mL of chlorosulfonyl isocyanate to a three-necked round-bottom flask, add 200 mL of dichloromethane, and cool the system to 0°C. Dissolve 27 mL of formic acid in 50 mL of dichloromethane and slowly add the mixture to the system while maintaining the temperature at 0°C. After 30 minutes, warm to room temperature and stir overnight. Dissolve 36.3 mL of hydroxyacetone and 58 mL of pyridine in 1000 mL of dichloromethane and slowly add the mixture to the system at 0°C. After the addition is complete, warm the system to room temperature and stir overnight. The organic solvent in the system was evaporated, and the mixture was passed through a silica gel column using dichloromethane as the eluent to obtain the title compound 3C (36 g, 56%).

[0163] [Article 26, 08.04.2025] 1 H NMR (400MHz, CDCl3) δ5.06 (s, 2H), 2.42 (s, 3H).

[0164] [Article 26, 08.04.2025] Step 2:

[0165] [Article 26 of the Detailed Rules, 08.04.2025] Under nitrogen atmosphere, 3C (36 g, 267 mmol) was dissolved in 800 mL of methyl tert-butyl ether. The system was cooled to 0°C and a solution of 2-methylallylmagnesium chloride in tetrahydrofuran (0.55 L, 0.5 M) was added dropwise. After the disappearance of the starting material by spot plate detection, saturated aqueous ammonium chloride was added to quench the reaction. The mixture was extracted with ethyl acetate, dried by spin drying, and passed through a silica gel column to give the title compound 3D (43 g, 84%).

[0166] [Article 26, 08.04.2025] 1 H NMR (400MHz, CDCl3) δ5.06-5.01(m,1H),4.85-4.83(m,1H),4.59(s,1H),4.38(d, 1H),4.27(d,1H),2.57-2.50(m,1H),2.42-2.29(m,1H),1.84(s,3H),1.46(s,3H).

[0167] [Article 26, 08.04.2025] Step 3:

[0168] [Article 26 of the Detailed Rules, 08.04.2025] Under nitrogen, 3D (1.91 g, 10 mmol) was dissolved in 50 mL of tetrahydrofuran, and 15 mL of a 1 M solution of potassium tert-butoxide in tetrahydrofuran was added, followed by CbzCl (2.1 mL, 15 mmol). After the disappearance of the starting material by spot plate detection, saturated aqueous ammonium chloride was added to quench the reaction. The tetrahydrofuran in the system was evaporated, and the product was extracted with ethyl acetate. The product was passed through a silica gel column (petroleum ether:ethyl acetate = 10:1) to give the title compound 3E (2.6 g, 80%).

[0169] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 343.0 [M+NH4] + .

[0170] [Article 26, 08.04.2025] 120g of 3E was chirally prepared to give 55g of the target compound 3F.

[0171] [Article 26, 08.04.2025] Preparation method: Instrument: Waters SFC 150Mgm, Column: Daicel Chiralpak OJ (250 mm × 50 mm, 10 μm); Mobile phase: A for CO2 and B for MeOH (Base); Gradient: 10% B; Flow rate: 130 mL / min, Back pressure: 100 bar; Column temperature: 35°C; Wavelength: 220 nm; Cycle time: 4.5 min; Sample preparation: Sample concentration: 157.5 mg / mL, ethanol solution; Injection: 0.8 mL per sample. After separation, the fraction was dried on a rotary evaporator at 40°C to yield compound 3F (Retention time: 0.680 min).

[0172] [Article 26, 08.04.2025] Step 4:

[0173] [Article 26, 08.04.2025] Dissolve compound 3F (5 g, 15.4 mmol) in 500 mL of methanol and add 50 mg of 10% palladium-on-carbon catalyst to replace the hydrogen atmosphere. After the fluorescence disappears as detected by spot plate analysis, the palladium-on-carbon in the system is removed by filtration. The resulting filtrate is spin-dried to give the crude title compound 3G, which is directly used in the next step.

[0174] [Article 26, 08.04.2025] Step 5:

[0175] [Article 26, 08.04.2025] Compound 3G was dissolved in 150 mL of tetrahydrofuran. Lithium aluminum tetrahydride (1.8 g, 47.4 mmol) was added portionwise at 0°C and allowed to warm to room temperature and stir overnight. 1.8 mL of water, 3.6 mL of 10% aqueous sodium hydroxide solution, and 5.4 mL of water were added. After stirring for 1 hour, the solid was removed by filtration. The filtrate was dried to give the crude product of intermediate 3, which was used directly in the next reaction.

[0176] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 130.1 [M+H] + .

[0177] [Article 26, 08.04.2025] Intermediate 4:

[0178] [Article 26, 08.04.2025] Step 1:

[0179] [Article 26 of the Detailed Rules, 08.04.2025] Compound 4A (10 g, 57.8 mmol) was dissolved in 200 mL of acetone. Meta-chloroperbenzoic acid (11 g, 63.6 mmol) was dissolved in 200 mL of acetone at room temperature and added. The mixture was stirred for 5 min to generate a large amount of solid. The solid was filtered and washed with acetone. After drying, the crude product of compound 4B (10.7 g, 98%) was obtained.

[0180] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 189.0 and 191.0 [M+H] + .

[0181] [Article 26, 08.04.2025] Step 2:

[0182] [Article 26, 08.04.2025] Dissolve 10.7 g of crude compound 4B in 200 mL of trimethyl orthoformate, add 1.25 mL of boron trifluoride etherate, heat the system to 105°C and react overnight. The organic phase in the system is dried and separated by column chromatography to obtain compound 4C (9.1 g, 69%).

[0183] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 231.0 and 233.0 [M+H] + .

[0184] [Article 26, 08.04.2025] Step 3:

[0185] [Article 26, 08.04.2025] Compound 4C (3.5 g, 15.1 mmol), Xphos PdG2 (600 mg, 0.76 mmol, CAS: 1310584-14-5), Xphos (700 mg, 1.47 mmol, CAS 564483-18-7), potassium acetate (4.5 g, 45.8 mmol), and pinacol diboron (6 g, 23.6 mmol) were placed in a round-bottom flask and dissolved in 250 mL of ethanol. The atmosphere was purged with nitrogen, and the temperature was raised to 80°C and the reaction was allowed to proceed overnight. The ethanol was evaporated, and the mixture was extracted with ethyl acetate to obtain the title compound, intermediate 4 (4 g).

[0186] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 197.1 [M+H] + .

