Crystal form of a heterocyclic compound and pharmaceutical compositions thereof

Abstract

The present application belongs to the field of pharmaceutical chemistry, and particularly relates to a crystal form of a heterocyclic compound and a pharmaceutical composition thereof. Specifically, the present application relates to a crystal form of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiolane-2-formamidyl)pyridine-2-formamide and a pharmaceutical composition thereof. The crystal form of the present application has good crystallinity, excellent properties in terms of chemical stability and crystal form stability, high process safety, and good pharmacological effects in various animal models.

Description

[0001] This application is a divisional application of application number 202411718609.2, filed on November 28, 2024, entitled "Crystal form of heterocyclic compound and pharmaceutical composition thereof". Technical Field

[0002] This invention belongs to the field of pharmaceutical chemistry and relates to the crystal form of heterocyclic compounds and their pharmaceutical compositions. Specifically, this invention relates to the crystal form of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiocyclopentane-2-carboxamido)pyridine-2-carboxamide and its pharmaceutical compositions. Background Technology

[0003] Opioids are alkaloids and their analogues that interact with specific receptors in the central nervous system. They are commonly used for pain relief in pain disorders, but repeated use can lead to tolerance and addiction. While effective, opioids are often accompanied by numerous problems related to safety, tolerability, and addiction. Therefore, the development of non-opioid acute pain medications has enormous potential market potential.

[0004] In recent years, researchers have discovered that NaV1.8 is an important sodium ion channel related to pathology and pain, playing a crucial role in pain signal transduction in the peripheral nervous system, particularly in chronic neuropathic and chronic inflammatory pain. This mechanism of action is completely different from opioids; it does not act on the brain but directly blocks pain in the peripheral nervous system. It has a relatively high safety profile, is not addictive, and lacks the gastrointestinal and cardiovascular side effects of nonsteroidal anti-inflammatory drugs (NSAIDs). It can also be used in combination with other analgesics to enhance efficacy and reduce side effects.

[0005] Researchers have discovered a series of NaV1.8 blockers, including the small-molecule NaV1.8 blocker VX-548 from Vertex Technologies in the United States. Vertex Technologies has also made structural modifications based on VX548, such as WO2022256676. We have also discovered a class of NaV1.8 blockers, which are described in Chinese patent application CN202311551257.1.

[0006] The regulatory mechanism of Nav1.8 expression remains unclear, but further research is needed to address its clinical needs, which include high selectivity, strong efficacy, low side effects, and non-addictiveness.

[0007] Through extensive research, this application has discovered that the compounds of the present invention possess properties that are more suitable for medical and pharmaceutical applications under certain conditions, such as better solubility, bioavailability, physical and / or chemical stability, purity and impurity characteristics, filtration properties, drying properties, hygroscopicity, or easier processing. In addition to facilitating industrial production, the compounds of the present invention exhibit excellent efficacy and good safety in various models. Summary of the Invention

[0008] The first aspect of the present invention provides crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carbamate.

[0009] A second aspect of the invention provides a pharmaceutical composition comprising the crystal form A and one or more pharmaceutically acceptable carriers.

[0010] A third aspect of the invention provides a formulation comprising the crystal form A and one or more pharmaceutically acceptable carriers.

[0011] A fourth aspect of the invention provides a medicine box containing the crystal form A or the pharmaceutical composition or formulation, optionally further comprising one or more other therapeutic agents, and instructions for use.

[0012] The fifth aspect of the invention provides the use of the above-described crystal form A, pharmaceutical composition, formulation, or cassette in the preparation of a medicament for the prevention and / or treatment of NaV1.8-related diseases, preferably, said NaV1.8-related diseases being pain.

[0013] A sixth aspect of the present invention provides the above-described crystal form A, pharmaceutical composition, formulation, or cassette for the prevention and / or treatment of NaV1.8-related diseases, preferably, said NaV1.8-related diseases being pain.

[0014] A seventh aspect of the invention provides a method for preventing and / or treating NaV1.8-related diseases, comprising administering to an individual a therapeutically effective amount of the aforementioned crystal form A, pharmaceutical composition, formulation, or kit; preferably, the NaV1.8-related disease is pain.

[0015] The present invention has the following beneficial effects: Compound 1 of the present invention has crystal form A, high crystallinity, high purity, uniform particle size distribution, no hygroscopicity, excellent thermal stability and high process safety, simple operation, and is suitable for industrial production. The compound of the present invention has good solubility and bioavailability. The compound of the present invention also shows excellent efficacy and good safety in multiple models. Attached Figure Description

[0016] Figure 1 The image shows the XRPD pattern of crystal form A of compound 1. Detailed Implementation

[0017] The present invention will be further illustrated by the following embodiments. The embodiments of the present invention are only used to illustrate the technical solutions of the present invention and are not intended to limit the scope of the present invention. Those skilled in the art can make some non-essential improvements and adjustments, which still fall within the protection scope of the present invention.

[0018] definition

[0019] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. In case of any conflict, the definitions herein shall prevail. When a quantity, concentration, or other value or parameter is expressed as a range, preferred range, or preferred upper and lower limits of a numerical value, it should be understood that this is equivalent to specifically disclosing any range by combining any pair of upper or preferred values ​​with any lower or preferred values ​​of a range. Unless otherwise stated, the numerical ranges listed herein are intended to include the endpoints of the range and all integers and fractions (decimals) within that range.

