Salts of endothelin A (ETA) receptor antagonist compounds, methods for producing the same, and pharmaceutical uses

Abstract

The present invention belongs to the technical field of chemical drugs, and provides a salt of an endothelin A (ETA) receptor antagonist compound, a method for producing the same, and uses thereof.

Description

Technical Field

[0001] The present invention belongs to the technical field of chemical drugs, and provides a salt of an endothelin A (ETA) receptor antagonist compound, a method for producing the same, and uses thereof.

Background Art

[0002] Atrasentan (CAS: 173937-91-2) is a potent and selective endothelin A (ETA) receptor antagonist, and its structural formula is as follows.

Chemical formula

[0003] Atrasentan hydrochloride (CAS: 195733-43-8) is used clinically, and its structure is as follows.

Chemical formula

[0004] Previously, its use in the treatment of prostate cancer has been evaluated in clinical trials, and currently its use in the treatment of chronic kidney disease related to type II diabetes is being evaluated in clinical trials. Also, it has been proven to reduce albuminuria in patients with diabetic nephropathy, but currently there are no commercially available products.

[0005] Nearly half of drug molecules exist in the form of salts, but salt formation may improve undesirable physical, chemical, and biological properties of the drug. Therefore, it is very important to develop salts of endothelin A (ETA) receptor antagonist compounds having more excellent properties in terms of physical, chemical, or pharmaceutical properties.

Summary of the Invention

Problems to be Solved by the Invention

[0006] In view of the problems existing in the prior art, the present invention provides a salt of an endothelin A (ETA) receptor antagonist compound with a novel structure, a method for producing the same, and uses thereof.

Means for Solving the Problems

[0007] Specifically, the present invention provides a salt of a compound represented by formula (I), and as shown in Figure 1, its structural formula is as follows.

Chemical formula

[0008] As a preferred technical solution of the present invention, the inorganic acid is selected from sulfuric acid, hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, carbonic acid, or nitric acid, The organic acid is selected from benzoic acid, 2,5-dihydroxybenzoic acid, 4-acetaminobenzoic acid, 4-aminobenzoic acid, oxalic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, hexanoic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, octanoic acid, decanoic acid, cinnamic acid, citric acid, aspartic acid, gluconic acid, glutamic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, succinic acid, formic acid, fumaric acid, gentisic acid, glutaric acid, valeric acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, nicotinic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, palmitic acid, pamoic acid, trifluoroacetic acid, thiocyanic acid, p-toluenesulfonic acid, and L-malic acid.

[0009] As a preferred technical solution of the present invention, the inorganic acid is selected from phosphoric acid.

[0010] As a preferred technical solution of the present invention, the inorganic acid is selected from phosphoric acid, where n = 3 and x = 3 to 6.

[0011] As a preferred technical solution of the present invention, the inorganic acid is selected from phosphoric acid, where n = 3, x = 3, or n = 3, x = 4, or n = 3, x = 5, or n = 3, x = 6.

[0012] As a preferred technical solution of the present invention, the organic acid is selected from oxalic acid.

[0013] As a preferred technical solution of the present invention, the organic acid is selected from oxalic acid, where n = 1, x = 1.

[0014] The present invention further provides a pharmaceutical composition containing a salt of the compound represented by the formula (I) and one or more pharmaceutically acceptable carriers.

[0015] The present invention further provides the use of the compound represented by the formula (I) in the manufacture of a drug for treating and / or preventing endothelin A (ETA) receptor-related diseases.

[0016] As a preferred technical solution of the present invention, the diseases include chronic kidney disease, IgA, FSGS, Alport, and hypertension diseases.

[0017] The present invention further provides a method for producing a salt of the compound represented by the formula (I), which is produced by mixing the compound A with an acid.

Chemical formula

[0018] As a preferred technical solution of the present invention, the molar ratio of the compound to the acid molecule is 1:1 to 2.

[0019] As a preferred technical solution of the present invention, Step 1) Weigh an appropriate amount of free alkali and dissolve it in a good solvent; Step 2) Weigh an appropriate amount of ionic acid, add it to Step 1 and react to form a salt; Step 3) of reverse titrating the reaction solution of Step 2 into a poor solvent and stirring to precipitate, and Step 4) of obtaining the salt of the compound by high-speed centrifugation or standing still, are included.

