HK40082657B - Pharmaceutical composition, formulation thereof, preparation method therefor and use thereof - Google Patents

Description

A pharmaceutical composition, formulation, preparation method thereof and application

This application claims priority to the earlier application filed on June 10, 2021, with China National Intellectual Property Administration, patent application number 202110651136.9, entitled "A pharmaceutical composition, formulation and preparation method thereof and application thereof". The entire contents of the earlier application are incorporated herein by reference.

Technical Field

This invention belongs to the field of pharmaceutical compositions, specifically relating to a pharmaceutical composition, formulation, preparation method thereof, and application.

Background Technology

ATP receptors are classified into two main families based on their molecular structure, transduction mechanism, and pharmacological properties: P2Y- and P2X-purine receptors. P2X-purine receptors are a family of ATP-gated cation channels, and several subtypes have been cloned, including: six homopolymers, P2X1, P2X2, P2X3, P2X4, P2X5, and P2X7; and three heteropolymers, P2X2/3, P2X4/6, and P2X1/5. Studies have found that P2X3 receptors are particularly expressed in primary afferent nerve fibers of "hollow viscera," such as the lower urinary tract and respiratory tract.

Coughing is a major symptom of respiratory diseases, affecting 70% to 80% of patients in respiratory clinics. With the increasing prevalence of conditions like COPD and IPF, and given that coughing is a primary symptom of most exhaled airway diseases, the demand for treatment is also growing. As a defensive reflex, coughing helps clear respiratory secretions and harmful agents; however, frequent and severe coughing can seriously impact a patient's work, daily life, and social activities.

There are not many P2X3 antagonists specifically developed for the indication of cough. The project that is progressing the fastest is Roche's AF-219 project. Its latest completed Phase II clinical trial showed good efficacy for refractory cough, but the taste disorder problem is serious.

Currently, there are no marketed drugs that inhibit the P2X3 pathway to treat many conditions, including chronic cough. Therefore, developing new drugs that can inhibit P2X3 activity is of great significance for the treatment of these diseases.

Summary of the Invention

This invention provides a pharmaceutical composition comprising an active ingredient and pharmaceutically acceptable excipients; the active ingredient comprising a compound of formula A:

The excipients are selected from one, two or more of the following excipients, including but not limited to: diluents, disintegrants, binders, flow aids and lubricants.

According to the technical solution of the present invention, the compound represented by formula A is selected from one, two or more of crystal form I, crystal form III and crystal form V.

According to the technical solution of the present invention, the crystal form I is subjected to Cu-Kα radiation, and the X-ray powder diffraction, expressed in 2θ angle, has characteristic peaks at 8.56°±0.20°, 12.48°±0.20° and 22.13±0.20°.

Furthermore, the crystal form I, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 8.56°±0.20°, 12.48±0.20°, 22.13°±0.20°, 13.53°±0.20°, 14.25°±0.20°, 25.18°±0.20°, and 26.07°±0.20° in 2θ angles.

Furthermore, the crystal form I, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 2θ at 8.56°±0.20°, 12.48±0.20°, 22.13°±0.20°, 13.53°±0.20°, 14.25±0.20°, 25.18°±0.20°, 26.07°±0.20°, 22.32°±0.20°, 23.23°±0.20°, and 23.42°±0.20°.

Preferably, the crystal form I has an XRPD pattern as shown in Figure 3.

Preferably, the differential scanning calorimetry spectrum of crystal form I has an endothermic peak at about 152°C and a melting enthalpy of about 44±2 J/g.

Preferably, the thermogravimetric analysis spectrum and differential scanning calorimetry spectrum of crystal form I are basically as shown in Figure 4;

Preferably, the polarized light microscope pattern of crystal form I is basically as shown in Figure 5.

According to the technical solution of the present invention, the crystal form III uses Cu-Kα radiation, and the X-ray powder diffraction, expressed in 2θ angle, has characteristic peaks at 12.91°±0.20°, 16.77±0.20°, 19.27°±0.20° and 22.80°±0.20°.

Furthermore, the crystal form III, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 12.91°±0.20°, 16.77°±0.20°, 19.27°±0.20°, 22.80°±0.20°, 13.75°±0.20°, 14.46°±0.20°, and 20.86°±0.20° in 2θ angles.

Furthermore, the crystal form III, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 2θ at 12.91°±0.20°, 16.77°±0.20°, 19.27°±0.20°, 22.80°±0.20°, 13.75°±0.20°, 14.46°±0.20°, 20.86°±0.20°, 21.08°±0.20°, 23.75°±0.20°, and 24.05°±0.20°.

Preferably, the crystal form III has an XRPD pattern as shown in Figure 10.

Preferably, the crystal form III contains 0.4 equivalents of water.

Preferably, in the thermogravimetric analysis spectrum of crystal form III, the weight loss gradient in the range of room temperature to 100°C is approximately 1.5%;

Preferably, the first endothermic peak in the differential scanning calorimetry spectrum of crystal form III is due to the removal of 0.4 water molecules.

Preferably, the thermogravimetric analysis spectrum and differential scanning calorimetry spectrum of crystal form III are basically as shown in Figure 11.

Preferably, the polarized microscope pattern of crystal form III is basically as shown in Figure 12.

According to the technical solution of the present invention, the crystal form V is subjected to Cu-Kα radiation, and the X-ray powder diffraction, expressed in 2θ angle, has characteristic peaks at 8.38°±0.20°, 9.15°±0.20°, 13.52°±0.20° and 18.44±0.20°.

Furthermore, the crystal form V, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 8.38°±0.20°, 9.15°±0.20°, 13.52°±0.20°, 18.44°±0.20°, 16.26°±0.20°, 16.89°±0.20°, and 17.86°±0.20° in 2θ angles.

Furthermore, the crystal form V, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at angles of 2θ at 8.38°±0.20°, 9.15°±0.20°, 13.52°±0.20°, 18.44°±0.20°, 16.26°±0.20°, 16.89°±0.20°, 17.86°±0.20°, 22.35°±0.20°, 23.56°±0.20°, and 24.74°±0.20°.

Preferably, the crystal form V has an XRPD pattern as shown in Figure 16.

Preferably, the differential scanning calorimetry spectrum of crystal form V has an endothermic peak at about 166°C and a melting enthalpy of about 70±2 J/g.

Preferably, the thermogravimetric analysis spectrum and differential scanning calorimetry spectrum of crystal form V are basically as shown in Figure 17.

Preferably, the polarized microscope pattern of the crystal type V is basically as shown in Figure 18.

According to the technical solution of the present invention, the particle size of the compound shown in Formula A is 1-40 μm, for example 1.5-35 μm.

According to the technical solution of the present invention, the D10 particle size of the compound shown in Formula A is 1-5 μm, for example 1.5-4 μm, and exemplary values are 2 μm, 2.21 μm, 2.5 μm, and 3 μm.

Furthermore, the D50 particle size of the compound shown in Formula A is 6-15 μm, for example 8-12 μm, with exemplary values of 9 μm, 10.2 μm, 11 μm, and 11.5 μm.

Furthermore, the D90 particle size of the compound shown in Formula A is 20-40 μm, for example 25-35 μm, with exemplary values of 27 μm, 29.1 μm, and 30 μm.

According to the technical solution of the present invention, the bulk density (bulk density) of the compound represented by Formula A is 0.2-0.3 g/mL, for example 0.22-0.28 g/mL, and exemplary values are 0.23 g/mL, 0.24 g/mL, 0.25 g/mL, 0.26 g/mL, 0.27 g/mL, and 0.29 g/mL.

According to the technical solution of the present invention, the actual density (bulk density) of the compound shown in Formula A is 0.32-0.5 g/mL, for example 0.35-0.45 g/mL, and exemplary values are 0.33 g/mL, 0.36 g/mL, 0.38 g/mL, 0.40 g/mL, 0.42 g/mL, 0.44 g/mL, and 0.48 g/mL.

The pharmaceutically acceptable excipients are preferably non-chemically reactive or inert to the active ingredient.

According to the technical solution of the present invention, the diluent may be selected from one, two or more of the following substances: lactose, microcrystalline cellulose, sucrose, glucose, mannitol, sorbitol, calcium sulfate, calcium gluconate, calcium hydrogen phosphate, calcium phosphate, calcium carbonate, calcium bicarbonate, starch, carboxymethyl starch, pregelatinized starch, etc., for example, lactose and/or microcrystalline cellulose; further, the lactose is lactose monohydrate.

For example, the diluent includes a first diluent and a second diluent, which are different from each other and are independently selected from one of the diluents mentioned above. Preferably, the first diluent is microcrystalline cellulose and the second diluent is lactose monohydrate.

According to the technical solution of the present invention, the disintegrant may be selected from one, two or more of the following substances: croscarmellose sodium, pregelatinized starch, microcrystalline cellulose, alginate, lignocellulose, sodium carboxymethyl starch, guar gum, croscarmellose, etc., for example, croscarmellose sodium.

According to the technical solution of the present invention, the adhesive may be selected from one, two or more of the following substances: hydroxypropyl cellulose, gelatin, dextrin, maltodextrin, sucrose, gum arabic, polyvinylpyrrolidone, methylcellulose, carboxymethyl cellulose, ethylcellulose, polyvinyl alcohol, polyethylene glycol and hydroxypropyl methylcellulose, for example hydroxypropyl cellulose, and may further be low-substituted hydroxypropyl cellulose or high-substituted hydroxypropyl cellulose.

According to the technical solution of the present invention, the flow aid can be selected from one, two or more of the following substances: colloidal silica, silica, talc, calcium silicate, magnesium silicate and calcium hydrogen phosphate, etc., for example, colloidal silica.

According to the technical solution of the present invention, the lubricant may be selected from one, two or more of the following substances: magnesium stearate, calcium stearate, zinc stearate, talc, glyceryl monostearate, polyethylene glycol (e.g., polyethylene glycol 4000, polyethylene glycol 6000, polyethylene glycol 8000), sodium benzoate, adipic acid, fumaric acid, boric acid, sodium chloride, sodium oleate, glyceryl triacetate, polyoxyethylene monostearate, monolauric sucrose ester, sodium chloride, sodium lauryl sulfate and magnesium lauryl sulfate, etc., for example selected from magnesium stearate, calcium stearate and/or zinc stearate.

According to the technical solution of the present invention, the excipients may further include flavoring agents. For example, the flavoring agents may be selected from one, two or more of the following substances: stevioside, fructose, glucose, high-fructose corn syrup, honey, aspartame, protein sugar, xylitol, mannitol, lactose, sorbitol, flavorings and maltitol, etc.

According to one embodiment of the present invention, the diluent is lactose monohydrate and microcrystalline cellulose, the disintegrant is croscarmellose sodium, the binder is hydroxypropyl cellulose, the flow aid is colloidal silica, and the lubricant is magnesium stearate.