[0020] As used in this article, “Compound 1” refers to compound 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carbamate.

[0021] When used with a numerical variable, the term "about" usually means that the value of the variable and all values ​​of the variable are within the experimental error (e.g., within a 95% confidence interval for the mean) or within ±20%, ±10%, ±5%, or ±2% of the specified value.

[0022] The term "comprising" or similar expressions such as "including," "containing," and "having" are open-ended and do not exclude additional unlisted elements, steps, or components. The expression "consisting of" excludes any unspecified elements, steps, or components.

[0023] The term "substantially composed of" refers to a scope limited to the specified elements, steps, or ingredients, plus optional elements, steps, or ingredients that do not substantially affect the essential and novel features of the claimed subject matter. It should be understood that the term "comprising" and similar terms encompass the terms "substantially composed of" and "composed of".

[0024] As used herein, the terms “optional” or “optionally” mean that the event or situation subsequently described may or may not occur, including both the occurrence and non-occurrence of the event or situation.

[0025] Unless otherwise stated, all percentages, parts, etc. in this document are by weight.

[0026] As used herein, the term “crystal form” or “crystal” refers to any solid substance exhibiting a three-dimensional arrangement, as opposed to amorphous solid substances, which produce characteristic XRPD patterns with clearly defined peaks.

[0027] As used herein, the term "seed crystal" refers to an additive that can form a crystal nucleus in a crystallization process, thereby accelerating or promoting the growth of enantiomers with the same crystal form or stereoconfiguration.

[0028] As used herein, the terms “X-ray powder diffraction pattern” or “XRPD pattern” refer to an experimentally observed diffraction pattern or the parameters, data, or values ​​derived from it. XRPD patterns are typically characterized by peak position (x-axis) and / or peak intensity (y-axis).

[0029] As used herein, the terms “diffraction angle” or “2θ” refer to the peak position, expressed in degrees (°), based on the setup of an X-ray diffraction experiment, and are typically the horizontal axis unit in a diffraction pattern. If the incident beam is diffracted when it forms an angle θ with a lattice plane, the experimental setup requires recording the reflected beam at a 2θ angle. It should be understood that specific 2θ values ​​for a particular crystal form mentioned herein are intended to represent 2θ values ​​(expressed in degrees) measured using the X-ray diffraction experimental conditions described herein. For example, as described herein, using… Monochromatic radiation. The XRPD spectra in this paper were preferably acquired using a PANalytacal X'Pert3 Powder X-ray powder diffractometer, and the transmission mode was preferably acquired using a PANalytacal X'Pert3 Powder X-ray powder diffractometer.

[0030] As used herein, the terms "substantially identical" or "substantially as shown in Figure ×" for X-ray diffraction peaks mean that representative peak positions and intensity variations are taken into account. For example, those skilled in the art will understand that peak positions (2θ) will show some variation, typically up to 0.1 to 0.2 degrees, and that the instrument used to measure diffraction will also cause some variation. Furthermore, those skilled in the art will understand that relative peak intensities will vary due to differences between instruments, as well as the degree of crystallinity, preferred orientation, the surface of the prepared sample, and other factors known to those skilled in the art.

[0031] Similarly, as used herein, the phrase “basically as shown in Figure ×” for DSC and TGA spectra is intended to cover variations associated with these analytical techniques known to those skilled in the art. For example, for well-defined peaks in DSC spectra, there can typically be variations of up to ±5°C, and even greater variations (e.g., up to ±10°C) for broad peaks.

[0032] As used in this article, the term "room temperature" refers to 20℃±5℃.

[0033] As used in this article, the term “prevention” includes suppressing and delaying the onset of disease, and includes not only prevention before the development of disease, but also prevention of recurrence of disease after treatment.

[0034] As used herein, the term “treatment” means reversing, alleviating, or eliminating a targeted disease or symptom. A subject is considered successfully “treated” if, upon receiving a therapeutic amount of a salt of a compound of the present invention or a compound in its crystalline form, or a pharmaceutical composition of the present invention, at least one indicator and symptom of the subject shows observable and / or detectable relief and / or improvement. It is understood that treatment includes not only complete treatment but also the achievement of some biologically or medically relevant outcome without achieving complete treatment. Specifically, “treatment” means that a salt of a compound of the present invention or a compound in its crystalline form, or a pharmaceutical composition of the present invention, can achieve at least one of the following effects: (1) inhibiting disease (i.e., preventing further development of pathology and / or symptomology) in animals experiencing or exhibiting disease pathology or symptomology; (2) improving disease (i.e., reversing pathology and / or symptomology) in animals experiencing or exhibiting disease pathology or symptomology. Invention Details

[0036] Compound 1, crystal form A

[0037] The present invention provides crystalline form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxamide, wherein the XRPD pattern of crystalline form A includes characteristic peaks at diffraction angles (2θ) at 12.8±0.2°, 14.5±0.2° and / or 21.9±0.2°.

[0038] In some embodiments, the XRPD pattern of crystal form A has a diffraction peak intensity of 100% at a diffraction angle (2θ) of 14.5° ± 0.2° and a diffraction peak intensity of ≥ 70% at a diffraction angle (2θ) of 12.8° ± 0.2°.

[0039] In some embodiments, the XRPD pattern of crystal form A includes characteristic peaks at diffraction angles (2θ) at 12.8±0.2°, 13.2±0.2°, 14.5±0.2°, 19.3±0.2° and / or 21.9±0.2°.