[0187] [Article 26, 08.04.2025] Intermediate 5:

[0188] [Article 26, 08.04.2025] Step 1:

[0189] [Article 26 of the detailed rules, 08.04.2025] The raw material 5A (5.00 g, 24.51 mmol) was dissolved in 100 mL of dichloromethane, cooled to -20°C, DAST (6.5 mL, 49.02 mmol) was added, and the temperature was slowly raised to room temperature for 2 h. After the raw material disappeared after spot plate detection, the reaction was quenched with saturated sodium bicarbonate aqueous solution, extracted with dichloromethane, and the organic phase was dried and passed through a silica gel column (petroleum ether: ethyl acetate = 20:1) to obtain intermediate 5 (5.00 g, 90.27%).

[0190] [Article 26, 08.04.2025] 1 H NMR (400MHz, CDCl3) δ8.40(dd,1H),8.15(dt,1H),6.95-6.67(m,1H).

[0191] [Article 26, 08.04.2025] 1.1 Preparation of Compound 1

[0192] [Article 26, 08.04.2025] Step 1:

[0193] [Article 26 of the detailed rules, 08.04.2025] 3D (8 g, 42 mmol) was dissolved in 500 mL of tetrahydrofuran, and the system was cooled to 0°C. Lithium aluminum tetrahydride (3.99 g, 105 mmol) was slowly added, and then the temperature was raised to room temperature for 6 h. 4 mL of water, 8 M NaOH aqueous solution, and 12 mL of water were added in sequence, and the mixture was stirred for 1 h. The solid was removed by filtration, and the filtrate was dried to give the crude product target compound 1b (9 g), which was directly used in the next step without purification.

[0194] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 130.2 [M+H] + .

[0195] [Article 26, 08.04.2025] Step 2:

[0196] [Article 26, 08.04.2025] Crude product 1b (2 g) was added to 27 mL of potassium tert-butoxide in tetrahydrofuran and stirred at room temperature for 5 minutes. Intermediate 1 (4 g, 18 mmol) was then added. After nitrogen was replaced, the mixture was heated to 80°C and allowed to react overnight. The organic phase was evaporated to dryness and purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to afford the title compound 1c (1.1 g, 35%).

[0197] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 335.1 and 337.1 [M+H] + .

[0198] [Article 26, 08.04.2025] Step 3:

[0199] [Article 26, 08.04.2025] Intermediate 2 (1.1 g, 3.3 mmol), 1c (880 mg, 5 mmol), potassium phosphate (9.2 g, 43 mmol), Xphos PdG2 (500 mg, 0.63 mmol, CAS: 1310584-14-5), and Xphos (650 mg, 1.36 mmol, CAS 564483-18-7) were added to a sealed tube. 30 mL of tetrahydrofuran was added, the atmosphere was replaced with nitrogen, and the temperature was raised to 80°C for 5 h. After the disappearance of the starting material by spot plate detection, the solid was removed by filtration and washed with methanol. The filtrate was spin-dried and purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to obtain the title compound 1d (360 mg, 29%).

[0200] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 384.2 [M+H] + .

[0201] [Article 26, 08.04.2025] 1 H NMR(400MHz,DMSO-d6)δ8.80-8.76(m,1H),8.42-8.36(m,1H),8.32(s,1H),8.24-8.18(m,1H),7.84-7.79 (m,1H),7.42-6.88(m,2H),4.87(s,1H),4.72(s,1H),3.88(s,2H),2.22(s,2H),1.78(s,3H),1.15(s,3H).

[0202] [Article 26, 08.04.2025] Step 4:

[0203] [Article 26, 08.04.2025] Dissolve 1d (360 mg, 0.94 mmol) in 20 mL of dichloromethane, cool to -60°C, and introduce ozone. After the disappearance of the starting material by a spectrophotometer, add 1 g of triphenylphosphine and warm to room temperature with stirring for 15 minutes. The organic phase is then dried and purified by silica gel column chromatography (dichloromethane:methanol = 10:1) to yield the title compound 1e (300 mg, 83%).

[0204] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 386.2 [M+H] + .

[0205] [Article 26, 08.04.2025] Step 5:

[0206] [Article 26, 08.04.2025] Under nitrogen atmosphere, 1e (300 mg, 0.78 mmol) was dissolved in 20 mL of tetrahydrofuran and the system was cooled to 0°C. Methylmagnesium bromide in THF (1 mL, 3 M) was added and the mixture was slowly warmed to room temperature and then tested on a plate. After the disappearance of the starting material, saturated aqueous ammonium chloride was added to quench the reaction. The mixture was extracted with dichloromethane and the organic phase was dried to give the title compound 1f (240 mg, 0.6 mmol), which was directly used in the next step.

[0207] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 402.2 [M+H] + .

[0208] [Article 26, 08.04.2025] Step 6:

[0209] [Article 26 of the detailed rules, 08.04.2025] Under a nitrogen atmosphere, 1f (240 mg, 0.6 mmol) was dissolved in 15 mL of dichloromethane and cooled to -78°C. DAST (0.4 mL, 2.8 mmol) was added and the system was slowly warmed to room temperature. After the disappearance of the starting material by spot plate detection, saturated aqueous sodium bicarbonate solution was added to quench the reaction. The reaction was extracted with dichloromethane, and the organic phase was spin-dried and the resulting product was separated by HPLC and freeze-dried to obtain the title compound 1 (110 mg, 42%).

[0210] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 404.2 [M+H] + .

[0211] [Article 26, 08.04.2025] 1 H NMR(400MHz,DMSO-d6)δ8.82-8.76(m,1H),8.42-8.36(m,1H),8.32(s,1H),8.23-8.18(m,1H),7.8 1-7.75(m,1H),7.41-6.89(m,2H),3.95(s,2H),1.94-1.86(m,2H),1.49-1.36(m,6H),1.23(s,3H).

[0212] [Article 26, 08.04.2025] 1.2 Preparation of Compounds 2 and 3

[0213] [Article 26, 08.04.2025] 1d (80 mg) was chirally resolved to give compound 2 (33.7 mg) and compound 3 (25.3 mg).

[0214] [Article 26, 08.04.2025] Preparation method:

[0215] [Article 26, 08.04.2025] Instrument: SHIMADZU LC-20AP, Column: DAICEL CHIRALPAK IG (250 mm × 30 mm, 10 μm); Mobile Phase: A: n-hexane, B: ethanol (0.1% NH3·H2O); Gradient: 8% B gradient elution; Flow rate: 120 mL / min, Column temperature: 25°C, Wavelength: 254 nm, Cycle time: 16 min; Sample preparation: Sample concentration: 1.5 mg / mL, ethanol solution; Injection: 2 mL per sample. After separation, the fractions were dried on a rotary evaporator at 40°C to yield P1 (retention time: 2.658 minutes, assumed to be compound 2) and P2 (retention time: 4.205 minutes, assumed to be compound 3).

