Oral pharmaceutical composition of pde4b inhibitor compound and preparation method therefor

Abstract

NOT VISIBLE DUE TO STATUS OF PATENT

Description

An oral pharmaceutical composition of a PDE4B inhibitor compound and its preparation method Technical Field

[0001] The present invention relates to the technical field of chemical medicines, and in particular to an oral pharmaceutical composition of a PDE4B inhibitor compound and a preparation method thereof. Background Art

[0002] Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, fibrotic interstitial lung disease with lesions localized to the lungs. It predominates in middle-aged and elderly individuals. Pulmonary histology and / or chest high-resolution CT (HRCT) findings are characteristically of usual interstitial pneumonia (UIP). The etiology remains unclear. The disease course can be acute, subacute, or chronic, and is mostly sporadic. Statistics show an annual prevalence of approximately 2 to 29 cases per 100,000 people in the general population, with a steadily increasing trend, estimated at 11% per year. Approximately 100,000 people in the United States and 110,000 in the European Union have IPF, with 35,000 new cases reported annually in the EU. The annual prevalence of IPF in Japan is approximately 2.23 to 10 cases per 100,000 people, but the actual number is far higher. my country, a country with a rapidly aging population, also sees an increasing number of IPF cases, conservatively estimated to be at least 500,000. As a chronic interstitial lung disease, IPF has an insidious onset and gradually worsens, but can also present with acute exacerbations. The average survival after IPF diagnosis is only 2.8 years, and the mortality rate is higher than that of most cancers. IPF has been called a "tumor-like disease."

[0003] Current treatment options are limited. Glucocorticoids or combined cytotoxic drugs are customarily used for treatment, and the dosage and course of treatment depend on the patient's specific condition. The currently recommended treatment regimen is glucocorticoids combined with cyclophosphamide or azathioprine. Other therapeutic drugs include N-acetylcysteine, gamma-interferon and pirfenidone (TNF-α inhibitor), colchicine, penicillamine, etc. The clinical efficacy of these drugs needs further demonstration. The present technical solution provides an oral pharmaceutical composition of a phosphodiesterase (PDEs) inhibitor compound and a preparation method thereof, which is only used to treat idiopathic pulmonary fibrosis (IPF) and other related diseases.

[0004] Phosphodiesterases (PDEs) are a family of enzymes that hydrolyze cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP), playing a crucial role in regulating cAMP and cGMP levels. PDEs are divided into three categories: the first category specifically hydrolyzes cAMP, including PDE4, PDE7, and PDE8; the second category specifically hydrolyzes cGMP, including PDE5, PDE6, and PDE9; and the third category, which can hydrolyze both cAMP and cGMP, includes PDE1, PDE2, PDE3, PDE10, and PDE11.

[0005] PDE4 is widely expressed in various tissues. PDE4 enzymes are generally divided into four subtypes (PDE4A, PDE4B, PDE4C, and PDE4D). Their functions include regulating brain function, regulating monocyte and macrophage function, and regulating vascular smooth muscle cell proliferation. Although PDE4 is expressed in most cells, immune cells primarily rely on PDE4 to catalyze the hydrolysis of cAMP. Therefore, PDE4 inhibitors may be used to treat a variety of inflammatory diseases, including psoriasis, neuroinflammation, chronic obstructive pulmonary disease (COPD), asthma, ankylosing spondylitis (AS), inflammatory bowel disease (IBD), atopic dermatitis, rheumatoid arthritis (RA), and systemic lupus erythematosus (SLE).

[0006] The present invention relates to the field of pharmaceutical preparations, and more particularly to an oral pharmaceutical composition of a small molecule phosphodiesterase (PDEs) inhibitor compound (I). The present invention also relates to a method for preparing the pharmaceutical composition, and the use of the pharmaceutical composition in a medicament for treating diseases or conditions associated with phosphodiesterase (PDEs) receptors. The formulation technology employs specific solubilization technology, compositions, and preparation methods to obtain a formulation with high bioavailability, good content uniformity, good stability, and industrial production capabilities, intended to treat idiopathic pulmonary fibrosis and other related diseases, enriching the current treatment options in this disease field. Summary of the Invention

[0007] The present invention provides an oral pharmaceutical composition comprising:

[0008] An active ingredient, wherein the active ingredient is a compound of formula (I) or a pharmaceutically acceptable salt thereof,

[0009] The mass percentage of the active ingredient in the pharmaceutical composition is 0.1%-10%, preferably 0.5%-6.5%.

