WO2026012382A1 - Pharmaceutical formulation of thiocyclopentyl derivative and pharmaceutical use thereof - Google Patents

Abstract

A pharmaceutical formulation of a thiocyclopentyl derivative and pharmaceutical use thereof. The pharmaceutical formulation comprises a therapeutically effective amount of an active ingredient M and a pharmaceutical excipient. The active ingredient M is selected from a compound represented by general formula (I) or a stereoisomer thereof, a tautomer thereof, a deuterated compound thereof, a solvate thereof, a prodrug thereof, a metabolite thereof, a pharmaceutically acceptable salt thereof, or a eutectic thereof. The pharmaceutical formulation comprises 1-800 mg of the active ingredient M. The use is use of the pharmaceutical formulation in the preparation of related drugs for treating pain.

Description

A pharmaceutical formulation of a thiocyclopentyl derivative and its pharmaceutical application Technical Field

This invention belongs to the field of pharmaceutical formulations, specifically relating to a pharmaceutical formulation comprising a therapeutically effective amount of an active ingredient M and a pharmaceutical excipient, wherein the active ingredient M is selected from compounds of general formula (I) or their stereoisomers, tautomers, or pharmaceutically acceptable salts. This invention also relates to the use of the pharmaceutical formulation in the preparation of pain-related medications.

Background Technology

Nerve impulses (action potentials) are transmitted via afferent nerve fibers to the cell body of the dorsal root ganglion (DRG), ultimately reaching higher nerve centers and causing pain. The generation and conduction of action potentials in neurons depend on voltage-gated sodium channels (VGSCS) on the cell membrane. When the cell membrane depolarizes, sodium channels are activated, opening and causing an influx of sodium ions, further depolarizing the cell membrane and leading to the generation of action potentials.

VGSCS consists of a porous α-subunit (approximately 260 kDa) and an associated smaller β-subunit (30–40 kDa). The associated α-subunit family comprises 10 members, nine of which (Nav1.1–1.9) are voltage-gated. Nav1.8 is encoded by the gene SCN10A and is preferentially expressed in peripheral sensory neurons. It has been shown to shape action potentials in these neurons. Nav1.8 transcripts and proteins have been found in dorsal root ganglion (DRG) neurons. Nav1.8 has not been detected in non-neuronal tissues (e.g., heart and skeletal muscle) or the central nervous system (including the brain and spinal cord).

The crucial role of Nav1.8 in pain signal transduction has been supported by multiple pieces of evidence. Based on a series of animal experiments and human genetic evidence, selective inhibition of Nav1.8 has the potential to become a novel analgesic therapy. Currently, drugs targeting this target have entered clinical trials.

Summary of the Invention

The present invention aims to provide a pharmaceutical preparation comprising a therapeutically effective amount of an active ingredient M and a pharmaceutical excipient, wherein the active ingredient M is selected from compounds of general formula (I) or their stereoisomers, tautomers, or pharmaceutically acceptable salts, and the pharmaceutical preparation has a dosage strength of 1-800 mg. The present invention also relates to the use of the pharmaceutical preparation in the preparation of pain-related medications.

The pharmaceutical formulation of this invention has the advantages of stable quality, good dissolution, good absorption, low irritation, good safety, high bioavailability, oral absorption, weak inhibition of CYP enzymes, weak CYP3A4 induction, lower risk of drug-drug interactions, weak UGT1A1 inhibition, and lower risk of hepatotoxicity.

This invention relates to a pharmaceutical formulation comprising a therapeutically effective amount of an active ingredient M and a pharmaceutical excipient. The pharmaceutical formulation may be in unit dosage form.

This invention relates to a pharmaceutical formulation comprising an active ingredient M and a pharmaceutical excipient, wherein the active ingredient M is selected from compounds of general formula (I) or their stereoisomers, tautomers, or pharmaceutically acceptable salts.

_Q1 is selected from Preferably, _Q1 is selected from The _Q1 can be optionally replaced by 1 to 5 ^Rq ;

B is selected from Or phenyl, wherein the B is optionally substituted with 1 to 4 ^RBs ;

R ^Q1 is selected from H, COOH, NR ^q1 R ^q2 , -C(=O)NR ^q1 R ^q2 , -S(=O) ₂ NR ^q1 R ^q2 , OH, =O, -OR ^q1 , -C(=O)R ^q1 , -S(=O) ₂ R ^q1 , -S(=O)(=NR ^q1 )R ^q2 or -P(=O)R ^q1 R ^q2 ;

R ^q1 and R ^q2 are each independently selected from H, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, or cyclopentyl, wherein the methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, or cyclopentyl group is optionally substituted by 1 to 4 R ^k groups.

R ^_q and R_^B are each independently selected from H, F, Cl, Br, cyano, CH _₂ F, CH_{_F2}, CF_₃ , -OCH_₂F, -OCHF_₂, -OCF_₃, -OCD₃, methyl, -S-methyl, -S-CF₃, ethyl, isopropyl, ethynyl, methoxy, ethoxy, isopropyloxy, propyloxy, cyclopropyl, -O-cyclopropyl, -P(=O)(CH _₃ )_₂, -P(=O)(CH _₂CH₃)_₂, -P(=O)(CH _₃)(cyclopropyl), wherein the methyl, ethyl, isopropyl, ethynyl, methoxy, ethoxy, isopropyloxy, propyloxy, and cyclopropyl groups are optionally substituted by 1 to 4 R_^kgroups;

R ^k is selected from deuterium, =O, F, Cl, Br, I, CN, OH, NH ₂ , NH(CH ₃ ), NH(CH ₂ CH ₃ ), N(CH ₃ ) ₂ , N(CH ₂ CH ₃ ) ₂ , methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, -O-cyclopropyl, -NH-cyclopropyl, -CH ₂ -cyclopropyl, -CH ₂ -cyclobutyl, -CH ₂ -cyclopentyl, -CH ₂ -cyclohexyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, wherein the methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl groups are optionally substituted by 1 to 4 substituents selected from deuterium, halogen, CN, OH, NH ₂ , C _1-4 alkyl, and C _1-4 alkoxy groups;

In some embodiments, the compound of general formula (I) Selected from _Q1 can be optionally replaced by 1 to 3 ^Rq ;

^RQ1 is selected from H, COOH, _NH2 , -C(＝O) _NH2 , -S(＝O _{)2NH2 ,} OH, ＝O, -S(＝O) _2CH3 , _-S (＝O) ₂ -cyclopropyl, -S(＝O)(＝NH) _CH3 , -P(＝O)( _CH3 ) ₂ , -P(＝O)( _{CH2CH3 ) 2} , -P(＝O)( _CH3 )(cyclopropyl);

R ^qa is selected from _-CH₂OH , _{-CF₂CH₂OH} , _NH₂ , -P(＝O)( _CH₃ ) _₂ , -P(＝O)( _CH₂CH₃ ) _₂ , -P(＝O)( _CH₃ )( _cyclopropyl ), preferably R _qa is selected from ^... -CH ₂ OH;

Preferably, Selected from _Q1 can be optionally replaced by 1 to 3 ^Rq ;

B is selected from Preferred

R and ^q are each independently selected from H, F, Cl, Br, cyano, _CH₂F , _CHF₂ , _CF₃ , _-OCH₂F , _-OCHF₂ , _-OCF₃ , _-OCD₃ , methyl, -S-methyl, -S- _CF₃ , ethyl, isopropyl, ethynyl, methoxy, ethoxy, isopropyloxy, propyloxy, cyclopropyl, -O-cyclopropyl, _-CH₂OH , _{-CF₂CH₂OH} ; preferably from F, Cl, Br, cyano, _CH₂F , _CHF₂ , _{CF₃ ;}

In some embodiments, the compound of general formula (I) Selected from R ^Q1 is selected from COOH, -C(＝O)NH ₂ , -S(＝O) _{2NH 2} , -S(＝O) _{2CH 3} , -S(＝O)(＝NH)CH ₃ , -P(＝O)(CH ₃ ) ₂ , and R ^qa is selected from... _-CH₂OH , ^Rq is selected from F, Cl or methyl;

In some embodiments, in the compound of general formula (I), R and ^B are each independently selected from F, Cl, Br, cyano, _CH₂F , _CHF₂ , _CF₃ , _-OCH₂F , _-OCHF₂ , -OCF₃, _-OCD₃ , methyl, ethyl, isopropyl _, methoxy, ethoxy, isopropyloxy, propyloxy, cyclopropyl, -O-cyclopropyl;

In some embodiments, the structure of the compound of general formula (I) is selected from one of the structures shown in Table S-1;

Table S-1

In some embodiments, the compound of general formula (I) is selected from the following structures:

The total content of all components in any of the pharmaceutical preparations described in this invention is 100%.

The pharmaceutical preparations described in any of the present invention are solid dosage forms.

This invention relates to a pharmaceutical formulation comprising the aforementioned active ingredient M (calculated as a free base, hereinafter the same) and a pharmaceutical excipient, wherein the pharmaceutical formulation comprises 1-800 mg of active ingredient M; wherein the pharmaceutical formulation comprises 1-400 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 5-350 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 5-300 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 5-250 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 10-200 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 15-150 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 20-150 mg of active ingredient M; in some embodiments, the pharmaceutical formulation comprises 50-100 mg of active ingredient M; and in some embodiments, the pharmaceutical formulation comprises 10-100 mg of active ingredient M.

In some embodiments, the amount of active ingredient M in a unit formulation of the pharmaceutical preparation includes, but is not limited to, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 125 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 300 mg, 400 mg, 500 mg, and 600 mg.