[0040] In some embodiments, the XRPD pattern of crystal form A includes characteristic peaks at diffraction angles (2θ) at 12.8±0.2°, 13.2±0.2°, 14.5±0.2°, 17.8±0.2°, 19.3±0.2° and / or 21.9±0.2°.

[0041] In some embodiments, the XRPD pattern of crystal form A includes characteristic peaks at diffraction angles (2θ) at 12.8±0.2°, 13.2±0.2°, 14.5±0.2°, 17.8±0.2°, 19.3±0.2°, 20.6±0.2°, 21.9±0.2°, 25.0±0.2°, 25.6±0.2° and / or 27.0±0.2°.

[0042] In some embodiments, the XRPD pattern of crystal form A includes characteristic peaks at diffraction angles (2θ) at 12.8±0.2°, 13.2±0.2°, 14.5±0.2°, 16.0±0.2°, 17.8±0.2°, 18.6±0.2°, 19.3±0.2°, 19.6±0.2°, 20.6±0.2°, 21.4±0.2°, 21.9±0.2°, 23.6±0.2°, 24.3±0.2°, 25.0±0.2°, 25.6±0.2°, 26.0±0.2°, 27.0±0.2°, 27.9±0.2°, 28.5±0.2°, 29.3±0.2°, and 36.3±0.2°.

[0043] In some embodiments, the XRPD pattern of crystal form A includes... Figure 1 The peaks shown are at essentially the same diffraction angle (2θ).

[0044] In some embodiments, the XRPD pattern of crystal form A is substantially as follows: Figure 1 As shown.

[0045] In some embodiments, the XRPD pattern of crystal form A is as follows: Figure 1 As shown.

[0046] Pharmaceutical compositions, formulations and packaging

[0047] The pharmaceutical composition provided by the present invention comprises crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carboxamido)pyridine-2-carboxamide, and one or more pharmaceutically acceptable carriers.

[0048] In some embodiments, the pharmaceutical composition comprises 0.01-10000 mg of crystal form A, and one or more pharmaceutically acceptable carriers.

[0049] In some embodiments, the pharmaceutical composition comprises 0.03-8000 mg of crystal form A, for example 0.05-8000 mg, 0.1-8000 mg, 10-8000 mg, 10-1000 mg, 10 mg, 20 mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, 90 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, etc. g, 250mg, 300mg, 350mg, 400mg, 450mg, 500mg, 550mg, 600mg, 650mg, 700mg, 750mg, 800mg, 850mg, 900mg, 950mg, 1000mg , 1500mg, 2000mg, 2500mg, 3000mg, 3500mg, 4000mg, 4500mg, 5000mg, 5500mg, 6000mg, 6500mg, 7000mg, 7500mg, 8000mg.

[0050] The formulation provided by the present invention comprises crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carboxamido)pyridine-2-carboxamide, and one or more pharmaceutically acceptable carriers.

[0051] In some embodiments, the formulation comprises 0.01-10000 mg of crystal form A, and one or more pharmaceutically acceptable carriers.

[0052] The medicine box provided by the present invention contains crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carbamate, or the pharmaceutical composition or the formulation thereof, optionally further comprising one or more other therapeutic agents.

[0053] Treatment methods and uses

[0054] This invention provides the use of crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxamide, pharmaceutical compositions, formulations, or kits in the preparation of medicaments for the prevention and / or treatment of NaV1.8-related diseases, preferably, said NaV1.8-related diseases being pain.

[0055] This invention provides crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxamide, pharmaceutical compositions, formulations, or kits for the prevention and / or treatment of NaV1.8-related diseases, preferably, said NaV1.8-related diseases being pain.

[0056] The present invention provides a method for preventing and / or treating NaV1.8-related diseases, comprising administering to an individual a therapeutically effective amount of crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carboxamido)pyridine-2-carboxamide, a pharmaceutical composition, a formulation, or a kit; preferably, the NaV1.8-related disease is pain.

[0057] In some implementations, the pain is selected from chronic pain, acute pain, inflammatory pain, cancer pain, ICU analgesia, fracture or postoperative pain (e.g., pain from bunion removal, hernia repair, and abdominoplasty), neuropathic pain (e.g., neuralgia from peripheral neuropathy, postherpetic neuralgia, peripheral neuropathy, small fiber neuropathy, trigeminal neuralgia, idiopathic small fiber neuralgia, or diabetic neuropathy), visceral pain (e.g., bowel pain), musculoskeletal pain, primary pain, idiopathic pain, osteoarthritis pain, gouty arthritis pain, rheumatoid arthritis pain, toothache, joint pain, labor pain, fibromyalgia, chronic low back pain, bladder pain syndrome, and sciatica.

[0058] In some embodiments of the present invention, the pain is selected from postoperative pain, neuropathic pain (e.g., postherpetic neuralgia, small fiber neuropathy pain, diabetic neuropathy), osteoarthritis pain, bladder pain syndrome, and cancer pain.

[0059] Crystal form preparation method

[0060] In some embodiments, the present invention provides a method for preparing crystal form A of compound 1, which includes chiral separation of methyl 4-(3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carbamate, mixing the product obtained by chiral separation with a good solvent, and then adding a poor solvent to separate the resulting solid.