[0010] In some embodiments, the composition further comprises at least one of a filler, a disintegrant, a binder, and a lubricant.

[0011] In some embodiments, the composition further comprises a glidant.

[0012] In some embodiments, the filler is selected from at least one of lactose, microcrystalline cellulose, mannitol, starch, sucrose, dextrin, calcium hydrogen phosphate, calcium sulfate, and calcium carbonate.

[0013] In some embodiments, the disintegrant is selected from at least one of cross-linked carboxymethyl cellulose sodium, cross-linked polyvinylpyrrolidone, sodium starch glycolate, and low-substituted hydroxypropyl cellulose.

[0014] In some embodiments, the binder is selected from at least one of hydroxypropyl cellulose, hypromellose, copovidone, povidone, ethyl cellulose, and methyl cellulose.

[0015] In some embodiments, the lubricant is selected from at least one of magnesium stearate, calcium stearate, sodium stearyl fumarate, and glyceryl behenate.

[0016] In some embodiments, the glidant is selected from at least one of silicon dioxide, talc, and micro-powdered silica gel.

[0017] In some embodiments, the mass percentage of the disintegrant in the pharmaceutical composition is 1%-15%, preferably 2%-5%.

[0018] In some embodiments, the binder accounts for 1% to 20% by weight in the pharmaceutical composition, preferably 2% to 5% by weight.

[0019] In some embodiments, the mass percentage of the lubricant in the pharmaceutical composition is 0.5%-3%.

[0020] In some embodiments, the mass percentage of the glidant in the pharmaceutical composition is 0%-5%, preferably 0%-1%.

[0021] The present invention provides an oral pharmaceutical composition comprising:

[0022] a) a compound of formula (I) or a pharmaceutically acceptable salt thereof,

[0023] as well as

[0024] Solubilizers and / or acidity regulators.

[0025] In some embodiments, the solubilizing agent is selected from an ionic surfactant, a nonionic surfactant, or a combination thereof, and the nonionic surfactant does not include polyethylene glycol-15-hydroxystearate.

[0026] In some embodiments, the ionic surfactant is selected from sulfated castor oil, sodium lauryl sulfate, sodium hexadecyl sulfate, sodium octadecyl sulfate.

[0027] In some embodiments, the nonionic surfactant is selected from Spans (Span 20, Span 40, Span 60, etc.), Tweens (Tween 60, Tween 80, etc.), Poloxamer (Poloxamer 40, Poloxamer 45, etc.), Benzyls (Benzyl 35, etc.) and Poloxamers (Poloxamer 188, Poloxamer 407, etc.).

[0028] In some embodiments, the solubilizing agent is selected from at least one of sodium cetyl sulfate, sodium lauryl sulfate, poloxamer 188, Tween 80, and Span 60.

[0029] In some embodiments, the acidity regulator is selected from at least one of citric acid, fumaric acid, citric acid, succinic acid, sorbic acid, oxalic acid, and ascorbic acid.

[0030] In some embodiments, the acidity regulator is selected from at least one of citric acid, fumaric acid, succinic acid, sorbic acid, oxalic acid, and ascorbic acid.

[0031] In some embodiments, the composition further comprises a filler, a disintegrant, a binder, a lubricant.

[0032] In some embodiments, the filler is selected from at least one of lactose, microcrystalline cellulose, mannitol, starch, sucrose, dextrin, calcium hydrogen phosphate, calcium sulfate, and calcium carbonate.

[0033] In some embodiments, the disintegrant is selected from at least one of cross-linked carboxymethyl cellulose sodium, cross-linked polyvinylpyrrolidone, sodium starch glycolate, and low-substituted hydroxypropyl cellulose.