In some embodiments, the formulation strengths of the pharmaceutical preparations described in this invention include, but are not limited to, 1-500mg, 2-500mg, 3-500mg, 5-500mg, 6-500mg, 10-500mg, 20-500mg, 25-500mg, 30-500mg, 40-500mg, 50-500mg, 60-500mg, 70-500mg, 75-500mg, 80-500mg, 90-500mg, 100-500mg, 200-500mg, 300-500mg, 400-500mg, 1-400mg, 2-400mg, 3-400mg, 5-400mg, 6-400mg, 10-400mg, 20-400mg, and 25-400mg. 0mg, 30-400mg, 40-400mg, 50-400mg, 60-400mg, 70-400mg, 75-400mg, 80-400mg, 90-400mg, 100-400mg, 200-400mg, 300-400mg, 1-300mg, 2-300mg, 5-300mg, 10-30 0mg, 20-300mg, 25-300mg, 30-300mg, 40-300mg, 50-300mg, 60-300mg, 70-300mg, 7 5-300mg, 80-300mg, 90-300mg, 100-300mg, 125-300mg, 150-300mg, 200-300mg, 25 0-300mg, 1-200mg, 2-200mg, 5-200mg, 10-200mg, 20-200mg, 25-200mg, 30-200mg , 40-200mg, 50-200mg, 60-200mg, 70-200mg, 75-200mg, 80-200mg, 90-200mg, 100- 200mg, 125-200mg, 150-200mg, 1-100mg, 2-100mg, 5-100mg, 10-100mg, 15-100mg, 20-100mg, 25-100mg, 30-100mg, 40-100mg, 50-100mg, 60-100mg, 70-100mg, 75-10 0 mg, 80-100 mg, 90-100 mg; in some embodiments, the formulation strengths of the drug preparation include, but are not limited to, 1 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 45 mg, 50 mg, 55 mg, 65 mg, 70 mg, 75 mg, 80 mg, 85 mg, 90 mg, 95 mg, 100 mg, 110 mg, 120 mg, 125 mg, 130 mg, 140 mg, 150 mg, 160 mg, 170 mg, 180 mg, 190 mg, 200 mg, 210 mg, 220 mg, 230 mg, 240 mg, 250 mg, 300 mg, 400 mg, 500 mg, 600 mg, and 800 mg.

The pharmaceutical formulation according to any one of the present invention comprises the aforementioned active ingredient M and a pharmaceutical excipient, wherein the content of active ingredient M is 1%-99%; in some embodiments, it is 1%-90%; in some embodiments, it is 1%-80%; in some embodiments, it is 1%-70%; in some embodiments, it is 1%-60%; in some embodiments, it is 1%-50%; in some embodiments, it is 1%-40%; in some embodiments, it is 1%-30%; in some embodiments, it is 1%-20%; in some embodiments, it is 1%-10%; in some embodiments, it is 5%-90%; in some embodiments, it is 5%-80%; in some embodiments, it is 5%-70%; in some embodiments... The percentages are as follows: 5%-60% in some embodiments; 5%-50% in some embodiments; 0.5%-47.5% in some embodiments; 5%-40% in some embodiments; 5%-30% in some embodiments; 5%-20% in some embodiments; 5%-10% in some embodiments; 10%-90% in some embodiments; 10%-80% in some embodiments; 10%-70% in some embodiments; 10%-60% in some embodiments; 10%-50% in some embodiments; 10%-40% in some embodiments; 10%-30% in some embodiments; 10%-20% in some embodiments.

In some embodiments, the weight ratio of the active ingredient M to the pharmaceutical excipient is 1:0.01 to 1:100, 1:0.5 to 1:10, 1:1.5 to 1:9, or 1:7 to 1:9;

In some embodiments, the pharmaceutical excipient comprises one or two of a filler and a disintegrant; in some embodiments, the pharmaceutical excipient comprises one or more of a filler, a disintegrant, a binder, a flow aid, a lubricant, and a pH adjuster; in some embodiments, the pharmaceutical excipient comprises one or more of a filler, a binder, a disintegrant, a flow aid, a lubricant, a solubilizer, a flavoring agent, an antioxidant, a preservative, a light-blocking agent, and a film-coating premix; in some embodiments, the pharmaceutical excipient comprises a filler, a disintegrant, a binder, a flow aid, and a lubricant; in some embodiments, the pharmaceutical excipient further comprises a pH adjuster; in some embodiments, the pharmaceutical excipient comprises a filler, a disintegrant, a binder, and a lubricant; in some embodiments, the pharmaceutical excipient comprises a filler and a disintegrant; in some embodiments, the pharmaceutical excipient further comprises one or more of a binder, a flow aid, and a lubricant; in some embodiments, the pharmaceutical excipient further comprises one or more of a binder, a flow aid, a lubricant, and a pH adjuster.

In some embodiments, in any of the foregoing embodiments, the filler is selected from one or more of microcrystalline cellulose, mannitol, lactose, sucrose, sorbitol, dextran, anhydrous dicalcium phosphate, pregelatinized starch, dicalcium phosphate, and starch; in some embodiments, the filler is selected from a mixture of microcrystalline cellulose and lactose.

In some embodiments, the filler comprises 50%-90% of the pharmaceutical formulation; in some embodiments, it comprises 60%-90%; in some embodiments, it comprises 70%-90%; in some embodiments, it comprises 50%-80%; in some embodiments, it comprises 60%-80%; in some embodiments, it comprises 55%-80%; in some embodiments, it comprises 70%-85%; and in some embodiments, it comprises 75%-82%.

In some embodiments, the content of microcrystalline cellulose in the pharmaceutical preparation is 30%-50% in any of the foregoing embodiments; in some embodiments, the content of microcrystalline cellulose in the pharmaceutical preparation is 30%-40%; in some embodiments, the lactose content is 30%-60% of the total content; in some embodiments, the lactose content is 30%-50% of the total content; in some embodiments, the lactose content in the pharmaceutical preparation is 40%-60%; in some embodiments, the lactose content in the pharmaceutical preparation is 45%-52%.

In some embodiments, the ratio of microcrystalline cellulose to lactose in any of the aforementioned embodiments is 1:0.5-1:2; in some embodiments, the ratio of microcrystalline cellulose to lactose is 1:1-1:1.8.

In some embodiments, in any of the foregoing embodiments, the disintegrant is selected from one or more of sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, crospovidone, crospovidone sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, and dry starch; in some embodiments, the disintegrant is selected from one or more of crospovidone, crospovidone sodium carboxymethyl cellulose, sodium carboxymethyl starch, and dry starch; in some embodiments, the disintegrant is selected from crospovidone sodium carboxymethyl cellulose.

In some embodiments, the disintegrant in any of the foregoing embodiments is present in a content of 1%-10% in the pharmaceutical preparation; in some embodiments, the content is 2%-8%; and in some embodiments, the content is 3%-7%.

In some embodiments, in any of the foregoing embodiments, the adhesive is selected from one or more of povidone, hydroxypropyl cellulose, hydroxypropyl methylcellulose, and methylcellulose; in some embodiments, the adhesive is selected from povidone.

In some embodiments, the adhesive content in the pharmaceutical preparation is 1%-5% in any of the foregoing embodiments; in some embodiments, the content is 1%-3%; and in some embodiments, the content is 3%-5%.

In some embodiments, in any of the foregoing embodiments, the flow aid is selected from one or more of talc, silica, micronized silica gel, polyethylene glycol, and magnesium dodecyl sulfate; in some embodiments, the flow aid is selected from silica.

In some embodiments, the gliding agent in the pharmaceutical preparation is 0.1%-3%; in some embodiments, the content is 0.1%-2%; in some embodiments, the content is 0.1%-1%; in some embodiments, the content is 0.5%-2%; in some embodiments, the content is 0.5%-1%; and in some embodiments, the content is 1%-3%.

In some embodiments, in any of the foregoing embodiments, the lubricant is selected from magnesium stearate, calcium stearate, stearic acid, and sodium stearate fumarate; in some embodiments, the lubricant is sodium stearate fumarate; in some embodiments, in any of the foregoing embodiments, the lubricant content in the pharmaceutical preparation is 0.1%-3%; in some embodiments, the lubricant content is 0.5%-3%; in some embodiments, the lubricant content is 0.5%-1%; in some embodiments, the lubricant content is 0.5%-2%; in some embodiments, the lubricant content is 1%-2%; in some embodiments, the lubricant content is 1%-3%.

The pharmaceutical formulation according to any one of the present invention comprises the active ingredient M in any of the foregoing embodiments and a pharmaceutical excipient, wherein the pharmaceutical excipient comprises povidone, silicon dioxide, croscarmellose sodium, microcrystalline cellulose, lactose, and sodium stearate fumarate.

In some embodiments, in any of the foregoing embodiments, the pH adjuster is selected from fumaric acid; in any of the foregoing embodiments, the pH adjuster has a content of 0%-10% or 1%-10% in the pharmaceutical preparation.

This invention provides a pharmaceutical formulation comprising the active ingredient M from any of the foregoing embodiments and a pharmaceutical excipient, wherein:

(i) The active ingredient M is preferably a compound of formula Ia, IIa, IIIa, IVa, or Va, and the content of active ingredient M is 0.5%-99%, 0.5%-47.5%, or 5%-35%, respectively; and optionally,

(ii) Pharmaceutical excipients include one or more of the following: fillers, binders, wetting agents, disintegrants, flow aids, and lubricants.

Preferably, the filler includes, but is not limited to, one or more of microcrystalline cellulose, mannitol, lactose, sucrose, sorbitol, dextran, pregelatinized starch, dicalcium phosphate, and starch;

Preferably, the adhesive includes, but is not limited to, one or more of polyvinylpyrrolidone, hydroxypropyl cellulose, hydroxypropyl methylcellulose, and methylcellulose;

Preferably, the wetting agent includes, but is not limited to, one or more of water and ethanol;

Preferably, the disintegrant includes, but is not limited to, one or more of sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, crospovidone, crospovidone sodium carboxymethyl cellulose, and calcium carboxymethyl cellulose;

Preferably, the gliding agent includes, but is not limited to, one or more of talc, silica, polyethylene glycol, and magnesium dodecyl sulfate;

Preferably, the lubricant includes, but is not limited to, magnesium stearate, calcium stearate, stearic acid, sodium stearate fumarate, and hydrogenated castor oil;

Optionally, the pharmaceutical formulation further includes one or more of the following: flavoring agents, antioxidants, preservatives, light-blocking agents, and film-coating premixes.

This invention provides a pharmaceutical formulation comprising the active ingredient M, a filler, a binder, a wetting agent, a disintegrant, a flow aid, and a lubricant as described in any of the foregoing embodiments, wherein the content of the active ingredient M is 0.5%-47.5%, the content of the filler is 50%-90%, the content of the disintegrant is 1%-10%, the content of the binder is 1%-5%, the content of the lubricant is 0.5%-3%, and the content of the flow aid is 0%-3%.

