Patent Application
20230060174
The present application claims the benefit and priority to the Chinese Patent Application No. 202010042865.X filed to China National Intellectual Property Administration on Jan. 15, 2020, which is incorporated herein by reference in its entirety.
The present application relates to the technical field of pharmaceutics, specifically to a pharmaceutical composition of a tricyclic PDE3/PDE4 dual inhibitor compound, a method for preparing the same and use thereof, and more specifically to a pharmaceutical composition of a compound of formula (I) or a pharmaceutically acceptable salt thereof, a method for preparing the same and use thereof.
Phosphodiesterases (PDEs) are a superfamily of enzyme systems including 11 members that participate in different signaling pathways and regulate different physiological processes. Among them, PDE3 is a major phosphodiesterase in human airway smooth muscle (ASM), and inhibition of PDE3 increases intracellular cAMP concentration and thus slackens bronchial smooth muscle. PDE4 plays a major regulatory role in the expression of proinflammatory and anti-inflammatory mediators, and a PDE4 inhibitor can inhibit the release of harmful mediators from inflammatory cells. Thus, in theory, an inhibitor that inhibits both PDE3 and PDE4 would have both the bronchodilation of a beta-adrenoreceptor agonist and the anti-inflammatory action of an inhaled glucocorticoid. The functional complementation of dual targeting is theoretically more effective than a sole targeting, providing a therapeutic effect which can be achieved only by a combination at present is achieved through a monotherapy and thus eliminating the defect that the physicochemical properties of the ingredients of medicaments used in a combination cannot be completely matched. In this way, the administration is simplified, and is convenient for a fixed dose regimen. Victoria Boswell et al, I Pharmaco. Experi. Therap., 2006, 318:840-848 and W0200005830 reported that compounds RPL554 and RPL565 have long-acting bronchodilator and anti-inflammatory effect, as well as poor solubility, high plasma clearance and other physicochemical properties, and are suitable for inhalational administration. But the data also showed that the PDE4 inhibitory activity is unsatisfactory, and the anti-inflammatory effect is insufficient. Thus there's still a need for developing a compound having good PDE3/4 inhibitory activity.
In one aspect, the present application provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof, and a surfactant,
In some embodiments, the pharmaceutical composition further comprises a metal chelating agent (or metal complexing agent).
In some embodiments, the pharmaceutical composition further comprises a buffering agent. In some embodiments, the pharmaceutical composition further comprises a diluent.
In some embodiments, the pharmaceutical composition further comprises an osmotic pressure regulator.
In some embodiments, the pharmaceutical composition comprises the compound of formula (I) or the pharmaceutically acceptable salt thereof, the surfactant and the buffering agent. In some embodiments, the pharmaceutical composition comprises the compound of formula (I) or the pharmaceutically acceptable salt thereof, the surfactant, the buffering agent and the osmotic pressure regulator.
In some embodiments, the pharmaceutical composition comprises the compound of formula (I) or the pharmaceutically acceptable salt thereof, the surfactant, the buffering agent and the metal chelating agent.
In some embodiments, the pharmaceutical composition comprises the compound of formula (I) or the pharmaceutically acceptable salt thereof, the surfactant, the buffering agent, the osmotic pressure regulator and the metal chelating agent.
In some embodiments, the pharmaceutical composition comprises the compound of formula (I) or the pharmaceutically acceptable salt thereof, the surfactant, the buffering agent, the osmotic pressure regulator, the metal chelating agent and the diluent.
In some embodiments, the pharmaceutical composition comprises the compound of formula (I) or the pharmaceutically acceptable salt thereof, the surfactant, and at least one of the buffering agent, the osmotic pressure regulator, the metal chelating agent and the diluent.
In some embodiments, the pharmaceutically acceptable salt is selected from the group consisting of maleate, sulfate, methanesulfonate, or p-toluenesulfonate.
In some embodiments, in the pharmaceutically acceptable salt of the compound of formula (I), the molar ratio of the compound of formula (I) to the acid radical ion forming the pharmaceutically acceptable salt can be 1:1 to 1:2, e.g., 1:1 or 1:2.
In some embodiments, in the pharmaceutical composition, “the compound of formula (I) or the pharmaceutically acceptable salt thereof′ can be used interchangeably with “the compound of formula (I)”.
In some embodiments, the pharmaceutical composition comprises the compound of formula (I), the surfactant, the buffering agent, the osmotic pressure regulator, the metal chelating agent and the diluent.
In some embodiments, in the pharmaceutical composition, the compound of formula (I) or the pharmaceutically acceptable salt thereof, based on the compound of formula (I), has a concentration of about 0.001 mg/mL to about 80 mg/mL, preferably 0.002 mg/mL to 50 mg/mL, and more preferably 0.1 mg/mL to 20 mg/mL.
The surfactant of the present application is a pharmaceutically acceptable surfactant, e.g., a wetting agent. The surfactant can be a non-ionic surfactant, an anionic surfactant, a cationic surfactant, or a zwitterionic surfactant. Preferably, one or more of the surfactants are non-ionic surfactants. In some embodiments, the surfactant is one or more selected from the group consisting of polyoxyethylene glycol, polypropylene glycol alkyl ether, alkyl polyglucoside, octylphenol polyoxyethylene ether, alkylphenol polyoxyethylene ether, glycerin alkyl ester, polyoxyethylene sorbitan fatty acid ester (polysorbate), sorbitan alkyl ester, sorbitan fatty acid ester, cocamide MEA, cocamide DEA, dodecyldimethylamine oxide, a block copolymer of polyethylene glycol and polypropylene glycol (poloxamer), and polyethoxylated tallow amine (POEA).