[0061] In some embodiments, the preparation method of crystal form A of compound 1 specifically includes the following steps: the product obtained by chiral separation of methyl 4-(3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxylate is mixed with a good solvent, then cooled to 50-60°C, a poor solvent is added, and then cooled to 0-30°C to separate the obtained solid.

[0062] In some embodiments, the preparation method of crystal form A of compound 1 specifically includes the following steps: the product obtained by chiral separation of methyl 4-(3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carbamate is mixed with methanol, heated to dissolve, then cooled to 50-60°C, water is added, and then cooled to 0-30°C to crystallize, and the obtained solid is separated.

[0063] In some embodiments, in the method for preparing crystal form A of compound 1, the mass-to-volume ratio of the product obtained by chiral separation of methyl 4-(3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxylate to a good solvent, in g / ml, is 1:2-30, for example 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:21, 1:22, 1:23, 1:24, 1:25, 1:26, 1:27, 1:28, 1:29, 1:30.

[0064] In some embodiments, in the method for preparing crystal form A of compound 1, the mass-to-volume ratio (g / ml) of the product obtained by chiral separation of methyl 4-(3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxylate to the unsuitable solvent is 1:5-30, for example, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:21, 1:22, 1:23, 1:24, 1:25, 1:26, 1:27, 1:28, 1:29, 1:30.

[0065] In some embodiments, the chiral separation conditions are an AD-H chromatographic column, a column temperature of 20-30°C, and a mobile phase selected from a mixed solution of n-hexane, anhydrous ethanol, isopropanol, and diethylamine.

[0066] In some embodiments, the flow rate for the chiral separation is selected from 1-10 mL / min.

[0067] In some embodiments, the volume ratio of n-hexane-anhydrous ethanol-isopropanol-diethylamine in the chiral separated mobile phase is selected from (70-90):(10-20):(1-10):(0.01-1).

[0068] In some embodiments, the volume ratio of n-hexane-anhydrous ethanol-isopropanol-diethylamine in the chiral separated mobile phase is 80:16:4:0.05.

[0069] In some embodiments, the chiral separation product is selected as a component with a retention time of approximately 5.3 min, such as 5.2 min, 5.3 min, and 5.4 min.

[0070] In some embodiments, the crystal form A of compound 1 provided by the present invention can be prepared using the preparation method described above.

[0071] The crystalline or amorphous forms prepared in the examples were characterized by X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), polarized light microscopy (PLM), dynamic moisture adsorption (DVS), and X-ray single crystal diffraction.

[0072] (1) X-ray powder diffraction (XRPD)

[0073] XRPD patterns of crystalline or amorphous materials were acquired using an X'Pert3 Powder Diffractometer irradiated with Cu-palladium. Absolute scanning was performed at room temperature. The detection range was 3.5° to 40°, with a step size of 0.013°, a dwell time of 50 s, and one scan per cycle.

[0074] (2) Differential Scanning Calorimetry (DSC)

[0075] The DSC spectra of the crystal form were acquired using a TA DSC 2500 differential scanning calorimeter. The test temperature range was 35℃ to 250℃, and the heating rate was 10℃ / min.

[0076] (3) Thermogravimetric analysis (TGA)

[0077] The TGA spectra of the crystal form were acquired using a METTLER TOLEDO thermogravimetric analyzer. The test temperature range was 35℃ to 500℃, and the heating rate was 10℃ / min.

[0078] (4) Dynamic Water Adsorption (DVS)

[0079] The DVS spectrum of the crystal form was tested using DVS Intrinsic (SMS) at a temperature of 25°C in DMDT mode.

[0080] Example 1:

[0081] Step 1: Synthesis of methyl 4-(3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxylate

[0082] 3-(3,4-Difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carboxylic acid (837 mg, 2.35 mmol), DMF (30 mL), NMI (1.17 g, 11.4 mmol), and methyl 4-aminopyridinecarboxylate (550 mg, 3.64 mmol) were added in an ice bath with TCFH (2.08 mg, 7.2 mmol). After allowing the mixture to return to room temperature, stirring was continued for 1 h. The reaction was quenched with water (20 mL), and the mixture was extracted with ethyl acetate (30 mL × 3). The organic phases were combined, washed with water and saturated brine, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the crude product was separated by Pre-HPLC to obtain the target compound (910 mg). MS: m / z = 491.1, [M+H] + .

[0083] Step 2: Synthesis of 4-(3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxamide

[0084] The product from step one (910 mg) and a 2 M ammonia methanol solution (15 mL) were mixed, heated to 60 °C and stirred. The system was then cooled to room temperature and concentrated under reduced pressure. The crude product was separated by Pre-HPLC to obtain the target compound (711 mg), MS: m / z = 476.3, [M+H]. + , 1 HNMR (400MHz, DMSO) δ10.81(s,1H),8.46(d,J=5.5Hz,1H),8.15(d,J=1.7Hz,1H),8.03(s,1H),7.67(dd,J=5.5,2.0Hz,1H),7.6 0(s,1H),7.28-7.09(m,2H),4.61(d,J=10.6Hz,1H),4.39-7.18(m,1H),3.97(d,J=1.5Hz,3H),2.47-2.13(m,2H),1.74(s,3H).