[0216] [Article 26, 08.04.2025] 1.3 Preparation of Compound 4

[0217] [Article 26, 08.04.2025] Step 1:

[0218] [Article 26 of the detailed rules, 08.04.2025] Intermediate 4 (500 mg, 1.8 mmol), intermediate 1 (500 mg, 2.2 mmol), Xphos PdG2 (200 mg, 0.25 mmol, CAS: 1310584-14-5), Xphos (250 mg, 0.52 mmol, CAS 564483-18-7), and potassium phosphate (4.5 g, 21.2 mmol) were added to a sealed tube. 20 mL of tetrahydrofuran was added and the nitrogen atmosphere was replaced. The system was heated to 80°C for 3 h, the sample was mixed with silica gel, and compound 4a (197 mg, 37%) was separated by column chromatography.

[0219] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 298.1 [M+H] + .

[0220] [Article 26, 08.04.2025] Step 2:

[0221] [Article 26, 08.04.2025] Compound 4a (197 mg, 0.66 mmol), intermediate 3 (90 mg, 0.7 mmol), and 1 mL of potassium tert-butoxide (1 M in THF) were added to a sealed tube. After nitrogen displacement, the system was heated to 80°C and reacted for 3 h. The reaction solution was concentrated to dryness and purified by preparative separation to yield the title compound 4 (30 mg, 11%).

[0222] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 407.1 [M+H] + .

[0223] [Article 26, 08.04.2025] 1 H NMR(400MHz,DMSO-d6)δ10.23(s,1H),8.49(s,1H),8.38-8.33(m,1H),8.18-8.13(m,1H),7.79-7.75(m,1H),7.69-7.63 (m,1H),7.40-7.08(m,1H),4.86(s,1H),4.71(s,1H),3.88(s,2H),3.71(s,3H),2.23(s,2H),1.78(s,3H),1.14(s,3H).

[0224] [Article 26, 08.04.2025] 1.4 Preparation of Compound 5

[0225] [Article 26, 08.04.2025] Step 1:

[0226] [Article 26, 08.04.2025] 5a (1.5 g, 7.89 mmol), intermediate 4 (2.3 g, 11.84 mmol), potassium phosphate (21.8 g, 102.57 mmol), Xphos PdG2 (1.24 g, 1.58 mmol, CAS: 1310584-14-5), and Xphos (1.5 g, 3.16 mmol, CAS 564483-18-7) were added to a sealed tube. 60 mL of tetrahydrofuran was added, and the atmosphere was replaced with nitrogen. The temperature was raised to 80°C and the reaction was allowed to react for 5 h. After the disappearance of the starting material by spot plate detection, the solid was removed by filtration and washed with methanol. The filtrate was dried and passed through a silica gel column (dichloromethane:methanol = 10:1) to obtain the title compound 5b (1.4 g, 68%).

[0227] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 262.0 [M+H] + .

[0228] [Article 26, 08.04.2025] Step 2:

[0229] [Article 26, 08.04.2025] Intermediate 3 (495 mg, 3.83 mmol) was added to 15 mL of DMF solution. NaH (275 mg, 11.49 mmol) was added under ice-cooling and stirred for 10 min. Compound 5b (1 g, 3.83 mmol) was then added. After nitrogen was replaced, the reaction was incubated at 0°C for 1 h. The reaction was quenched with water and extracted with ethyl acetate. The organic phase was dried and passed through a column (dichloromethane:methanol = 10:1) to afford compound 5 (110 mg).

[0230] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 371.2 [M+H] + .

[0231] [Article 26, 08.04.2025] 1H NMR (400MHz, DMSO-d6) δ10.16(s,1H),8.47(s,1H),8.29(d,1H),7.80(d,1H),7.62(dd,1H),7.40(d,1H),4.86( s,1H),4.70(s,1H),3.75(s,2H),3.70(s,3H),2.50(s,3H),2.23(s,2H),1.78(s,3H),1.58(s,2H),1.14(s,3H).

[0232] [Article 26, 08.04.2025] 1.5 Preparation of Compound 6

[0233] [Article 26, 08.04.2025] Step 1:

[0234] [Article 26 of the Detailed Rules, 08.04.2025] Intermediate 3 (1 g, 7.7 mmol), intermediate 1 (1.6 g, 7.1 mmol) and 12 mL of potassium tert-butoxide (1 M in THF) were added to a sealed tube. After nitrogen was replaced, the system was heated to 80°C and reacted for 3 h. The system was cooled to room temperature, mixed with silica gel, and separated by column chromatography (petroleum ether: ethyl acetate = 1:1 to ethyl acetate) to obtain the target compound 6a (500 mg, 21%).

[0235] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 355.1 [M+H] + .

[0236] [Article 26, 08.04.2025] Step 2:

[0237] [Article 26 of the Detailed Rules, 08.04.2025] Compound 6a (500 mg, 1.5 mmol), intermediate 2 (620 mg, 3.6 mmol), Xphos PdG2 (200 mg, 0.25 mmol, CAS: 1310584-14-5), Xphos (250 mg, 0.52 mmol, CAS 564483-18-7), and potassium phosphate (4.5 g, 21.2 mmol) were added to a sealed tube. 20 mL of tetrahydrofuran was added and the nitrogen atmosphere was replaced. The system was heated to 80°C for 3 h, the sample was mixed with silica gel, and the reaction was separated by column chromatography (petroleum ether: ethyl acetate = 1:1 to ethyl acetate) to give compound 6b (350 mg, 61%).

[0238] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 384.2 [M+H] + .

[0239] [Article 26, 08.04.2025] Step 3:

[0240] [Article 26 of the Detailed Rules, 08.04.2025] Compound 6b (350 mg, 0.91 mmol) was dissolved in 20 mL of dichloromethane, and the system was cooled to -78°C. Ozone was introduced, and after the disappearance of the starting material on the spot plate detection, an excess of triphenylphosphine was added, and the mixture was slowly warmed to room temperature. The sample was mixed with silica gel and separated by column chromatography (petroleum ether: ethyl acetate = 1:1 to ethyl acetate) to obtain compound 6c (310 mg, 89%).

[0241] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 386.1 [M+H] + .

[0242] [Article 26, 08.04.2025] Step 4:

[0243] [Article 26, 08.04.2025] Compound 6c (160 mg, 0.42 mmol) was dissolved in 10 mL of tetrahydrofuran and the atmosphere was replaced with nitrogen. 1.4 mL of methylmagnesium chloride (3 M in THF) was added at 0°C. The mixture was slowly warmed to room temperature and quenched by adding saturated aqueous ammonium chloride. The organic phase was dried and extracted with dichloromethane. The organic phase was dried and freeze-dried to give compound 6 (30 mg, 18%).