According to the technical solution of the present invention, the pharmaceutical composition comprises the following components: the compound shown in Formula A, lactose monohydrate, microcrystalline cellulose, croscarmellose sodium, hydroxypropyl cellulose, colloidal silica, and magnesium stearate;

In the pharmaceutical composition described herein, the content of each component can be selected as needed. For example, the content of the compound shown in Formula A can be present in the pharmaceutical composition at a therapeutically effective amount.

According to the technical solution of the present invention, the pharmaceutical composition comprises 10-40 parts, for example 15-35 parts, of the compound shown in Formula A, in parts by weight, such as 12 parts, 18 parts, 20 parts, 23 parts, 25 parts, 27 parts, 30 parts, 32 parts, and 38 parts, by weight.

According to the technical solution of the present invention, the pharmaceutical composition comprises 50-80 parts by weight of a diluent, for example 55-75 parts, such as 52 parts, 58 parts, 60 parts, 63 parts, 65 parts, 70 parts, 72 parts, and 78 parts. Further, the weight ratio of the first diluent to the second diluent is (10-25):(40-55), for example (15-20):(40-55), such as 17.4:49.6.

According to the technical solution of the present invention, the pharmaceutical composition contains 0.5-6 parts, for example 1-5 parts, of disintegrant by weight, such as 0.75 parts, 1.5 parts, 2 parts, 2.5 parts, 3 parts, 3.5 parts, 4 parts, 4.5 parts, and 5.5 parts.

According to the technical solution of the present invention, the pharmaceutical composition comprises 0.5-6 parts, for example 1-5 parts, of the binder, in parts by weight, such as 0.75 parts, 1.5 parts, 2 parts, 2.5 parts, 3 parts, 3.5 parts, 4 parts, 4.5 parts, and 5.5 parts.

According to the technical solution of the present invention, the pharmaceutical composition contains 0.1-3 parts by weight of a gliding agent, for example 0.3-2 parts, such as 0.5 parts, 1 part, 1.2 parts, 1.5 parts, and 2.5 parts.

According to the technical solution of the present invention, the pharmaceutical composition contains 0.1-3 parts by weight of lubricant, for example 0.3-2 parts, and exemplary parts are 0.5 parts, 1 part, 1.2 parts, 1.5 parts, and 2.5 parts.

The total weight of each component in the pharmaceutical composition is 100 parts.

According to the technical solution of the present invention, the pharmaceutical composition comprises the following components in parts by weight: 10-40 parts of the compound shown in Formula A, 50-80 parts of lactose monohydrate and microcrystalline cellulose, 0.5-6 parts of croscarmellose sodium, 0.5-6 parts of hydroxypropyl cellulose, 0.1-3 parts of colloidal silica, and 0.1-3 parts of magnesium stearate;

According to an exemplary embodiment of the present invention, the pharmaceutical composition comprises the following components in parts by weight: 25 mg of the compound shown in Formula A, 49.6 mg of lactose monohydrate, 17.4 mg of microcrystalline cellulose, 3.0 mg of croscarmellose sodium, 3.0 mg of hydroxypropyl cellulose, 1.0 mg of colloidal silica, and 1.0 mg of magnesium stearate;

According to an exemplary embodiment of the present invention, the pharmaceutical composition comprises the following components in parts by weight: 100 mg of the compound shown in Formula A, 198.4 mg of lactose monohydrate, 69.6 mg of microcrystalline cellulose, 12.0 mg of croscarmellose sodium, 12.0 mg of hydroxypropyl cellulose, 4.0 mg of colloidal silica, and 4.0 mg of magnesium stearate.

According to an embodiment of the present invention, the pharmaceutical composition is in solid form, such as a powder solid form.

According to embodiments of the present invention, the pharmaceutical composition can be prepared into a dosage form suitable for administration.

The present invention also provides a pharmaceutical formulation, such as a solid dosage form, comprising the pharmaceutical composition. Exemplarily, the solid dosage form may be a tablet, capsule, or granule.

According to an exemplary embodiment of the present invention, the tablet is a coated tablet, comprising a core and a coating layer. Preferably, the core contains the pharmaceutical composition.

According to an exemplary embodiment of the present invention, there are no particular limitations on the composition of the coating layer. For example, a commercially available known gastric-soluble film coating premix can be used, or it can be prepared according to a known method. For example, the coating layer may also contain one, two, or more of the following: polyvinyl alcohol, polyethylene glycol, hydroxypropyl methylcellulose, hydroxypropyl cellulose, acrylic resin VI, polyvinylpyrrolidone, propylene glycol, castor oil, silicone oil, triglycerides, talc, titanium dioxide, and colorants. The film coating material may also be commercially available, such as the Opadry gastric-soluble coating series or the Easily Dissolved gastric-soluble coating series.

According to one technical solution of the present invention, the core of the coated tablet comprises the following components in parts by weight: 10-40 parts of the compound shown in Formula A, 50-80 parts of lactose monohydrate and microcrystalline cellulose, 0.5-6 parts of croscarmellose sodium, 0.5-6 parts of hydroxypropyl cellulose, 0.1-3 parts of colloidal silica, and 0.1-3 parts of magnesium stearate;

The film coating material of the coating layer is the Opadry gastric-soluble coating series;

Preferably, the compound represented by Formula A exists in the chip core in crystal forms I, III, and V.

For example, the core of the coated tablet contains the following components: 25 mg of the compound shown in Formula A, 49.6 mg of lactose monohydrate, 17.4 mg of microcrystalline cellulose, 3.0 mg of croscarmellose sodium, 3.0 mg of hydroxypropyl cellulose, 1.0 mg of colloidal silica, and 1.0 mg of magnesium stearate;

Preferably, the compound represented by formula A exists in the core in crystal forms I, III, and V;

The film coating material of the coating layer is the Opadry gastric-soluble coating series.

For example, the core of the coated tablet contains the following components: 100 mg of the compound shown in Formula A, 198.4 mg of lactose monohydrate, 69.6 mg of microcrystalline cellulose, 12.0 mg of croscarmellose sodium, 12.0 mg of hydroxypropyl cellulose, 4.0 mg of colloidal silica, and 4.0 mg of magnesium stearate;

Preferably, the compound represented by formula A exists in the core in crystal forms I, III, and V;

The film coating material of the coating layer is the Opadry gastric-soluble coating series.

The present invention further provides a method for preparing the pharmaceutical composition, comprising mixing the components therein. Preferably, the prescribed amounts of the compound of formula A, the gliding agent, and the first diluent are first sieved (e.g., through a 60-mesh sieve) before being mixed with the other components.

The present invention also provides a method for preparing the tablet, comprising compressing the pharmaceutical composition into tablets, for example by wet granulation tableting, and optionally performing or not performing coating.

Preferably, the wet granulation and tableting method includes: wet granulating the pharmaceutical composition excluding the lubricant, sizing, drying, and granulating again to obtain dry granules; mixing the dry granules with the lubricant and tableting.

According to an embodiment of the present invention, in the drying step of the wet granulation and tableting method, the material drying loss is controlled at 1.5%-2.5%. Preferably, in the drying step, when the material drying loss is between 1.0% and 2.5%, the inlet air temperature is turned off and drying is stopped. Preferably, the equipment used in the drying step is a fluidized bed.

According to an embodiment of the present invention, in the wet granulation step, the stirring speed is 200-400 rpm, preferably 250.0 rpm; the cutting speed is 1000.00-2000.00 rpm, preferably 1500.0 rpm; and the granulation time is 30 seconds to 2 minutes, preferably 60 seconds.

Furthermore, the wet granulation process includes: mixing the compound shown in Formula A, a flow aid (e.g., colloidal silica), a diluent (e.g., microcrystalline cellulose and lactose monohydrate), and a disintegrant (e.g., croscarmellose sodium), spraying an adhesive solution (e.g., hydroxypropyl cellulose) into the mixture, and optionally adding water or not after the adhesive solution has been sprayed, granulating.

Preferably, the adhesive solution is an aqueous solution of the adhesive, and its concentration is further 5-15%, for example 9%.

Preferably, the preparation of the mixture includes:

The compound shown in Formula A, a flow aid (e.g., colloidal silica), and a first diluent (e.g., microcrystalline cellulose) are mixed and sieved to obtain mixture 1;

The second diluent (e.g., lactose monohydrate) and the disintegrant (e.g., sodium croscarmellose) are mixed and sieved to obtain mixture 2;

The mixture 1 and mixture 2 are mixed to obtain a mixture. Preferably, the method for preparing the coated tablets includes the following steps:

(1) The above mixture is wet-granulated, sizing, dried, and granulated again to obtain dry granules;

(2) The dry granules are mixed with the lubricant (e.g., magnesium stearate) and compressed into tablets to obtain tablet cores;

(3) Spray coating liquid onto the core to obtain the coated sheet.

The present invention also provides a method for storing the pharmaceutical composition or the preparation, comprising storing the pharmaceutical composition or the preparation in the dark. Further, the storage conditions also include dry storage.

The present invention also provides the use of the pharmaceutical composition in the preparation of P2X3 inhibitors.

In the described applications, the P2X3 inhibitor can be used in mammalian organisms; it can also be used in vitro, primarily for experimental purposes, such as providing a standard or control sample for comparison, or preparing a kit according to conventional methods in the art to provide rapid detection of the inhibitory effect of P2X3.

The present invention also provides the use of the pharmaceutical composition in the preparation of pharmaceutical formulations, such as solid dosage forms, further such as tablets, particularly in the preparation of tablets by direct compression.

According to the technical solution of the present invention, the pharmaceutical preparation is a drug for protecting against, treating, or alleviating at least partially P2X3-mediated or activity-related diseases in animals; or, the drug is a drug for treating pain, itching, endometriosis, urinary tract diseases, or respiratory diseases.

In some implementations, the disease includes pain; the pain includes, but is not limited to: inflammatory pain, surgical pain, visceral pain, toothache, premenstrual pain, central pain, pain caused by burns, migraine, or cluster headache.

In some implementations, the diseases include urinary system diseases; the urinary tract diseases include: urinary incontinence, overactive bladder, dysuria, cystitis, prostatitis, prostatodynia, and benign prostatic hyperplasia;

The urinary incontinence includes uncontrolled urinary incontinence, which is associated with urge incontinence, cough incontinence, stress incontinence, overflow incontinence, functional incontinence, neurogenic incontinence, postprostatectomy incontinence, urinary urgency, nocturia, and enuresis.

In some implementations, the disease includes respiratory diseases, including but not limited to: respiratory disorders, including idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, bronchospasm, acute cough, or chronic cough.

The chronic cough is defined as a cough that lasts for more than eight weeks and has a serious adverse effect on social, psychological and physical well-being.