Preferably, the surfactant is one or more selected from the group consisting of polyoxyethylene sorbitan fatty acid ester (e.g., a Tween) and sorbitan fatty acid ester (e.g., a Span). In some embodiments, the polyoxyethylene sorbitan fatty acid ester is one or more selected from the group consisting of polysorbate 20 (polyoxyethylene sorbitan laurate; Tween 20), polysorbate 40 (polyoxyethylene sorbitan monopalmitate), polysorbate 60 (polyoxyethylene sorbitan stearate) and polysorbate 80 (polyoxyethylene sorbitan monooleate; Tween 80).
In some embodiments, the sorbitan fatty acid ester is one or more selected from the group consisting of sorbitan monolaurate (Span 20), sorbitan monopalmitate, sorbitan monostearate, sorbitan tristearate and sorbitan monooleate.
More preferably, the surfactant is one or more selected from the group consisting of a Tween and a Span.
In some specific embodiments, the surfactant is one or more selected from the group consisting of Tween 20, Tween 80 and Span 20.
In some embodiments, the surfactant has a concentration of about 0.01 mg/mL to about 8 mg/mL. More typically, the concentration of the surfactant in the pharmaceutical composition is about 0.01 mg/mL to 5 mg/mL, preferably about 0.02 mg/mL to 3 mg/mL, more preferably about 0.05 mg/mL to 2 mg/mL, and even more preferably about 0.1 mg/mL to 1 mg/mL.
In some embodiments, in the pharmaceutical composition, the mass ratio of the compound of formula (I) or the pharmaceutically acceptable salt thereof (based on the compound of formula (I)) to the surfactant is about 1:200 to 100:1, preferably about 1:150 to 50:1, more preferably about 1:50 to 25:1, and even more preferably about 1:1 to 15:1, for example, about 10:1.
In some embodiments, the buffering agent is a pharmaceutically acceptable buffering agent. The buffering agent can be any buffer suitable for use in a liquid pharmaceutical composition suitable for inhalation. The buffering agent is generally one or more selected from the group consisting of sulfuric acid, hydrochloric acid, sodium hydroxide, citric acid, sodium citrate, lactic acid, sodium lactate, acetic acid, sodium acetate, trisodium phosphate, sodium dihydrogen phosphate, disodium hydrogen phosphate, potassium dihydrogen phosphate, tartaric acid, sodium tartrate, glycine, boric acid and phthalic acid. The preferred number of the buffering agent is 2 or more than 2, and the preferred type of the buffering agent is citrate buffered saline or phosphate buffered saline, more preferably the sodium salt of citric acid or phosphoric acid. The citrate buffered saline includes citric acid, sodium citrate, and a mixture thereof The phosphate buffered saline includes phosphoric acid, monosodium phosphate (i.e., sodium dihydrogen phosphate), disodium hydrogen phosphate, and a mixture thereof.
In some embodiments, the buffering agent is selected from the group consisting of citric acid, a citrate (e.g., sodium citrate), tartaric acid, a tartrate (e.g., sodium tartrate), phosphoric acid and a phosphate (e.g., sodium dihydrogen phosphate and disodium hydrogen phosphate).
In some embodiments, the buffering agent is selected from the group consisting of a citrate (e.g., sodium citrate), a tartrate (e.g., sodium tartrate) and a phosphate (e.g., sodium dihydrogen phosphate and disodium hydrogen phosphate).
In some embodiments, the buffering agent has a concentration of about 0.01 mg/mL to about 50 mg/mL, preferably about 0.05 mg/mL to about 40 mg/mL, more preferably 0.1 mg/mL to about 25 mg/mL, and even more preferably 0.5 mg/mL to about 6 mg/mL.
In some embodiments, the buffering agent is used to control the pharmaceutical composition at pH between about 3.0 and about 8.5, preferably between about 5 and about 7.
In some embodiments, the osmotic pressure regulator is generally one or more selected from the group consisting of a simple non-toxic salt, such as sodium chloride, potassium chloride and the like, or a saccharide, such as one or more of glucose, mannitol or xylitol and the like. In some embodiments, the osmotic pressure regulator is sodium chloride.
The concentration of the osmotic pressure regulator depends on the amount needed to achieve a desired isotonicity, e.g., isotonic to plasma or lung fluid. The concentration of the osmotic pressure regulator is typically about 0.01 mg/mL to about 10 mg/mL, and more typically about 5 mg/mL to 9 mg/mL.
In some embodiments, the metal chelating agent is one or more selected from the group consisting of edetic acid and an edetate such as disodium edetate, calcium disodium edetate, and the like. Edetates (e.g., calcium salts, sodium salts) are preferred, and disodium edetate (EDTA-2Na) is particularly preferred.
The concentration of the metal chelating agent depends on the amount of metal ions that can be introduced during the preparation of the pharmaceutical composition and is generally about 0.01 mg/mL to about 40 mg/mL, preferably about 0.01 mg/mL to about 20 mg/mL, more preferably about 0.01 mg/mL to about 5 mg/mL, and even more preferably about 0.01 mg/mL to about 2 mg/mL.
In some embodiments, in the pharmaceutical composition, the diluent can be any pharmaceutically acceptable diluent. The diluent is suitable for inhalational administration. Generally, the diluent is one or more selected from the group consisting of water, ethanol and glycerol. A preferred diluent is water, and a more preferred diluent is sterile water.
In some embodiments, in the pharmaceutical composition, the amount of the diluent used can be an appropriate amount, such that the concentration of the compound of formula (I) or the pharmaceutically acceptable salt thereof or an excipient in the pharmaceutical composition is within a certain range.