[0020] The acid is selected from M, where M is an inorganic acid or an organic acid. The inorganic acid is selected from sulfuric acid, hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, carbonic acid, or nitric acid. The organic acid is benzoic acid, 2,5-dihydroxybenzoic acid, 4-acetaminobenzoic acid, 4-aminobenzoic acid, oxalic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, hexanoic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, octanoic acid, decanoic acid, cinnamic acid, citric acid, aspartic acid, gluconic acid, glutamic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, succinic acid, formic acid, fumaric acid, gentisic acid, glutaric acid, valeric acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, nicotinic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, palmitic acid, pamoic acid, trifluoroacetic acid, thiocyanic acid, p-toluenesulfonic acid, and L-malic acid.

[0021] In the present disclosure, the solvent used to form the salt is at least one selected from ethyl acetate, methanol, n-propanol, isopropanol, isopropyl ether, tetrahydrofuran, isopropyl acetate, acetone, methyl tert-butyl ether, acetonitrile, ethanol, 1,4-dioxane, n-hexane, and isopropyl ether.

[0022] Furthermore, in any embodiment, the method for producing the aforementioned pharmaceutically acceptable salt further includes steps such as solvent evaporation or stirring for crystallization, filtration, and drying.

[0023] Unless otherwise specified, the following terms and phrases used in this specification shall have the following meanings. Specific terms and phrases should not be considered uncertain or unclear when there is no specific definition, but should be understood in their ordinary meanings. When a trade name appears in this specification, it is intended to refer to the corresponding product or its active ingredient. As used herein, the term "pharmaceutically acceptable" relates to compounds, materials, compositions and / or dosage forms suitable for contact use with human and animal tissues within the scope of reliable medical judgment, without undue toxicity, irritation, allergic reaction or other problems or complications and corresponding to a reasonable benefit / risk ratio.

[0024] As used herein, "pharmaceutically acceptable salt" refers to a derivative of a compound of the present invention, wherein the parent compound is modified by salt formation with an acid or salt formation with a base.

[0025] The prodrugs of the compounds described herein are readily chemically transformed under physiological conditions to convert to the compounds of the present invention. Further, the prodrugs can be converted to the compounds of the present invention by chemical or biochemical means in an in vivo environment.

[0026] Certain compounds of the present invention may exist in unsolvated forms or solvated forms including hydrated forms. Generally, solvated forms and unsolvated forms are equivalent to each other and all are included within the scope of the present invention.

[0027] The atoms of the compound molecules of the present invention are isotopes, and isotope derivatization can usually extend the half-life, reduce the clearance rate, enhance the metabolic stability and improve the in vivo activity. Embodiments in which at least one atom is replaced with an atom having the same number of atoms (number of protons) and a different mass number (sum of protons and neutrons) are also included. Examples of isotopes contained in the compounds of the present invention include hydrogen atoms, carbon atoms, nitrogen atoms, oxygen atoms, phosphorus atoms, sulfur atoms, fluorine atoms, chlorine atoms, respectively 2 H, 3 H, 13 C, 14C, 15 N, 17 O, 18 O, 31 P, 32 P, 35 S, 18 F, and 36 Cl. In particular, radioisotopes such as 3H and 14C that emit radiation when decaying can be used for examining the local anatomical structures of pharmaceutical preparations and compounds in vivo. Stable isotopes do not decay or change according to the amount and are not radioactive, so they can be used safely. When the atoms constituting the molecule of the compound of the present invention are isotopes, the isotopes can be converted according to a conventional method by replacing the reagents used in the synthesis with reagents containing the corresponding isotopes.

[0028] The compounds of the present invention may contain non-natural proportions of atomic isotopes on one or more atoms constituting the compound. For example, the compound can be labeled with radioisotopes such as deuterium ( 2 H), iodine-125 ( 125 I) or C-14 ( 14 C). All changes in the isotope composition of the compounds of the present invention are included within the scope of the present invention, whether radioactive or not. Furthermore, when one or more hydrogen atoms of the compound of the present invention are replaced with the isotope deuterium ( 2 H), the deuterated compound of the present invention has effects such as extending the half-life, reducing the clearance rate, enhancing the metabolic stability, and improving the in vivo activity. As a method for producing the isotope derivatives, a phase transfer catalyst method is usually included. For example, a preferred deuteration method utilizes a phase transfer catalyst (e.g., tetraalkylammonium salt, NBu4HSO4). When the methylene protons of a diphenylmethane compound are exchanged using a phase transfer catalyst, a higher level of deuterium is introduced than by reduction with a deuterated silane (e.g., triethylsilane-d) in the presence of an acid (e.g., methanesulfonic acid) or reduction with sodium borohydride-d in the presence of a Lewis acid such as aluminum trichloride.