In some implementations, the disease includes respiratory diseases including acute or chronic cough. The cough is either acute or chronic and is associated with a condition selected from chronic obstructive pulmonary disease, asthma, tuberculosis, bronchitis, bronchiectasis, purulent lung disease, respiratory malignancies, allergic reactions, cystic fibrosis, pulmonary fibrosis, respiratory tract inflammation, emphysema, pneumonia, lung cancer, lung tumor formation, sore throat, common cold, influenza, respiratory infection, bronchoconstriction, sarcoidosis, viral or bacterial infection of the upper airway, angiotensin-converting enzyme (ACE) inhibitor therapy, smoker's cough, chronic dry cough without sputum, neoplastic cough, cough caused by gastroesophageal reflux, and diseases, symptoms, or conditions caused by inhalation of irritants, smoke, fog, dust, or air pollution.

In some implementations, itching is associated with inflammatory skin diseases, infectious skin diseases, autoimmune skin diseases, or pregnancy-related skin diseases.

In some implementations, itching is associated with an inflammatory skin disease selected from atopic dermatitis, allergic dermatitis, irritant contact dermatitis, xeritis, dysplastic dermatitis, lichen planus, lichen sclerosus, polymorphic psoriasis, Grignard disease, myxoid degeneration, mastocytosis, and urticaria.

In some implementations, itching is associated with an infectious dermatitis selected from fungal infections, bacterial and viral infections, scabies, athlete's foot, insect bites, and folliculitis.

In some implementations, itching is associated with autoimmune skin diseases selected from herpetic dermatitis (Dullin's disease), bullous pemphigus; genotypic skin diseases, Darriee's disease, and Helle-Helle's disease.

In some implementations, the itching is associated with pregnancy-related dermatitis selected from polymorphic dermatitis of pregnancy (PEP), atopic dermatitis of pregnancy, pemphigoid pregnancy, tumor formation, and cutaneous T-cell lymphoma.

In some implementations, itching is associated with nodular prurigo.

In some implementations, itching is associated with kidney disease or the treatment process for kidney disease.

In some implementations, itching is associated with chronic kidney disease.

In some implementations, the itching is associated with a treatment process for kidney disease, which may be selected from hemodialysis and peritoneal dialysis.

In some implementations, itching is associated with a medical procedure or treatment.

In some implementations, the itching is associated with medical treatment using drugs selected from opioids, antimalarial drugs, anticancer therapies, and epidermal growth factor receptor inhibitors.

In some implementations, the P2X3-mediated or activity-related disease is endometriosis. Endometriosis-related symptoms are selected from dysmenorrhea, dyspareunia, urinary retention, and schizophrenia.

The present invention also provides the use of the above-described pharmaceutical compositions or preparations in the treatment and/or prevention of diseases.

This invention also provides the use of the above-described pharmaceutical compositions or formulations in the protection, treatment, therapy, or relief of at least a portion of P2X3-mediated or activity-related diseases in animals (e.g., humans). These diseases include, but are not limited to, respiratory diseases, cough, chronic cough, idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, pain, urinary incontinence, autoimmune diseases, overactive bladder, dysuria, inflammation, Alzheimer's disease, Parkinson's disease, sleep disorders, epilepsy, mental illness, arthritis, neurodegeneration, traumatic brain injury, myocardial infarction, rheumatoid arthritis, stroke, thrombosis, atherosclerosis, colitis, inflammatory bowel disease, gastrointestinal diseases; gastrointestinal dysfunction, respiratory failure, sexual dysfunction, cardiovascular diseases, heart failure, hypertension, urinary incontinence, cystitis, arthritis, endometriosis, hematologic disorders, musculoskeletal and connective tissue developmental disorders, and systemic disorders.

In some implementations, the disease includes pain; the pain includes, but is not limited to: inflammatory pain, surgical pain, visceral pain, toothache, premenstrual pain, central pain, pain caused by burns, migraine, or cluster headache.

In some implementations, the disease includes urinary tract diseases.

In some implementations, the disease includes respiratory diseases, including but not limited to: respiratory disorders, including idiopathic pulmonary fibrosis, chronic obstructive pulmonary disease, asthma, bronchospasm, or chronic cough.

The chronic cough is defined as a cough that lasts for more than eight weeks and has a serious adverse effect on social, psychological and physical well-being.

In some implementations, the disease includes pruritus, which is associated with inflammatory skin diseases, infectious skin diseases, autoimmune skin diseases, or pregnancy-related skin diseases.

In some implementations, itching is associated with an inflammatory skin disease selected from atopic dermatitis, allergic dermatitis, irritant contact dermatitis, xeritis, dysplastic dermatitis, lichen planus, lichen sclerosus, polymorphic psoriasis, Grignard disease, myxoid degeneration, mastocytosis, and urticaria.

In some implementations, itching is associated with an infectious dermatitis selected from fungal infections, bacterial and viral infections, scabies, athlete's foot, insect bites, and folliculitis.

In some implementations, itching is associated with autoimmune skin diseases selected from herpetic dermatitis (Dullin's disease), bullous pemphigus; genotypic skin diseases, Darriee's disease, and Helle-Helle's disease.

In some implementations, the itching is associated with pregnancy-related dermatitis selected from polymorphic dermatitis of pregnancy (PEP), atopic dermatitis of pregnancy, pemphigoid pregnancy, tumor formation, and cutaneous T-cell lymphoma.

In some implementations, itching is associated with nodular prurigo.

In some implementations, itching is associated with kidney disease or the treatment process for kidney disease.

In some implementations, itching is associated with chronic kidney disease.

In some implementations, the itching is associated with a treatment process for kidney disease, which may be selected from hemodialysis and peritoneal dialysis.

In some implementations, itching is associated with a medical procedure or treatment.

In some implementations, the itching is associated with medical treatment using drugs selected from opioids, antimalarial drugs, anticancer therapies, and epidermal growth factor receptor inhibitors.

In some implementations, the P2X3-mediated or activity-related disease is endometriosis. Endometriosis-related symptoms are selected from dysmenorrhea, dyspareunia, urinary retention, and schizophrenia.

The present invention also provides a method for treating or preventing disease by administering to a patient a preventive and/or therapeutically effective amount of the pharmaceutical composition or preparation.

By administering the pharmaceutical composition, the side effects of treatment-related taste disturbances are reduced.

Terminology Definitions and Explanations

The various terms and phrases used in this invention have their general meanings known to those skilled in the art. Nevertheless, this invention still intends to provide a more detailed description and explanation of these terms and phrases. In the event of any inconsistency between the terms and phrases mentioned and their known meanings, the meanings expressed in this invention shall prevail.

The polymorphs of the compounds represented by Formula A of the present invention include the nonsolvent (anhydrous) form and the solvate form of the compounds represented by Formula A.

The polymorph of the compound shown in Formula A of the present invention has X-ray powder diffraction characteristic peaks represented by an angle of 2θ, wherein "±0.20°" is the allowable measurement error range.

The polymorph of the compound shown in Formula A of the present invention can be used in combination with other active ingredients, as long as it does not produce other adverse effects, such as allergic reactions.

As used in this invention, the term "composition" means a product comprising specified amounts of each specified ingredient, and any product derived directly or indirectly from a combination of specified amounts of each specified ingredient.

Those skilled in the art can use known drug carriers to prepare suitable pharmaceutical compositions from the polymorphs of the compounds of Formula A of the present invention. The pharmaceutical compositions can be specifically formulated for oral, parenteral, or rectal administration in solid or liquid form. The pharmaceutical compositions can be formulated into various dosage forms to facilitate administration, such as oral formulations (e.g., tablets, capsules, solutions, or suspensions); injectable formulations (e.g., injectable solutions or suspensions, or injectable dry powders that can be used immediately after addition of a pharmaceutical solvent prior to injection).

As used in this invention, the term "therapeutic and/or preventive effective amount" is the amount of a drug or pharmaceutical preparation that elicits a biological or medical response in a tissue, system, animal, or human sought by a researcher, veterinarian, physician, or other person.

When used for the above-described therapeutic and/or preventative purposes, the total daily dosage of the compound (including its crystal form) and pharmaceutical composition of Formula A of the present invention must be determined by the attending physician within the bounds of reliable medical judgment. For any given patient, the specific therapeutically effective dose level must be determined based on a number of factors, including the disorder being treated and its severity; the activity of the specific compound used; the specific composition used; the patient's age, weight, general health condition, sex, and diet; the timing, route of administration, and excretion rate of the specific compound used; the duration of treatment; drugs used in combination with or concurrently with the specific compound used; and similar factors known in the medical field. For example, it is practiced in the art to start the dose of the compound below the level required to achieve the desired therapeutic effect and gradually increase the dose until the desired effect is achieved.

Beneficial effects

The pharmaceutical compositions and formulations of this invention exhibit good safety and/or stability, as well as high P2X3 antagonistic activity, with minimal impact on taste. Studies have found that the active ingredients in the pharmaceutical compositions of this invention, in crystal forms I, III, and IV, all possess good solid-state properties. Crystal form III exhibits excellent physicochemical properties, physical and chemical stability, and compared to crystal forms I and IV, it demonstrates better PK advantages. Furthermore, its solubility in biological media is not significantly different from that of crystal forms I and V, minimizing the risk of crystal transformation during storage and production. It is also easy to prepare and suitable for industrial-scale production, especially in the development of solid dosage forms. This facilitates maintaining the stability, efficacy, safety, and quality control of the API during formulation, resulting in better pharmaceutical prospects.

Attached Figure Description

Figure 1 shows the amorphous DSC spectrum;

Figure 2 shows the amorphous TGA spectrum;

Figure 3 shows the XRPD diagram of crystal form I;

Figure 4 is a TGA/DSC overlay image of crystal form I;

Figure 5 shows the PLM diagram of crystal form I;

Figure 6 shows the XRPD diagram of crystal form II;

Figure 7 shows the ^1H NMR spectrum of crystal form II;

Figure 8 shows the TGA/DSC overlay diagram of crystal form II;

Figure 9 shows the PLM diagram of crystal form II;

Figure 10 shows the XRPD diagram of crystal form III;

Figure 11 is a TGA/DSC overlay image of crystal form III;

Figure 12 shows the PLM diagram of crystal form III;

Figure 13 shows the XRPD diagram of crystal form IV;

Figure 14 is a TGA/DSC overlay image of crystal form IV;

Figure 15 shows the PLM diagram of crystal form IV;

Figure 16 shows the XRPD diagram of crystal form V;

Figure 17 is a TGA/DSC overlay image of crystal form V;

Figure 18 is the PLM diagram of crystal form V;

Figure 19 shows the XRPD diagram of crystal form VI;

Figure 20 shows the XRPD diagram of crystal form VII;

Figure 21 shows the residual ethylene glycol ^1H NMR spectrum of crystal form VII;

Figure 22 is a TGA/DSC overlay diagram of crystal form VII;

Figure 23 shows the XRPD diagram of crystal form VIII;

Figure 24 is a TGA/DSC overlay diagram of crystal form VIII;

Figure 25 shows the ^1H NMR spectra of crystal form VIII and the raw material;

Figure 26 is a TGA/DSC overlay image of crystal form IX;

Figure 27 shows the ^1H NMR spectrum of crystal form IX;

Figure 28 shows the XRPD diagram of crystal form IX;

Figure 29 shows the XRPD overlay images of crystal form I under ambient humidity, low humidity, and high humidity conditions;

Figure 30 shows the DVS test results for crystal form I;

Figure 31 shows the XRPD overlay results before and after the DVS test of crystal form I;

Figure 32 is an XRPD overlay of crystal form III before and after heating to dehydration;

Figure 33 shows the DVS test results for crystal form III;

Figure 34 shows the XRPD overlay images of crystal form III before and after DVS testing;

Figure 35 shows the XRPD overlay before and after dehydration of crystal form IV;

Figure 36 shows the XRPD overlay images of crystal form I, crystal form VI, and crystal form VI after being placed under ambient humidity for several minutes.