In some embodiments, the pharmaceutical composition comprises the compound of formula (I), and the Tween or the Span; furthermore, the Tween or the Span is one or more selected from the group consisting of Tween 20, Tween 80, and Span 20.
In some embodiments, the pharmaceutical composition further comprises the phosphate; furthermore, the phosphate can be selected from the group consisting of sodium dihydrogen phosphate or monohydrate thereof, and disodium hydrogen phosphate; in some embodiments, the pharmaceutical composition further comprises sodium citrate or sodium tartrate.
In some embodiments, the pharmaceutical composition comprises the compound of formula (I), a mixture of the Span and the Tween (such as Tween 80, Tween 20 or Span 20), and water. In some embodiments, the pharmaceutical composition comprises the compound of formula (I), the mixture of the Span and the Tween (such as Tween 80, Tween 20 or Span 20), sodium citrate or sodium tartrate, and water.
In some embodiments, the pharmaceutical composition comprises the compound of formula (I), the Tween (such as Tween 80 or Tween 20), sodium dihydrogen phosphate or monohydrate thereof, disodium hydrogen phosphate, disodium edetate and water.
In some embodiments, the pharmaceutical composition comprises the compound of formula (I), the Tween (such as Tween 80 or Tween 20), sodium dihydrogen phosphate or monohydrate thereof, disodium hydrogen phosphate, sodium chloride, disodium edetate and water.
In some embodiments, the pharmaceutical composition comprises the compound of formula (I), the Tween (such as Tween 80 or Tween 20), sodium citrate or sodium tartrate, sodium chloride, and water.
In some embodiments, the pharmaceutical composition comprises the compound of formula (I), the mixture of the Span and the Tween (such as Tween 80, Tween 20 or Span 20), sodium citrate or sodium tartrate, sodium chloride and water.
In some embodiments, the pharmaceutical composition comprises the compound of formula (I), the surfactant, the buffering agent, the osmotic pressure regulator, the metal chelating agent and the diluent, wherein the compound of formula (I) has a concentration of 0.002 mg/mL to 50 mg/mL, the surfactant has a concentration of 0.02 mg/mL to 3 mg/mL, the buffering agent has a concentration of 0.1 mg/mL to about 25 mg/mL, the osmotic pressure regulator has a concentration of 5 mg/mL to 9 mg/mL, and the metal chelating agent has a concentration of 0.01 mg/mL to about 5 mg/mL. In some embodiments, in the pharmaceutical composition, the surfactant, the buffering agent, osmotic pressure regulator, the metal chelating agent and the diluent are as defined above. In some embodiments, the pharmaceutical composition comprises the compound of formula (I), the surfactant, the buffering agent, the osmotic pressure regulator, the metal chelating agent and the diluent, wherein the compound of formula (I) has a concentration of 0.002 mg/mL to 50 mg/mL, the surfactant has a concentration of 0.02 mg/mL to 3 mg/mL, the buffering agent has a concentration of 0.1 mg/mL to about 25 mg/mL, the osmotic pressure regulator has a concentration of 5 mg/mL to 9 mg/mL, and the metal chelating agent has a concentration of 0.01 mg/mL to about 5 mg/mL; the surfactant is one or more of Tween 20, Tween 80 and Span 20, the buffering agent is one or more of sodium dihydrogen phosphate or monohydrate thereof, disodium hydrogen phosphate, tartaric acid and citric acid, the osmotic pressure regulator is sodium chloride, the metal chelating agent is one or more of disodium edetate and calcium disodium edetate, and the diluent is water.
In some embodiments, the pharmaceutical composition comprises the compound of formula (I), Tween 80, sodium dihydrogen phosphate or monohydrate thereof, disodium hydrogen phosphate, sodium chloride, disodium edetate and water; wherein the compound of formula (I) has a concentration of 0.002 mg/mL to 50 mg/mL, Tween 80 has a concentration of 0.02 mg/mL to 3 mg/mL, sodium dihydrogen phosphate and disodium hydrogen phosphate have a concentration of 0.1 mg/mL to 25 mg/mL, sodium chloride has a concentration of 5 mg/mL to 9 mg/mL, and disodium edetate has a concentration of 0.01 mg/mL to about 5 mg/mL.
In some embodiments, in the pharmaceutical composition of the compound of formula (I) or the pharmaceutically acceptable salt thereof, the compound of formula (I) is in a solid form; in some embodiments, the compound of formula (I) is a crystalline form of the compound of formula (I). In some embodiments, in the pharmaceutical composition of the compound of formula (I) or the pharmaceutically acceptable salt thereof, the compound of formula (I) is a product obtained by subjecting the crystalline form of the compound of formula (I) to particle size control. In some embodiments, the crystalline form of the compound of formula (I) has diffraction peaks in an X-ray powder diffraction pattern using Cu Ka radiation at the following 20: 5.81±0.2°, 13.96±0.2°, 15.01±0.2°, 17.95±0.2° and 24.73±0.2°.
In some embodiments of the present application, the crystalline form of the compound of formula (I) has diffraction peaks in an X-ray powder diffraction pattern using Cu Ka radiation at the following 20: 5.81±0.2°, 8.38±0.2°, 11.16±0.2°, 13.96±0.2°, 14.47±0.2°, 15.01±0.2°, 17.95±0.2°, 24.73±0.2° and 26.13±0.2°.
In some embodiments, the crystalline form of the compound of formula (I) has diffraction peaks in an X-ray powder diffraction pattern using Cu Ka radiation at the following 20: 5.81±0.2°, 8.38±0.2°, 11.16±0.2°, 13.96±0.2°, 14.47±0.2°, 15.01±0.2°, 16.76±0.2°, 17.95±0.2°, 20.83±0.2°, 24.73±0.2° and 26.13±0.2°.