This invention provides a pharmaceutical formulation comprising the active ingredient M, a filler, a binder, a wetting agent, a disintegrant, a flow aid, and a lubricant as described in any of the foregoing embodiments, wherein the content of the active ingredient M is 5%-35%, the content of the filler is 50%-85%, the content of the disintegrant is 1%-10%, the content of the binder is 1%-5%, the content of the lubricant is 0.5%-3%, and the content of the flow aid is 0%-3%.

This invention provides a pharmaceutical formulation comprising the active ingredient M from any of the foregoing embodiments and a pharmaceutical excipient, wherein the pharmaceutical excipient comprises a filler, a disintegrant, a binder, a flow aid, and a lubricant.

The active ingredient M is preferably a compound of formula Ia, IIa, IIIa, IVa, or Va, and the content of active ingredient M is 0.5%-99%, 0.5%-47.5%, or 5%-35%, respectively.

The filler is a mixture of microcrystalline cellulose and lactose or microcrystalline cellulose, with a filler content of 50%-90% or 60%-80%; optionally, the ratio of microcrystalline cellulose to lactose is 1:0.5-1:2.

The disintegrant is croscarmellose sodium, and the content of the disintegrant is 1%-10% or 1%-5%.

The adhesive is polyvinyl acetate, and the adhesive content is 1%-5% or 1%-4%.

The lubricant is sodium stearate fumarate, with a lubricant content of 0.5%-3% or 1%-3%.

The gliding agent is silica, and its content is 0%-3%.

This invention provides a pharmaceutical formulation comprising the aforementioned active ingredient M, microcrystalline cellulose, lactose, croscarmellose sodium, povidone, sodium stearate, and silicon dioxide; in some embodiments, the pharmaceutical formulation comprises the aforementioned active ingredient M, microcrystalline cellulose, povidone K30, croscarmellose, and magnesium stearate; in some embodiments, the pharmaceutical formulation comprises the aforementioned active ingredient M, microcrystalline cellulose, lactose, croscarmellose sodium, povidone K30, and sodium stearate.

The present invention provides a pharmaceutical preparation comprising 5%-35% of active ingredient M, 30%-40% of microcrystalline cellulose, 30%-50% of lactose, 1%-5% of croscarmellose sodium, 1%-4% of povidone, 1%-3% of sodium stearate fumarate, and 0%-3% of silica.

Optionally, the pharmaceutical preparation described in any of the above embodiments can be prepared as a formulation selected from oral tablets, capsules, granules, and powders.

The pharmaceutical preparation of the present invention is prepared by one or more of the following processes: direct mixing, wet granulation, dry granulation, fluidized bed granulation, spray drying, and hot melt extrusion. The preferred processes are direct mixing, wet granulation, fluidized bed granulation, dry granulation, and spray drying. The more preferred processes are wet granulation and spray drying.

Optionally, in the pharmaceutical formulations described above, the binder may be added in a solution state or in a powder state; the disintegrant may be added internally, externally, or both.

The present invention also provides an application in the preparation of a pain-related medicament, wherein the administration method includes: oral administration, once a day, twice a day, three times a day, once a week, once every two weeks, or once a month.

This invention provides the use of a pharmaceutical preparation in the preparation of a medicament for pain, wherein the use is achieved by administering to a subject a pharmaceutical preparation comprising the aforementioned compound, wherein the dosage of the compound of formula (I) is selected from 1-2000 mg/dose, 1-1000 mg/dose, 1-800 mg/dose, 1-600 mg/dose, 1-400 mg/dose, 1-350 mg/dose, 1-300 mg/dose, 1-5 mg/dose, 5-10 mg/dose, etc. g/dose, 10-20mg/dose, 20-25mg/dose, 25-50mg/dose, 50-75mg/dose, 75-100mg/dose, 100-125mg/dose, 125-150mg/dose, 150-175mg/dose, 175-200mg/dose, 200-225mg/dose, 225-250mg/dose, 250-275mg/dose, 275-300mg/dose.

A method for treating a disease in a mammal, the method comprising administering an active ingredient M to a subject at a daily dose of 1-5000 mg/day, said daily dose being a single dose or divided doses, and in some embodiments, the daily dose including but not limited to 10-5000 mg/day, 25-5000 mg/day, 50-5000 mg/day, 100-4500 mg/day, 100-4000 mg/day, 100-3000 mg/day, 50-3000 mg/day, 50-2500 mg/day, 50-2000 mg/day, 50-1500 mg/day, 20-1500 mg/day, 50-1000 mg/day, 100-1000 mg/day, 10 0-800mg/day, 200-800mg/day, 25-400mg/day, 50-400mg/day, 100-400mg/day, 200-400mg/day. In some embodiments, the daily dose includes, but is not limited to, 10mg/day, 20mg/day, 25mg/day, 50mg/day, 100mg/day, 120mg/day, 125mg/day, 150mg/day, 200mg/day, 240mg/day, 400mg/day, 600mg/day, 800mg/day, 900mg/day, 1000mg/day, 1500mg/day, 2000mg/day, 3500mg/day, 4500mg/day, and 4000mg/day.

Unless otherwise stated, the terms used in this specification and claims have the following meanings.

"Product specification" refers to the weight of the active pharmaceutical ingredient (M) contained in each vial, tablet, or other unit of preparation.

"Excipients" refers to substances that are not therapeutic agents themselves, but are used as diluents, excipients, binders and/or mediators to be added to pharmaceutical preparations to improve their disposal or storage properties or to allow or facilitate the formation of unit dosage forms of compounds or pharmaceutical preparations for administration.

"Stereoisomers" are isomers that are produced by different spatial arrangements of atoms in a molecule, including cis-trans isomers, enantiomers, and conformational isomers.

The carbon, hydrogen, oxygen, sulfur, nitrogen, or F, Cl, Br, I involved in the groups and compounds described in this invention include their isotopes, and the carbon, hydrogen, oxygen, sulfur, or nitrogen involved in the groups and compounds described in this invention may optionally be further replaced by one or more of their corresponding isotopes, wherein the isotopes of carbon include ^12C , ^13C , and ^14C , the isotopes of hydrogen include protium (H), deuterium (D, also called heavy hydrogen), and tritium (T, also called superheavy hydrogen), the isotopes of oxygen include ^16O , ^17O , and ^18O , the isotopes of sulfur include ^32S , ^33S , ^34S , and ^36S , the isotopes of nitrogen include ^14N and ^15N , the isotopes of fluorine include ^17F and ^19F , the isotopes of chlorine include ^35Cl and ^37Cl , and the isotopes of bromine include ^79Br and ^81Br .

"alkyl" refers to a substituted or unsubstituted straight-chain or branched saturated aliphatic hydrocarbon group, including but not limited to alkyl groups with 1 to 20 carbon atoms, alkyl groups with 1 to 8 carbon atoms, alkyl groups with 1 to 6 carbon atoms, and alkyl groups with 1 to 4 carbon atoms. Non-limiting examples include methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, and their various branched isomers; the alkyl group can be monovalent, divalent, trivalent, or tetravalent.

"Alkoxy" refers to a substituted or unsubstituted -O-alkyl group. Non-limiting examples include methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, n-hexoxy, cyclopropoxy, and cyclobutoxy.

"Substituted" or "substituted" means substituted by one or more (including but not limited to 2, 3, 4, or 5) substituents, including but not limited to H, F, Cl, Br, I, alkyl, cycloalkyl, alkoxy, haloalkyl, thiol, hydroxyl, nitro, mercapto, amino, cyano, isocyano, aryl, heteroaryl, heterocyclic, bridged cycloyl, spirocyclic, fused cycloyl, hydroxyalkyl, =O, carbonyl, aldehyde, carboxylic acid, formate, -( _CH2 ) _m -C(=O) ^-Ra , -O-( _CH2 ) _m -C(=O) ^-Ra , -( _CH2 ) _m - ^C (=O) ^-NRbRc , -( _CH2 ) _mS (=O) _nRa , -( _CH2 ) _m - ^alkenyl - ^Ra , ^ORd , or -( _CH2 ) _m -alkynyl- ^Ra. (where m and n are 0 ^, 1, or 2), arylthio, thiocarbonyl, silyl, or ^-NRbRc groups, wherein ^Rb and ^Rc are independently selected from H, hydroxyl, amino, carbonyl, alkyl, alkoxy, cycloalkyl, heterocyclic, aryl, heteroaryl, sulfonyl, trifluoromethanesulfonyl, and alternatively, ^Rb and ^Rc can form five- or six-membered cycloalkyl or heterocyclic groups, and ^Ra and ^Rd are each independently selected from aryl, heteroaryl, alkyl, alkoxy, cycloalkyl, heterocyclic, carbonyl, ester, bridged cycloalkyl, spirocyclic, or fused cycloalkyl groups.

"1 to X substituents selected from..." means substituted by 1, 2, 3...X substituents selected from..., where X is any integer between 1 and 10. For example, "1 to 4 ^Rk substitutions" means substituted by 1, 2, 3, or 4 ^Rk substituents. Similarly, "1 to 5 substituents selected from..." means substituted by 1, 2, 3, 4, or 5 substituents selected from... For example, "the heterobridged ring is optionally substituted by 1 to 4 substituents selected from H or F" means the heterobridged ring is optionally substituted by 1, 2, 3, or 4 substituents selected from H or F.

Unless otherwise specified, the “content” of a substance in the pharmaceutical preparation of this application refers to the percentage of the weight of the substance in the total weight of the pharmaceutical preparation.

Detailed Implementation

The following embodiments illustrate the technical solution of the present invention in detail, but the scope of protection of the present invention includes, but is not limited to, these embodiments.

The compounds used in the reactions described herein were prepared according to organic synthesis techniques known to those skilled in the art, and were derived from commercially available chemicals and/or compounds described in chemical literature. “Commercially available chemicals” are obtained from legitimate commercial sources, and suppliers include: Titan Technology, Energetic Chemicals, Shanghai Demo, Chengdu Kelon Chemical, Shaoyuan Chemical Technology, Nanjing Yaoshi, WuXi AppTec, and Bailingwei Technology, among others.

The structure of the compounds was determined by nuclear magnetic resonance (NMR) and/or mass spectrometry (MS). NMR shifts (δ) are given in units of ^10⁻⁶ (ppm). NMR measurements were performed using Bruker Avance III 400 and Bruker Avance 300 NMR spectrometers in the following solvents: deuterated dimethyl sulfoxide (DMSO- _d₆₆ ), deuterated chloroform ( _CDCl₃ ), and deuterated methanol ( _CD₃OD ), with tetramethylsilane (TMS) as the internal standard.