[0029] The term "pharmaceutically acceptable carrier" refers to any formulation carrier or medium that can deliver an effective amount of the active substance of the present invention, does not interfere with the biological activity of the active substance, and causes no toxicity or side effects to the host or patient. Representative carriers include water, oils, vegetables and minerals, cream bases, lotion bases, ointment bases, etc. These bases include suspending agents, thickening agents, transdermal penetration enhancers, etc. These formulations are well known to those skilled in the art of cosmetics or topical medications. Other information regarding carriers can be found in Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams & Wilkins (2005), the content of which is incorporated herein by reference.

[0030] The term "excipient" generally refers to the carrier, diluent, and / or medium necessary for preparing an effective pharmaceutical composition.

[0031] The terms "effective amount" or "therapeutically effective amount" with respect to a drug or pharmacologically active agent refer to an amount of the drug or agent that is non-toxic but sufficient to achieve the desired effect. With respect to the oral dosage forms of the present invention, the "effective amount" of the active substance in the composition refers to the amount necessary to achieve the desired effect when used in combination with another active substance in the composition. The determination of the effective amount varies from person to person, depends on the age and general condition of the recipient, also depends on the specific active substance, and the appropriate effective amount in an individual case can be determined by those skilled in the art based on routine tests.

[0032] The terms "active ingredient", "therapeutic agent", "active substance", or "active agent" refer to a chemical substance that is effective in the treatment of a target disorder, disease, or medical condition.

[0033] The term "tautomer" or "tautomeric form" refers to structural isomers having different energies that can interconvert via a low energy barrier. When tautomerism is possible (e.g., in solution), a chemical equilibrium of tautomers can be achieved. For example, proton tautomers (also called prototropic tautomers) include interconversions by proton transfer such as keto - enol isomerization and imine - enol isomerization. Valence tautomers include interconversions by partial rearrangement of bonding electrons. Keto - enol tautomerism. Another example of tautomerization is phenol - keto tautomerization. Unless otherwise indicated, all tautomers of the compounds of the present invention are within the scope of the present invention.

[0034] The compounds of the present invention may exist in the form of specific geometric isomers or stereoisomers. The present invention contemplates cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and all such compounds including their racemic mixtures and other mixtures, such as mixtures rich in enantiomers or diastereomers. All of these mixtures are within the scope of the present invention. Other asymmetric carbon atoms may be present in substituents such as alkyl. All of these isomers, and their mixtures, are included within the scope of the present invention.

[0035] The optically active (R)- and (S)-isomers, as well as D and L isomers, can be produced by chiral synthesis or chiral reagents or other conventional techniques. When one enantiomer of the compounds of the present invention is contemplated, it can be produced by asymmetric synthesis or derivatization using a chiral auxiliary, where the resulting mixture of diastereomers is separated and the auxiliary group is cleaved to obtain the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (e.g., amino) or an acidic functional group (e.g., carboxyl), a diastereomeric salt is formed with a suitable optically active acid or base, and then the diastereomers are resolved by conventional methods known in the art, and thereafter, the pure enantiomer is recovered. Further, the separation of enantiomers and diastereomers is usually carried out using chromatography with a chiral stationary phase, optionally in combination with chemical derivatization methods (e.g., formation of carbamates from amines).

[0036] "Optional" or "optionally" means that the event or situation described thereafter may occur, but need not necessarily occur, and the description includes both the case where the described event or situation occurs and the case where it does not occur.

[0037] The compounds of the present invention can be produced by various synthetic methods well known to those skilled in the art, including the specific embodiments listed below, the embodiments formed by combining them with other chemical synthesis methods, and equivalent alternative methods well known to those skilled in the art. Preferred embodiments include, but are not limited to, the examples of the present invention. [[Effect of the Invention]]

[0038] Compared with the prior art, the beneficial effects of the present invention are as follows.