In some embodiments, the crystalline form of the compound of formula (I) has diffraction peaks in an X-ray powder diffraction pattern using Cu Ka radiation at the following 20: 5.81±0.2°, 8.38±0.2°, 9.13±0.2°, 11.16±0.2°, 11.60±0.2°, 12.82±0.2°, 13.96±0.2°, 14.47±0.2°, 15.01±0.2°, 16.76±0.2°, 17.95±0.2°, 18.91±0.2°, 20.83±0.2°, 24.36±0.2°, 24.73±0.2°, 25.78±0.2° and 26.13±0.2°.
In some embodiments, the crystalline form of the compound of formula (I) comprises 5, 6, 7, 8, 9, 10, 11, 12 or more diffraction peaks in an X-ray powder diffraction pattern using Cu Ka radiation selected from the following 20 angles: 5.81±0.2°, 8.38±0.2°, 9.13±0.2°, 11.16±0.2°, 11.60±0.2°, 12.82±0.2°, 13.96±0.2°, 14.47±0.2°, 15.01±0.2°, 16.76±0.2°, 17.95±0.2°, 18.91±0.2°, 20.83±0.2°, 24.36±0.2°, 24.73±0.2°, 25.78±0.2° and 26.13±0.2°.
In some embodiments, the crystalline form of the compound of formula (I) comprises 5, 6, 7, 8, 9, 10 or 11 diffraction peaks in an X-ray powder diffraction pattern using Cu Ka radiation selected from the following 20 angles: 5.81±0.2°, 8.38±0.2°, 11.16±0.2°, 13.96±0.2°, 14.47±0.2°, 15.01±0.2°, 16.76±0.2°, 17.95±0.2°, 20.83±0.2°, 24.73±0.2° and 26.13±0.2°. In some embodiments, the crystalline form of the compound of formula (I) comprises 5, 6, 7, 8 or 9 diffraction peaks in an X-ray powder diffraction pattern using Cu Ka radiation selected from the following 20 angles: 5.81±0.2°, 8.38±0.2°, 11.16±0.2°, 13.96±0.2°, 14.47±0.2°, 15.01±0.2°, 17.95±0.2°, 24.73±0.2° and 26.13±0.2°.
In some embodiments, the crystalline form of the compound of formula (I) has diffraction peaks in an XRPD pattern using Cu Ka radiation with peak positions and relative intensities as shown in Table 1 below:
In some embodiments, the crystalline form of the compound of formula (I) has diffraction peaks in an XRPD pattern using Cu Ka radiation as shown in
In some embodiments, the crystalline form of the compound of formula (I) has an exothermic peak in a differential scanning calorimetry curve at 247.70° C.±2° C.
In some embodiments, the crystalline form of the compound of formula (I) has a differential scanning calorimetry pattern as shown in
In some embodiments, the crystalline form of the compound of formula (I) has a weight loss of 0.4870% at 155.75±2° C. and a weight loss of 7.287% at 155.75±2° C. to 262.18±2° C. in a thermogravimetric analysis curve.
In some embodiments, the crystalline form of the compound of formula (I) has a thermogravimetric analysis pattern as shown in
In some embodiments, in the pharmaceutical composition, the compound of formula (I) or the pharmaceutically acceptable salt thereof has a particle size of X50<10 μm, preferably between 0.1 pm and 8 pm.
In some embodiments, in the pharmaceutical composition, the compound of formula (I) or the pharmaceutically acceptable salt thereof has a particle size of X50<5 μm and X90<10 pm. The pharmaceutical compound disclosed herein may be in a variety of dosage forms suitable for oral or inhalational administration to human, for example, a solution.
In some embodiments, the pharmaceutical composition disclosed herein is administered by inhalation.
In some embodiments, the pharmaceutical composition disclosed herein is administered by oral or nasal inhalation.
In some embodiments, the pharmaceutical composition disclosed herein is a solution for inhalation. In some embodiments, the pharmaceutical composition disclosed herein is in the form of a suspension.
In some embodiments, the pharmaceutical composition disclosed herein is in the form of a suspension for inhalation.
In another aspect, the present application provides a method for preparing the pharmaceutical composition comprising: mixing the surfactant and the compound of formula (I) or the pharmaceutically acceptable salt thereof. Preferably, the method comprises mixing the surfactant, the compound of formula (I) or the pharmaceutically acceptable salt thereof, and at least one selected from the group consisting of: the metal chelating agent, the buffering agent, the diluent, and the osmotic pressure regulator. More preferably, the present application provides a method for preparing the pharmaceutical composition comprising: mixing the wetting agent, the buffering agent, the osmotic pressure regulator, the metal chelating agent, the compound of formula (I) or the pharmaceutically acceptable salt thereof, and the diluent.
In some embodiments, the method for preparing the pharmaceutical composition comprises: 1) mixing the wetting agent, the buffering agent, the osmotic pressure regulator, the metal chelating agent and the diluent to give a solution,
In some embodiments, the method further comprises step 3): homogenizing the product obtained in step 2).
In some embodiments, after the homogenizing procedure, the compound of formula (I) or the pharmaceutically acceptable salt thereof has a particle size of X50<5 μm and X90<10 μm. In some embodiments, the method further comprises a filling step.
In yet another aspect, the present application further provides a method for preventing or treating a condition associated with PDE3 and/or PDE4 in a mammal, comprising administering to a mammal, preferably a human, in need thereof a therapeutically effective amount of the pharmaceutical composition.