Figure 37 shows the XRPD overlay of crystal form VIII and the dried sample;

Figure 38 shows the XRPD overlay of the crystal form I stability test sample;

Figure 39 shows the TGA results of crystal form I after being placed in a 60℃ (closed) stability chamber for seven days;

Figure 40 shows the XRPD overlay of the crystal form V stability test sample;

Figure 41 shows the XRPD overlay of the crystal form III stability test (7 days);

Figure 42 shows the XRPD overlay of the crystal form III stability test (3 months);

Figure 43 shows the solubility data for crystal forms I, III, and V;

Figure 44 is an XRPD overlay of the remaining solid after the solubility test of crystal form I;

Figure 45 shows the XRPD overlay of the remaining solid after the solubility test of crystal form V;

Figure 46 shows the XRPD overlay of the remaining solid after the solubility test of crystal form III;

Figure 47 is a schematic diagram of the water absorption and dehydration operation procedure in the dynamic water adsorption experiment of crystal form III;

Figure 48 shows the DVS curve of dynamic water adsorption experiment for crystal form III;

Figure 49 shows the XRPD overlay before and after the dynamic water adsorption experiment of crystal form III;

Figure 50 shows the in-situ XRPD test results for crystal form III under varying humidity;

Figure 51 shows the in-situ XRPD test results of crystal form III under varying humidity (partial magnified view);

Figure 52 is a superimposed image of XRPD crystal forms during the formulation process.

Figure 53 shows the XRPD overlay of the crystal form of the coated tablets with that of the API and blank excipients.

Figure 54 shows the crystal stability results of 100mg tablet cores.

Figure 55 shows the crystal stability results of 100mg coated tablets.

Detailed Implementation

The technical solution of the present invention will be further described in detail below with reference to specific embodiments. It should be understood that the following embodiments are merely illustrative and explanatory of the present invention, and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are covered within the scope of protection intended by the present invention.

Unless otherwise stated, the raw materials and reagents used in the following examples are commercially available products or can be prepared by known methods.

Experimental instruments and parameter testing:

XRPD (X-ray Powder Diffraction)

The crystal form of the solid obtained in the experiment was tested and analyzed using a PANalytical Empyrean equipped with a PIXcel ^1D detector. The instrument parameters were: scan range 3-40° (2θ), step size 0.013° (2θ), phototube voltage 45KV, and phototube current 40mA.

Variable humidity X-ray powder diffractometer (VH-XRPD)

The VH-XRPD was used to test and analyze the crystal form III sample (batch number: A10230-047P1). This work was performed by WuXi AppTec-WuXi Pharma. The instrument parameters are shown below:

The humidity change program settings are as follows:

TGA (Thermogravimetric Analysis)

Thermogravimetric analysis (TGA) of the samples was performed using a Discovery TGA 55 (TA Instruments, US). 2-3 mg of sample was placed in a pre-peeled open aluminum dish. The sample mass was automatically weighed inside the TGA furnace, and then heated to 250 °C at a heating rate of 10 °C/min under dry _N₂ protection.

DSC (Differential Scanning Calorimetry)

Solid samples were analyzed using a TA Instrument Q200 differential scanning calorimeter and a Discovery DSC 250. Samples were weighed and the values recorded, then placed in the sample chamber. Samples were heated from 25°C to different endpoint temperatures at a rate of 10°C/min.

PLM (Polarizing Light Microscopy) analysis

Place a small amount of powder sample on a glass slide, add a small amount of silicone oil to better disperse the powder sample, cover with a coverslip, and then place the sample on the stage of the Polarizing Microscope ECLIPSE LV100POL (Nikon, JPN). Select an appropriate magnification to observe the morphology of the sample and take a picture.

Dynamic moisture adsorption meter (DVS)

Water vapor adsorption/desorption data of the samples were collected using a ProUmid GmbH & Co. KG adsorption instrument manufactured in Germany. Typically, ~100 mg of sample was placed in a tare sample dish, and the instrument software recorded the weight change of the sample during humidity variations. For non-crystalline I and V samples, the following parameters were used for testing.

Sample temperature: 25℃ Cycle time: 10 minutes Minimum time to balance: 50 minutes Maximum balance duration: 120 minutes Weight balance: 100% Equilibrium condition: 0.01%/45 minutes Environmental Cycle #1: 0% to 0%, Step 1, 40℃, 3 hours Environmental Cycle #2: 0% to 90%, 9 steps, 25℃ Environmental Cycle #3: 80% to 0%, 8 steps, 25℃ Adsorption: 0, 10, 20, 30, 40, 50, 60, 70, 80, 90 Desorption: 80, 70, 60, 50, 40, 30, 20, 10, 0

The initial humidity for crystal form III testing was 50% (ambient humidity). Specific test parameters are shown in the table below:

Sample temperature: 25℃ Cycle time: 10 minutes Minimum time to balance: 50 minutes Maximum balance duration: 120 minutes Weight balance: 100% Equilibrium condition: 0.01%/45 minutes Environmental Cycle #1: 50% to 50%, 1 step, 25℃, 3 hours Environmental Cycle #2: 50% to 90%, 4 steps, 25℃ Environmental Cycle #3: 80% to 0%, 8 steps, 25℃ Environmental Cycle #4: 0% to 90%, 9 steps, 25℃ Environmental Cycle #5: 80% to 50%, 3 steps, 25℃ Adsorption: 50, 60, 70, 80, 90 Desorption: 80, 70, 60, 50, 40, 30, 20, 10, 0 Adsorption: 0, 10, 20, 30, 40, 50, 60, 70, 80, 90 Desorption: 80, 70, 60, 50

Hydrogen nuclear magnetic resonance (1H-NMR)

1H-NMR was performed on an AVANCE III HD 300 equipped with a SampleXpress 60 autosampler.

High-performance liquid chromatography (HPLC)

The instrument used for liquid chromatography analysis was an Agilent HPLC 1260 series, and the analytical methods are shown in the table below.

HPLC method used in solubility testing

Example 1: Preparation of the compound shown in Formula A

Step 1: Preparation of tert-butyl (S)-2-((2-(4-bromo-2,6-difluorophenyl)-7-chloroimidazolo[1,2-a]pyridin-3-yl)methyl)morpholine-4-carboxylic acid

In a 100 mL round-bottom flask, (S)-2-ethynylmorpholine-4-carboxylic acid tert-butyl ester (3.1 g, 1.0 eq, intermediate 1-4), 4-bromo-2,6-difluorobenzaldehyde (2.76 g, 1.0 eq, compound 172-1), 4-chloropyridin-2-amine (1.61 g, 1.0 eq, compound 172-2), CuCl (0.37 g, 0.3 eq), Cu(OTf)₂ (1.36 g, 0.3 eq), and isopropanol (50 mL) were added sequentially. The mixture was purged with nitrogen three times and heated overnight in an oil bath at 80 °C. TLC analysis showed that the starting material, compound 172-2, had disappeared. The isopropanol was evaporated to dryness, and the mixture was extracted sequentially with EA and ammonia. The EA phase was collected, washed sequentially with saturated brine and citric acid, dried over anhydrous sodium sulfate, and evaporated to dryness before column chromatography to obtain intermediate 172-3, a white solid (3.0 g, purity 78%). LC-MS:[M+H]+＝542.2.

Preparation of step 2(S)-2-((2-(4-bromo-2,6-difluorophenyl)-7-chloroimidazolo[1,2-a]pyridin-3-yl)methyl)morpholine

Intermediate 172-3 (2.67 g) was dissolved in dichloromethane (24 mL), followed by the addition of dioxane hydrochloride (24 mL). The mixture was stirred at room temperature for 1.0 h, and the reaction was monitored by LC-MS until complete. The reaction solution was evaporated to dryness, and water (15 mL) and dichloromethane (15 mL) were added to the solution. After extraction, the aqueous phase was discarded, and the pH was adjusted to a weakly alkaline state (pH = 8–9) with sodium bicarbonate solution. The dichloromethane phase was separated, and the aqueous phase was extracted again with dichloromethane (10 mL × 2). The dichloromethane phases were combined, washed with saturated brine, and evaporated to dryness to obtain intermediate 172-4, a white solid (1.70 g, purity 88.6%). LC-MS: [M+H]+ = 442.1.

Step 3. Preparation of methyl (S)-2-((2-(4-bromo-2,6-difluorophenyl)-7-chloroimidazolo[1,2-a]pyridin-3-yl)methyl)morpholine-4-carboxylate

Intermediate 172-4 (1.4 g, 1.0 eq) was dissolved in dichloromethane (10 mL), and triethylamine (480 mg, 1.5 eq) was added, followed by dropwise addition of methyl chloroacetate (388 mg, 1.3 eq). After 1.0 h of reaction, LC-MS showed product formation. After the reaction was complete, water (10 mL) was added and the mixture was stirred for 30 min. The dichloromethane phase was separated, and the aqueous phase was extracted again with dichloromethane (10 mL × 2). The combined dichloromethane phases were washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain intermediate 172-5, a white solid (1.01 g, purity 93.02%). LC-MS: [M+H]+ = 499.8.

Step 4: Preparation of methyl morpholine-4-carboxylate (S)-2-((2-(4-(benzylthio)-2,6-difluorophenyl)-7-chloroimidazolo[1,2-a]pyridin-3-yl)methyl)morpholine-4-carboxylate

Intermediate 172-5 (0.73 g, 1.0 eq) was dissolved in dioxane (4 mL), and BnSH (0.24 g, 1.3 eq), Pd2(dba)3 (0.04 g, 0.03 eq), Xantphos (0.04 g, 0.05 eq), and DIEA (0.60 g, 3.0 eq) were added. N2 was replaced three times, and the reaction was carried out overnight at 80 °C. LC-MS was used to monitor complete disappearance of the starting material. Dichloromethane (10 mL) and water (10 mL) were added to the reaction mixture. The dichloromethane phase was separated, and the aqueous phase was extracted again with dichloromethane (10 mL × 2). The combined dichloromethane phases were washed with saturated sodium chloride, dried over anhydrous sodium sulfate, and purified by column chromatography to obtain intermediate 172-6, a white solid (0.82 g, purity 91.53%). LC-MS: [M+H]+ = 544.2.