[0039] The compound of the present invention is an endothelin A (ETA) receptor antagonist compound, and this prodrug is completely converted to atrasentan in human liver microsomes, indicating that it has the same technical effect as atrasentan in pharmacokinetic studies.

[0040] The salts of the compounds represented by formula (I) according to the present invention have improved physical and chemical properties compared to the free compounds represented by formula (I) and are more convenient for pharmaceutical use.

Brief Description of Drawings

[0041]

Figure 1

Figure 2

Figure 3

Modes for Carrying Out the Invention

[0042] Hereinafter, the present invention will be described in more detail with reference to examples and drawings, but the embodiments of the present invention are not limited thereto.

[0043] The structure of the compound is determined by nuclear magnetic resonance (NMR) or mass spectrometry (MS). The NMR shift (δ) is given in units of 10 -6 (ppm). NMR is measured using a Bruker AVANCE-III nuclear magnetic spectrometer, and the measurement solvent is deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), and the internal standard is tetramethylsilane (TMS).

[0044] MS is measured using an ISQ EC mass spectrometer (manufacturer: Thermo, model: ISQ EC).

[0045] High-performance liquid chromatography (HPLC) analysis is performed using a Thermo U3000 HPLC DAD high-performance liquid chromatograph.

[0046] The CombiFlash high-performance liquid preparative chromatograph uses Combi Flash Rf+LUMEN (TELEDYNE ISCO).

[0047] Silica gel plates for thin-layer chromatography are HSGF 254 or GF 254 manufactured by Yantai Yinlong Co., Ltd. The specifications of the silica gel plates used in thin-layer chromatography (TLC) are 0.17 mm to 0.23 mm, and the specifications used for the separation and purification of products by thin-layer chromatography are 0.4 mm to 0.5 mm.

[0048] In silica gel column chromatography, generally, silica gel with a mesh size of 100 - 200 manufactured by Yushan Shangbang Co., Ltd. is used as the carrier.

[0049] The present invention relates to the following reagents. DMF (N,N-dimethylformamide), KI (potassium iodide), Cs2CO3 (cesium carbonate), DCM (dichloromethane), n-hexane, EA (ethyl acetate), Py (pyridine), CF3COOH (trifluoroacetic acid), IPA (isopropanol), isopropyl ether, acetone.

[0050] Example 1 1-[(ethoxycarbonyl)oxy]methyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

Chemical formula

[0051] Example 2 1-[(Ethoxycarbonyl)oxy]ethyl (2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

Chemical formula

Chemical formula

[0052] Example 3 1-[(Isopropoxycarbonyl)oxy]ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate [Chemical Structure] Step A: Synthesis of 1-[(Isopropoxycarbonyl)oxy]ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate [Chemical Structure] At room temperature, atrasentan (2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidone-3-carboxylic acid (500 mg, 0.98 mmol), 1-chloroethyl isopropyl carbonate (325 mg, 1.96 mmol), cesium carbonate (640 mg, 1.96 mmol), and potassium iodide (325 mg, 1.96 mmol) were added to 10 ml of dry DMF, the temperature was raised to 65 °C, and the mixture was reacted for 2 hours. After completion of the reaction, the temperature was lowered to room temperature, the reaction mixture was poured into 40 ml of ice-water solution, and extracted with dichloromethane (20 ml × 3). The combined organic phases were washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure to dryness, and the crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 1 / 1) to obtain 482 mg of 1-[(isopropoxycarbonyl)oxy]ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidone-3-carboxylate as a colorless oily product (yield: 76.9%). LC-MS: RT = 2.29 min, [M+H] + = 641.48. 11H NMR (400 MHz, DMSO) δ 7.25 (t, J = 9.0 Hz, 2H), 7.07-7.04 (m, 1H), 6.93-6.89 (m, 2H), 6.85-6.78 (m, 2H), 6.59-6.54 (m, 1H), 5.99 (d, J = 6.2 Hz, 2H), 4.79-4.69 (m, 1H), 3.76-3.70 (m, 4H), 3.55-3.44 (m, 1H), 3.33-3.23 (m, 4H), 3.21-3.13 (m, 1H), 3.02-2.90 (m, 3H), 2.80-2.75 (m, 1H)2.71 (d, J = 13.9 Hz, 1H), 1.39-1.33 (m, 1H), 1.32 (d, J = 5.4 Hz, 2H), 1.26 (d, J = 5.4 Hz, 2H), 1.22-1.14 (m, 9H), 1.00-0.94 (m, 2H), 0.82 (t, J = 7.3 Hz, 3H), 0.72 (t, J = 7.3 Hz, 3H).