In yet another aspect, the present application further provides use of the pharmaceutical composition in preparing a medicament for preventing or treating a condition associated with PDE3 and/or PDE4.
In yet another aspect, the present application further provides use of the pharmaceutical composition in preventing or treating a condition associated with PDE3 and/or PDE4. In yet another aspect, the present application further provides the pharmaceutical composition for use in preventing or treating a condition associated with PDE3 and/or PDE4.
In some embodiments of the present application, the condition associated with PDE3 and/or PDE4 is selected from the group consisting of asthma and chronic obstructive pulmonary disease (COPD). Technical Effects The compound of formula (I) and the pharmaceutical composition thereof disclosed herein have remarkable dual inhibitory effect on PDE3 and PDE4, have significant inhibitory effect on TNF-α in human peripheral blood mononuclear cells (hPBMCs), and also show excellent anti-inflammatory effect in rat acute lung injury model induced by lipopolysaccharide (LPS). The compound has high in vivo plasma clearance, low systemic exposure in plasma by oral administration and low oral bioavailability, and good safety in administration via a local route. Its inhibitory effect is low on 5 isozymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) of human liver microsomal cytochrome P450, and the risk of drug-drug interaction is avoided. Besides, the compound reduces the total white blood cells in BALF, has remarkable anti-inflammatory effect, takes effect at a low dose, and reduces the airway resistance index Penh. The crystalline form of the compound of formula (I) and the pharmaceutically acceptable salt thereof of the present application have advantages in terms of pharmaceutical activity, pharmacokinetics, bioavailability, hygroscopicity, melting point, stability, solubility, purity, ease of preparation, etc., to meet the requirements of pharmaceutics in terms of production, storage, transportation, formulation, etc. The crystal form of the compound of formula (I) has low hygroscopicity, and is favorable for the absorption of inhaled compound.
The pharmaceutical composition of the compound of formula (I) has good stability without increase of impurities and shows pharmaceutically acceptable level of impurities. The pharmaceutical composition has good dispersibility and stable dynamics, demonstrated no significant particle precipitations and no increase in particle size, and is within the range required for effective delivery of approved inhalation products. In addition, the composition has uniform and moderate particle size and high absorption rate. The composition has good delivery rate, an accurate dosage, a high proportion of inhalable aerosol particles, and a high amount of inhalable fine particles.
Unless otherwise required in the present application, throughout the whole specification and the claims which follow, the word “comprise”, and variants thereof such as “comprises” and “comprising”, or equivalents shall be construed in an open-ended, inclusive sense, i.e., “includes but is not limit to”, indicating that in addition to the listed elements, components and procedures, other unspecified elements, components and procedures may also be encompassed. “One embodiment”, “an embodiment”, “in another embodiment” or “in some embodiments” used in the specification means that a specific reference element, structure or feature described in connection with the embodiment is included in at least one embodiment. Thus, the phrases “in one embodiment”, “in an embodiment”, “in another embodiment” and “in some embodiments” in various places throughout the specification are not necessarily all referring to the same embodiment. Furthermore, the specific elements, structures, or features may be combined in any suitable manner in one or more embodiments.
It should be understood that, unless otherwise specified clearly, the singular forms “a,” “an,” and “the” used in the specification and the appended claims of the present application include plural referents; in other words, singular terms encompass plural terms herein, and vice versa. Thus, for example, the mentioned reaction including “a catalyst” includes one catalyst, or two or more catalysts. It should be understood that, unless otherwise specified clearly, the term “or” is generally employed in its sense including “and/or”.
The term “treat” or “treatment” means administering the compound or formulation described in the present application to ameliorate or eliminate a disease or one or more symptoms associated with the disease, and includes:
The term “prevent” or “prevention” means administering the compound or formulation of the present application to prevent a disease or one or more symptoms associated with the disease, and includes: preventing the occurrence of the disease or disease state in a mammal, particularly when such a mammal is predisposed to the disease state but has not yet been diagnosed with it. The term “therapeutically effective amount” refers to an amount of the compound of the present application for (i) treating or preventing a specific disease, condition or disorder; (ii) alleviating, ameliorating or eliminating one or more symptoms of a specific disease, condition or disorder, or (iii) preventing or delaying onset of one or more symptoms of a specific disease, condition or disorder described herein. The amount of the compound of the present application composing the “therapeutically effective amount” varies dependently on the compound, the disease state and its severity, the mode of administration, and the age of the mammal to be treated, but can be determined routinely by those skilled in the art in accordance with their knowledge and the present disclosure.
Typically, the particle size is quantified by measuring the characteristic equivalent spherical diameter (referred to as the volume diameter) via laser diffraction, e.g., via a laser particle size analyzer.
In the present application, the particle size distribution is expressed in terms of volume diameter (VD).
The term “Xio” refers to the particle size corresponding to a cumulative volume distribution percentage of 10%, which physically means that particles with particle sizes less than it make up 10% of the total volume.
The term “Xso” refers to the particle size corresponding to a cumulative volume distribution percentage of 50%, which is referred to as the volume median diameter and physically means that particles with particle sizes less than it make up 50% of the total volume.
The term “X9o” refers to the particle size corresponding to a cumulative volume distribution percentage of 90%, which physically means that particles with particle sizes less than it make up 90% of the total volume.
Unless otherwise indicated herein, parameter values (including 20 values, reaction conditions) are to be construed as modified by the term “about” to reflect the measurement error and the like existing in the values, e.g., there is an error of ±5% relative to the given value. All patents, patent applications and other identified publications are expressly incorporated herein by reference for the purpose of description and disclosure. These publications are provided solely because they were disclosed prior to the filing date of the present application. All statements as to the dates of these documents or description as to the contents of these documents are based on the information available to the applicant and do not constitute any admission as to the correctness of the dates or the content of these documents. Moreover, in any country or region, any reference to these publications herein is not to be construed as an admission that the publications form part of the commonly recognized knowledge in the art.