MS determination was performed using (Agilent 6120B (ESI) and Agilent 6120B (APCI));

HPLC determinations were performed using an Agilent 1260DAD high-performance liquid chromatograph (Zorbax SB-C18 100×4.6mm, 3.5μM).

Thin-layer chromatography silica gel plates are Yantai Huanghai HSGF ₂₅₄ or Qingdao GF _254. The silica gel plates used for thin-layer chromatography (TLC) are 0.15mm-0.20mm in diameter, and the silica gel plates used for thin-layer chromatography separation and purification are 0.4mm-0.5mm in diameter.

Column chromatography typically uses Yantai Huanghai silica gel 200-300 mesh silica gel as the carrier;

Synthesis Examples

Example 1: Preparation of compounds 1-2a and 1-2b

Step 1: Preparation of compounds 1g-2a and 1g-2b

A mixture of 1f-2a and 1f-2b (0.148 g, 0.38 mmol) was dissolved in methanol (10 mL), and palladium on carbon (0.15 g, 1.41 mmol) was added. After the addition was complete, the mixture was pressurized to 2.5 MPa and reacted at 90 °C for 24 h under a hydrogen atmosphere. The reaction mixture was filtered and the filtrate was concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography to obtain a mixture of 1f-2a and 1f-2b (0.052 g, yield 34.35%).

Step 2: Preparation of compounds 1h-2a and 1h-2b

Under a nitrogen atmosphere and in an ice bath, a mixture of 1 g⁻²a and 1 g⁻²b (0.052 g, 0.13 mmol) was dissolved in tetrahydrofuran (5 mL), pre-cooled for 15 minutes, and potassium tert-butoxide (0.048 g, 0.43 mmol) was slowly added dropwise to the system (internal temperature <13 °C). After the addition was complete, the reaction was carried out in an ice bath for 2 h. Under an ice bath, 1 N hydrochloric acid was slowly added dropwise to the system (internal temperature <13 °C) until pH = 1, water (5 mL) was added, and the mixture was extracted with ethyl acetate (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a mixture of 1 h⁻²a and 1 h⁻²b.

LC-MS m/z = 369.0 [MH] ^-

Step 3: Preparation of compounds 1i-2a and 1i-2b

A mixture of 1h-2a and 1h-2b (0.048 g, 0.13 mmol) was dissolved in DMF (3 mL), followed by the sequential addition of 1h-2 (0.03 g, 0.20 mmol), TCFH (0.073 g, 0.26 mmol), and N-methylimidazole (0.022 g, 0.26 mmol). The reaction was carried out under nitrogen atmosphere at room temperature for 18 h. The reaction was quenched by adding water (10 mL), and the mixture was extracted with ethyl acetate (10 mL × 3). The organic phases were combined, washed with saturated brine (30 mL), collected, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by silica gel column chromatography to obtain a mixture of 1i-2a and 1i-2b (0.01 g, yield 15.25%).

LC-MS m/z = 505.2[M+H] ⁺

Step 4: Preparation of compounds 1-2a and 1-2b

A mixture of 1i-2a and 1i-2b (0.01 g, 0.02 mmol) was dissolved in 7 M ammonia-methanol solution (3 mL) and reacted at room temperature for 8 h. The reaction system was concentrated to obtain a crude product, which was then subjected to silica gel column chromatography to obtain a mixture of compounds 1-2a and 1-2b (0.006 g, yield 61.91%).

LC-MS m/z = 490.2[M+H] ⁺ ;

¹ H NMR (400MHz, DMSO-d ₆ ): δ10.92(s,1H),8.47(d,1H),8.17(d,1H),8.06-8.00(m,1H),7.70-7.64(m,1H),7.63-7.57(m,1H),7.22-7 .06(m,2H),4.86(d,1H),4.54-4.46(m,1H),4.01(d,3H),2.66-2.58(m,1H),1.85(s,3H),0.83-0.75(m,3H).

SFC preparation conditions:

Instrumentation: Waters 150Prep-SFC C; Preparative column: Chiralcel OX Column. Preparation method: The crude product was dissolved in acetonitrile to prepare a 10 mg/ml sample solution. Mobile phase: Carbon dioxide/methanol, methanol content 30%; Elution time: 8 min.

A mixture of 504 mg of compounds 1-2a and 1-2b was prepared by SFC, and after lyophilization, compounds 1-2P1 (193.8 mg, retention time: 1.137 min) and 1-2P2 (218 mg, retention time: 1.526 min) were obtained. Chirality test method: (Instrument: SHIMADZU LC-30AD sf, chiral column: Chiralcel OX Column. Preparation method: The crude product was dissolved in acetonitrile to prepare the sample solution. Mobile phase system: carbon dioxide/0.05% DEA in methanol solution. Elution gradient: 5%-40%; flow rate: 3.0 mL/min, elution time: 3 min).

Compound 1-2P1 has the structure of either formula 1-2a or 1-2b; and it is an enantiomer of compound 1-2P2, meaning that when compound 1-2P1 has the structure of formula 1-2a, compound 1-2P2 has the structure of formula 1-2b; and when compound 1-2P1 has the structure of formula 1-2b, compound 1-2P2 has the structure of formula 1-2a. Compounds 1-2P1 and 1-2P2 show no difference in NMR and mass spectrometry, and are consistent with mixtures of compounds 1-2a and 1-2b.

Example 2: Preparation of compounds 1-2a

Step 1: Preparation of compound 1h-2a

A mixture of 3.0 g of compounds 1h-2a and 1h-2b was prepared by SFC, and after lyophilization, compound 1h-2a (1.28 g, chiral HPLC retention time: 0.760 min) and compound 1h-2b (1.11 g, chiral HPLC retention time: 0.966 min) were obtained. Chiral HPLC test method: (Instrument: SHIMADZU LC-30AD, chiral column: Chiralcel IG column. Preparation method: The crude product was dissolved in acetonitrile to prepare the sample solution. Mobile phase system: carbon dioxide/0.05% DEA in ethanol solution. Elution gradient: 5%-40%; elution time: 3 min).

SFC preparation conditions: Instrument: Waters 150Prep-SFC A; Preparative column: Chiralcel IG column. Preparation method: The crude product was dissolved in acetonitrile to prepare a sample solution with a concentration of 2 mg/mL. Mobile phase system: carbon dioxide/ethanol, ethanol content 10%; flow rate: 100 mL/min; elution time: 2 min.

Compound 1h-2a:

¹ HNMR (400MHz, CDCl ₃ ): δ6.91-6.78(m,2H),4.53(d,1H),4.46-4.34(m,1H),4.02(d,3H),2.68-2.54(m,1H),1.85(s,3H),0.88-0.75(m,3H).

Step 2: Preparation of compound 1i-2a

1h-2a (1.0 g, 2.7 mmol) was dissolved in tetrahydrofuran (20 mL), followed by the addition of triethylamine (1.64 g, 16.20 mmol) and T3P (50% wt in EtOAc 6.88 g, containing 3.44 g, 10.79 mmol of 1-propylphosphonic anhydride). The mixture was stirred at room temperature for 15 minutes, then 1h-2 (0.62 g, 4.07 mmol) was added, and the reaction was carried out at room temperature under a nitrogen atmosphere for 18 hours. The mixture was then extracted with 40 mL of saturated sodium bicarbonate aqueous solution and 30 mL × 3 ethyl acetate. The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to give compound 1i-2a (0.919 g, yield 67.47%).

Step 3: Preparation of compounds 1-2a

Compound 1-2a (0.87 g) was obtained by referring to step 4 of Example 1;

The absolute configurations of compounds 1-2a were verified by Micro-ED.

Example 3: Preparation of compounds 27-1a and 27-1b

A mixture of 1h-2a and 1h-2b (0.052 g, 0.14 mmol) was dissolved in ethyl acetate (3 mL). Triethylamine (0.085 g, 0.84 mmol) and T3P (50% wt in EtOAc 0.36 g, containing 0.18 g, 0.56 mmol of 1-propylphosphonic anhydride) were added sequentially. 27-1a-1 (0.040 g, 0.21 mmol) was added to the system, and the reaction was carried out at room temperature under a nitrogen atmosphere for 18 h. The reaction was quenched by adding sodium bicarbonate aqueous solution (10 mL), extracted with ethyl acetate (10 mL × 3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was purified by prep-HPLC (condition 1) to obtain a mixture of 27-1a and 27-1b trifluoroacetate.

LCMS m/z = 541.1[M+H] ⁺

¹ H NMR (400MHz, DMSO-d ₆ )δ10.72(s,1H),8.17-8.09(m,1H),7.81-7.72(m,1H),7.37(t,1H),7.22-7.14(m,1H),7.11-7.04(m,1H),4 .83(d,1H),4.53-4.45(m,1H),4.00(d,3H),3.19(s,3H),2.65-2.57(m,1H),1.85(s,3H),0.83-0.74(m,3H).

Example 4: Preparation of compound 27-1a (compound of formula VIa)

Using 1h-2a as a substrate, 27-1a was synthesized under the same conditions as in Example 3.

LCMS m/z = 541.1[M+H] ⁺

Example 5: Preparation of compounds 3-1 and 3-2 (compounds IVa and Va)

Step 1: Synthesis of compound 3b

Substrate a (6005.69 mg, 20.7 mmol) was dissolved in dry 1,4-dioxane (65 mL), followed by the sequential addition of sodium methanethiol solid (2176.29 mg, 31.05 mmol), _Pd₂ (dba) _₃ (1895.54 mg, 2.07 mmol), Xant Phos (2395.49 mg, 4.14 mmol), and DIPEA (8025.80 mg, 62.10 mmol). After the addition was complete, the mixture was heated to 110 °C under a nitrogen atmosphere and reacted for 18 hours. The reaction mixture was cooled to room temperature, filtered, and the filter cake was washed with dichloromethane (10 mL × 3). The filtrate was collected and concentrated to obtain the crude product, which was then purified by silica gel column chromatography to obtain the target product 3b (3.4 g, yield 63.83%).

LCMS m/z = 202.0[M-55] ⁺

¹ H NMR (400MHz, CDCl ₃ ): δ7.40-7.34(m,1H),7.08-7.01(m,1H),6.94(t,1H),6.40(s,1H),2.47(s,3H),1.52(s,9H).