[0085] Step 3: Preparation of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carboxamido)pyridine-2-carboxamide

[0086] The product from step two was separated using the following chiral separation method: AD-H column, column temperature 30℃, mobile phase (n-hexane-anhydrous ethanol-isopropanol-diethylamine = 80:16:4:0.05 (volume ratio)), flow rate 1 mL / min, and the fraction with a retention time of approximately 5.3 min was dried.

[0087] Add 25 ml of methanol to 5 g of the product obtained by the above method, heat and stir to reflux temperature to dissolve and obtain a clear solution, then cool to 50 °C, add 100 mL of purified water dropwise to the clear solution; after the addition is complete, cool to room temperature to grow crystals, then filter, and then vacuum dry at 45 °C and -0.07 MPa to obtain 4.05 g of the target compound.

[0088] The XRPD spectrum of the obtained compound is as follows: Figure 1 As shown, the obtained compound is compound 1, crystal form A.

[0089] Experimental Example 1: Monitoring the effect of test substance on the current of stably overexpressed Nav1.8 channel using manual patch-clamp technique. Experimental method:

[0090] 1. Reagent Preparation

[0091] The test compound is soluble in dimethyl sulfoxide (DMSO).

[0092] The extracellular fluid consisted of 140 mM NaCl, 3.5 mM KCl, 1 mM MgCl₂·6H₂O, 2 mM CaCl₂·2H₂O, 10 mM Md-Glucose, 10 mM HEPES, and 1.25 mM NaH₂PO₄·2H₂O, with pH adjusted to 7.4 using NaOH. The intracellular fluid consisted of 50 mM CsCl, 10 mM NaCl, 10 mM HEPES, 60 mM CsF, and 20 mM EGTA, with pH adjusted to 7.2 using CsOH.

[0093] 2. Experimental Materials and Instruments

[0094] 1) Patch clamp amplifier: EPC 10 (HEKA)

[0095] 2) Micromanipulator: MP225 (Sutter Instrument)

[0096] 3) Inverted microscope: MF53 (Mshot)

[0097] 4) Microelectrode pulling instrument: P97 (Sutter Instrument)

[0098] 5) Capillary glass tube: BF150-86-10 (Sutter Instrument)

[0099] 3 Experimental Steps

[0100] 1) After the compound is prepared into a solution of a specified concentration, it is added to the drug delivery system tubing in sequence and labeled.

[0101] 2) Place the cell slide in the recording chamber, select suitable cells under an inverted microscope, and adjust the position of the drug delivery head.

[0102] 3) The capillary glass tube is drawn into a suitable recording electrode using a microelectrode drawing device. The electrode, filled with intracellular fluid, is then inserted into the microelectrode holder. Under an inverted microscope, the micromanipulator is adjusted to bring the recording electrode into contact with the cell. Negative pressure is applied to the electrode to create a high-resistance seal. Fast capacitance compensation is then performed, followed by continued application of negative pressure to rupture the cell membrane, establishing a whole-cell recording mode. Finally, slow capacitance compensation is performed, and relevant parameters are recorded.

[0103] 4) Once the cell current has stabilized, begin drug administration. Each drug concentration is administered for five minutes or until the current stabilizes, then monitor the next concentration. The drug solution is administered to the cells sequentially from low to high concentration through the recording bath by gravity, and a peristaltic pump is used for fluid replacement during the recording process.

[0104] 4. Test voltage procedure (resting state) and results

[0105] After whole-cell sealing was achieved, the cell voltage was clamped at -120 mV, then depolarized to 0 mV using a 50 ms square wave pulse to obtain the Nav1.8 current. This procedure was repeated every 20 s, monitoring the maximum current induced by the square wave, and the test compound was introduced after it stabilized. The strength of the current blocking was calculated after the reaction stabilized.

[0106] The IC50 of the compound in Example 1 was tested. 50 ≤0.06nM.

[0107] Experimental Example 2: Pharmacological Effects of Compounds in an In vivo Incisional Pain Model in SD Rats / ICR Mice

[0108] Experimental animals: SD rats, male, 180-220g; ICR mice, male, 28-35g.

[0109] Compound preparation: First, add 5% DMSO to dissolve the compound to be tested, then add 10% polyoxyethylene castor oil (Cremophor EL) and 85% physiological saline (Saline) to the compound in sequence.

[0110] 1. Model Establishment

[0111] Rats / mice were placed in an anesthesia induction box and given 3%-4% isoflurane for induction anesthesia. After induction anesthesia, a breathing mask was connected, and the isoflurane concentration was adjusted to 1%-2% to prevent rats / mice from suffering pain during the operation. The rats / mice were placed in a supine position, and the surgical site was disinfected with alcohol and povidone-iodine. Under aseptic conditions, a 0.5cm longitudinal incision was made with a blade 0.2cm from the heel of the left hind foot toward the fingertips. The skin and fascia were cut open, the plantar muscles were separated, slightly elevated, and longitudinally cut. The skin was then sutured and disinfected.

[0112] 2. Drug administration test

[0113] A Von Frey test was performed before surgery. The day after animal modeling, a second test was conducted. Animals meeting baseline requirements were selected for enrollment and randomly divided into 6 groups of 10 animals each, based on 50% PWT. The drug administration was performed in a double-blind manner. 50% PWT was measured before administration (baseline) and at 0.5h, 2h, 4h, or 6h after administration, using a single dose. The model control group received saline, while the compound group received the compound according to the dosage shown in the table below.