The following specific examples are intended to allow those skilled in the art to clearly understand and implement the present application. These specific examples should not be considered as limit to the scope of the present application, but merely as exemplary description and representative of the present application.
Synthesis of intermediate BB-1
Step 1: synthesis of compound BB-1-2 A mixture of compound BB-1-1 (21.10 g) and ethyl cyanoacetate (11.00 g, 10.38 mL) was stirred at 100° C. for 16 h in nitrogen atmosphere. After completion of the reaction, the mixture was cooled to 70° C., ethanol (30 mL) was slowly and dropwise added, and a large amount of solid was precipitated. The resulting mixture was filtered, and the filter cake was dried under reduced pressure to give product BB-1-2.
1H NMR (400 MHz, DMSO-d6) 6=8.26 (t, J=5.2 Hz, 1H), 6.86 (d, J=8.0 Hz, 1H), 6.79 (br s, 1H), 6.71 (d, 8.0 Hz, 1H),4.00 (q, J=6.8 Hz, 2H), 3.72 (s, 3H), 3.59 (s, 2H), 3.31-3.23 (m, 2H), 2.64 (t, J=7.2 Hz, 2H), 1.32 (t, J=6.8 Hz, 3H). MS-ESI m/z: 263.1[M+H]+.
Step 2: synthesis of compound BB-1-3 Phosphorus oxychloride (379.50 g, 230.00 mL) was heated to 85° C. in nitrogen atmosphere and compound BB-1-2 (26.00 g) was added in portions. The reaction mixture was stirred at 85° C. for 2 h. After the reaction was completed, most of the phosphorus oxychloride was removed by reduced pressure distillation. To the residue was added dichloromethane (200 mL) and the mixture was washed with water (100 mL×2). The organic phase was dried over anhydrous sodium sulfate, filtered to remove the desiccant, and then concentrated under reduced pressure. The resulting crude product was purified by slurrying with ethyl acetate (20 mL) to give compound BB-1-3.
1H NMR (400 MHz, CD3OD) 6=7.16 (s, 1H), 6.83 (s, 1H), 4.62 (s, 1H), 4.12 (q, J=6.8 Hz, 2H), 3.86 (s, 3H), 3.35 (d, J=6.4 Hz, 2H), 2.84 (t, J=6.4 Hz, 2H), 1.44 (t, J=6.8 Hz, 3H). MS-ESI m/z: 245.1[M+H]+.
Step 3: synthesis of compound BB-1-4 Compound BB-1-3 (1.00 g) was added to 98% concentrated sulfuric acid (12.88 g, 128.69 mmol, 7.00 mL) in portions at 0° C. The reaction mixture was stirred at 27° C. for 3 h. After the reaction was completed, the mixture was added to cold water (15 mL), and then aqueous sodium hydroxide solution (4 mol/L, 32 mL) was added dropwise to adjust the pH to neutral, followed by extraction with ethyl acetate (100 mL×3). Then the organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and concentrated under reduced pressure to give compound BB-1-4.
MS-ESI m/z: 263.1[M+H]+.
Step 4: synthesis of compound BB-1-5 Sodium (2.42 g) was added in portions to ethanol (80 mL) at 0° C. After the mixture was stirred at 28° C. for 0.5 h, compound BB-1-4 (6.90 g) was added to the solution in portions, and the mixture was stirred at 80° C. for 0.5 h. Then, diethyl carbonate (9.32 g, 9.51 mL) was added in one portion, and the mixture was stirred for 5 h at 80° C. After the reaction was completed, the mixture was cooled to room temperature, ice water (30 mL) was slowly added, and then diluted hydrochloric acid (2 mol/L, 53 mL) was added to adjust the mixture to a neutral pH. A large amount of solid was precipitated. The mixture was filtered, and the resulting filter cake was purified by slurrying with ethanol (10 mL) to give compound BB-1-5.
1H NMR (400 MHz, DMSO-d6) 6=11.22 (br s, 1H), 7.35 (s, 1H), 6.95 (s, 1H), 6.22 (s, 1H), 4.09 (q, J=6.8 Hz, 2H), 3.90 (br s, 2H), 3.83 (s, 3H), 2.89 (br s, 2H), 1.35 (t, J=6.8 Hz, 3H). MS-ESI
Step 5: synthesis of compound BB-1-6 Compound BB-1-5 (5.00 g) was dissolved in phosphorus oxychloride (30 mL) at room temperature. The reaction mixture was stirred at 100° C. for 16 h in nitrogen atmosphere. After the reaction was completed, most of the solvent was removed by reduced pressure distillation. Water (100 mL) was added and the resulting mixture was extracted with dichloromethane (150 mL×2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and concentrated under reduced pressure to give compound BB-1-6. MS—ESI m/z: 306.9[M+H]+. Step 6: synthesis of compound BB-1 Compound BB-1-6 (925.67 mg) was dissolved in isopropanol (8 mL) at room temperature, and then 2,4,6-trimethylaniline (2.10 g) was added. The reaction mixture was stirred at 90° C. for 15 h in nitrogen atmosphere. After the reaction was completed, the mixture was cooled to room temperature and concentrated under reduced pressure, and the resulting residue was purified by slurrying with ethanol (6 mL) to give compound BB-1.