[0034] In some embodiments, the binder is selected from at least one of hydroxypropyl cellulose, hypromellose, copovidone, povidone, ethyl cellulose, and methyl cellulose.

[0035] In some embodiments, the lubricant is selected from at least one of magnesium stearate, calcium stearate, sodium stearyl fumarate, and glyceryl behenate.

[0036] In some embodiments, the composition further comprises a glidant, and the glidant is selected from at least one of silicon dioxide, talc, and micro-powdered silica gel.

[0037] In some embodiments, the mass percentage of the compound of formula (I) in the pharmaceutical composition is 0.1%-10%, preferably 0.5%-7%.

[0038] In some embodiments, the mass percentage of the compound of formula (I) in the pharmaceutical composition is 0.1%-10%, preferably 0.5%-6.5%.

[0039] In some embodiments, the mass ratio of the solubilizer to the compound of formula (I) is 1:1 to 50:1, preferably 1:1 to 20:1.

[0040] In some embodiments, the weight percentage of the acidity regulator in the pharmaceutical composition is 1%-20%.

[0041] In some embodiments, the mass percentage of the disintegrant in the pharmaceutical composition is 1%-15%, preferably 2%-5%.

[0042] In some embodiments, the binder accounts for 1% to 20% by weight in the pharmaceutical composition, preferably 2% to 5% by weight.

[0043] In some embodiments, the mass percentage of the lubricant in the pharmaceutical composition is 0.5%-3%.

[0044] In some embodiments, the mass percentage of the glidant in the pharmaceutical composition is 0%-5%, preferably 0%-1%.

[0045] In some embodiments, the oral pharmaceutical composition is formulated into a pharmaceutical preparation.

[0046] In some embodiments, the amount of the compound of formula (I) in the unit preparation is 0.1 mg to 100 mg.

[0047] In some embodiments, the amount of the compound of formula (I) in the unit preparation is selected from 0.5 mg, 1 mg, 2 mg, 4 mg, 8 mg or 10 mg, preferably 0.5 mg or 4 mg.

[0048] In some embodiments, the formulation is selected from a tablet, a capsule, a granule, or a dispersible tablet.

[0049] In some embodiments, the preparation is a tablet, and the method for preparing the tablet comprises the following steps:

[0050] (1) uniformly mixing the compound of formula (I) and the auxiliary materials;

[0051] (2) Press into tablets to obtain the product.

[0052] In some embodiments, the method for preparing the tablet comprises the following steps:

[0053] (1) Weighing: Weigh the raw and auxiliary materials for granulation according to the prescribed dosage.

[0054] (2) Mixing: Take some of the auxiliary materials and raw materials manually, then place the remaining auxiliary materials in the granulator, mix the blank auxiliary materials for 2 minutes, then add the raw materials and stir and mix for 2 minutes, and then start shear mixing for 3 minutes.

[0055] (3) Granulation: Start stirring and shearing at the same time, add appropriate amount of wetting agent, and granulate for 2 minutes before taking out.

[0056] (4) Wet granulation: Use a 20-mesh sieve for wet granulation.

[0057] (5) Drying: Dry in an oven at 60°C for 1-3 hours and then take out.

[0058] (6) Dry granulation: Use a 24-mesh sieve for dry granulation.

[0059] (7) Total mixing: Weigh silicon dioxide and magnesium stearate, add them to the dry granules at once, mix well, and take out.

[0060] (8) Tabletting: Control the tableting parameters so that the tablet weight is about 60 mg or about 70 mg and the tablet hardness is greater than 25N.

[0061] The optional main processes of the present invention include: wet granulation, dry granulation and powder direct compression.

[0062] The process characteristics of the present invention are:

[0063] (1) A specific mixing process is used to mix low-dose drugs, thus avoiding the problem of uniformity of low-dose drug content.

[0064] (2) The raw materials are pre-treated (micronized).

[0065] (3) Process preparation: raw material micronization - raw material and auxiliary material mixing - wet granulation - drying - granulation - total mixing - tableting.