Step 2: Synthesis of compound 3c

Substrate 3b (3.4 g, 13.21 mmol) was dissolved in methanol (35 mL), and then ammonium carbamate (1.96 g, 25.10 mmol) and iodobenzene acetate (6.38 g, 19.82 mmol) were added sequentially under ice bath conditions. After the addition was complete, the reaction was continued at room temperature for 18 hours. The crude product was concentrated under reduced pressure and purified by silica gel column chromatography to obtain the target product 3c (2.23 g, yield 58.5%).

LCMS m/z = 289.1[M+1] ⁺

¹ H NMR (400MHz, CDCl ₃ ): δ7.85-7.80(m,1H),7.80-7.72(m,1H),7.16(t,1H),6.74(s,1H),3.27(s,3H),1.52(s,9H).

SFC preparation conditions:

Instrumentation: SFC Prep 150AP; Preparative column: IG (19mm × 250mm). Preparation method: The crude product was dissolved in methanol and filtered through a 0.45μm filter to prepare the sample solution. Mobile phase system: carbon dioxide/methanol (0.05% ammonia), methanol content 18%; flow rate: 43mL/min.

Compound 3c (2.23 g) was prepared by SFC and concentrated to obtain compound 3c-1 (1.07 g, retention time: 7.43 min) and compound 3c-2 (1.12 g, retention time: 11.9 min).

Step 3: Preparation of compound 3d-1

The substrate 3c-1 (430 mg, 1.5 mmol) was dissolved in 1,4-dioxane (2.5 mL), and then a solution of 1,4-dioxane in 4 M hydrochloric acid (2.5 mL) was added dropwise. The mixture was heated to 55 °C and reacted for 2 hours. The system was concentrated under reduced pressure to obtain crude 3d-1 (335 mg, 99.99% yield). The crude product was used directly in the next step without any purification.

LCMS m/z = 189.1[M+H] ⁺

Step 4: Preparation of compounds 3-1 and 3-2

Under ice bath conditions, a mixture of substrate 3d (370.33 mg, 1.0 mmol) was dissolved in DCM (5 mL), followed by the sequential addition of oxalyl chloride (253.86 mg, 2.0 mmol) and DMF (7.31 mg, 0.10 mmol). The mixture was stirred under ice bath conditions for 1 hour (LCMS analysis showed complete reaction of the starting materials). The system was concentrated to obtain crude acyl chloride. Then, 3d-1 (282.33 mg, 1.5 mmol) was dissolved in DCM (3 mL), followed by the sequential addition of triethylamine (505.95 mg, 5 mmol). Finally, acyl chloride dissolved in DCM (2 mL) was added dropwise to the system. After the addition was complete, the mixture was reacted at room temperature under a nitrogen atmosphere for 18 hours. The reaction was quenched by adding sodium bicarbonate aqueous solution (20 mL), extracted with ethyl acetate (10 mL x 3), the organic phases were combined, dried over anhydrous sodium sulfate, filtered and concentrated to obtain crude product, and purified by silica gel column chromatography to obtain compound 3-1 (235.0 mg, yield 43.48%).

LCMS m/z = 541.2[M+H] ⁺ ;

¹ H NMR (400MHz, DMSO-d ₆ )δ10.70(s,1H),8.13-8.07(m,1H),7.81-7.75(m,1H),7.36(t,1H),7.23-7.14(m,1H),7.12-7.04(m,1H),4.85(d ,1H),4.64(s,1H),4.53-4.45(s,1H),4.1(d,3H),3.15(s,3H),2.67-2.58(m,1H),1.86(s,3H),0.84-0.74(m,3H).

Crude 3d-2 (336 mg, 99.70% yield) was obtained from substrate 3c-2 (432.51 mg, 1.5 mmol) using the same synthetic method as 3d-1. The crude product was used directly in the next reaction without any purification.

LCMS m/z = 189.1[M+H] ⁺

Compound 3-2 (320.0 mg, 59.2% yield) was obtained from substrate 3d-2 (370.33 mg, 1.0 mmol) using the same synthetic method as compound 3-1.

LCMS m/z = 541.2[M+H] ⁺ ;

¹ H NMR (400MHz, DMSO-d ₆ )δ10.70(s,1H),8.16-8.11(m,1H),7.77-7.71(m,1H),7.35(t,1H),7.22-7.13(m,1H),7.10-7.04(m,1H),4.84(d ,1H),4.64(s,1H),4.52-4.45(s,1H),4.0(d,3H),3.14(s,3H),2.65-2.57(m,1H),1.85(s,3H),0.82-0.74(m,3H).

One of the structures in compound 3c-1 or compound 3c-2 is 3c-PA, and the other structure is 3c-PB.

One of the structures in compound 3d-1 or compound 3d-2 is 3d-PA, and the other structure is 3d-PB.

One of the structures in compound 3-1 or compound 3-2 is 3-A, and the other structure is 3-B.

The compound 3-1 was determined to be 3-A using MicroED technology.

Example 6: Preparation of compounds 8-1a and 8-1b

Preparation of compounds 8-1a and 8-1b

A mixture of 1h-2a and 1h-2b (0.050 g, 0.14 mmol) was dissolved in ethyl acetate (3 mL). Triethylamine (0.042 g, 0.42 mmol) and T3P (50% wt in EtOAc 0.35 g, containing 0.175 g, 0.55 mmol of 1-propylphosphonic anhydride) were added sequentially. 8-1-1 (0.040 g, 0.21 mmol, reference: WO 2020/028724) was added, and the reaction was carried out at room temperature under nitrogen atmosphere for 18 h. The reaction was quenched by adding sodium bicarbonate aqueous solution (10 mL), extracted with ethyl acetate (10 mL × 3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain a crude product. The crude product was prepared by prep-HPLC (condition 1) and lyophilized to obtain a mixture of 8-1a and 8-1b (0.007 g, yield 9.22%).

LCMS m/z = 543.2[M+H] ⁺

¹ H NMR (400MHz, CDCl ₃ )δ8.11(s,1H),7.98-7.92(m,1H),7.81-7.76(m,1H),7.17(t,1H),7.05-6.98(m,1H),6.91-6.83(m,1H), 5.04(s,2H),4.57(d,1H),4.46(dd,1H),4.04(d,3H),2.68-2.59(m,1H),1.92(s,3H),0.91-0.85(m,3H).

Example 7: Preparation of compound 8-1a (compound of formula Ia)

Compound 8-1a (95 mg, 62.9%) was purified by silica gel column chromatography using the synthetic method described in Example 6 for 1h-2a (100 mg, 0.27 mmol).

LCMS m/z = 543.2[M+H] ⁺

¹ H NMR (400MHz, CDCl ₃ ): δ8.54(s,1H),8.14-8.05(m,1H),7.74-7.65(m,1H),7.13(t,1H),7.04-6.95(m,1H),6.89-6.79(m,1H),5 .33(s,2H),4.67(d,1H),4.59-4.47(m,1H),4.04(d,3H),2.69-2.55(m,1H),1.91(s,3H),0.93-0.85(m,3H).

Example 8: Preparation of compounds 36-2a and 36-2b

Step 1: Preparation of compounds 36-1a and 36-1b

A mixture of 1h-2a and 1h-2b (50 mg, 0.14 mmol) was dissolved in a 2 M, 2 mL solution of oxalyl chloride in dichloromethane. DMF (0.002 g, 0.028 mmol) was added dropwise under ice bath conditions, and the reaction was carried out at room temperature for 2 h. After concentration, the crude product was added to a 2 mL solution of 4-amino-2-methoxypyridine (26 mg, 0.21 mmol) and triethylamine (42 mg, 0.42 mmol) in dichloromethane, and the reaction was carried out overnight at room temperature. The crude product was concentrated and purified by silica gel column chromatography to give a mixture of 36-1a and 36-1b (55 mg, 82.45% yield).

LCMS m/z = 477.1[M+H] ⁺

Step 2: Preparation of compounds 36-2a and 36-2b

A mixture of 36-1a and 36-1b (45 mg, 0.10 mmol) was dissolved in THF (1 mL) and HCl (1 mL) and reacted at 60 °C for 18 h. The reaction system was concentrated to obtain a crude product, which was then prepared by prep-HPLC (condition 1) to obtain a mixture of compounds 36-2a and 36-2b (4 mg, yield 8.65%).

LCMS m/z = 463.1[M+H] ⁺

Example 9: Preparation of compound 36-2a (compound of formula IIIa)

Using 1h-2a as a substrate, compound 36-2a was synthesized according to the synthesis method of Example 8.

LCMS m/z = 463.1[M+H] ⁺

Example 10: Preparation of compound 77 (compound of formula IIa)

Step 1: Preparation of compound 77b

77a (3.04 g, 20.0 mmol), (Boc)₂O (4.8 g, 22 mmol), and 4-dimethylaminopyridine (244 mg, 2.0 mmol) were added to dichloromethane (70 mL) and reacted at room temperature for 16 hours. The reaction solution was concentrated under reduced pressure and purified by column chromatography to obtain 77b (2.38 g, yield 33.8%).

LCMS m/z = 353.4[M+H] ⁺

Step 2: Preparation of compound 77c

77b (352 mg, 1.0 mmol) was added to 10 mL of dichloromethane and placed in an ice bath. Then, m-chloroperoxybenzoic acid (516 mg, 3.0 mmol) was added, and the reaction was continued at room temperature for 12 hours. The reaction solution was concentrated under reduced pressure and purified by silica gel column chromatography to obtain 77c (211 mg, 57.3% yield).

LCMS m/z = 369.2[M+H] ⁺

Step 3: Preparation of compound 77d

77c (134 mg, 0.364 mmol) was added to dichloromethane (2 mL), followed by 1,4-dioxane-hydrochloride solution (4 M, 4 mL). The mixture was reacted at room temperature for 2 hours. The reaction solution was then concentrated under reduced pressure, and dichloromethane (2 mL) was added again. The 1,4-dioxane-hydrochloride solution (4 M, 4 mL) was then added again, and the mixture was reacted for another 2 hours. The reaction solution was then concentrated under reduced pressure to obtain 77d (71 mg).