Step 5: Preparation of methyl morpholine-4-carboxylate (S)-2-((7-chloro-2-(4-(chlorosulfonyl)-2,6-difluorophenyl)imidazo[1,2-a]pyridin-3-yl)methyl)morpholine-4-carboxylate

Intermediate 172-6 (510 mg) was added to a reaction flask, dissolved in acetonitrile (3 mL), followed by glacial acetic acid (281 mg, 5.0 eq). SO2Cl2 (506 mg, 4.0 eq) was then added dropwise under ice bath conditions. The reaction was carried out at 0 °C for 1 h. LC-MS showed the disappearance of the starting material and the formation of intermediate 172-7. No further treatment was performed, and the reaction solution was used directly in the next step.

Step 6: Preparation of methyl morpholine-4-carboxylate (S)-2-((7-chloro-2-(2,6-difluoro-4-aminosulfonylphenyl)imidazo[1,2-a]pyridin-3-yl)methyl)morpholine-4-carboxylate

At 0°C, 2 mL of ammonia solution was diluted with 1 mL of acetonitrile and added dropwise to the above reaction solution, and the reaction was carried out at room temperature for 0.5 h. LC-MS showed that the starting material disappeared completely and the target product was formed. The reaction solution was extracted twice with water and ethyl acetate, washed with saline solution, dried over anhydrous sodium sulfate, concentrated, and purified using a C18 column (water/acetonitrile, RRt = 22.5 min). Amorphous compound A (compound A is the compound shown in formula A) was obtained as a white solid (185 mg, purity 99.74%). LC-MS: [M+H] ⁺ = 501.1.

¹ H NMR (400MHz, DMSO-d ₆ )δ＝8.11(d,J＝7.4,1H),7.29(d,J＝1.6,1H),7.22(s,2H),7.14(d,J＝6.6,2H),6.60(dd,J＝7.4,2.1,1H),3.33(d,J＝12.8,1H ),3.13(d,J＝11.3,2H),3.07(s,3H),2.97(d,J＝7.8,1H),2.77–2.69(m,1H),2.69–2.61(m,1H),2.53(dd,J＝15.5,8.3,1H).

Raw material post-processing

11.1 g of compound A was prepared according to the method shown in Formula A. 20 mL of acetone was added, and the mixture was refluxed at 65°C (under nitrogen protection) for 2.0 h. The acetone was directly evaporated to dryness, and the product was then vacuum dried at 40°C for 12 h. NMR showed approximately 1% acetone residue. The product was then vacuum dried again at 80°C for 12 h, and NMR still showed acetone residue. 5.3 g of this product was then vacuum dried again at 80°C for 12 h, and NMR still showed acetone residue. 16 mL of acetonitrile was added to this batch of product, and the mixture was refluxed at 85°C (under nitrogen protection) for 2.0 h. The acetonitrile was directly evaporated to dryness, and the product was then vacuum dried at 80°C for 12 h. NMR showed no solvent residue, and 5.2 g was stored. The product purity was 99.29%, and it was a white powder.

PLM and XRPD results show that the raw material is an irregularly shaped crystal with a diameter of 10-50 μm, exhibiting moderate crystallinity and an amorphous state. As shown in Figure 1, the DSC spectrum reveals two consecutive endothermic peaks between approximately 150-180 °C, with peak temperatures of 164±2 °C and 173±2 °C, respectively. As shown in Figure 2, the TGA spectrum shows that the sample exhibits virtually no weight loss before 230 °C.

Example 2: Preparation and Characterization of Amorphous Forms

Compound A was dissolved in a certain amount of THF and concentrated to dryness under reduced pressure to obtain an amorphous sample. XRPD characterization is shown in Figure 1.

Example 3: Preparation and Characterization of Crystal Form I

3.1 Nine solvents were selected: EtOH, IPA, NBA, MEK, ACN, acetone, EA, IPAc, and Hept. The mixtures were suspended and pulped at room temperature to a pulping concentration of 60 mg/mL to obtain crystal form I.

3.2 Six solvents were selected: IPA, NBA, MEK, acetone, Tol, and EA. The mixtures were suspended and pulped at 50°C with a pulping concentration of 100 mg/mL to obtain crystal form I.

3.3 Solvent used: IPAc. The mixture was suspended and pulped at 50°C to a pulping concentration of 50 mg/mL to obtain crystal form I.

3.4 Crystallization by slow cooling in methanol and ethanol, with cooling temperatures ranging from 50°C to RT, yields crystal form I.

3.5 Tetrahydrofuran was used as a good solvent to dissolve the sample to form a sample solution of a certain concentration. Tol, Hept, and water were slowly added dropwise to the sample solution as antisolvents, with a volume ratio of 1:10, to increase the supersaturation and thus precipitate the solid to obtain crystal form I.

3.6 EtOH was selected as the solvent for evaporation crystallization test, and crystal form I was obtained.

The XRPD of crystal form I, represented by a 2θ angle, is shown in Figure 3. In the DSC spectrum of crystal form I, an endothermic peak is observed at 152℃, with a melting enthalpy of 44±2 J/g. In the TGA spectrum of crystal form I, no weight loss is observed within the temperature range of RT-230℃. Its TGA and DSC spectra are shown in Figure 4. Combining the DSC and TGA spectra, it can be determined that this product is amorphous. In the PLM spectrum of crystal form I, the crystal form is an irregular crystal of approximately 5 μm, as shown in Figure 5.

Example 4: Preparation and Characterization of Crystal Form II

4.1 In MTBE, suspension and pulping at room temperature yielded crystal form II:

4.2 Solvent used: MTBE, suspended and pulped at 50℃, pulping concentration of 100mg/mL, to obtain crystal form II;

4.3 Tetrahydrofuran was used as a good solvent to dissolve the sample to form a sample solution of a certain concentration. MTBE was slowly added dropwise to the sample solution as an antisolvent at a volume ratio of 1:10 to increase the supersaturation, thereby causing the solid to precipitate and obtain crystal form II.

The X-ray powder diffraction pattern of crystal form II, expressed at a 2θ angle, is shown in Figure 6. In the 1H NMR spectrum of crystal form II, residual MTBE solvent signals are observed at chemical shifts of 1.10 and 3.08; its molar ratio is 0.39, and its HNMR spectrum is shown in Figure 7. In the TGA spectrum of crystal form II, a weight loss of 3.5% is observed in the temperature range of 100-160℃, and a weight loss of 2.9% is observed in the temperature range of 160-200℃. In the DSC spectrum of crystal form II, two adjacent endothermic peaks are observed, and its TGA and DSC spectra are shown in Figure 8. This is comparable to the residual MTBE 1H spectrum shown in Figure 7, which indicates that this product is an MTBE solvate. In the PLM of crystal form II, the crystal form is an irregular crystal of approximately 2 μm, and its polarized light microscope image is shown in Figure 9.

Example 5: Preparation and Characterization of Crystal Form III

5.1 Solvent used: water. The mixture was suspended and pulped at 50°C to a pulping concentration of 50 mg/mL to obtain crystal form III.

5.2 Crystal form III was prepared by suspending and pulping an amorphous sample in water at 40°C; specifically, the sample was dissolved in methanol, filtered to obtain a sample solution, and then rotary evaporated to obtain 250 mg of amorphous sample. Crystal form III was obtained after suspending and pulping in water for 20 hours.

5.3 At room temperature, the amorphous sample was dissolved in DMSO and filtered to obtain a DMSO solution. The resulting solution was slowly added dropwise to water containing seed crystals of crystal form III. As the amount of DMSO solution added increased, the amount of solid precipitated gradually increased (DMSO/water 1:2, volume ratio). After magnetic stirring at room temperature for 16 hours, stirring was stopped. The upper liquid was basically transparent, and the solid quickly settled to the bottom, yielding crystal form III. The XRPD pattern of crystal form III, expressed at a 2θ angle, is shown in Figure 10. In the TGA spectrum of crystal form III, the weight loss gradient in the RT-100℃ range is 1.5%, where "%" refers to weight percentage. In the DSC spectrum of crystal form III, the first endothermic peak is due to the removal of 0.4 water molecules, and the second endothermic peak is attributed to the melting endothermic peak after sample dehydration. Its TGA and DSC spectra are shown in Figure 11. In the PLM of crystal form III, the crystal form is an irregular crystal of about 2 μm with agglomeration of 20-50 μm, and its PLM diagram is shown in Figure 12.

Example 6: Preparation and Characterization of Crystal Form IV

Crystal form I product slurry is pulped in 50-95% water/acetone (V/V) to obtain crystal form IV.

The XRPD pattern of crystal form IV, expressed at a 2θ angle, is shown in Figure 13. In the TGA spectrum of crystal form IV, the weight loss is 1.2% within the RT-60℃ temperature range. In the DSC spectrum of crystal form IV, there are two endothermic peaks. The first broad endothermic peak is presumably due to dehydration, and the subsequent endothermic peak is a melting peak. Its TGA and DSC spectra are shown in Figure 14. Combining the DSC and TGA spectra, it can be determined that this product is a hydrate crystal form, containing approximately 0.34 water molecules. In the PLM spectrum of crystal form IV, the crystal structure is an irregular crystal of approximately 5 μm, as shown in Figure 15.

Example 7: Preparation and Characterization of Crystal Form V

7.1 Solvent used: water. The mixture was suspended and pulped at 50°C to a pulping concentration of 50 mg/mL to obtain crystal form V.

7.2 Solvent selected: ACN; suspension and pulping at 50℃; pulping concentration: 3.0 mg/mL; to obtain crystal form V.

7.3 Using MeOH as the solvent, a volatile crystallization experiment was conducted to obtain crystal form V.

The XRPD diagram of crystal form V, expressed at a 2θ angle, is shown in Figure 16. The TGA spectrum of crystal form V shows no weight loss within the RT-230℃ temperature range. The DSC spectrum of crystal form II shows an endothermic peak at 166℃±2℃, with a melting enthalpy of 70±2 J/g; its TGA and DSC spectra are shown in Figure 17. Combining the DSC and TGA spectra, this product is an amorphous form. In the PLM diagram of crystal form V, the crystal structure is an irregular crystal of approximately 5 μm; its PLM diagram is shown in Figure 18.

Example 8: Preparation and Characterization of Crystal Form VI

Crystal form VI is obtained by pulping crystal form I or crystal form V in a water/acetone mixed solvent with a water content of 10% (by volume) at 60°C.

The X-ray powder diffraction pattern of crystal form VI, expressed in terms of 2θ angle, is shown in Figure 19.

Example 9: Preparation and Characterization of Crystal Form VII

9.1 Solvent used: ethylene glycol. The mixture was suspended and pulped at 90°C to a pulping concentration of 320 mg/mL to obtain crystal form VII.

9.2 Crystallization was carried out by slow cooling in ethylene glycol, with the cooling temperature increasing from 50°C to RT, to obtain crystal form VII.