[0053] Example 4 1-[(Methoxycarbonyl)oxy]ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

Chemical formula

Chemical formula

[0054] Example 5 1 - ((((2 - (methylamino)ethoxy)carbonyl)oxy)ethyl) - (2R,3R,4S) - 4 - (benzo[d][1,3]dioxolan - 5 - yl) - 1 - [2 - (dibutylamino) - 2 - oxoethyl] - 2 - (4 - methoxyphenyl)pyrrolidone - 3 - carboxylate

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0055] Example 6 1-((((S)-2,3-dihydroxypropoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidone-3-carboxylate [Chemistry] Step A: 1-Chloroethyl ((S)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl carbonate [Chemistry] Under an ice bath, (S)-(+)-1,2-sor ketal (315 mg, 2.38 mmol) and pyridine (225 mg, 2.85 mmol) were added to 10 ml of dry dichloromethane. Under the ice bath, 1-chloroethyl chloroformate (408 mg, 2.85 mmol) was added dropwise. After the addition was complete, the temperature was raised to room temperature and the reaction was carried out for 2 hours. After the reaction was completed, it was poured into 40 ml of ice water solution, extracted with dichloromethane (20 ml × 3), washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure until dry, and the crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 10 / 1) to obtain 250 mg of 1-chloroethyl ((S)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl carbonate as a colorless oily product (yield: 44.2%). Step B: Synthesis of 1-(((((S)-2,2-dimethyl-1,3-dioxolan-4-yl)methoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidone-3-carboxylate [Chemistry] At room temperature, atrasentan (2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidone-3-carboxylic acid (100 mg, 0.20 mmol), 1-chloroethyl(((S)-2,2-dimethyl-1,3-dioxolan-4-yl)methyl) carbonate (72 mg, 0.3 mmol), cesium carbonate (130 mg, 0.4 mmol), and potassium iodide (66.4 mg, 0.4 mmol) were added to 5 ml of dry DMF, heated to 65 °C, and reacted for 4 hours. After completion of the reaction, the temperature was lowered to room temperature, poured into 40 ml of ice aqueous solution, extracted with dichloromethane (20 ml × 3), the organic phases were combined, washed with saturated brine (50 ml), dried over anhydrous magnesium sulfate, filtered, concentrated under reduced pressure to dryness, and the crude compound was purified by silica gel column chromatography (eluent: n-hexane / ethyl acetate = 3 / 1) to obtain 85 mg of 1-(((((S)-2,2-dimethyl-1,3-dioxolan-4-yl)methoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidone-3-carboxylate (yield: 59.9%) as a colorless oily product. LC-MS: RT = 2.21 min, [M+H] + = 713.46. Step C: Synthesis of 1-((((S)-2,3-dihydroxypropoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidone-3-carboxylate

Chemical Structure

[0056] Example 7 1-[(Cyclopropyloxycarbonyl)oxy]ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidine-3-carboxylate

Chem.

Chem.

[0057] Example 8 1-(((2-Hydroxyethoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidone-3-carboxylate

Chem.

Chem.

Chemical formula

Chemical Structure

[0058] Example 9 1-(((((R)-1,4-dioxan-2-yl)methoxy)carbonyl)oxy)ethyl-(2R,3R,4S)-4-(benzo[d][1,3]dioxolan-5-yl)-1-[2-(dibutylamino)-2-oxoethyl]-2-(4-methoxyphenyl)pyrrolidone-3-carboxylate

Chemical Structure

Chemical Structure

[0059] Example 10 1 - ((((Oxetan - 3 - yl)oxy)carbonyl)oxy)ethyl - (2R,3R,4S) - 4 - (benzo[d][1,3]dioxolan - 5 - yl) - 1 - [2 - (dibutylamino) - 2 - oxoethyl] - 2 - (4 - methoxyphenyl)pyrrolidine - 3 - carboxylate

Chem.

Chem.

Chem.