LCMS m/z = 169.1 [M+H] ⁺

Step 4: Preparation of compound 77e

77d (71 mg) was dissolved in tetrahydrofuran (8 mL), followed by the sequential addition of triethylamine (121.4 mg, 1.2 mmol) and T3P (50% wt in EtOAc 509 mg, containing 254.5 mg, 0.8 mmol of 1-propylphosphonic anhydride). Finally, substrate 1h-2a (74 mg, 0.2 mmol) was added to the system. After the addition was complete, the reaction was carried out at room temperature under a nitrogen atmosphere for 20 hours. The reaction was quenched by adding sodium bicarbonate aqueous solution (20 mL), extracted with ethyl acetate (20 mL × 3), and the organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude product. The crude product was purified by silica gel column chromatography to obtain 77e (81 mg, yield 77.8%).

LCMS m/z = 521.1[M+H] ⁺

Step 5: Preparation of Compound 77

Substrate 77e (81 mg, 0.155 mmol) was dissolved in 7 M ammonia-methanol solution (10 mL) and reacted at room temperature for 2 hours. The reaction system was concentrated to obtain a crude product, which was then subjected to silica gel column chromatography to obtain compound 77 (42 mg, yield 53.6%).

LCMS m/z = 506.1[M+H] ⁺ .

Reference compound 1:

Formulation Examples

1. Formulations 1-1 to 1-5

The above prescription preparations 1-5 use the following processes:

1) Weighing: Weigh each raw material and excipient according to the prescription (converted content of raw drug).

2) Mixing: Mix the active ingredient, binder, glidant, disintegrant, and filler for 5 minutes, then add the lubricant and mix for 2 minutes.

3) Tableting: The total mixed powder from step 2 is compressed into tablets using a shallow concave punch with a diameter of φ11mm, with the tablet weight controlled at 500mg±5% and the hardness at 80±30N/ ^mm² .

2. Formulations 1-6 to 1-10:

The prescription process is as follows:

Premix: Mix API, microcrystalline cellulose, and crospovidone in a wet granulator for 3 minutes to obtain a mixture;

Adhesive preparation: Prepare a 10% solids content polyvinyl ketone/(75% ethanol/water) solution (w/w);

Granulation: Run a wet granulator, slowly add 10% povidone solution, and continue stirring for about 120 seconds to make a soft material.

Granulation and drying: The soft material is wet-granulated by passing it through a 20-mesh sieve using a gyratory pelletizer to obtain wet pellets; the wet pellets are dried in an oven at 80℃, with the moisture content controlled below 5%, and then dry-granulated by passing it through a 20-mesh sieve using a gyratory pelletizer to obtain dry pellets for later use.

Total Mixture: Take whole dry granules, place them in a hopper mixer, add magnesium stearate, and mix for 5 minutes to obtain total mixed granules.

Tableting: Using a φ10mm shallow concave punch, tablets are pressed according to the theoretical tablet weight, controlling the tablet weight to be within ±5% of the theoretical tablet weight, with a hardness of 70N～130N, to obtain unprocessed tablets.

3. Formulations 1-11 to 1-15:

Preparation process:

1) Weighing: Weigh each raw material and excipient according to the prescription (converted content of raw drug).

2) Mixing: Mix the active ingredient, binder, glidant, disintegrant, and filler for 5 minutes, then add lubricant and mix for 2 minutes.

3) Tableting: The total mixed powder from step 2 is compressed into tablets using a shallow concave punch with a diameter of φ8mm, with the tablet weight controlled at 200mg±5% and the hardness at 70±30N/ ^mm² .

4. Formulations 1-16 to 1-20:

The preparation process is as follows:

Premix: Mix API with microcrystalline cellulose, croscarmellose sodium cellulose and povidone in a wet granulator for 3 minutes to obtain a mixture;

Granulation: Run the wet granulator, slowly add an appropriate amount of water, and continue stirring for about 120 seconds to make a soft material.

Granulation and drying: The soft material is wet-granulated by passing it through a 20-mesh sieve using a gyratory pelletizer to obtain wet pellets; the wet pellets are dried in an oven at 80℃, with the moisture content controlled below 5%, and then dry-granulated by passing it through a 20-mesh sieve using a gyratory pelletizer to obtain dry pellets for later use.

Total Mixture: Take whole dry granules, place them in a hopper mixer, add magnesium stearate, and mix for 5 minutes to obtain total mixed granules.

Tableting: Use a 20*10mm shaped punch to compress tablets according to the theoretical tablet weight, controlling the tablet weight to be within ±5% of the theoretical tablet weight, with a hardness of 110N～160N, to obtain unprocessed tablets.

Biological test example 1:

1.1 Nav1.8 Manual Patch Clamp Test

(1) Cell Culture

CHO cell lines stably expressing human Nav1.8 were cultured in Ham's F-12 medium containing 10% fetal bovine serum and 10 μg/mL Blasticidin, 200 μg/mL Hygromycin B, and 100 μg/mL Zeocin. The cell culture temperature was 37°C and the carbon dioxide concentration was 5%. After removing the old medium and washing once with PBS, 1 mL of 0.25% Trypsin-EDTA solution was added, and the cells were incubated at 37°C for approximately 1.5 min. Once the cells detached from the bottom of the dish, preheated complete medium (37°C) was added. The cell suspension was gently pipetted to separate aggregated cells. The cell suspension was transferred to sterile centrifuge tubes and centrifuged at 1000 rpm for 5 min to collect the cells. Cells were seeded in 6 cm cell culture dishes at a seeding density of 2.5 × ^10⁵ cells per dish (final volume 5 mL) for expansion or maintenance culture. To maintain cell electrophysiological activity, the cell density should not exceed 80%. Before patch-clamp assay, cells were separated with 0.25% Trypsin-EDTA, and 6.5 × ^10³ cells were seeded onto coverslips and cultured in 24-well plates (final volume 500 μL). Assay was performed 18 hours later.

(2) Compound preparation

The compound was dissolved in dimethyl sulfoxide (DMSO) to prepare a 30 mM DMSO stock solution. The stock solution was diluted to the test concentration with extracellular fluid (140 mM NaCl, 3.5 mM KCl, ₁ mM MgCl₂· _6H₂O , ₂ mM CaCl₂· _2H₂O , 10 mM D-Glucose, 10 mM HEPES and 1.25 mM _NaH₂PO₄ · _2H₂O , pH adjusted to 7.4 with NaOH) _. The final DMSO concentration of all test samples was 0.1%.

(3) Electrophysiological tests

First, a capillary glass tube was drawn into a recording electrode using a microelectrode drawing device. Then, the electrode, filled with intracellular fluid (50 mM CsCl, 10 mM NaCl, 10 mM HEPES, 60 mM CsF, and 20 mM EGTA, pH adjusted to 7.2 with CsOH), was placed into a microelectrode holder. Under an inverted microscope, the microelectrode manipulator was used to immerse the electrode in the extracellular fluid, and the electrode resistance (Rpip) was recorded. Next, the electrode was slowly brought into contact with the cell surface, and negative pressure was applied to aspirate and form a GΩ seal. Fast capacitance compensation was then performed, and negative pressure was continued to rupture the cell membrane, establishing a whole-cell recording mode. Finally, slow capacitance compensation was performed, and experimental parameters such as series resistance (Rs) were recorded. No leakage compensation was applied. Once the Nav1.8 current recorded in the whole cell stabilized, drug administration began, with each drug concentration acting for approximately 5 minutes (or until the current stabilized). A coverslip containing cells was placed in a recording bath under an inverted microscope. Blank control solution and the working solution of the test compound were perfused through the recording bath by gravity to act on the cells, with fluid exchange facilitated by a peristaltic pump. The current detected in the cells in the solution without the compound served as a control group. All electrophysiological experiments were performed at room temperature. The inhibitory rate of the compound on Nav1.8 was determined by calculating the relative percentage of peak currents generated before and after cell treatment.

The voltage stimulation protocol for whole-cell patch-clamp recording of Nav1.8 sodium current was as follows: After whole-cell sealing, the cell voltage was clamped at -120 mV. First, the voltage was stepped from -110 mV to -30 mV in 10 mV increments, maintained for 5 s, and then a 0 mV depolarization pulse was applied to obtain the half-inactivation voltage (Vhalf). Then, Vhalf was used as the stimulation voltage and maintained for 5 s. Next, the voltage was restored to -120 mV and maintained for 20 ms, followed by a depolarization pulse (TP2) to 0 mV for 50 ms to detect the sodium current in the half-inactivated state. Finally, the clamp voltage was restored to -120 mV, and data were repeatedly acquired every 20 ms to observe the effect of the drug on the peak sodium current. Experimental data were acquired using an EPC 10 amplifier (HEKA) and stored in PatchMaster (HEKA) software.

Table 1. _IC50 of the tested compounds against hNav1.8 inhibitory activity

Conclusion: The compounds of the present invention, such as the compounds in the examples, have good Nav1.8 inhibitory activity.

1.2 CYP3A4 Induction Activity Assay

1. Cell inoculation

1) DPX2 cells were cultured in growth medium containing 10% fetal bovine serum.

2) DPX2 cells were cultured in T-75 culture flasks in an incubator at 37°C, 5% _CO2 and 95% relative humidity. The cells were digested when they reached 80-90% confluence with the bottom of the culture flask.

3) Wash the surface of T-75 cultured cells with 10mL PBS, remove the PBS, add 3-5mL trypsin, digest at 37℃ for 5 minutes or until the cells are digested and suspended, and add excess culture medium containing fetal bovine serum to stop the trypsin digestion.

4) Transfer the cell suspension to a conical-bottom centrifuge tube and centrifuge at 150g for 5 minutes at room temperature. Carefully aspirate the supernatant, resuspend the cells in treatment medium, and adjust the concentration to 3.2 × ^10⁵ cells/mL (incubation time: 24 hours, seeding density: 4.0 × ^10⁵ cells/mL). Add 25 μL of cell suspension to each well of a 384-well cell culture plate. Incubate the cell culture plate at 37°C, 5% _CO₂ , and 95% relative humidity for 24 hours.

2. Compound preparation

1) Prepare stock solutions of the test compound, positive control (rifampin), and negative control (propranolol) at a concentration of 1000× using DMSO. The final concentrations of the positive control (rifampin) are 1 μM and 10 μM, and the final concentration of the negative control (propranolol) is 10 μM. The final concentrations of the test compound are 10, 1, 0.1 μM or 30, 10, 3, 1, 0.3, 0.1 μM. The final concentration of DMSO is 0.1%.