1H NMR (400 MHz, DMSO-d6) 6=8.85 (br s, 1H), 7.27 (s, 1H), 6.97 (s, 1H), 6.90 (s, 2H), 6.45 (s, 1H), 4.10 (q, J=6.8 Hz, 2H), 3.90 (t, J=6.0 Hz, 2H), 3.86 (s, 3H), 2.87 (t, J=6.0 Hz, 2H), 2.45 (s, 3H), 2.11 (s, 6H), 1.37 (t, J=6.8 Hz, 3H). MS—ESI m/z: 406.2[M+H]+.
Synthesis of compound BB-4
Step 1: synthesis of compound BB-4-1 Compound BB-1 (1.00 g) was dissolved in 2-butanone (35 mL) at room temperature, and 2-(2-bromoethyl)isoindoline-1,3-dione (3.76 g), potassium carbonate (3.07 g) and sodium iodide (2.22 g) were added successively. The reaction mixture was stirred at 85° C. for 72 h in nitrogen atmosphere. After the reaction was completed, the mixture was concentrated to remove most of the organic solvent before water (30 mL) and ethyl acetate (25 mL×3) were added for extraction. The organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant and concentrated under reduced pressure. The resulting residue was purified by flash silica gel column chromatography (eluent: petroleum ether/ethyl acetate=15/1-3/1) to give compound BB-4-1.
MS—ESI m/z: 579.3[M+H]+.
Step 2: synthesis of compound BB-4 Compound BB-4-1 (500.00 mg) was dissolved in trichloromethane (3 mL) and ethanol (3 mL) at room temperature, and hydrazine hydrate (152.67 mg, 85% purity) was added. The mixture was stirred at 28° C. for 16 h in nitrogen atmosphere. After the reaction was completed, the mixture was concentrated to remove most of the organic solvent before water (15 mL) and dichloromethane (15 mL x 3) were added for extraction. Then the organic phases were combined, dried over anhydrous sodium sulfate, filtered to remove the desiccant, and concentrated under reduced pressure to give compound BB-4.
1H NMR (400 MHz, DMSO-d6) 6=6.95 (s, 1H), 6.85 (br s, 2H), 6.66 (s, 1H), 5.31 (s, 1H), 4.14 (t, J=6.8 Hz, 2H), 4.05 (q, J=6.8 Hz, 2H), 3.91 (t, J=6.4 Hz, 2H), 3.62 (s, 3H), 2.90-2.86 (m, 4H), 2.22 (s, 3H), 1.95 (br s, 6H), 1.33 (t, J=6.8 Hz, 3H). MS—ESI m/z: 449.2[M+H]+.
5-Hydroxy-3-methyl-1,2,3-triazole-4-carboxylic acid (18.50 mg) was dissolved in DCM (1 mL) at 20° C. HATU (8.80 mg) and triethylamine (57.40 μL) were added and the mixture was stirred for 2 h, followed by addition of compound BB-4 (50 mg) and stirring at the temperature for 16 h. The mixture was diluted to 10 mL with DCM, washed with water (30 mL×3), dried over anhydrous sodium sulfate, and filtered to remove the desiccant, and the filtrate was concentrated under reduced pressure to remove the solvent to give a crude product. The crude product was separated and purified by prep-HPLC to give the target compound of formula (I) in the form of a yellow solid.
1H NMR (400 MHz, CD3OD) 6=6.94 (s, 2H), 6.87 (s, 1H), 6.77 (s, 1H), 5.52 (s, 1H), 4.48 (t, J=6.0 Hz, 2H), 4.15 (s, 3H), 4.12-4.08 (m, 2H), 4.01 (t, J=6.0 Hz, 2H), 3.87 (t, J=6.0 Hz, 2H), 3.69 (s, 3H), 2.94 (t, J=6.0 Hz, 2H), 2.29 (s, 3H), 2.06 (s, 6H), 1.41 (t, J=6.8 Hz, 3H). MS m/z [M+1-1]+574.1.
50 mg of the compound of formula (I) was added to a 4-mL glass bottle, 1 mL of anhydrous ethanol and 0.2 mL of water were added, and the mixture was heated to 40° C. and stirred for 48 h. The mixture was naturally cooled to room temperature, centrifuged to separate the solid, and dried in vacuum to give 46 mg of solid crystalline form. The XRPD pattern is shown in
Procedures:
The amounts of the specific excipients and the products are shown in the following Table 2.
Experimental Example 1: Stability study of solid crystalline form of the compound of formula (I)
High performance liquid chromatography (HPLC)
Preparation of sample: the sample was dissolved with a mixed solvent of acetonitrile and water (acetonitrile: water=50:50 (v/v))
Solid stability stakeout method
The stability of the compound in the following conditions was examined, and samples were taken at different time points to detect the content. About 5 mg of the crystalline form of the compound of formula (I) prepared in Example 2 was accurately weighted in duplicate, transferred to a dry and clean glass bottle, spread into a thin layer as test samples, and placed in experimental conditions of influential factors ((60° C.), (relative humidity 92.5%), illumination (total illumination of 1.2×106 Luxhr/near UV energy of 200 whr/m2), (40° C., relative humidity 75%), or (60° C., relative humidity 75%)). The samples were covered with aluminum foils having holes, and thus completely exposed to the conditions. Sampling analysis was performed at 5 days, 10 days, 1 month, 2 months and 3 months. The samples were completely exposed to illumination (visible light of 1200000 Lux, UV of 200 W) at room temperature. The experimental results are shown in Table 3.
As can be seen, the crystalline form of the compound of formula (I) of the present application has good stability in the conditions of high temperature, high humidity or illumination without the increase of impurities during the test.