The XRPD of crystal form VII, expressed at a 2θ angle, is shown in Figure 20. The ^1H NMR spectrum of crystal form VII shows residual ethylene glycol at chemical shifts ^δ of 3.39 and 4.44, as shown in Figure 21. The TGA spectrum of crystal form VII shows a 25.7% weight loss in the rt-120℃ range. The DSC spectrum of crystal form VII shows two broad endothermic peaks; the first endothermic peak is presumably due to solvent desolvation, and its TGA and DSC spectra are shown in Figure 22. Combining the DSC and TGA spectra, this product is a solvate containing 2.79 molecules of ethylene glycol.

Example 10 Preparation and characterization of crystal form VIII

Crystal form III was used as a seed crystal. Water was added dropwise to a saturated 50% THF/water solution at 40°C as an antisolvent, resulting in an oily state. After cooling to room temperature, the mixture was further pulped to obtain crystal form VIII (wet filter cake).

The XRPD pattern of crystal form VIII, expressed at a 2θ angle, is shown in Figure 23. In the TGA spectrum of crystal form VIII, a weight loss of 5.7% occurs within the temperature range of Rt-160℃. The DSC spectrum of crystal form VIII shows only one endothermic peak, which is the melting peak after solvent removal. Therefore, crystal form VIII is a solvate, and its TGA and DSC spectra are shown in Figure 24. In the ^1H NMR spectrum of crystal form VIII, THF residues are observed at chemical shifts δ of ^1.76 and 3.60, as shown in Figure 25. Crystal form VIII contains 0.42 molecules of THF.

Example 11 Preparation and characterization of crystal form IX

Crystal form IX is prepared by pulping crystal form III in a 50% DMSO/water saturated solution at 40°C.

In the TGA spectrum of crystal form IX, the weight loss is 18.23% in the rt-160℃ temperature range. The DSC spectrum of crystal form IX shows a corresponding endothermic peak for the TGA weight loss. The TGA and DSC spectra are shown in Figure 26. In the ^1H NMR spectrum of crystal form IX, the chemical shift δ at 2.68 indicates residual DMSO solvent. The ^1H NMR spectrum of the residual DMSO is shown in Figure 27. Combining the DSC and TGA spectra, this product is a DMSO solvate. The XRPD pattern of crystal form IX, expressed at a 2θ angle, is shown in Figure 28.

Example 12: List of XRPD characteristic peaks of the crystal form of the present invention

The table below shows the positions of diffraction peaks with a relative intensity greater than 10% and ranking among the top ten in relative peak intensity in the XRPD images of nine different crystal forms of this invention.

Table A. Summary of the top ten peaks of XRPD for nine different crystal forms

Example 13: Crystal form screening and performance study of the present invention

13.1 Summary of Crystal Form Characterization and Performance Studies of the Invention

To further develop a suitable pharmaceutical form for compound A of this invention, nine different crystal forms were obtained through screening experiments, including two amorphous forms (crystal forms I and V), three hydrated forms (crystal forms III, IV, and VI), and four solvent compounds (crystal forms II, VIII, and IX). Specific characterization data and performance comparisons are summarized in Table B below. Comparative analysis showed that crystal forms I, III, and V exhibited better solid-state properties compared to other crystal forms, and their stability and solubility in biological media will be subsequently tested.

Table B

13.2 Crystal Form I

Crystal form I can be obtained by the preparation method in Example 3. The study found that crystal form I absorbs moisture and transforms into hydrate crystal form IV under high humidity conditions, but after drying in a vacuum drying oven at 30°C, it transforms back into the initial crystal form I (Figure 29). Combined with the DVS test results (Figure 30), it can be seen that the interconversion between crystal forms I and IV is reversible. Crystal form I exhibits hygroscopicity (6.8%, 80% RH), but the crystal form remains unchanged after DVS testing (Figure 31).

13.3 Crystal Form III

Crystal form III was obtained using the preparation method described in Example 5. Experimental results (Figure 32) show that after dehydration, crystal form III quickly reabsorbs moisture and reverts to crystal form III under ambient humidity. DVS results (Figure 33) show a sudden weight jump in the crystal form III sample at 10% RH, presumably corresponding to the removal and gain of water of crystallization, respectively; and the water content of the sample does not change significantly over a wide humidity range. As shown in Figure 34, there is no significant change in XRPD before and after the DVS test.

13.4 Crystal form IV

Crystal form IV is a product of hygroscopic formation of crystal form I under high humidity conditions. Crystal form IV can also be obtained by pulping crystal form I or V in a water/acetone mixed solvent with a water content of 50-95%. Crystal form IV is only stable under high humidity conditions and transforms into amorphous crystal form I after vacuum drying and dehydration (Figure 35).

13.5 Crystal Form V

Crystal form V is an amorphous form, obtained only by pulping the first batch of raw materials in water at 60°C or by suspending and pulping crystal form I and V in equal proportions at 60°C, but subsequent preparations have failed to be replicated.

13.6 Crystal form VI

Crystal form VI was obtained by pulping either crystal form I or crystal form V in a water/acetone mixed solvent with a water content of 10% (volume ratio) at 60°C. As shown in Figure 36, this crystal form sample transformed into crystal form I after being placed at ambient humidity (35% RH) for a few minutes. This indicates that crystal form VI may be a highly unstable hydrate.

13.7 Crystal form VIII

Crystal form VIII is a hydrate, but this hydrate is unstable. After dehydration (vacuum drying at 40°C for 3 hours), it transforms into crystal form I (Figure 37).

13.8 Stability Study of Crystal Forms I, III, and V

The physical and chemical stability of crystal forms I, III, and V was investigated after 7 days at 60℃ (closed) and 40℃/75%RH (open). The results are shown in Table C and Figures 38-41. Crystal forms III and V did not change in crystal form after 7 days under the above test conditions, and their chemical purity did not decrease significantly. This indicates that crystal forms III and V have good physical and chemical stability. However, crystal form I underwent hygroscopic transformation at 40℃/75%RH (open) to become hydrated crystal form IV, a result consistent with the DVS results.

Crystal form III was selected as the target crystal form for further development, and its stability was further confirmed. Crystal form III was placed at 60℃ (closed) and 40℃/75%RH (open) for three months to examine its physical and chemical stability. The results are shown in Table D and Figure 42. Crystal form III did not change its crystal form after three months under the above test conditions, indicating good physical stability.

Table C. Stability evaluation results for crystal forms I, III, and V

Table D. Three-month stability evaluation results of crystal form III

13.9 Solubility test of crystal forms I, III, and V in biological media

The solubility test results of crystal forms I, III, and V in biological media of SGF, FaSSIF, and FeSSIF are shown in Table E and Figure 43. The solubility of the three crystal forms in different biological media is not significantly different, and the solubility in SGF is higher than 5 mg/mL. The solubility in FeSSIF is approximately three times that in FaSSIF, indicating that food may aid drug absorption. The pH of the biological media buffer did not change significantly during the test.

As shown in Figure 44, crystal form I transformed into hydrate crystal form IV after 0.5 hours in FaSSIF and FeSSIF buffer solutions. Crystal forms III and V did not change their crystal form during the test; the results are shown in Figures 45 and 46.

Table E. Solubility test results of crystal forms I, III, and V in biological solvents

"Soluble": This indicates that the sample's solubility in the medium is greater than 5 mg/mL, as observed by the naked eye.

“N/A”: Indicates that no solid sample was used for XRPD testing.

The above experimental results show that crystal form III exhibits good physicochemical properties, physical and chemical stability, and its solubility in biological media is not significantly different from that of crystal forms I and V. Since crystal form I readily transforms into crystal form IV at RH > 40%, posing a risk of transformation during storage and production, and crystal form V is difficult to prepare and unsuitable for large-scale production, this invention prefers crystal form III for formulation development.

Example 14: Scale-up preparation of crystal form III

1) Solution preparation:

Under nitrogen protection, add DMSO (4V) to reactor A, start stirring, add API to the reactor, rinse the reactor with DMSO (0.8V), adjust the temperature to 25±5℃, and stir for at least 0.5h.

2) Crystal transformation:

Add purified water (20V) to reactor B through a microfiltration filter, adjust the temperature to 40±5℃, and add the DMSO solution from reactor A dropwise to reactor B through a microfiltration filter. Rinse the microfiltration filter with DMSO (0.2V). Continue adding the solution for at least 2 hours and stirring for at least 4 hours. Take a sample for XRPD analysis. The standard is that the XRPD spectrum is consistent with the reference standard (crystal form III). If it does not meet the standard, maintain the temperature at 40±5℃ and continue stirring for at least 4 hours. Take a sample for XRPD analysis until it meets the standard. Cool to 25±5℃ and stir for at least 2 hours. Centrifuge using a 300-mesh filter bag, rinse the filter cake with purified water (2V), and collect the filter cake. Under nitrogen protection, purified water (10V) was added to reactor B through an empty filter. Stirring was started, and the above wet filter cake was added to the reactor. The temperature was controlled at 25±5℃, and the mixture was stirred for at least 0.5h. The mixture was centrifuged with a 300-mesh filter bag, and the filter cake was rinsed with purified water (5V) and collected.

3) Drying:

After drying the filter cake in a vacuum oven at 45±5℃ for at least 4 hours, samples are sent for GC analysis, with a standard of DMSO ≤ 0.5000%. If the standard is not met, samples are sent for GC analysis at least every 2 hours until the standard is met. Samples are then sent for KF analysis, with a standard of 2.0%–3.0%. If the KF is higher than 3.0%, the filter cake is dried at 45±5℃ for at least 1 hour, and samples are sent for KF analysis until the standard is met. If the KF is lower than 2.0%, a stream of moist nitrogen is blown into the oven for at least 0.5 hours, and samples are sent for KF analysis until the standard is met.

The basic information of the multiple batches of products obtained is summarized in Table F:

Table F. Summary of Moisture and Particle Size Data for Each Batch

Example 15: Stability Study of Crystal Form III Formulation

In specific formulations and manufacturing processes, crystal forms are prone to transformation, leading to changes in drug properties and compatibility, which in turn affects the stability, efficacy, safety, and quality control of the formulation. Therefore, further research is needed on the stability properties of crystal form III in the formulation of this invention.

15.1 Dynamic water adsorption experiment of crystal form III,

The experimental procedure is shown in Figure 47, its DVS curve is shown in Figure 48, and the XRPD overlay before and after the DVS test is shown in Figure 49.

The results showed that the adsorption and desorption curves of crystal form III almost completely overlapped, indicating that crystal form III did not form hydrates or formed non-stoichiometric hydrates during the hygroscopic process. Combined with the low dehydration temperature in the aforementioned TGA spectrum (Figure 11), crystal form III is a non-stoichiometric hydrate.

The hygroscopic curves in Figure 48 show that the weight change is abrupt in the range of 0-10% RH, and the sample rapidly adsorbs water. Subsequently, the weight gain slows down. It is inferred that the dehydrated crystal form III is very easy to return to a watery state under low humidity conditions, and crystal form III is slightly hygroscopic. In the range of 10-80% RH, the adsorbed water content is ~1.2%.