2) Remove the cell culture plate from the incubator and add 25 nL of positive and negative control drugs or test compound stock solutions directly using Echo, setting up three replicates for each concentration. Place the cell culture plate back into the incubator and continue incubation for 48 hours.

3) Before starting experiments using the substrate, check cell morphology and monolayer integrity to ensure that the monolayer has acceptable research quality.

3. Quantitative detection of PXR activation

1) After 48 hours of drug treatment, the culture can be used for quantitative detection of PXR activation.

2) Equilibrate the CellTiter-Fluor ^™ Cell Viability Assay Kit and One-Glo Luciferase reagent to room temperature. Add 10 μL of GF-AFC substrate to 10 mL of Assay Buffer to form a 2X reagent, then dilute with 10 mL of PBS to form a 1X reagent. Add ONE-Glo Luciferase substrate to ONE-Glo Luciferase Assay Buffer.

3) Remove the culture plate from the incubator, discard the culture medium, pour 1X CellTiter-Fluor ^™ reagent into the sample loading tank, add 25μL of reagent to each well of the culture plate using a pipette, and then incubate in the incubator for 30 minutes.

4) Remove the cell culture plate from the incubator, let it cool slightly to room temperature, and measure the fluorescence value using a fully automated quantitative microplate reader. The excitation light is 400nm and the emission light is 505nm.

5) Pour ONE-Glo reagent into the sample loading tank, add 25 μL to each well, gently mix the plate, incubate at room temperature for 5 minutes, and measure the luminescence value.

4. Data Analysis

All data was calculated using Microsoft Excel.

1) The activity of luciferase is represented by RFU/RLU, where RLU is the average luminescence intensity value of three parallels for each concentration of each compound, and RFU is the average fluorescence intensity value of three parallels for each concentration of each compound.

The activation fold of mRNA is calculated using the following formula:

Fold activation＝(RLU test/RFU test)/(RLUvehicle/RFUvehicle)

2) The cell viability percentage of the compound is calculated using the following formula:

Cell Viability%=(RFUtest/RFUvehicle)×100

3) The percentage relative to the positive control drug is calculated using the following formula:

Percent of positive control(%)=(Fold activation test/Fold activationPositive control)×100

The experimental results are shown in Table 2:

Table 2

Conclusion: The compounds of the present invention, such as the compounds in the examples, have no or weak CYP3A4 induction activity, and therefore have a lower risk of drug-drug interactions.

1.3 UGT1A1 Inhibitory Activity Assay

This experiment evaluated the inhibitory potential of test substances against UGT1A1 using recombinant human UGT1A1 enzyme. Bilirubin, a probe substrate for UGT1A1 enzyme, was co-incubated with recombinant human UGT1A1 enzyme and different concentrations (0–10 μM) of test substances. Uridine diphosphate glucuronide (UDPGA) was added to initiate the reaction. After the reaction, the samples were processed, and specific metabolites produced by bilirubin were quantitatively detected using liquid chromatography-tandem mass spectrometry (LC-MS/MS). Changes in UGT1A1 enzyme activity were measured, and the _IC50 value was calculated to evaluate the inhibitory potential of the test substances against each UGT1A1 enzyme.

The experimental results are shown in Table 3:

Table 3

Conclusion: The compounds of the present invention, such as the compounds in the examples, have weaker UGT1A1 inhibition and lower risk of hepatotoxicity compared to the control compounds.

Biological Test Example 2: Rat Pharmacokinetic Test

Experimental animals: Male SD rats, approximately 220g, 6-8 weeks old, 6 rats/compound.

Experimental design: On the day of the experiment, SD rats were randomly divided into groups according to their body weight. They were fasted for 12-14 hours before administration but allowed free access to water, and were fed 4 hours after administration.

Table 4. Dosage Information

Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Gavage administration solvent: 0.5% MC

(DMA: dimethylacetamide; Solutol: polyethylene glycol-15-hydroxystearate; Saline: physiological saline; MC: methylcellulose)

Blood samples of 0.10 mL were collected via the orbital cavity before and after isoflurane anesthesia, placed in EDTAK2 centrifuge tubes, and centrifuged at 5000 rpm for 10 min at 4°C to collect plasma. Blood collection time points for both the intravenous and gavage groups were 0, 5, 15, 30 min, 1, 2, 4, 6, 8, and 24 h. All samples were stored at -80°C before analysis and quantitative analysis was performed using LC-MS/MS.

Conclusion: The compounds of the present invention, such as the compounds in the examples, have good oral performance in rats.

Biological Test Example 3: Mouse Pharmacokinetic Test

Experimental animals: C57 mice, 22-25g, 6 mice/compound.

Experimental design: On the day of the experiment, C57 mice were randomly divided into groups according to body weight. They were fasted for 12-14 hours before administration but allowed free access to water, and were fed 4 hours after administration.

Table 5. Dosage Information

Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Gavage administration solvent: 0.5% MC

Blood samples of 0.06 mL were collected via the orbital cavity before and after isoflurane anesthesia, placed in EDTAK2 centrifuge tubes, and centrifuged at 5000 rpm for 10 min at 4°C to collect plasma. Blood collection time points for both the intravenous and gavage groups were 0, 5, 15, 30 min, 1, 2, 4, 7, 24, and 48 h. All samples were stored at -80°C before analysis and quantitative analysis was performed using LC-MS/MS.

Conclusion: The compounds of the present invention, such as the compounds in the examples, have good oral performance in mice.

Biological Test Example 4: Pharmacokinetic Test in Beagle Dogs

Experimental animals: Male beagles, approximately 8–11 kg, 6 per compound.

Experimental Methods: On the day of the experiment, beagles were randomly grouped according to their body weight. They were fasted for 12–14 hours prior to administration but allowed free access to water. Food was given 4 hours after administration. Administration was performed according to Table 6.

Table 6. Dosage Information

Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Gavage administration solvent: 0.5% MC

(DMA: dimethylacetamide; Solutol: polyethylene glycol-15-hydroxystearate; Saline: physiological saline; MC: methylcellulose solution;)

Blood samples (1 ml) were collected via jugular or limb veins before and after drug administration and placed in EDTAK2 centrifuge tubes. Plasma was collected by centrifugation at 5000 rpm and 4°C for 10 min. Blood collection time points for both the intravenous and gavage groups were: 0, 5, 15, 30 min, 1, 2, 4, 6, 8, 10, 12, 24, 48, and 72 h. All samples were stored at -80°C before analysis and quantitative analysis was performed using LC-MS/MS.

Table 7. Pharmacokinetic parameters of the tested compounds in beagle dog plasma.

Conclusion: The compounds of the present invention, such as the compounds in the examples, have good oral performance in dogs.

Biological Test Example 5: Monkey Pharmacokinetic Test

Experimental animals: male cynomolgus monkeys, 3-5 kg, 3-6 years old, 6 per compound.

Experimental method: On the day of the experiment, monkeys were randomly divided into groups according to their body weight. They were fasted for 14-18 hours before administration but allowed free access to water. They were fed 4 hours after administration.

Table 8. Drug Administration Information

Note: Intravenous administration solvent: 5% DMA + 5% Solutol + 90% Saline; Gavage administration solvent: 0.5% MC (containing 0.5% Tween)
80);

*Dosage is calculated based on free base.

Blood samples of 1.0 mL were collected from venous sites in the extremities before and after drug administration and placed in EDTAK2 centrifuge tubes. Plasma was collected by centrifugation at 5000 rpm and 4°C for 10 min. Blood collection time points for both the intravenous and gavage groups were: 0, 5 min, 15 min, 30 min, 1, 2, 4, 6, 8, 10, 12, and 24 h. All samples were stored at -80°C before analysis and quantitative analysis was performed using LC-MS/MS.

Conclusion: The compounds of the present invention, such as the compounds in the examples, have good oral performance in monkeys.

Biological Test Example 6: CYP450 Enzyme Inhibition Test

The aim of this study was to evaluate the effects of test substances on the activities of five isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4) of human liver microsomal cytochrome P450 (CYP) using an in vitro assay system. Specific probe substrates for CYP450 isoenzymes were co-incubated with human liver microsomes and different concentrations of the test substances. Reduced nicotinamide adenine dinucleotide phosphate (NADPH) was added to initiate the reaction. After the reaction, the metabolites produced by the specific substrates were quantitatively detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS) after sample processing. Changes in CYP enzyme activity were measured, _IC50 values were calculated, and the inhibitory potential of the test substances against each CYP enzyme isoform was evaluated.

Conclusion: The compounds of the present invention, such as the compounds in the examples, have weak CYP inhibition, specifically compounds 3-1, 3-2, and 77, with IC50 _≥20 μM for CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A4.

Biological Test Example 7: CaCO2 Permeability Test

The experiment used monolayers of Caco-2 cells, incubated in triple parallel in 96-well Transwell plates. A transport buffer solution (HBSS, 10 mM HEPES, pH 7.4 ± 0.05) containing either the compound of the present invention (2 μM) or the control compounds digoxin (10 μM), naldolol (2 μM), and metoprolol (2 μM) was added to the dosing well on the apical or basal side. A transport buffer solution containing DMSO was added to the corresponding receiving well. After incubation at 37 ± 1 °C for 2 hours, the cell plate was removed, and appropriate amounts of sample were transferred from both the apical and basal sides to new 96-well plates. Acetonitrile containing an internal standard was then added to precipitate the protein. The samples were analyzed using LC MS/MS to determine the concentrations of the compound of the present invention and the control compounds. The concentration data were used to calculate the apparent permeability coefficients for transport from the apical to the basal side of the monolayer cells, and from the basal side to the apical side, thereby calculating the efflux rate. Leakage of fluorescein was used to evaluate the integrity of the monolayer cells after 2 hours of incubation.

Conclusion: The compounds of the present invention, such as the compounds in the examples, have good permeability.