[0114] Table 1: PWT 50% values ​​(g, Mean±SEM, n=10) at different time points in different groups of mice. Table 2: PWT 50% values ​​(g, Mean±SEM, n=10) at different time points in different groups of rats.

[0115]

[0116] In mouse and rat models of incisional pain, the compound of Example 1 exhibited good analgesic activity and a relatively long duration of analgesia at a dose of 20 mg / kg.

[0117] Experimental Example 3: Pharmacological Effects of Compounds in a Rat Spinal Nerve Ligation (SNL) Model

[0118] Experimental animals: SD rats, male, 180-220g.

[0119] Compound preparation: First, add 5% DMSO to dissolve the compound to be tested, then add 10% polyoxyethylene castor oil (Cremophor EL) and 85% physiological saline (Saline) to the compound in sequence.

[0120] 1. Model Establishment

[0121] SD rats were anesthetized with isoflurane inhalation. The rats were placed in a prone position, and the surgical site was shaved. The site was disinfected with alcohol and povidone-iodine. Under aseptic conditions, a 3-4 cm longitudinal incision was made at the level of the lower lumbar region / upper sacrum, exposing the left paravertebral muscles. Using blunt scissors, the paravertebral muscles were bluntly dissected, and the L6 transverse process was removed. The L4 and L5 spinal nerves were exposed, and the L5 spinal nerve was ligated with 6-0 silk suture. Ceftriaxone was used to treat the wound and prevent infection. The muscles and skin were sutured layer by layer, and disinfected with povidone-iodine. After surgery, the animals were kept warm on a warm electric blanket until fully recovered, after which they were returned to their cages for postoperative behavioral observation.

[0122] 2. Drug administration test

[0123] A Von Frey test was performed before surgery. Leg-raising training was conducted on days 5 and 6 post-surgery. Animals with a baseline weight of 2-5g were enrolled and divided into groups of 10 animals each, based on their 50% post-weighted total weight (PWT). On the second day, a single, blinded administration was administered. The 50% PWT of the animals was measured before administration (baseline) and at 2, 4, 6, or 8 hours post-administration. The model control group received saline, while the compound group received the compound at the dosages shown in the table below.

[0124] Table 3: PTW 50% values ​​(g, Mean±SEM, n=10) at different time points in different groups of rats containing the compound

[0125] In a rat spinal nerve ligation model, the compound of Example 1 exhibited good analgesic activity and a relatively long duration of analgesia at a dose of 20 mg / kg.

[0126] Experimental Example 4: Pharmacological Effects of Compounds in a CFA-Induced Rats with Inflammatory Pain

[0127] Experimental animals: SD rats, male, 180-220g.

[0128] Compound preparation: First, add 5% DMSO to dissolve the compound to be tested, then add 10% polyoxyethylene castor oil (Cremophor EL) and 85% physiological saline (Saline) to the compound in sequence.

[0129] 1. Model Establishment

[0130] SD rats were anesthetized with isoflurane, and their right hind paw was disinfected. A needle was inserted subcutaneously into the posterior half of the sole, and 50 μL of Freund's complete adjuvant (CFA, 1 mg / mL) was injected subcutaneously into the middle of the sole. The needle was then slowly rotated out to avoid leakage. After modeling, local redness and swelling of the plantar surface of the paw were observed, along with difficulty walking and retraction of the affected paw.

[0131] 2. Drug administration test

[0132] A Von Frey test was performed before surgery. The day after modeling, animals meeting baseline requirements were selected for enrollment and randomly divided into 6 groups of 10 animals each, based on 50% PWT. A double-blind drug administration test was conducted, with 50% PWT measured before administration (baseline), and at 0.5h, 2h, and 8h after administration. The model control group received saline, while the compound group received the compound at the dosages shown in the table below.

[0133] Table 4: PWT 50% values ​​(g, Mean±SEM, n=10) of rats at different time points for the compounds of this invention

[0134] The compound in Example 1 showed good analgesic activity in a CFA-induced rat model of inflammatory pain.

[0135] Experimental Example 5: Study on the pharmacological effects of compounds in a mouse acetic acid-induced writhing model

[0136] Experimental animals: ICR mice, male, 20-24g.

[0137] Compound preparation: First, add 5% DMSO to dissolve the compound to be tested, then add 10% polyoxyethylene castor oil (Cremophor EL) and 85% physiological saline (Saline) to the compound in sequence.

[0138] 1. Model establishment: Animals were randomly divided into groups of 10 each according to their body weight. They were injected intraperitoneally with 0.8% (v / v) acetic acid solution at a volume of 10 mL / kg.

[0139] 2. Drug administration test

[0140] The drug was administered 60 minutes before modeling. After injection of acetic acid solution, the number of writhing movements in mice was observed. A complete writhing response was defined as an episode of abdominal concavity, trunk and hind limb extension, and raised buttocks. The number of writhing movements in mice was observed and recorded within 0-15 minutes and 15-30 minutes, for a total observation period of 30 minutes. Inhibition rate = (number of writhing movements in control group - number of writhing movements in drug-treated group) / number of writhing movements in control group × 100%. The model control group received physiological saline, while the other groups received the compound according to the dosages shown in the table below.

[0141] Table 5: Number of writhing responses in mice of each compound group (Mean±SD, n=10)

[0142] Group Dosage (mg / kg) Number of twists Inhibition rate (%) Model control group / 49±16 - Example 1 Compound 20mg / kg 30±14 38.35

[0143] The compound of Example 1 of the present invention can effectively inhibit the writhing effect at a low dose of 20 mg / kg.