[0244] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 402.2 [M+H] + .

[0245] [Article 26, 08.04.2025] 1 H NMR (400MHz, DMSO-d6) δ8.81-8.78(m,1H),8.42-8.37(m,1H),8.32(s,1H),8.23-8.19(m,1H),7.81-7.74(m, 1H),7.41-6.88(m,2H),4.02-3.91(m,2H),1.71(d,1H),1.60(d,1H),1.27(s,3H),1.23(s,3H),1.16(s,3H).

[0246] [Article 26, 08.04.2025] 1.6 Preparation of Compound 7

[0247] [Article 26, 08.04.2025] Step 1:

[0248] [Article 26 of the detailed rules, 08.04.2025] 7a (900 mg), intermediate 1 (633 mg, 2.8 mmol), Xphos PdG2 (250 mg, 0.32 mmol), Xphos (500 mg, 1.05 mmol), and potassium phosphate (6.0 g, 28.3 mmol) were added to a sealed tube. 30 mL of tetrahydrofuran was added and the nitrogen atmosphere was replaced. The system was heated to 80°C for 3 h. The sample was mixed with silica gel and separated by column chromatography to obtain compound 7b (428 mg, 54%).

[0249] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 282.2 [M+H] + .

[0250] [Article 26, 08.04.2025] Step 2:

[0251] [Article 26, 08.04.2025] Compound 7b (200 mg, 0.71 mmol), intermediate 3 (100 mg, 0.77 mmol), and 2.5 mL of potassium tert-butoxide (1 M in THF) were added to a sealed tube. After nitrogen displacement, the system was heated to 80°C and reacted for 3 h. Preparative isolation and purification yielded compound 7 (89 mg, 32%).

[0252] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 391.1 [M+H] + .

[0253] [Article 26, 08.04.2025] 1 H NMR (400MHz, DMSO-d6) δ10.54(s,1H),8.68(s,1H),8.41-8.37(m,1H),8.16-8.10(m,1H),7.81-7.75(m,1H),7.72-7.67 (m,1H),7.38-7.07(m,1H),4.86(s,1H),4.71(s,1H),3.88(s,2H),2.23(s,2H),2.12(s,3H),1.78(s,3H),1.14(s,3H).

[0254] [Article 26, 08.04.2025] 1.7 Preparation of Compound 8

[0255] [Article 26, 08.04.2025] Step 1:

[0256] [Article 26, 08.04.2025] Ferric nitrate nonahydrate (133 mg, 0.33 mmol) was dissolved in 3 mL of water, nitrogen was replaced, and then cooled to 0°C. A selective fluorine reagent (117 mg, 0.33 mmol) and 3 mL of acetonitrile were added, and compound 7 (35 mg, 0.09 mmol) was dissolved in 3 mL of acetonitrile and added to the system. After stirring for 5 minutes, sodium borohydride (40 mg, 1.05 mmol) was added in batches. The reaction was maintained at 0°C for 30 minutes. 1 mL of ammonia was added to quench the reaction, and the mixture was extracted with a mixed solvent of dichloromethane and methanol (10:1). The mixture was spin-dried and purified by HPLC to obtain compound 8 (10 mg, 28%).

[0257] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 411.3 [M+H] + .

[0258] [Article 26, 08.04.2025] 1 H NMR(400MHz,DMSO-d6)δ10.55(s,1H),8.69(s,1H),8.43-8.35(m,1H),8.18-8.10(m,1H),7.78-7.73(m,1H),7.7 2-7.65(m,1H),7.40-7.05(m,1H),3.94(s,2H),2.12(s,3H),1.95-1.86(m,2H),1.50-1.36(m,6H),1.23(s,3H).

[0259] [Article 26, 08.04.2025] 1.8 Preparation of Compound 9

[0260] [Article 26, 08.04.2025] Step 1:

[0261] [Article 26, 08.04.2025] Ferric nitrate nonahydrate (324 mg, 0.8 mmol) was dissolved in 7 mL of water, nitrogen was replaced, and then cooled to 0°C. A selective fluorine reagent (284 mg, 0.8 mmol) and 7 mL of acetonitrile were added, and compound 4 (81 mg, 0.2 mmol) was dissolved in 7 mL of acetonitrile and added to the system. After stirring for 5 minutes, sodium borohydride (100 mg, 2.6 mmol) was added portionwise. The reaction was maintained at 0°C for 30 minutes. 2.5 mL of ammonia was added to quench the reaction, and the mixture was extracted with a mixed solvent of dichloromethane:methanol (10:1). The mixture was spin-dried and purified by HPLC to obtain compound 9 (9 mg, 11%).

[0262] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 427.2 [M+H] +.

[0263] [Article 26, 08.04.2025] 1 H NMR(400MHz,DMSO-d6)δ10.25(s,1H),8.49(s,1H),8.38-8.33(m,1H),8.19-8.13(m,1H),7.78-7.72(m,1H),7.6 8-7.64(m,1H),7.38-7.09(m,1H),3.94(s,2H),3.71(s,3H),1.95-1.86(m,2H),1.50-1.37(m,6H),1.23(s,3H).

[0264] [Article 26, 08.04.2025] 1.9 Preparation of Compound 10

[0265] [Article 26 of the detailed rules, 08.04.2025] Ferric nitrate nonahydrate (88 mg, 0.22 mmol) was dissolved in water (2 mL), sonicated for 5 min, cooled to 0°C, and then a solution of a selective fluorine reagent (76 mg, 0.22 mmol) in 2 mL of acetonitrile was added, followed by a solution of compound 5 (20 mg, 0.05 mmol) in 2 mL of acetonitrile, and then sodium borohydride (30 mg, 0.79 mmol) was added in batches. The reaction was allowed to proceed for 1 h. LC-MS showed that the reaction of the raw material was complete. The mixture was diluted with water and extracted with dichloromethane. The organic phases were combined, dried, and concentrated to obtain a crude product, which was then purified by preparative separation to obtain compound 10 (10 mg, 47%).

[0266] [Article 26, 08.04.2025] LC-MS (ESI): m / z = 391.3 [M+H] + .

[0267] [Article 26, 08.04.2025] 1 H NMR(400MHz,CD3OD)δ8.42(s,1H),8.27(d,1H),7.76(d,1H),7.61(d,1H),7.40(d,1H),3.98(s, 2H),3.80(s,3H),2.57(s,3H),2.09(s,1H),2.04(s,1H),1.50(d,3H),1.45(d,3H),1.40(s,3H).