[0066] The oral pharmaceutical composition of the PDE4B inhibitor compound provided by the present invention has advantages such as high bioavailability, good content uniformity, good stability, and easy industrialization.

[0067] The solubility test method of the present invention comprises the following steps: using purified water as a medium, using different solubilizers to prepare different concentrations of the medium, adding an excess of the API to allow visible precipitation, ultrasonicating for 10 minutes, preparing a sample, and detecting the concentration by HPLC.

[0068] The dissolution curve detection method of the present invention is as follows: paddle method, 900 ml of dissolution medium, rotation speed 50 rpm, temperature 37.5 ° C, dissolution rate detection by HPLC. Sampling time points: 5 / 10 / 15 / 20 / 30 / 45 / 60 min. DETAILED DESCRIPTION

[0069] The following embodiments illustrate the technical problems, technical solutions and beneficial effects to be solved by the present invention. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the present invention.

[0070] Example 1

[0071] The solubility of the drug substance in solubilizer solutions with different concentrations was investigated, including sodium dodecyl sulfate (SDS), fumaric acid and citric acid.

[0072] Fumaric acid was selected as the solubilizer, samples were prepared, and the effect of fumaric acid dosage on the preparation was investigated.

[0073] Sodium lauryl sulfate was selected as the solubilizer to prepare samples and investigate the effect of its dosage on the preparation.

[0074] Dissolution curve data of prescription 1-prescription 5

[0075] Conclusion: The solubilizer sodium lauryl sulfate and the acid regulator fumaric acid significantly improved the dissolution curve of the preparation and greatly increased the solubility of the preparation. The greater the amount of fumaric acid, the greater the solubility. The sodium lauryl sulfate dosage within the range of 2.5:1-5:1 had a good solubility.

[0076] Example 2 Pharmacokinetic Study in Dogs

[0077] Prescribing Information:

[0078] In vivo PK test data in dogs

[0079] Prescription 9: Raw materials: sodium lauryl sulfate = 1:5, disintegration time 30min

[0080] Prescription 10: Raw materials: sodium lauryl sulfate = 1:5, raw material is not crushed, disintegration time is 16-18 minutes

[0081] Prescription 11: Raw materials: sodium lauryl sulfate = 1:2.5, disintegration time 16-17 minutes

[0082] Prescription 12: Raw materials: sodium lauryl sulfate = 1:5, disintegration time is less than 15 minutes

[0083] Prescription 6: No sodium lauryl sulfate, raw materials are directly encapsulated

[0084] Prescription 7: Raw material: sodium lauryl sulfate = 1:10

[0085] Prescription 8: Raw materials: sodium lauryl sulfate = 1:20

[0086] Note: APIs not otherwise specified are those that have undergone airflow milling.

[0087] Analysis of dog in vivo PK test data:

[0088] (1) The bioavailability of the drug substance in dogs was the lowest when it was not micronized.

[0089] (2) Formulation 12, which had the fastest disintegration time, did not have the best bioavailability in dogs;

[0090] (3) Formulation 9 without disintegrant had a longer disintegration time but the best bioavailability in dogs;

[0091] (4) Sodium lauryl sulfate significantly improves the bioavailability in dogs. Its dosage within the range of raw material: SDS = 1:2.5-1:20 can effectively improve the bioavailability.

[0092] Example 3: Preparation of compound of formula (I)

[0093] Step 1: Dissolve compound 31C (1.0 g, 3.63 mmol) in 1,4-dioxane (30 mL), add compound 47B (880 mg, 4.36 mmol) and N,N-diisopropylethylamine (1.40 g, 10.86 mmol), and stir overnight at 90°C under nitrogen. After completion of the reaction, the reaction mixture was concentrated and purified by silica gel column chromatography (dichloromethane / methanol (v / v) = 95 / 5) to afford compound 84B (1.6 g, 94%).