15.2 Stability Test Experiment II for Crystal Form III

Based on the test results of Example 13, the stability of crystal form III under the dehydration-hygroscopic cycle conditions of 25°C, 40%RH-0%RH-40%RH was investigated in order to confirm the control strategy of crystal form III.

The results showed (see Table G): Under 25℃ conditions, in the dehydration-hygroscopic cycle of 40%RH-0%RH-40%RH, it was found that under 0%RH conditions, complete dehydration transformed into a new crystal form, amorphous crystal form X, which is reversibly interconvertible with crystal form III. Combined with the complete match of the DVS adsorption-desorption curve of crystal form III, it was determined that crystal form III is a non-stoichiometric hydrate.

Within the 5%RH to 20%RH range, the position of ~12.9° (2θ) varies slightly under different humidity conditions, but the crystal form spectrum (see Figure 50 - in-situ XRPD test results of crystal form III under humidity and Figure 51 - in-situ XRPD test results of crystal form III under humidity (partial magnified view)) both show the characteristic diffraction peaks of crystal form III. When the RH is 40%, the sample completely transforms into crystal form III. Combined with the DVS spectrum showing the water content at 40%RH and the TGA results of crystal form III, crystal form III can be stably maintained when the water content is controlled above 1.6%.

Table 1 Summary of VH-XRPD Results for Crystal Form III G

Note: This crystal form is defined as a new crystal form X because it has a diffraction peak at 12.9. However, due to the extreme conditions, it was impossible to obtain and accurately characterize this crystal form.

Example 16 Pharmacokinetic Analysis of Different Crystal Forms in Animals

In this invention, the crystal forms I, III, and V of the compounds in the above embodiments were preferably used in pharmacokinetic experiments in beagle dogs. The relevant experimental details are as follows:

Animals: 9 male beagles, divided into three groups of 3 each, weighing between 10-13 kg (Source & Certificate No.: Beijing Mas Biotechnology Co., Ltd., No. 1103182011000161);

Administration route and dosage: PO: 10 mg/kg

Drug preparation: 0.5% CMC-Na was used as the solvent. 400 mg of different crystal forms of compound A were weighed and added to 79 mL of the solvent. After sonication for 20 min, a stir bar was added and the mixture was stirred thoroughly for 3 hours (the time from compound preparation to animal administration was less than 6 hours at room temperature). After the drug solution was observed to be a uniform and fine suspension, the solvent was gradually added until the specified volume was reached to achieve the target concentration. The preparation was stirred at room temperature for 10 minutes before administration and continued to be stirred during administration.

Sampling points: Blood was collected from the PO group at 0.167, 0.5, 1, 2, 4, 6, 8, and 24 hours.

Animals in the PO group were fasted overnight before administration but allowed free water. Food was resumed 4 hours after administration.

Sample processing:

1. After administration to both groups of beagle dogs, blood was collected from the jugular vein at each sampling time point. Approximately 1.0 mL of blood was collected at each time point into an anticoagulant EP tube (containing 10 μL of EDTA-K2, 375 mg/mL). The tube was slowly inverted three times and stored in an ice box (not exceeding 30 minutes). The tube was centrifuged at 3200g at 4℃ for 10 min. The plasma was then stored in an ultra-low temperature freezer until testing.

2. Sample pretreatment: Take 40 μL of sample, add 160 μL of acetonitrile containing 0.1% FA and 200 ng/mL mixed standard solution to precipitate protein, vortex mix thoroughly, and centrifuge the sample at 13000 pm for 10 minutes at 4℃. Take 100 μL of the supernatant into another 96-well plate, add 100 μL of methanol:water (1:3, v:v) solution containing 0.1% FA, and vortex mix for 10 minutes.

3. Sample testing:

(1) Mass spectrometry conditions

1. Mass spectrometry parameters

Ion source: Ion Electrospray (ESI)

Ionization mode: Positive ion mode

Detection Mode: Multiple Response Monitoring (MRM)

Electrospray voltage: 5500V

Ion spray temperature (Turbo Ion Spray Temp): 550℃

Curtain Gas Type: Nitrogen Setting: 35

Collision cell gas type (CAD Gas Type): Nitrogen Setting: Medium

Nebulizing Gas Type (Gas 1): Nitrogen Setting: 55.00

Auxiliary Gas Type (Gas 2): Nitrogen Setting: 55.00

(2), liquid phase conditions

Column: Poroshell 120EC-C18 (4.6×50mm, 2.7μm)

Mobile Phase A: 0.1% aqueous solution of formic acid

Mobile Phase B: 0.1% formic acid in methanol solution

Rinse Port Wash Solution: 50% methanol

Column temperature: 40℃

Flow rate: 0.55 mL/min

Automated sampler temperature (Sample Tray Temp): 10℃

Injection volume: 3 μL

Needle Stroke: 49mm

Automated Sampler Cleaning Settings (Rinse Pump Setting): Rinse Port Only

Rinse Mode (Before aspiration)

Rinse Volume for Autosampler: 500 μL

Rinse Dip Time during syringe cleaning: 2 seconds

Elution gradient:

Time (min) Module Function Value (%) 0.01 Pumps Pump B Conc. 30 0.20 Pumps Pump B Conc. 30 1.20 Pumps Pump B Conc. 98 2.40 Pumps Pump B Conc. 98 2.41 Pumps Pump B Conc. 30 3.00 System Controller Stop  

Data Analysis:

The data will be analyzed using WinNonlin (version 5.2.1 Pharsight, Mountain View, CA) through a non-compartmental model to obtain PK parameters (C _max , T _max , AUC _last , T _1/2 , etc., selected according to different routes of administration). The results are shown in the table below.

Example 17: Formulation and Process of the Present Invention

Example 17.1 Experimental Materials

Table H-1. Experimental Materials

17.2 The formulations of 25mg and 100mg coated tablets are shown in Table H-2.

Table H-2. Formulation Prescription

17.3 Preparation process of 25mg coated tablets and 100mg coated tablets

(1) Weighing

Equipment: Balance

Steps: Weigh the raw and auxiliary materials and set aside.

(2) Pretreatment

Equipment: 60 mesh sieve, 40 mesh sieve

Steps: Mix the compound shown in Formula A with colloidal silica 200 in a low-density polyethylene bag for 1 min, pass through a 60-mesh sieve, rinse the used 60-mesh sieve with microcrystalline cellulose 102, and place it in the same low-density polyethylene bag as mixture 1.

Mix lactose monohydrate and croscarmellose sodium in a low-density polyethylene bag for 1 minute, and then pass through a 40-mesh sieve to obtain mixture 2.

Magnesium stearate was sieved once through the same 40-mesh sieve and then placed in another pharmaceutical low-density polyethylene bag.

(3) Adhesive formulation

Equipment: Top-mounted electric stirrer, pneumatic stirrer

Steps: Weigh the purified water, slowly add hydroxypropyl cellulose EXF to the purified water to dissolve while stirring until completely dissolved, and prepare a 9% concentration adhesive solution.

(4) Wet granulation

Equipment: Wet granulation machine;

Step 1: Mixing: Place approximately half of Mixture 2 and Mixture 1 into a wet granulation pan. Rinse the low-density polyethylene bag containing Mixture 1 with the remaining Mixture 2, then add it to the wet granulation pan. Set the stirring speed to 250.0 rpm, the cutting speed to 450.0 rpm, and the mixing time to 5 min. After mixing, measure the loss on drying of the material and, if necessary, test the moisture content.

Step 2: Spraying: Set the stirring speed to 250.0 rpm and the cutting speed to 1500.0 rpm, and spray the prepared binder into the wet granulation pot. Adjust the spraying speed of the wet granulation to approximately 200–347 g/min.

Step 3: Add water: After adding the adhesive, observe the granulation process. If water needs to be added, add an appropriate amount of water.

Step 4: Granulation: Set the stirring speed to 250.0 rpm, the cutting speed to 1500.0 rpm, and the granulation time to 60 s. Obtain wet granules.

IPC: Mixed powder: Loss on drying, moisture content to be tested as needed.

(5) Wet whole grain

Equipment: Granulator, 6x6mm screen

Steps: Place the wet granules into a granulator and granulate them through a 6x6mm mesh screen. The granulator speed is 1000.0 rpm.

(6) Drying

Equipment: Fluidized bed;

Step 1: Preheating: Set the equipment parameters as follows: inlet air temperature: 50-60℃, inlet air volume: 20-150 ^m³ /h. When the outlet air temperature reaches approximately 35℃ or higher, add the wet granules to begin drying.

Step 2: Drying: Inlet air temperature: 50-60℃, inlet air volume: 20-150 ^m³ /h, filter bag shaking time: 1±0.8s, filter bag shaking frequency: 8±5s, backflushing pressure: 200-350 kPa. Record equipment parameters every 10 minutes. When the product temperature reaches approximately 30℃, take a 2-3g sample to determine the material's loss on drying. When the material's loss on drying is between 1.0% and 2.5%, turn off the inlet air temperature, stop drying, and record the final product's loss on drying value. Detect moisture content as needed.

IPC: Loss on drying; moisture content is measured as needed.

(7) Dry granulation

Equipment: Granulator, with round hole screen

The dry granules are placed in a granulator and granulated through a perforated screen. The granulator rotates at 1000 rpm.

(8) Lubrication

Equipment: Hopper mixer

Steps: Take a portion of the dry granules, magnesium stearate and the remaining dry granules and put them into a 15L hopper for total mixing. The mixing speed is 20.0 r/min and the mixing time is 5 min.

IPC: Mixing uniformity

(9) Tableting

Equipment: Tableting machine, rotary tablet sieve machine, metal detector

Steps: Add the material to the hopper of the tablet press for tableting.

25mg specification: 6mm shallow concave round punch, target weight of single core 100.0mg, single core weight controlled within ±7.5%, single core hardness controlled within 35-90N;

100mg specification: 10mm shallow concave round punch, target weight of single core 400.0mg, single core weight controlled within ±5.0%, single core hardness controlled within 70-130N;

IPC: Tablet weight, hardness, disintegration time, friability, and appearance.

(10) Coating

Equipment: Coating machine, top-mounted electric stirrer

Step 1: Preparation of coating solution: Weigh purified water and turn on the stirrer. Slowly add the coating powder to the purified water at room temperature. After it is completely added, continue stirring for at least 45 minutes to ensure that the coating solution is evenly dispersed, thus obtaining a coating solution with a 15% solid content.

Step 2: Preheating: Set the inlet air temperature to 50-70℃, the inlet air volume to 400± ^100m³ /h, and the coating pan speed to 2-5rpm to preheat the two types of cores. When the exhaust air temperature reaches about 45℃, start coating.