[0268] [Article 26, 08.04.2025] Example 2: In vitro AAK1 enzyme activity assay

[0269] [Article 26, 08.04.2025] A 10 mM stock solution of compound (dissolved in DMSO) was diluted to 0.2 mM with DMSO. This solution was then diluted 5-fold with DMSO to obtain 10 concentrations of compound solution. Each concentration of compound was then diluted 50-fold with 1× kinase reaction buffer (containing 40 mM Tris, 20 mM MgCl2, 0.1% BSA, and 0.5 mM DTT) for later use. AAK1 (Signalchem, Cat#A01-11G-10) was diluted to twice the final concentration (30 nM and 28 nM, respectively) with 1× kinase reaction buffer. AAK1 was added at 2 μL / well to a 384-well white plate. Compound was then added at 1 μL / well. The plate was sealed with a film sealer and centrifuged at 1000 rpm for 30 seconds. The plate was then incubated at room temperature for 10 minutes. A mixture of ATP (Promega, Cat#V914B) and substrate Micro2 (GenScript, Cat#PE0890) was prepared at 4 times the final concentration (the final ATP concentrations for AAK1 were 15 μM and 5 μM, respectively, and the final concentration of Micro2 was 0.1 mg / mL). 1 μL / well of the ATP-substrate mixture was added to the reaction plate. The plate was sealed with a sealing film and centrifuged at 1000 rpm for 30 seconds. The reaction was allowed to react at room temperature for 60 minutes (AAK1). Transfer 4 μL / well ADP-Glo ​​(Promega, Cat# V9102) to a 384-well plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 40 minutes; transfer 8 μL / well Detection solution to a 384-well plate, centrifuge at 1000 rpm for 1 minute, and incubate at 25°C for 40 minutes; read the RLU (Relative luminescence unit) signal value using a Biotek multi-function microplate reader, and calculate the percentage inhibition rate according to the following formula: [1-(LUM 化合物 -LUM 阳性对照 ) / (LUM 阴性对照 -LUM 阳性对照 )] × 100. IC was calculated using a four-parameter nonlinear fitting equation in Graphpad 7.0 software. 50 The specific results are shown in Table 1.

[0270] [Article 26, 08.04.2025] Table 1 AAK1 inhibitory activity

[0271] [Article 26, 08.04.2025] Conclusion: The compounds of the present invention showed high inhibitory activity against AAK1 receptor.

[0272] [Article 26, 08.04.2025] Example 3: Canine Pharmacokinetics

[0273] [Article 26, 08.04.2025] Experimental animals: Male beagle dogs, approximately 8–11 kg, 6 per compound, purchased from Beijing Mas Biotechnology Co., Ltd.

[0274] [Article 26, 08.04.2025] Method: On the day of the experiment, 12 beagle dogs were randomly divided into groups based on body weight. Food and water were withheld for 12–14 hours prior to dosing. Food was resumed 4 hours after dosing. Dosing was performed according to Table 2.

[0275] [Article 26, 08.04.2025] Table 2. Dosage Information

[0276] [Article 26, 08.04.2025] Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Oral administration solvent: 0.5% MC

[0277] [Article 26, 08.04.2025] (DMA: dimethylacetamide; Solutol: polyethylene glycol-15-hydroxystearate; Saline: normal saline; MC: methylcellulose)

[0278] [Article 26, 08.04.2025] Before and after administration, 1 ml of blood was collected from the jugular vein or limb vein and placed in an EDTAK2 centrifuge tube. The blood was centrifuged at 5000 rpm at 4°C for 10 min, and plasma was collected. Blood was collected at the following time points: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 10, 12, and 24 h for both the intravenous and oral gavage groups of LX9211; and at the following time points: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 10, 12, 24, and 48 h for both the intravenous and oral gavage groups of Compound 9. All samples were stored at -80°C prior to analysis and quantitative analysis was performed using LC-MS / MS. The results are shown in Table 3.

[0279] [Article 26, 08.04.2025] Table 3. Pharmacokinetic parameters of test compounds in beagle dog plasma

[0280] [Rule 26, 08.04.2025]-: Not applicable.

[0281] [Article 26, 08.04.2025] Note: The structure of LX-9211 is

[0282] [Article 26, 08.04.2025] Conclusion: The compounds of the present invention have good pharmacokinetic characteristics.

[0283] [Article 26, 08.04.2025] Example 4: hERG potassium channel effect test

[0284] [Article 26, 08.04.2025] Experimental platform: Electrophysiology manual patch clamp system

[0285] [Article 26, 08.04.2025] Cell line: Chinese hamster ovary (CHO) cell line stably expressing the hERG potassium channel

[0286] [Article 26, 08.04.2025] Experimental Methods: hERG potassium channel currents were recorded using the whole-cell patch-clamp technique at room temperature using CHO (Chinese Hamster Ovary) cells stably expressing the hERG potassium channel. Glass microelectrodes were pulled from glass electrode blanks (BF150-86-10, Sutter) using a puller. After perfusion with electrode solution, the tip resistance was approximately 2-5 MΩ. The microelectrode was connected to the patch-clamp amplifier by inserting it into the amplifier headstage. Clamping voltage and data recording were controlled and recorded by a computer using pClamp 10 software, with a sampling frequency of 10 kHz and a filter frequency of 2 kHz. After whole-cell recordings were obtained, cells were clamped at -80 mV. The hERG potassium current (I hERG ) was evoked by applying a 2-s depolarizing voltage step from -80 mV to +20 mV, followed by a repolarization voltage to -50 mV for 1 s before returning to -80 mV. The voltage stimulation was given every 10 seconds, and the administration process was started after the hERG potassium current was confirmed to be stable (at least 1 minute). Each compound was administered for at least 1 minute at each test concentration, and at least 2 cells (n≥2) were tested at each concentration.

[0287] [Article 26, 08.04.2025] Data processing: Data were analyzed and processed using pClamp 10, GraphPad Prism 5, and Excel software. The degree of inhibition of hERG potassium current (peak hERG tail current induced at -50 mV) by different compound concentrations was calculated using the following formula: Inhibition % = [1-(I / Io)] × 100%

[0288] [Article 26, 08.04.2025] In which, Inhibition% represents the percentage of inhibition of hERG potassium current by the compound, and I and Io represent the amplitude of hERG potassium current after and before drug addition, respectively.

[0289] [Article 26, 08.04.2025] Compound IC50 was calculated using GraphPad Prism 5 software by fitting the following equation: Y = Bottom + (Top - Bottom) / (1 + 10^((LogIC50 - X) × HillSlope))

[0290] [Article 26, 08.04.2025] Where X is the Log value of the test sample concentration, Y is the inhibition percentage at the corresponding concentration, and Bottom and Top are the minimum and maximum inhibition percentages, respectively.