[0094] LC-MS (ESI): m / z = 441.6 [M+H] +

[0095] Step 2: Compound 84B (1.6 g, 3.63 mmol) was dissolved in dichloromethane (5 mL), trichloroacetyl isocyanate (820 mg, 4.36 mmol) was added under ice-cooling, and the mixture was stirred under ice-cooling for one hour. The reaction mixture was concentrated to obtain compound 84C (2.28 g, 100%).

[0096] Step 3: Dissolve compound 84C (2.28 g, 3.63 mmol) in methanol (20 mL). Add potassium carbonate (1.51 g, 10.89 mmol) and water (20 mL) under ice-cooling, and stir at room temperature for 2.5 hours. The reaction mixture is diluted with water and extracted with dichloromethane. The combined organic phases are dried, filtered, and concentrated. Chiral separation by SFC affords compound (I) (1.3 g, 74%).

[0097] Preparation method: Instrument: Waters 150Prep-SFC E, Column: Chiralcel OX column (250 mm × 30 mm, 10 μm), Mobile phase: (Phase A: CO2, Phase B: EtOH (0.1% NH3·H2O)); Gradient: Isocratic elution with 50% mobile phase B; Flow rate: 100 mL / min, Back pressure: 100 bar, Column temperature: 25°C, Wavelength: 220 nm, Cycle time: 7.0 min. Sample preparation: Sample concentration: 10 mg / mL, acetonitrile solution injection: 8 mL per injection. Retention time: 1.499 minutes. After separation, the fractions were dried on a rotary evaporator in a 35°C water bath to obtain the product. The solvent was dried in a lyophilizer at -80°C to obtain the final product, compound of formula (I).

[0098] 1 H NMR(400MHz,DMSO-d6)δ8.54(s,2H),7.19(d,1H),6.70-6.40(m,3H),4.46(s,2H),4.30(s,2H),4.00(t,2H),3.48-3.38(m,1H),3.28- 3.23(m,1H),3.01-2.92(m,1H),2.92-2.82(m,1H),2.67(s,2H),1.98-1.88(m,1H),1.46(d,6H),1.10-0.96(m,2H),0.91-0.77(m,2H).

[0099] LC-MS (ESI): m / z = 484.2 [M+H] +

[0100] Preparation of compound 31C:

[0101] Step 1: Compound 31A (2.0 g, 9.64 mmol) was dissolved in acetonitrile (20 mL), and 2-amino-2-methylpropan-1-ol (950.0 mg, 10.64 mmol) and triethylamine (2.93 g, 28.92 mmol) were added. The mixture was heated to 70°C for 14 h, concentrated, and purified by column chromatography (petroleum ether / ethyl acetate = 1 / 1) to give compound 31B (600 mg, 24%).

[0102] LC-MS (ESI): m / z = 260.1 [M+1] +

[0103] Step 2: Compound 31B (600.0 mg, 2.31 mmol) was dissolved in dichloromethane (10 mL), and m-chloroperbenzoic acid (600.0 mg, 3.49 mmol) was added and stirred for 16 hours. The mixture was quenched with saturated aqueous sodium thiosulfate solution, and the aqueous phase was extracted with dichloromethane, dried, concentrated, and purified by column chromatography (dichloromethane / methanol = 10 / 1) to give compound 31C (632.0 mg, 99%).

[0104] LC-MS (ESI): m / z = 276.1 [M+H] +

[0105] Step 3: Dissolve compound 31C (632.0 mg, 2.29 mmol) in 1,4-dioxane (10.0 mL), add compound 9D (540.0 mg, 2.75 mmol) and N,N-diisopropylethylamine (890 mg, 6.87 mmol), heat to 90°C for 12 h, cool to room temperature, concentrate, and perform column chromatography (dichloromethane / methanol = 10 / 1) to obtain compound 31 (440.0 mg, 44%).

[0106] LC-MS (ESI): m / z = 435.1 [M+H] +

[0107] 1 H NMR(400MHz,DMSO-d6)δ8.90(s,2H),7.38-7.16(m,1H),6.23(s,1H),5.15(t,1H),4.49(s,2H),4.00(t ,2H),3.51(d,2H),3.48-3.33(m,2H),3.01-2.91(m,1H),2.91-2.82(m,1H),2.67(s,2H),1.43(s,6H).