Step 3: Coating: Set the inlet air temperature to 50-70℃, reactor rotation speed to 5-15 rpm, inlet air volume to 400±100 ^m³ /h, pump flow rate to 4-20 ml/min, atomization pressure to 1.5±1.0 bar, and atomization angle control pressure to 2.0±1.0 bar. During the coating process, monitor the reactor temperature, atomization coating conditions, and ensure there is no coating sticking. Control the coating weight gain to 2.0-4.0%.

Step 4: Drying and Cooling: After spraying, maintain an air inlet temperature of 50-70℃, adjust the coating pan speed by 2-5 rpm, and the air inlet volume to 400±100 ^m³ /h. Dry for 5 minutes. Stop heating, maintain an air inlet volume of 400±100 ^m³ /h, and cool for at least 5 minutes before discharging the material and placing it in a pharmaceutical low-density polyethylene bag.

IPC: Coating for Weight Gain

(11) Packaging and labeling

Equipment: Electromagnetic induction aluminum foil sealing machine, electronic automatic counting machine

Procedure: Pack 25mg coated tablets into 45mL high-density polyethylene (HDPE) bottles for oral solid medication, with a packaging specification of 30 tablets/bottle. Affix a label to each bottle.

100mg coated tablets are packaged into 75mL high-density polyethylene (HDPE) bottles for oral solid medication, with a packaging specification of 30 tablets per bottle. Each bottle is labeled.

Example 18: Stability study of formulation process on crystal form III

To investigate the effect of the formulation preparation process on the crystal form, samples were taken and analyzed at different stages of the preparation process of the coated tablets according to Example 17.3. The analysis results are shown in Table I, and the summary results of the XRPD diagrams of the products at each sampling point are shown in Figure 52. The results show that the crystal form of the active pharmaceutical ingredient remains stable during the wet granulation process and the tableting step.

Table I. Statistical Table of Crystal Forms in Formulation Process

sampling Crystal form raw materials Crystal form III Chip Crystal form III Blank supplementary materials —— Granulation of granules Crystal form III Mixing powder before granulation Crystal form III

Example 19: Dissolution detection of coated tablets and investigation of API crystal changes:

The results are shown in Table J, and the XRPD results of the coated tablets, API, and blank excipients are shown in Figure 53. As can be seen from Table J and Figure 53, the 25 mg and 100 mg coated tablets dissolved quickly and completely, and the API crystal form remained unchanged after coating.

Table J. Dissolution results of coated tablets

Example 20: Study on tablet stability

Stability studies were conducted on 25 mg and 100 mg tablet cores and coated tablets. Results on related substances are shown in Tables K-1 and K-2, results on dissolution tests are shown in Tables K-3 and K-4, and results on crystal form stability are shown in Figures 54 and 55.

Table K-1. Stability of related substances in the chip core

Note: Under conditions of 25°C/92.5% RH/open/30 days, the chip core developed mold, therefore it was not tested.

Table K-2. Stability of related substances in coated tablets

Note: Under conditions of 25°C/92.5% RH/open-face/30 days, the coated sheets developed mold, therefore no testing was conducted.

The results above show that the 25mg tablet cores showed no significant changes in related substances after 10 days of storage under the following conditions: 60℃/open, 25℃/92.5%RH/open, 40℃/75%RH/closed, and light/closed. However, after 10 and 20 days of storage under light/open conditions, the related substances increased significantly. In contrast, the coated tablets showed no significant changes, indicating that coating can effectively avoid the influence of light on the product.

No significant changes in related substances were observed in 25mg and 100mg coated tablets after 30 days of storage at 60℃/open and 40℃/75%RH/closed conditions. Under light exposure, no significant changes were observed in related substances after 20 days of open storage and 10 days of closed storage. No significant changes were observed in related substances after 10 days of storage at 25℃/92.5%RH/open conditions (mildew occurred after 30 days, but was not detected). These results indicate that the coated tablets have good stability.

Table K-3. Dissolution stability of 100mg tablet cores

Table K-4. Dissolution stability of 25mg and 100mg coated tablets

The results above show that the dissolution rate of 100mg tablet cores did not change significantly after 10 days of storage under the following conditions: 60℃/open, 40℃/75%RH/closed, 25℃/92.5%RH/open, and light/open. The dissolution rate of 25mg and 100mg coated tablets did not change significantly after 30 days of storage under the following conditions: 60℃/open, 40℃/75%RH/closed. The dissolution rate of 100mg coated tablets did not change significantly after 10 days of storage under the following conditions: 25℃/92.5%RH/open, and light/open. These data indicate that the product has good dissolution stability.

As shown in Figures 54 and 55, the crystal form stability of the 100mg tablet core and coated tablets did not change after being placed under the conditions of 60℃/open, 40℃/75%RH/closed, and 25℃/60%RH/open for 30 days, indicating that the product has good physical stability.

Experiments have demonstrated that the pharmaceutical compositions and formulations of the present invention have good safety and/or stability, high P2X3 antagonistic activity, and minimal impact on taste.

The embodiments of the present invention have been described above. However, the present invention is not limited to the above embodiments. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims (20)

1. A pharmaceutical composition, said pharmaceutical composition being a tablet, comprising, by weight: 10-40 parts of the compound shown in Formula A; 50-80 parts of diluent; Disintegrant 0.5-6 parts; 0.5-6 parts adhesive; 0.1-3 parts of gliding agent; Lubricant 0.1-3 parts; The sum of the weight parts of each component in the pharmaceutical composition is 100 parts; The structure of the compound shown in formula A is as follows: The compound represented by Formula A is selected from crystal form III. Crystal form III exhibits characteristic peaks in X-ray powder diffraction at 12.91°±0.20°, 16.77°±0.20°, 19.27°±0.20°, 22.80°±0.20°, 13.75°±0.20°, 14.46°±0.20°, and 20.86°±0.20° when irradiated with Cu-Kα radiation and expressed in 2θ angles. The diluent is selected from one or two of the following substances: lactose monohydrate, microcrystalline cellulose; The disintegrant is selected from croscarmellose sodium cellulose; The adhesive is selected from one, two or more of the following substances: hydroxypropyl cellulose, methylcellulose, carboxymethyl cellulose, ethylcellulose and hydroxypropyl methylcellulose; The gliding agent is selected from one, two or more of the following substances: colloidal silica, silica, talc; The lubricant is selected from one, two or more of the following substances: magnesium stearate, calcium stearate, and zinc stearate. 2. The pharmaceutical composition according to claim 1, characterized in that, The crystal form III, when subjected to Cu-Kα radiation, exhibits characteristic peaks in X-ray powder diffraction at 12.91°±0.20°, 16.77°±0.20°, 19.27°±0.20°, 22.80°±0.20°, 13.75°±0.20°, 14.46°±0.20°, 20.86°±0.20°, 21.08°±0.20°, 23.75°±0.20°, and 24.05°±0.20° in 2θ angles. 3. The pharmaceutical composition according to claim 1, characterized in that, The crystal form III has an XRPD pattern as shown in Figure 10. 4. The pharmaceutical composition according to claim 1, characterized in that: The particle size of the compound shown in Formula A is 1-40 μm. 5. The pharmaceutical composition according to claim 1, characterized in that: The D10 _particle size of the compound shown in Formula A is 1-5 μm; The D50 _particle size of the compound shown in Formula A is 6-15 μm; The compound shown in Formula A has a D ₉₀ particle size of 20-40 μm. 6. The pharmaceutical composition according to claim 1, characterized in that: The bulk density (bulk density) of the compound represented by Formula A is 0.2-0.3 g/mL. 7. The pharmaceutical composition according to claim 1, characterized in that: The actual density (bulk density) of the compound shown in Formula A is 0.32-0.5 g/mL. 8. The pharmaceutical composition according to any one of claims 1-7, characterized in that, The pharmaceutical composition also contains a flavoring agent. 9. The pharmaceutical composition according to any one of claims 1-7, characterized in that, The adhesive is hydroxypropyl cellulose, the flow aid is colloidal silica, and the lubricant is magnesium stearate. 10. The pharmaceutical composition according to any one of claims 1-7, characterized in that, The pharmaceutical composition comprises the following components in parts by weight: 15-35 parts of the compound shown in Formula A, 55-75 parts of lactose monohydrate and microcrystalline cellulose, 1-5 parts of croscarmellose sodium, 1-5 parts of hydroxypropyl cellulose, 0.3-2 parts of colloidal silica, and 0.3-2 parts of magnesium stearate. 11. The pharmaceutical composition according to any one of claims 1-7, characterized in that, The tablets are coated tablets, consisting of a core and a coating layer. 12. The pharmaceutical composition according to claim 11, characterized in that, The film coating material of the coating layer is the Opadry gastric-soluble coating series. 13. The pharmaceutical composition according to claim 11, characterized in that, The core of the coated tablet contains the following components: 25 mg of the compound shown in Formula A, 49.6 mg of lactose monohydrate, 17.4 mg of microcrystalline cellulose, 3.0 mg of croscarmellose sodium, 3.0 mg of hydroxypropyl cellulose, 1.0 mg of colloidal silica, and 1.0 mg of magnesium stearate. 14. The pharmaceutical composition according to claim 11, characterized in that, The core of the coated tablet contains the following components: 100 mg of the compound shown in Formula A, 198.4 mg of lactose monohydrate, 69.6 mg of microcrystalline cellulose, 12.0 mg of croscarmellose sodium, 12.0 mg of hydroxypropyl cellulose, 4.0 mg of colloidal silica, and 4.0 mg of magnesium stearate. 15. A method for preparing a pharmaceutical composition according to any one of claims 1-14, characterized in that, The pharmaceutical composition is prepared by wet granulation and tableting, comprising the following steps: wet granulation of the pharmaceutical composition excluding the lubricant, granulation, drying, and granulation again to obtain dry granules; mixing the dry granules with the lubricant and tableting. In the drying step of the wet granulation and tableting method, the material drying weight loss is controlled within 1.5%-2.5%. 16. The method according to claim 15, characterized in that, When the material's weight loss during drying is between 1.0% and 2.5%, turn off the air inlet and stop drying. 17. The method according to claim 15 or 16, characterized in that, The equipment used in the drying step is a fluidized bed. 18. The method according to claim 15 or 16, characterized in that, The wet granulation process includes: mixing the compound shown in Formula A, a flow aid, a diluent, and a disintegrant; spraying an adhesive solution into the mixture; and, after the adhesive solution has been sprayed, optionally adding water or not adding water, and then granulating. 19. The method according to claim 18, characterized in that, The tablets are prepared as coated tablets, and the method for preparing coated tablets includes the following steps: (1) The mixture is wet-granulated, sized, dried, and granulated again to obtain dry granules; (2) The dry granules are mixed with the lubricant and compressed into tablets to obtain tablet cores; (3) Spray coating liquid onto the core to obtain the coated sheet. 20. The use of the pharmaceutical composition according to any one of claims 1-14 in the preparation of a pharmaceutical formulation, characterized in that, The pharmaceutical preparation is a drug used to treat acute or chronic cough.