[0291] [Article 26, 08.04.2025] Experimental results: The IC50 values ​​of the test compounds for the inhibitory effect on hERG potassium channel current are shown in Table 4.

[0292] [Article 26, 08.04.2025] Table 4 Inhibition of hERG potassium channel current by test compounds

[0293] [Article 26, 08.04.2025] Example 5: Spinal Nerve Ligation (SNL)-Induced Neuropathic Pain Model in Mice

[0294] [Article 26, 08.04.2025] Male C57BL / 6J mice (8 weeks old) purchased from Jinan Pengyue Experimental Animal Breeding Co., Ltd. were adaptively raised for one week before model establishment. The specific establishment method is as follows:

[0295] [Article 26, 08.04.2025] 1) Sterilization of surgical instruments and ligatures;

[0296] [Article 26, 08.04.2025] 2) Anesthetize the mouse with isoflurane and place it in the prone position on the operating table;

[0297] [Article 26, 08.04.2025] 3) The mouse was skinned near the hip bone and an incision approximately 2 cm long was made along the spine;

[0298] [Article 26, 08.04.2025] 4) Separate the fascia along the spine and bluntly separate the muscles to expose the L5 transverse process;

[0299] [Article 26, 08.04.2025] 5) Carefully bite off the L5 transverse process with forceps to expose the L5 spinal nerve;

[0300] [Article 26, 08.04.2025] 6) Carefully separate the L5 nerve with a glass needle and ligate the L5 nerve with a 5-0 ligature.

[0301] [Article 26, 08.04.2025] 7) Suture the muscles and skin and disinfect with iodine;

[0302] [Article 26, 08.04.2025] The day after modeling, mice that failed to establish a model were eliminated (sign of successful modeling: the mouse's hind paw curled up). After modeling, the mice were stroked for 3 to 5 minutes every day to ensure that the animals were familiar with the experimenter. The mice were then placed on a metal pain test frame for 40 to 60 minutes to adapt to the environment. After the third day, the mice were acclimated to the environment using Von Frey fibers ( Pre-dose baseline values ​​(Ascending test) were obtained for test animals (0.16, 0.4, 0.6, 1.0, 1.4, and 2.0 g). Each animal was measured twice, with at least 5 minutes between measurements, and the average was calculated. Animals were then grouped according to baseline values ​​(10 animals per group). After grouping, LX-9211 (1 and 10 mg / kg), compound 2 (1 and 10 mg / kg), or vehicle (40% PEG-400 + 10% ethanol + 15% Tween 80 + 35% saline) was administered orally. Mechanical pain thresholds (MPTs) were measured 1, 3, and 6 hours after administration. Time-MPT curves were plotted and statistically analyzed using GraphPad 8.3.0.

[0303] [Article 26, 08.04.2025] Results and Conclusions: See Figure 1 for results. Both 10 mg / kg LX-9211 and Compound 2 effectively elevated the pain threshold of SNL-induced mice 1, 3, and 6 hours after a single dose. The analgesic efficacy of 10 mg / kg LX-9211 peaked 1 hour after administration and gradually declined thereafter. In contrast, the analgesic efficacy of 10 mg / kg Compound 2 peaked 3 hours after administration and remained stable from 1 to 6 hours, with superior efficacy to that of LX-9211 at 3 and 6 hours. These data demonstrate that Compound 2 has superior analgesic activity to LX-9211.

[0304] [Article 26, 08.04.2025] Example 6: Mouse brain-to-blood ratio test

[0305] [Article 26, 08.04.2025] 6.1 Experimental Animals: Male ICR mice, 20-25 g, 9 mice / compound. Purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0306] [Article 26, 08.04.2025] 6.2 Experimental Design: On the day of the experiment, 18 ICR mice were randomly divided into groups according to body weight. They were fasted (but not water) for 12–14 hours before dosing and fed 4 hours after dosing.

[0307] [Article 26, 08.04.2025] Table 5. Dosage Information

[0308] [Article 26, 08.04.2025] Note: Oral administration solvent: 40% PEG-400 + 10% Ethanol + 15% Tween 80 + 35% Saline;

[0309] [Article 26, 08.04.2025] (Saline: physiological saline; Ethanol: ethanol; Tween 80: Tween 80)

[0310] [Article 26, 08.04.2025] Whole blood and brain tissue were collected 0.5, 4, and 24 hours after oral administration. The whole blood was centrifuged to separate the plasma. The brain tissue was rinsed with cold saline to remove any residual blood, blotted dry, and homogenized. All samples were stored at -80°C prior to analysis and quantitative analysis was performed using LC-MS / MS.

[0311] [Article 26 of the Regulations, 08.04.2025] The test results are shown in Table 6.

[0312] [Article 26, 08.04.2025] Table 6. Pharmacokinetic parameters of the compounds in mouse plasma

[0313] [Rule 26, 08.04.2025]-: Not applicable.

[0314] [Article 26, 08.04.2025] Conclusion: The compounds of the present invention, especially compound 2, have high brain penetrance.

[0315] [Article 26, 08.04.2025] Example 7: Composition of Compound 3

[0316] [Article 26, 08.04.2025] 7.1 Prescriptions 1-6

[0317] [Article 26, 08.04.2025] Table 7. AAK1 inhibitor combination prescription (1)

[0318] [Article 26 of the Detailed Rules, 08.04.2025] Tablets containing a composition of Compound 3 were prepared according to the formula in the table: Compound 3 and a wetting agent were co-grinded and mixed for 15 minutes, followed by the addition of a diluent and a disintegrant in a mixer and mixing for 15 minutes. Finally, a lubricant was added and mixed for 3 minutes to obtain a composition containing Compound 3. The composition was compressed into tablets using a tablet press.

[0319] [Article 26, 08.04.2025] We observed whether the above formulation exhibited any agglomeration or adhesion to the equipment caused by softening or melting of Compound 3 during the manufacturing process. We also examined the content uniformity of the finished product.

[0320] [Article 26, 08.04.2025] Table 8. Test results

[0321] [Article 26, 08.04.2025] The results show that no agglomeration or adhesion to the equipment due to the softening or melting of compound 3 occurred, regardless of whether the wetting agent was added to the formulation.

[0322] [Article 26, 08.04.2025] 7.2 Prescriptions 7-12

[0323] [Article 26, 08.04.2025] Table 9. AAK1 inhibitor combination prescription (2)

[0324] [Article 26 of the Detailed Rules, 08.04.2025] Tablets containing a composition of Compound 3 were prepared according to the formulation in the table: Compound 3 and a wetting agent were first sieved and mixed five times, followed by mixing in a mixer for 15 minutes, followed by addition of a diluent and a disintegrant and mixing for 15 minutes, and finally, a lubricant and mixing for an additional 3 minutes to obtain a composition containing Compound 3. The composition was compressed using a tablet press to obtain tablets.