[0108] Biological testing:

[0109] 1. Effects of compounds on PDE4B2 activity

[0110] The effects of compounds on PDE4B2 activity were determined using a Fluorescence Polarization Assay Kit (BPS Bioscience, Catalog #60343). According to the kit instructions, a final concentration of 0.1 μM FAM-Cyclic-3',5'-AMP and 1 ng / well of PDE4B2 were added to each well (negative control wells were treated with PDE buffer). A serial dilution of compound was added (positive control wells were treated with PDE buffer containing 10% DMSO). The mixture was mixed thoroughly and incubated at room temperature for 1 hour. The binding agent was diluted 1:100 with binding agent diluent (cAMP). 50 μl / well of the binding agent diluent was added to the assay plate and incubated at room temperature for 20 minutes with slow shaking. After incubation, FP was detected using Envision (excitation 480 nm, emission 535 nm). FP is typically expressed as mP values.

[0111] III(S535): Fluorescence intensity in the parallel direction

[0112] I ┴ (P535): Fluorescence intensity in the vertical direction

[0113] G:G factor = 1

[0114] Calculation of inhibition rate (%Inhibition):

[0115] %Inhibition=[1-(mP(sample)-mP(negative control)) / (mP(positive control)-mP(negative control))]x100%

[0116] mP(sample): mP of the test compound reaction well

[0117] mP (negative control): negative control well mP

[0118] mP (positive control): positive control well mP

[0119] According to the calculated inhibition rate at each concentration, the IC of each compound was calculated using GraphPad Prism 8 software. 50 value.

[0120] The IC values ​​of the compounds of the present invention for PDE4B2 50 The value is <300 nM, preferably some compounds are <100 nM, more preferably some compounds are <50 nM, and further preferably some compounds are <10 nM.

[0121] IC of the compounds of the present invention against PDE4B2 50The value is less than 300nM, and the preferred compound IC 50 The IC values ​​of some specific compounds are less than 100 nM, more preferably less than 50 nM, and even more preferably less than 10 nM. 50 The values ​​are as shown in the following table, where A<10nM, 10nM≤B<50nM, and 50nM≤C<100nM.

[0122] PDE4B2 activity

[0123] 2. Effects of Compounds on PDE4D2 Activity

[0124] The effects of compounds on PDE4D2 activity were detected using a fluorescence polarization kit (BPS Bioscience, Catalog #60345). For the reaction, 12.5 μL of 0.272 ng / well enzyme (final concentration 0.068 ng / well) was preincubated with 12.5 μL of serially diluted compound (4% DMSO) at room temperature for 15 minutes. The same volume and concentration of enzyme and 12.5 μL of PDE buffer containing 4% DMSO were added to the positive control wells, and 25 μL of PDE buffer containing 2% DMSO was added to the negative control wells. Subsequently, 25 μL of 0.2 μM FAM-Cyclic-3',5'-AMP (final concentration 0.1 μM) was added to each well, mixed thoroughly, and incubated with slow shaking at room temperature for 30 minutes. Dilute the binding agent with Binding Agent Diluent (cAMP) at a ratio of 1:100. Add 100 μl of the diluent to all wells of the assay plate. Incubate at room temperature with slow shaking for 1 hour. After incubation, perform FP detection using a BMG LRBTECH microplate reader with excitation at 485 nm and emission at 520 nm. FP is typically expressed as milligrams (mP).

[0125] III(S520): Fluorescence intensity in the parallel direction

[0126] I ┴ (P520): Fluorescence intensity in the vertical direction

[0127] G:G factor = 1

[0128] Calculation of inhibition rate (%Inhibition):

[0129] %Inhibition=[1-(mP(sample)-mP(negative control)) / (mP(positive control)-mP(negative control))]x100%

[0130] mP(sample): mP of the test compound reaction well

[0131] mP (negative control): negative control well mP

[0132] mP (positive control): positive control well mP

[0133] According to the calculated inhibition rate at each concentration, the IC of each compound was calculated using GraphPad Prism 8 software. 50 value.