[0144] Experiment Example 6: Stability Experiment

[0145] The compound obtained in Example 1 of this invention was subjected to high temperature (60℃±2℃), high humidity (45℃±2℃ / 75%RH±5%RH), and light exposure (4500lx±500lx and ≥0.80×10⁻⁶ lx). 2 μW / cm 2 ), with packaging and illumination (4500lx±500lx and ≥0.80×10), 2 μW / cm 2 Stability experiments were conducted under the following conditions.

[0146] The results showed that the compounds of the present invention exhibited superior chemical stability and crystal form stability under light, packaged light, high temperature, high humidity and light-time conditions. The purity after 29 days was essentially unchanged from that after 0 days, and no crystal form change occurred. The HPLC purity was greater than 99.5%.

[0147] Experiment Example 7: Solubility Test

[0148] The solid form of the compound prepared in Example 1 of this invention was tested for solubility in different solutions. The results are as follows: The solubility of compound 1 in water and a series of pH buffer solutions within the physiological pH range is approximately 0.01–5.7 μg / mL. The solubility of crystal form A of compound 1 in FeSSIF (simulated postprandial intestinal fluid) and FaSSIF (simulated starvation intestinal fluid) is >55 μg / mL and ≥14 μg / mL, respectively. This indicates that compound 1 is a poorly soluble compound, and crystal form A of this invention can improve its solubility.

Claims

1. 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxamide, in crystal form A, characterized in that, The XRPD pattern of crystal form A includes characteristic peaks at diffraction angles (2θ) at 12.8±0.2°, 14.5±0.2° and / or 21.9±0.2°.

2. The crystal form A according to claim 1, characterized in that, In the XRPD pattern of crystal form A, the intensity of the diffraction peak at a diffraction angle (2θ) of 14.5°±0.2° is 100%, and the intensity of the diffraction peak at a diffraction angle (2θ) of 12.8°±0.2° is ≥70%.

3. The crystal form A according to claim 1 or 2, characterized in that, The XRPD pattern of crystal form A includes characteristic peaks at diffraction angles (2θ) at 12.8±0.2°, 13.2±0.2°, 14.5±0.2°, 19.3±0.2° and / or 21.9±0.2°.

4. The crystal form A according to claim 1 or 2, characterized in that, The XRPD pattern of crystal form A includes characteristic peaks at diffraction angles (2θ) at 12.8±0.2°, 13.2±0.2°, 14.5±0.2°, 17.8±0.2°, 19.3±0.2° and / or 21.9±0.2°.

5. Crystal form A according to claim 1 or 2, characterized in that, The XRPD pattern of crystal form A includes characteristic peaks at diffraction angles (2θ) at 12.8±0.2°, 13.2±0.2°, 14.5±0.2°, 17.8±0.2°, 19.3±0.2°, 20.6±0.2°, 21.9±0.2°, 25.0±0.2°, 25.6±0.2° and / or 27.0±0.2°.

6. The crystal form A according to claim 1 or 2, characterized in that, The XRPD pattern of crystal form A includes characteristic peaks at diffraction angles (2θ) at 12.8±0.2°, 13.2±0.2°, 14.5±0.2°, 16.0±0.2°, 17.8±0.2°, 18.6±0.2°, 19.3±0.2°, 19.6±0.2°, 20.6±0.2°, 21.4±0.2°, 21.9±0.2°, 23.6±0.2°, 24.3±0.2°, 25.0±0.2°, 25.6±0.2°, 26.0±0.2°, 27.0±0.2°, 27.9±0.2°, 28.5±0.2°, 29.3±0.2°, and 36.3±0.2°.

7. Crystal form A according to claim 1 or 2, characterized in that, The XRPD pattern of crystal form A includes peaks at essentially the same diffraction angle (2θ) as shown in Figure 1.

8. Crystal form A according to claim 1 or 2, characterized in that, The XRPD pattern of crystal form A is shown in Figure 1.

9. Crystal form A according to any one of claims 1-8, characterized in that, The crystal form A was prepared by the following method: the chiral separation product of methyl 4-(3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxylate was mixed with a good solvent, then cooled to 50-60°C, a poor solvent was added, and then cooled to 0-30°C.

10. Crystal form A according to any one of claims 1-8, characterized in that, Crystal form A was prepared by the following method: the chiral separation product of methyl 4-(3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxylate was mixed with methanol and heated to dissolve it. Then, the temperature was lowered to 50-60°C, water was added, and the temperature was lowered to 0-30°C to crystallize. The resulting solid was then separated.

11. A pharmaceutical composition, characterized in that, It comprises crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carboxamido)pyridine-2-carboxamide as described in any one of claims 1-10, and one or more pharmaceutically acceptable carriers.

12. The pharmaceutical composition of claim 11, characterized in that, It comprises 0.01-10000 mg of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carboxamido)pyridine-2-carboxamide as described in any one of claims 1-10, and one or more pharmaceutically acceptable carriers.