[0325] [Article 26, 08.04.2025] We observed whether the above formulation exhibited any agglomeration or adhesion to the equipment caused by softening or melting of Compound 3 during the manufacturing process. We also examined the content uniformity of the finished product.

[0326] [Article 26, 08.04.2025] Table 10. Test results

[0327] [Article 26, 08.04.2025] Conclusion: Adding a certain amount of wetting agent to the formulation can significantly improve the softening / melting phenomenon of compound 3 during the preparation process, and the content uniformity of the tablets is better.

[0328] [Article 26, 08.04.2025] 7.3 Prescriptions 13-18

[0329] [Article 26, 08.04.2025] Table 11. AAK1 inhibitor combination prescription (3)

[0330] [Article 26, 08.04.2025] Preparation method: Compound 3 and a wetting agent are mixed in a mixer for 15 minutes, followed by the addition of a diluent and a disintegrant and mixing for 15 minutes, and finally, magnesium stearate is added and mixed for 3 minutes to obtain a composition containing Compound 3. The composition containing Compound 3 is then filled into capsules using a capsule filling machine.

[0331] [Article 26, 08.04.2025] We observed whether the above formulation exhibited any agglomeration or adhesion to the equipment caused by softening or melting of Compound 3 during the manufacturing process. We also examined the content uniformity of the finished product.

[0332] [Article 26, 08.04.2025] Table 12. Test results

[0333] [Article 26, 08.04.2025] Conclusion: The addition of a wetting agent can significantly inhibit the softening / melting phenomenon of compound 3 during the preparation process.

[0334] [Article 26, 08.04.2025] 7.4 Prescriptions 19-24

[0335] [Article 26, 08.04.2025] Table 13. AAK1 inhibitor combination prescription (4)

[0336] [Article 26, 08.04.2025] Tablets containing the Compound 3 composition were prepared according to the following formulation: Compound 3 and the wetting agent were sieved and mixed five times, followed by mixing in a blender for 15 minutes. The diluent and disintegrant were then added and mixed for 15 minutes. The premix was then loaded into a roller compactor and compacted at 0.6 kN / cm. The resulting ribbon was passed through a 0.8 mm vibrating mill. The ground granules were then mixed with a lubricant for 3 minutes and compressed using a tablet press.

[0337] [Article 26, 08.04.2025] We investigated the dissolution behavior of the tablets prepared according to the above formulation and process in a pH 1.0 medium and the changes in their related substances after being stored at 40℃±2℃, 75%RH±5%RH for 6 months. See Table 14.

[0338] [Article 26, 08.04.2025] As shown in Figure 2, for formulations 19-21, as the proportion of compound 3 increases, the softening / melting of compound 3 causes AAK1 aggregation, further reducing its solubility. For formulations 22-24, the addition of a wetting agent prevents the agglomeration caused by the softening / melting of compound 3, thereby improving their dissolution behavior.

[0339] [Article 26, 08.04.2025] Table 14. Stability results

[0340] [Article 26, 08.04.2025] It can be seen from the table that the composition has good stability regardless of whether the composition contains a wetting agent or not.

[0341] [Article 26, 08.04.2025] 7.5 Prescriptions 25-28

[0342] [Article 26, 08.04.2025] Table 15. AAK1 inhibitor combination prescription (5)

[0343] [Article 26, 08.04.2025] Preparation method: Compound 3 was mixed with lactose, microcrystalline cellulose, and cross-linked sodium carboxymethyl cellulose in a mixer for 15 minutes, and then magnesium stearate was added and mixed for 3 minutes. The tablets were then compressed using a tablet press. The tablet hardness was controlled at 50-100N.

[0344] [Article 26, 08.04.2025] Table 16. AAK1 inhibitor combination prescription (6)

[0345] [Article 26, 08.04.2025] Preparation: Compound 3 was mixed with mannitol, microcrystalline cellulose, and low-substituted carboxymethyl cellulose sodium in a premixer for 15 minutes. The premix was then loaded into a roller compactor and compacted at 0.6 kN / cm. The resulting ribbon was passed through a 0.8 mm vibrating mill. The ground particles were then mixed with talc for 3 minutes and compressed using a tablet press. The tablet hardness was controlled at 50-100 N.

[0346] [Article 26, 08.04.2025] Table 17. AAK1 inhibitor combination prescription (7)

[0347] [Article 26, 08.04.2025] Preparation method: Compound 3 was mixed with pregelatinized starch, sodium carboxymethyl starch, and dextrin in a mixer for 15 minutes, and then magnesium stearate was added and mixed for 3 minutes. The mixture was then compressed using a tablet press. The tablet hardness was controlled at 50-100N.

[0348] [Article 26, 08.04.2025] Table 18. AAK1 inhibitor combination prescription (8)

[0349] [Article 26, 08.04.2025] Preparation method: Compound 3 was mixed with calcium phosphate, sucrose, crospovidone and talc in a mixer for 15 minutes, and then the mixture was filled into capsules using a capsule filling machine.

[0350] [Article 26, 08.04.2025] Dissolution profiles of tablets prepared according to prescription 25 were investigated in water at pH 1.0, pH 4.5, and pH 6.8 according to the second method (slurry method) in section 0931 of the 2015 edition of the Chinese Pharmacopoeia. The rotation speed was set at 50 rpm, and sampling was performed at 5, 10, 15, 20, 30, 45, and 60 minutes.

[0351] [Article 26 of the Detailed Rules, 08.04.2025] The results in Figure 3 show that: at pH = 1.0, the solubility of the tablets prepared with Prescription 25 can reach 90% in 5 minutes, and the dissolution is rapid; at 20 minutes, the solubility of the tablets prepared with Prescription 25 in water at pH 1.0, pH 4.5, and pH 6.8 can all reach 85%.

[0352] [Article 26, 08.04.2025] 7.6 Preparation Stability Study Methods and Results

[0353] [Article 26 of the Detailed Rules, 08.04.2025] The preparations prepared from Prescription 25, Prescription 26, Prescription 27 and Prescription 28 were packaged in aluminum-plastic blister packs, placed at 40℃±2℃, 75%RH±5%RH and the changes in their related substances were investigated.

[0354] [Article 26, 08.04.2025] Table 19. Stability test results

[0355] [Article 26 of the Detailed Rules, 08.04.2025] The results in Table 19 show that the preparations prepared by prescriptions 25-28 are relatively stable, and their stability is basically the same.