[0134] IC of some specific compounds of the present invention 50 The values ​​are as shown in the following table, where A<10nM, 10nM≤B<50nM, and 50nM≤C<100nM.

[0135] PDE4D2 activity

[0136] 3. Detection of the inhibitory activity of compounds on the release of tumor necrosis factor-α (TNF-α) from human peripheral blood mononuclear cells induced by lipopolysaccharide (LPS) in vitro

[0137] Normal human peripheral blood (citrate anticoagulated) was collected and hPBMCs were prepared using Ficoll-Paque PLUS (Cytiva, Cat#17144002, density 1.077 g / mL). The hPBMC cell concentration was adjusted to 0.25 x 10 6 / mL, and seeded in a 96-well plate with 50,000 cells per well. Subsequently, different concentrations of drugs were added for pre-incubation for 1 hour (the final concentration of DMSO was 0.1%, and the positive and negative control wells were equal volumes of RPMI1640 containing 0.1% DMSO). After the pre-incubation was completed, 100 ng / mL LPS (SIGMA, L2630) was added to the compound and positive control wells, and the negative control wells were equal volumes of RPMI1640 containing 0.1% DMSO. They were cultured in a 37°C, 5% CO2 incubator for 4 hours. The cell supernatant was collected, and the TNF-α content in the supernatant sample was detected using a human TNF-α Elisa quantitative detection kit (Sino Biological, Cat#KIT10602). GraphPad Prism software was used to calculate the IC 50 IC values ​​of some specific compounds of the present invention 50 The values ​​are as shown in the following table, where A<10nM, 10nM≤B<50nM, and 50nM≤C<100nM.

[0138] Inhibitory activity of compounds on LPS-induced TNF-α in vitro

[0139] Re IC 50 : Relative IC 50 .

[0140] 4 Pharmacokinetic test in rats

[0141] 4.1 Experimental Animals: Male SD rats, approximately 220 g, 6 to 8 weeks old, 6 rats per compound, purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0142] 4.2 Experimental Design: On the day of the experiment, six SD rats were randomly divided into groups according to body weight. They were fasted but not watered for 12-14 hours before administration and fed 4 hours after administration.

[0143] Dosing Information

[0144] Note: Intravenous administration solvent: 10% DMA + 10% Solutol + 80% Saline; Oral administration solvent: 0.5% MC

[0145] (DMA: dimethylacetamide; Solutol: polyethylene glycol-15-hydroxystearate; Saline: normal saline; MC: methylcellulose)

[0146] Before and after drug administration, 0.15 ml of blood was collected intraorbitally under isoflurane anesthesia. The blood was placed in an EDTAK2 centrifuge tube and centrifuged at 5000 rpm at 4°C for 10 minutes to collect plasma. Blood was collected from both the intravenous and oral gavage groups at 0, 5, 15, 30 minutes, and 1, 2, 4, 6, 8, and 24 hours. All samples were stored at -80°C prior to analysis and quantitative analysis was performed using LC-MS / MS.

[0147] Pharmacokinetic parameters of test compounds in rat plasma -:not applicable.

[0148] Conclusion: The compound of formula (I) of the present invention exhibits excellent pharmacokinetic properties in the mouse PK test.

Claims

1. An oral pharmaceutical composition, characterized in that It includes: An active ingredient, wherein the active ingredient is a compound of formula (I) or a pharmaceutically acceptable salt thereof, The mass percentage of the active ingredient in the pharmaceutical composition is 0.1%-10%, preferably 0.5%-6.5%.

2. The composition according to claim 1, characterized in that The composition further comprises at least one of a filler, a disintegrant, a binder, and a lubricant; optionally, the composition further comprises a glidant.