Experimental Example 2: Hygroscopicity study of crystalline form of the compound of formula (I)
Instrument model: SMS DVS Advantage
Test conditions: the sample (10-20 mg, the crystalline form prepared in Example 3) was placed in DVS sample tray for testing.
The detailed DVS parameters are as follows:
Temperature: 25° C.
Balancing: dm/dt=0.01%/min (shortest: 10 min, longest: 180 min)
Drying: drying at 0% RH for 120 min
RH (%) test gradient: 10%
Range of RH (%) test gradient: 0%-90%-0%. The resulting dynamic vapor sorption (DVS) plot is shown in
As can be seen from
Experimental Example 3: In vitro detection of the inhibitory activity of the compound against PDE 3A enzyme
Objective: to determine the AMP/GMP expression based on fluorescence polarization, i.e., to trace binding of AMP/GMP to antibody so as to indicate enzyme activity.
Reagents:
Buffer solution: 10 mM Tris—HCl (pH 7.5), 5 mM MgCl2, 0.01% Brij 35, 1 mM dithiothreitol (DTT), and 1% DMSO.
Enzyme: recombinant human PDE3A (Gene accession number: NM 000921; amino acid 669-end) was expressed by baculovirus in Sf9 insect cells using an N-terminal GST tag, with the molecular weight being 84 kDa.
Enzyme substrate: 1 μM cAMP Detection: TranscreenerOAMP2/GMP2 antibody and AMP2/GMP2 AlexaFluor633 tracer.
Procedures:
The results are shown in Table 4:
The active ingredient in the pharmaceutical composition of the present application has significant dual inhibitory effect on PDE3 and PDE4.
Experimental Example 5: Pharmacokinetic study in beagle dogs In this study, male beagle dogs were selected as test animals, and LC-MS/MS was used for quantitatively measuring the drug concentration in plasma of beagle dogs at different time points after intravenous injection or intragastric administration of the compound of formula (I) so as to evaluate the pharmacokinetics of the compound of formula (I) in beagle dogs.
The clear solution of the compound of formula (I) was injected into two beagle dogs of 10-12 kg via the cephalic vein or saphenous vein, and the clear solution of the compound of formula (I) was administered intragastrically to two beagle dogs of 10-12 kg (overnight fasted). The animals were all subjected to approximately 500-pt blood collection each time from peripheral veins at 0.0333, 0.0833, 0.25, 0.5, 1, 2, 4, 6, 8 and 24 h post-dose, and the blood was transferred into commercial centrifuge tubes containing 0.85-1.15 mg of K2 EDTA.2H20 anticoagulant, and plasma was separated by centrifugation at 3000 g for 10 min at 4° C. The plasma concentration was measured by LC-MS/MS, and the relevant pharmacokinetic parameters were calculated using WinNonlinTM Version 6.3 (Pharsight, Mountain View, Calif.), a pharmacokinetic software, with a non-compartmental model linear-log trapezoidal method.
The active ingredient in the pharmaceutical composition of the present application has high in vivo plasma clearance, low systemic exposure in plasma by oral administration and low oral bioavailability.
Experimental Example 6: Inhibitory effect on activity of isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) of human liver microsomal cytochrome P450 A total of 5 specific probe substrates of 5 isoenzymes of CYP, namely phenacetin (CYP1A2), diclofenac (CYP2C9), (S)-mephenytoin (CYP2C19), dextromethorphan (CYP2D6) and midazolam (CYP3A4) were each co-incubated with human liver microsomes and the compound of formula (I), and then reduced nicotinamide adenine dinucleotide phosphate (NADPH) was added to initiate the reaction. After the reaction was completed, the samples were treated, and the concentrations of 5 metabolites (acetaminophen, 4′-hydroxydiclofenac, 4′-hydroxymephenytoin, dextrorphan and l′-hydroxymidazolam) generated from the specific substrates were quantitatively detected by lC-MS/MS to calculate the correcnondino- half maximal inhibitory concentrationc (IC50)
The active ingredient in the pharmaceutical composition of the present application has low inhibitory effect on the 5 isoenzymes (CYP1A2, CYP2C9, CYP2C19, CYP2D6 and CYP3A4) of human liver microsomal cytochrome P450.
Experimental Example 7: Pharmacodynamic study in cigarette smoke-induced rat acute lung injury model
Animals
Male Sprague-Dawley rats (supplied by Shanghai SLAC Laboratory Animal Co., Ltd.), SPF grade, approximately 200 g.
Procedures
Administration frequency: the drug or solvent were given by atomization for 30 min in every morning before exposure to cigarette smoke, and were given before the inhalation of the atomized LPS on day 4.
The experimental results are shown in
As can be seen from
Experimental Example 8: In vitro detection of the inhibitory activity of the compound against TNF-α in human peripheral blood mononuclear cells Objective: to measure the anti-inflammatory activity at cellular level of the test compound based on the level of TNF-α in human peripheral blood mononuclear cells (hPBMCs).
Procedures:
The results are shown in Table 8:
Therefore, the active ingredient in the pharmaceutical composition of the present application shows potent anti-inflammatory activity, and has significant inhibitory effect on TNF-α in human peripheral blood mononuclear cells (hPBMCs).
Experimental Example 9: Stability Test
The product obtained in Example 28 was placed for 6 months in an accelerating condition (40° C. 2° C./RH 25%±5%) and a long-term condition (30° C.±2° C./RH 65%±5%), and the results are shown in Table 10.
As can be seen, the pharmaceutical composition of the present application shows good stability in the accelerating condition and the long-term condition without significant increase of impurities and particle size.
| Number | Date | Country | Kind |
|---|---|---|---|
| 202010042865.X | Jan 2020 | CN | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/CN2021/072145 | 1/15/2021 | WO |