[0356] [Article 26 of the Detailed Rules, 08.04.2025] The pharmaceutical compositions of compound 2 were prepared by referring to the prescriptions 1-6, 10-13, 16-28 and process of the pharmaceutical composition of compound 3. There was no agglomeration or adhesion to the equipment, the content uniformity (A+2.2S) was 3-5.5, and the pharmaceutical compositions were relatively stable.

Claims

1. A pharmaceutical composition, include: The active ingredient is selected from the compound of formula (I) or its stereoisomers and pharmaceutically acceptable salts: in, Z is selected from NH or O; R 1 , R 2 Each is independently selected from H, deuterium, halogen, amino, -COOH, cyano, sulfonyl, aminoacyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, wherein the alkyl group is optionally further substituted with 1 to 3 R A Substituent substitution; R 41 , R 42 Each independently selected from H, deuterium, amino, C 1-6 Alkyl, halogen, cyano, hydroxyl, halo 1-6 Alkyl, deuterated C 1-6 alkyl; R 51 , R 52 Each is independently selected from H, deuterium, amino, halogen; R 61 , R 62 , R 63 Each independently selected from H, deuterium, halogen, amino, cyano, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 alkyl; Or, R 51 and R 61 , or R 61 and R 62 Together with the carbon atoms to which they are attached, they form double bonds; R A Selected from deuterium, halogen, amino, cyano, hydroxyl, C 1-6 Alkyl, halogenated C 1-6 Alkyl, deuterated C 1-6 Alkyl, C 1-6 Alkoxy, halogenated C 1-6 Alkoxy, deuterated C 1-6 Alkoxy or hydroxyl C 1-6 alkyl; Provided that, when Z is selected from O, The following structure is not formed: b) inactive ingredients; The content of the active ingredient in the pharmaceutical composition is 5% to 90% w / w, preferably 5% to 80% w / w, and more preferably 10% to 80% w / w.

2. The pharmaceutical composition according to claim 1, in, Z is O; R1, R2 are selected from halogenated C1-2 alkyl; R51 and R52 are each independently selected from H and deuterium; R41 and R42 are each independently selected from amino and C1-2 alkyl; R61, R62, and R63 are each independently selected from H, deuterium, and C1-2 alkyl.

3. The pharmaceutical composition according to claim 1, wherein the compound of formula (I) is selected from one of the following structures:

4. The pharmaceutical composition according to claim 1, wherein the inactive ingredient is selected from diluents.

5. The pharmaceutical composition according to claim 4, wherein the diluent is selected from one or more of starch, pregelatinized starch, dextrin, lactose monohydrate, anhydrous lactose, sucrose, microcrystalline cellulose, inorganic salts, and sugar alcohols; preferably one or more of pregelatinized starch, dextrin, lactose, sucrose, microcrystalline cellulose, calcium sulfate, dibasic calcium phosphate dihydrate, anhydrous dibasic calcium phosphate, calcium phosphate, calcium carbonate, calcium stearate, magnesium oxide, aluminum hydroxide, mannitol, xylitol, and sorbitol; more preferably one or more of pregelatinized starch, lactose, sucrose, microcrystalline cellulose, dibasic calcium phosphate dihydrate, anhydrous dibasic calcium phosphate, calcium phosphate, mannitol, and dextrin.

6. The pharmaceutical composition according to claim 4, wherein the weight ratio of the active ingredient to the diluent is 1:0.05 to 1:10, preferably 1:0.1 to 1:8.5, and more preferably 1:0.18 to 1:8.

5.

7. The pharmaceutical composition according to claim 4, wherein the inactive ingredient further comprises one or more of a wetting agent, a disintegrant, and a lubricant; The wetting agent is selected from one or more silicates; preferably one or more of silicon dioxide, magnesium silicate, magnesium trisilicate, magnesium aluminum silicate and talc; more preferably one or more of fumed silica, precipitated silica, sol-gel silica, magnesium silicate, magnesium trisilicate, magnesium aluminum silicate and talc; The disintegrant is selected from one or more of cross-linked sodium carboxymethyl cellulose, cross-linked polyvinylpyrrolidone, starch and its derivatives, low-substituted hydroxypropyl cellulose, low-substituted sodium hydroxymethyl cellulose, surfactants, alginic acid and sodium alginate, and clays; preferably one or more of cross-linked sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, low-substituted sodium hydroxymethyl cellulose, cross-linked polyvinylpyrrolidone, starch, sodium carboxymethyl starch, hydroxypropyl starch, polysorbate 80, sodium lauryl sulfate, bentonite, and colloidal magnesium aluminum silicate; more preferably one or more of cross-linked sodium carboxymethyl cellulose, low-substituted hydroxypropyl cellulose, low-substituted sodium hydroxymethyl cellulose, sodium carboxymethyl starch, and cross-linked polyvinylpyrrolidone; The lubricant is selected from one or more of talc, stearic acid, metal stearate, stearic acid ester, glyceryl behenate, sodium lauryl sulfate or colloidal silicon dioxide; preferably one or more of talc, calcium stearate, magnesium stearate and zinc stearate, polyoxyethylene stearate, glyceryl monostearate and glyceryl palmitostearate; more preferably one or more of talc, calcium stearate, magnesium stearate and glyceryl monostearate.

8. The pharmaceutical composition according to claim 7, wherein the weight ratio of the active ingredient to the wetting agent is 1:0.05 to 1:

5. Preferably 1:0.08 to 1:3, more preferably 1:0.12 to 1:2; The weight ratio of the active ingredient to the disintegrant is 1:0.01 to 1:3, preferably 1:0.03 to 1:1.5, and more preferably 1:0.03 to 1:0.6; The weight ratio of the active ingredient to the lubricant is 1:0.001 to 1:2, preferably 1:0.006 to 1:0.

1.

9. A pharmaceutical preparation comprising the pharmaceutical composition according to any one of claims 1 to 8.

10. The pharmaceutical preparation according to claim 9, wherein the amount of the active ingredient in a unit preparation is 1 mg to 100 mg, preferably 5 mg, 10 mg, 20 mg, 25 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 mg, 100 mg.

11. The pharmaceutical preparation according to claim 9, which is in the form of tablets, granules, capsules, and soft capsules.

12. Use of the pharmaceutical composition according to any one of claims 1 to 8 or the pharmaceutical preparation according to any one of claims 9 to 11 in the preparation of drugs for treating diseases related to the inhibition or degradation of AAK1.

13. The use according to claim 12, wherein the disease is selected from pain, including inflammatory pain, postoperative pain, trigeminal neuralgia, acute postherpetic neuralgia and postherpetic neuralgia, diabetic peripheral neuropathy, causalgia, occipital neuralgia, fibromyalgia, phantom limb pain, burn pain and other forms of neuralgia, neuropathy and spontaneous pain syndrome.

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