Biological Test Example 8: Spinal Nerve Ligation (SNL) Induced Mouse Model of Neuropathic Pain

Male C57BL/6J mice purchased from Zhejiang Vital River Laboratory Animal Technology Co., Ltd. were acclimatized for one week before the model was established. The specific establishment method is as follows:

(1) Disinfection of surgical instruments and ligation sutures;

(2) Anesthetize the mice with isoflurane and place them in a prone position on the operating table;

(3) Clip the hair and prepare the skin near the hip bone of the mouse, and make an incision of about 2 cm along the spine;

(4) Dissect the fascia along the spine, bluntly dissect the muscles, and expose the L5 transverse process;

(5) Carefully bite off the L5 transverse process with tweezers to expose the L5 spinal nerve;

(6) Carefully separate the L5 nerve with a glass needle and ligate the L5 nerve with 5-0 ligation suture;

(7) Suture the muscles and skin, and disinfect with povidone-iodine;

Mice that failed to establish the model were culled the day after model establishment (successful model indicator: hind paw curled). After model establishment, mice were petted for 3-5 minutes daily to ensure familiarity with the experimenters, followed by placing them on a metal pain assessment frame for 40-60 minutes to acclimatize. After environmental acclimatization on day 3, Von Frey fibers (…) were used… The baseline values of the test animals (0.16, 0.4, 0.6, 1.0, 1.4, and 2.0 g) before drug administration (Ascending test method) were measured twice per animal, and the average value was taken, with an interval of at least 5 minutes between each measurement. The animals were grouped according to their baseline values (10 animals per group). After grouping, the test compound (3 and 30 mg/kg) or solvent (0.5% methylcellulose) was administered by gavage. The mechanical pain threshold (MPT) of the mice was measured at different time points after drug administration. Time-MPT curves were plotted using GraphPad 8.3.0, and statistical analysis was performed.

Conclusion: Based on the area under the time-MPT curve analysis, the compounds of the present invention, such as the compounds in the examples, have significant analgesic effects.

Claims (21)

A pharmaceutical formulation comprising an active ingredient M and a pharmaceutical excipient, wherein the active ingredient M is selected from compounds of general formula (I) or their stereoisomers, tautomers, or pharmaceutically acceptable salts.
_Q1 is selected from Preferably, _Q1 is selected from The _Q1 can be optionally replaced by 1 to 5 ^Rq ; B is selected from Or phenyl, wherein the B is optionally substituted with 1 to 4 ^RBs ; R ^Q1 is selected from H, COOH, NR ^q1 R ^q2 , -C(=O)NR ^q1 R ^q2 , -S(=O) ₂ NR ^q1 R ^q2 , OH, =O, -OR ^q1 , -C(=O)R ^q1 , -S(=O) ₂ R ^q1 , -S(=O)(=NR ^q1 )R ^q2 or -P(=O)R ^q1 R ^q2 ; R ^q1 and R ^q2 are each independently selected from H, methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, or cyclopentyl, wherein the methyl, ethyl, propyl, isopropyl, cyclopropyl, cyclobutyl, or cyclopentyl group is optionally substituted by 1 to 4 R ^k groups. ^Rq and ^RB are each independently selected from H, F, Cl, Br, cyano, _CH2F , _CHF2 , _CF3 , _-OCH2F , -OCHF2, _-OCF3 , _-OCD3 , methyl, -S- _methyl , -S- _CF3 , ethyl, isopropyl, ethynyl, methoxy, ethoxy, isopropyloxy, propyloxy, cyclopropyl, -O-cyclopropyl, -P(=O)( _CH3 ) ₂ , -P(=O)( _CH2CH3 ) ₂ or -P(=O)( _{CH3 )} (cyclopropyl), wherein the methyl, ethyl, isopropyl, ethynyl, methoxy, ethoxy, isopropyloxy, propyloxy, and cyclopropyl groups are optionally substituted by 1 to 4 ^{Rk groups} ; R ^k is selected from deuterium, =O, F, Cl, Br, I, CN, OH, NH ₂ , NH(CH ₃ ), NH(CH ₂ CH ₃ ), N(CH ₃ ) ₂ , N(CH ₂ CH ₃ ) ₂ , methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, -O-cyclopropyl, -NH-cyclopropyl, -CH ₂ -cyclopropyl, -CH ₂ -cyclobutyl, -CH ₂ -cyclopentyl, -CH ₂ -cyclohexyl, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl, wherein the methyl, ethyl, vinyl, ethynyl, methoxy, ethoxy, methylthio, cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl groups are optionally substituted by 1 to 4 substituents selected from deuterium, halogen, CN, OH, NH ₂ , C _1-4 alkyl or C _1-4 alkoxy groups; The pharmaceutical preparation contains 1-800 mg of active ingredient M; The excipients include one or both of fillers and disintegrants; The content (wt%) of the active ingredient M is 1%-99%; The total content of all components in the pharmaceutical preparation is 100%. The pharmaceutical preparation according to claim 1, wherein, Selected from _Q1 can be optionally replaced by 1 to 3 ^Rq ; ^RQ1 is selected from H, COOH, _NH2 , -C(＝O) _NH2 , -S(＝O _{)2NH2 ,} OH, ＝O, -S(＝O) _2CH3 , _-S (＝O) ₂ -cyclopropyl, -S(＝O)(＝NH) _CH3 , -P(＝O)( _CH3 ) ₂ , -P(＝O) _{( CH2CH3} ) ₂ or -P(＝O)( _CH3 )(cyclopropyl); R ^qa is selected from _-CH₂OH , _{-CF₂CH₂OH} , _NH₂ , -P(＝O)( _CH₃ ) _₂ , -P(＝O ₎ ( _CH₂CH₃ ) _₂ or -P(＝O)( _CH₃ )( _cyclopropyl ), preferably ^Rqa is selected from... or -CH ₂ OH; Preferably, Selected from _Q1 can be optionally replaced by 1 to 3 ^Rq ; B is selected from Preferred R and ^q are each independently selected from F, Cl, Br, cyano, _CH2F , _CHF2 , or _CF3 . According to claim 2, the structure of the compound of general formula (I) is selected from one of the structures shown in Table S-1. According to claim 3, the pharmaceutical formulation wherein the compound of general formula (I) is selected from the following structures:
The pharmaceutical preparation according to any one of claims 1-4, wherein the pharmaceutical preparation comprises 5-350 mg, 5-300 mg, 10-200 mg, 15-150 mg, 5-100 mg, 10-100 mg, 100-400 mg or 100-800 mg of active ingredient M, preferably 5 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, 50 mg, 55 mg, 60 mg, 65 mg, 100 mg, 120 mg, 150 mg, 200 mg, 250 mg, 300 mg or 400 mg. According to any one of claims 1-4, the weight ratio of the active ingredient M to the pharmaceutical excipient is 1:0.01 to 1:100, 1:0.5 to 1:10, or 1:1.5 to 1:9; or the content of the active ingredient M is 1%-80%, 1%-50%, 0.5%-47.5%, 1%-40%, 5%-40%, 5%-35%, 5%-30%, or 5%-20%. According to the pharmaceutical formulation of claim 6, optionally, the filler is selected from one or more of microcrystalline cellulose, mannitol, lactose, sucrose, sorbitol, dextran, anhydrous dicalcium phosphate, pregelatinized starch, dicalcium phosphate and starch, preferably a mixture of microcrystalline cellulose and lactose; The disintegrant is selected from one or more of sodium carboxymethyl starch, low-substituted hydroxypropyl cellulose, crospovidone, crospovidone sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, and dry starch, preferably from one or more of crospovidone, crospovidone sodium carboxymethyl cellulose, sodium carboxymethyl starch, and dry starch. According to claim 7, the content of the filler in the pharmaceutical formulation is 50%-90%, 70%-85%, 60%-80%, or 75%-82%. According to claim 7, the filler is a mixture of microcrystalline cellulose and lactose, wherein the content ratio of microcrystalline cellulose to lactose is 1:0.5-1:2, preferably 1:1-1:1.8, and optionally, the content of microcrystalline cellulose in the pharmaceutical preparation is 30%-50% or 30%-40%, and the content of lactose in the pharmaceutical preparation is 30%-60%, 40%-60%, or 45%-52%. According to claim 7, the disintegrant is present in the pharmaceutical preparation at a concentration of 0%-10%, 2%-8%, or 3%-7%. The pharmaceutical formulation according to any one of claims 1-4, wherein the pharmaceutical excipient further comprises one or more of a binder, a flow aid, a lubricant, and a pH adjuster. According to the pharmaceutical formulation of claim 11, optionally, the binder is selected from one or more of povidone, hydroxypropyl cellulose, hydroxypropyl methylcellulose and methylcellulose; The gliding agent is selected from one or more of talc, silica, micronized silica gel, polyethylene glycol, and magnesium dodecyl sulfate; The lubricant is selected from magnesium stearate, calcium stearate, stearic acid, and sodium stearate fumarate; The pH adjuster is selected from fumaric acid. According to claim 11, the content of the adhesive in the pharmaceutical preparation is 0%-5%, 1%-5%, 1%-3%, or 3%-5%. According to claim 11, the content of the gliding agent in the pharmaceutical preparation is 0%-3%, 0.1%-2%, 0.1%-1%, 0.5%-2%, 0.5%-1%, or 1%-3%. According to claim 11, the lubricant is present in the pharmaceutical preparation at a content of 0%-3%, 0.5%-2%, 1%-2%, or 1%-3%. According to claim 11, the pH adjuster is present in a concentration of 0%-10% or 1%-10% in the pharmaceutical preparation. A pharmaceutical formulation comprising the active ingredient M as described in any one of claims 1-4, a filler, a disintegrant, a binder, a flow aid, and a lubricant, wherein the content of the active ingredient M is 0.5%-47.5%, the content of the filler is 50%-90%, the content of the disintegrant is 1%-10%, the content of the binder is 1%-5%, the content of the lubricant is 0.5%-3%, and the content of the flow aid is 0%-3%. The pharmaceutical preparation according to claim 17, wherein, (i) Active ingredient M, with a content of 0.5%-47.5%; (ii) The filler is a mixture of microcrystalline cellulose and lactose, with a filler content of 50%-90%, preferably a microcrystalline cellulose to lactose content ratio of 1:0.5-1:2; (iii) Disintegrant croscarmellose sodium carboxymethyl cellulose, with a content of 1%-10%; (iv) Adhesive polyvinyl acetate, with a content of 1%-5%; (v) Lubricant: sodium stearate fumarate, content: 0.5%-3%; (vi) Flow aid silica, with a content of 0%-3%. The pharmaceutical formulation according to any one of claims 1-18, wherein the pharmaceutical excipient further comprises one or more of flavoring agents, antioxidants, preservatives, light-blocking agents, and film-coating premixes. The pharmaceutical preparation according to any one of claims 1-19 is in the form of oral tablets, capsules, granules or powders. The use of the pharmaceutical preparation according to any one of claims 1-20 in the preparation of a pain-related medicine.