13. The pharmaceutical composition of claim 12, characterized in that, It contains 0.03-8000mg, 0.05-8000mg, 0.1-8000mg, 10-8000mg, 10-1000mg, 10mg, 20mg, 30mg, 40mg, 50mg, 60mg, 70mg, 80mg, 90mg, 100mg, 110mg, 120mg, 125mg, 130mg, 140mg, 150mg, 160mg, 170mg, 180mg, 190mg, 200mg, 250mg, 300mg, 350mg, 400mg, 450mg, 500mg, 550mg, 600mg, 650mg, 700mg, 7 The crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carboxamid as described in any one of claims 1-10, in doses of 50 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1500 mg, 2000 mg, 2500 mg, 3000 mg, 3500 mg, 4000 mg, 4500 mg, 5000 mg, 5500 mg, 6000 mg, 6500 mg, 7000 mg, 7500 mg, or 8000 mg, and one or more pharmaceutically acceptable carriers.

14. A formulation, characterized in that, It comprises crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carboxamido)pyridine-2-carboxamide as described in any one of claims 1-10, and one or more pharmaceutically acceptable carriers.

15. The formulation of claim 14, characterized in that, It comprises 0.01-10000 mg of crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carboxamido)pyridine-2-carboxamide as described in any one of claims 1-8, and one or more pharmaceutically acceptable carriers.

16. The formulation of claim 15, characterized in that, It contains 0.03-8000mg, 0.05-8000mg, 0.1-8000mg, 10-8000mg, 10-1000mg, 10mg, 20mg, 30mg, 40mg, 50mg, 60mg, 70mg, 80mg, 90mg, 100mg, 110mg, 120mg, 125mg, 130mg, 140mg, 150mg, 160mg, 170mg, 180mg, 190mg, 200mg, 250mg, 300mg, 350mg, 400mg, 450mg, 500mg, 550mg, 600mg, 650mg, 700mg, 7 The crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carboxamid as described in any one of claims 1-8, in doses of 50 mg, 800 mg, 850 mg, 900 mg, 950 mg, 1000 mg, 1500 mg, 2000 mg, 2500 mg, 3000 mg, 3500 mg, 4000 mg, 4500 mg, 5000 mg, 5500 mg, 6000 mg, 6500 mg, 7000 mg, 7500 mg, or 8000 mg, and one or more pharmaceutically acceptable carriers.

17. A medicine box comprising crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxamide according to any one of claims 1-10, or a pharmaceutical composition according to any one of claims 11-13, or a formulation according to any one of claims 14-16, optionally further comprising one or more other therapeutic agents.

18. Use of crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxamide according to any one of claims 1-10, or the pharmaceutical composition according to any one of claims 11-13, or the formulation according to any one of claims 14-16, or the kit according to claim 17, in the preparation of a medicament for the prevention and / or treatment of NaV1.8-related diseases.

19. The use as described in claim 18, characterized in that, The disease associated with NaV1.8 is pain.

20. The use as described in claim 19, characterized in that, The pain described is selected from chronic pain, acute pain, inflammatory pain, cancer pain, ICU analgesia, fracture or postoperative pain, neuropathic pain, visceral pain, musculoskeletal pain, primary pain, idiopathic pain, osteoarthritis pain, gouty arthritis pain, rheumatoid arthritis pain, toothache, joint pain, childbirth pain, fibromyalgia, chronic low back pain, bladder pain syndrome, and sciatica.

21. The use as described in claim 20, characterized in that, The pain described is selected from inflammatory pain, postoperative pain, and visceral pain.

22. The use as described in claim 20, characterized in that, The postoperative pain is selected from pain from bunion removal surgery, hernia repair surgery, and abdominoplasty; the neuropathic pain is selected from neuralgia of peripheral neuropathy, postherpetic neuralgia, peripheral neuropathy, small fiber neuropathy, trigeminal neuralgia, idiopathic small fiber neuropathy, or diabetic neuralgia; the visceral pain is selected from intestinal pain.

23. The method for preparing crystal form A of 4-((2R,3S,5R)-3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxamide according to any one of claims 1-10, characterized in that, The process includes the following steps: mixing the chiral separation product of compound methyl 4-(3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxylate with a good solvent, then cooling to 50-60°C, adding a poor solvent, and then cooling to 0-30°C.

24. The preparation method according to claim 23, characterized in that, The good solvent is methanol; the poor solvent is water.

25. The preparation method according to claim 23 or 24, characterized in that, The mass-to-volume ratio (g / mL) of the product obtained by chiral separation of methyl 4-(3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxylate to a good solvent is 1:2-30, for example, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:21, 1:22, 1:23, 1:24, 1:25, 1:26, 1:27, 1:28, 1:29, and 1:

30.

26. The preparation method according to any one of claims 23-25, characterized in that, The mass-to-volume ratio (g / mL) of the product obtained by chiral separation of methyl 4-(3-(3,4-difluoro-2-methoxyphenyl)-5-methyl-5-(trifluoromethyl)thiacyclopentane-2-carbamate)pyridine-2-carboxylate to a poor solvent is 1:5-30, for example, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:16, 1:17, 1:18, 1:19, 1:20, 1:21, 1:22, 1:23, 1:24, 1:25, 1:26, 1:27, 1:28, 1:29, and 1:

30.

27. The preparation method according to any one of claims 23-26, characterized in that, The volume ratio of n-hexane-anhydrous ethanol-isopropanol-diethylamine in the chiral separation mobile phase is selected from (70-90):(10-20):(1-10):(0.01-1); preferably, the volume ratio of n-hexane-anhydrous ethanol-isopropanol-diethylamine in the chiral separation mobile phase is 80:16:4:0.05.

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