3. The composition according to claim 2, characterized in that The filler is selected from at least one of lactose, microcrystalline cellulose, mannitol, starch, sucrose, dextrin, calcium hydrogen phosphate, calcium sulfate, and calcium carbonate; The disintegrant is selected from at least one of cross-linked carboxymethyl cellulose sodium, cross-linked polyvinylpyrrolidone, sodium carboxymethyl starch, and low-substituted hydroxypropyl cellulose; The binder is selected from at least one of hydroxypropyl cellulose, hypromellose, copovidone, povidone, ethyl cellulose and methyl cellulose; The lubricant is selected from at least one of magnesium stearate, calcium stearate, sodium stearyl fumarate, and glyceryl behenate; The glidant is selected from at least one of silicon dioxide, talc, and micro-powdered silica gel.

4. The composition according to any one of claims 2 to 3, characterized in that The mass percentage of the disintegrant in the pharmaceutical composition is 1%-15%, preferably 2%-5%; The mass percentage of the binder in the pharmaceutical composition is 1%-20%, preferably 2%-5%; The mass percentage of the lubricant in the pharmaceutical composition is 0.5%-3%; The mass percentage of the glidant in the pharmaceutical composition is 0%-5%, preferably 0%-1%.

5. An oral pharmaceutical composition, characterized in that It includes: a) a compound of formula (I) or a pharmaceutically acceptable salt thereof, as well as b) Solubilizers and / or acidity regulators.

6. The composition according to claim 5, characterized in that The solubilizer is selected from an ionic surfactant, a nonionic surfactant, or a combination thereof, and the nonionic surfactant does not include polyethylene glycol-15-hydroxystearate; optionally, the ionic surfactant is selected from sulfated castor oil, sodium lauryl sulfate, sodium hexadecyl sulfate, sodium octadecyl sulfate; the nonionic surfactant is selected from Spans, Tweens, Poloxamer, benzyl sulfate, and poloxamer; further, the solubilizer is selected from at least one of sodium hexadecyl sulfate, sodium lauryl sulfate, poloxamer 188, Tween 80, and Span 60; Optionally, the acidity regulator is selected from at least one of citric acid, fumaric acid, citric acid, succinic acid, sorbic acid, oxalic acid, and ascorbic acid.

7. The composition according to any one of claims 5 to 6, characterized in that The composition further comprises at least one of a filler, a disintegrant, a binder, and a lubricant; and optionally, further comprises a glidant.

8. The composition according to claim 7, characterized in that The filler is selected from at least one of lactose, microcrystalline cellulose, mannitol, starch, sucrose, dextrin, calcium hydrogen phosphate, calcium sulfate, and calcium carbonate; The disintegrant is selected from at least one of cross-linked carboxymethyl cellulose sodium, cross-linked polyvinylpyrrolidone, sodium carboxymethyl starch, and low-substituted hydroxypropyl cellulose; The binder is selected from at least one of hydroxypropyl cellulose, hypromellose, copovidone, povidone, ethyl cellulose and methyl cellulose; The lubricant is selected from at least one of magnesium stearate, calcium stearate, sodium stearyl fumarate, and glyceryl behenate; The glidant is selected from at least one of silicon dioxide, talc, and micro-powdered silica gel.

9. The composition according to any one of claims 5 to 8, characterized in that The mass percentage of the compound of formula (I) in the pharmaceutical composition is 0.1%-10%, preferably 0.5%-6.5%; The mass ratio of the solubilizing agent to the compound of formula (I) is 1:1 to 50:1, preferably 1:1 to 20:1; The mass percentage of the acidity regulator in the pharmaceutical composition is 1%-20%; The mass percentage of the disintegrant in the pharmaceutical composition is 1%-15%, preferably 2%-5%; The mass percentage of the binder in the pharmaceutical composition is 1%-20%, preferably 2%-5%; The mass percentage of the lubricant in the pharmaceutical composition is 0.5%-3%; The mass percentage of the glidant in the pharmaceutical composition is 0%-5%, preferably 0%-1%.

10. A pharmaceutical preparation prepared from the oral pharmaceutical composition according to any one of claims 1 to 9, wherein the amount of the compound of formula (I) in the unit preparation is 0.1 mg to 100 mg, for example, selected from 0.5 mg, 1 mg, 2 mg, 4 mg, 8 mg or 10 mg.