[0060] Example 11 1 - ((((S)-1,4 - dioxan - 2 - yl)methoxy)carbonyl)oxy)ethyl - (2R,3R,4S)-4-(benzo[d][1,3]dioxolan - 5 - yl)-1-[2-(dibutylamino)-2 - oxoethyl]-2-(4 - methoxyphenyl)pyrrolidone - 3 - carboxylate

Chemical Structure

Chemical Structure

Chemical formula

[0061] Example 12 Study on Compound Microsomes (1) Experimental Materials Human liver microsomes were all purchased from Red Liver Disease Research (Shanghai). Reagents: DMSO (dimethyl sulfoxide), acetonitrile, formic acid, and propranolol (internal standard) are all commercially available. Equipment: Thermo Fisher's LC-MS (U300 UPLC, TSQ QUANTUMN ULTRA triple quadrupole mass spectrometer). (2) Experimental method A certain amount of the compound was accurately weighed and dissolved in DMSO to prepare a 10 mM stock solution. The stock solution was diluted with a diluent (ACN:H2O = 1:1) to a 100 μM working solution, and then diluted with 0.1 M potassium phosphate buffer to a 3 μM dosing solution for use. 75 μL of liver microsomes was added to 925 μL of 0.1 M potassium phosphate buffer, mixed well to obtain a 1.5 mg / mL liver microsome suspension, and pre-incubated at 37 °C for 10 min. At 0 h of preparation: Immediately after adding 6 mM NADPH solution to 15 μL of the above liver microsome suspension, 150 μL of propranolol acetonitrile solution was added to precipitate, and then 15 μL of the above dosing solution was added and mixed well for use. Preparation of samples at 20 min and 60 min: 15 μL of the dosing solution was taken, 15 μL of liver microsome suspension and 15 μL of 6 mM NADPH solution were added, mixed well, and incubated at 37 °C for 20 min and 60 min respectively. The above sample preparations were all carried out in parallel using duplicate wells. After incubating the above samples to the appropriate time point, 150 μL of propranolol acetonitrile solution was added to stop the reaction. All of the above samples were centrifuged at 4000 rpm for 5 min, 100 μL of the supernatant was added to 100 μL of ultrapure water, mixed well, and LC-MS / MS analysis was performed. The detection conditions of LC-MS / MS are as follows. Chromatography column: Waters ACQUITYTM PREMIER HSS T3, 50 * 2.1 mm, 1.8 μm. Mobile phase: water (0.1% formic acid) - acetonitrile (gradient elution is performed according to the following table) TIFF2025524934000037.tif48139(3) Data processing Taking the initial 0 hour as 100%, the relative residual amount of the drug at each time point was calculated, and taking each time point of atrasentan as 100%, the relative amount of atrasentan converted from the compound in the example was calculated. The results are shown in Table 1 and Table 2.

Table 1

Table 2

[0062] Example 13 Pharmacokinetic study of the compound in rats (1) Experimental materials SD rats: male, 200 - 300 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. Reagents: DMSO (dimethyl sulfoxide), PEG - 400 (polyethylene glycol 400), physiological saline, heparin, acetonitrile, formic acid, propranolol (internal standard) are all commercially available. Equipment: LC - MS / MS of Thermo Fisher (U300 UPLC, TSQ QUANTUMN ULTRA triple quadrupole mass spectrometer). (2) Experimental methods The compound was weighed, dissolved in a DMSO-PEG-400-physiological saline (5:60:35, v / v / v) system, and administered intragastrically to rats. After that, 200 μL of venous blood was collected into a heparin-added EP tube together with sodium fluoride at 15 min, 30 min, 1 h, 2 h, 5 h, 7 h, and 24 h, centrifuged at 12,000 rpm for 2 min, and the plasma was frozen and stored at -80 °C for the test. A certain amount of the test article was accurately weighed and dissolved in DMSO to a concentration of 2 mg / mL to obtain a stock solution. An appropriate amount of the compound stock solution was accurately taken, diluted by adding acetonitrile, and standard series solutions were prepared. 20 μL of each of the above standard series solutions was accurately taken, 180 μL of blank plasma was added, vortexed and mixed, and plasma samples with plasma concentrations of 1, 3, 5, 10, 30, 100, 300, 1000, and 3000 ng / mL were prepared. Two sample analyses were performed for each concentration, and a calibration curve was created. 30 μL of plasma was taken, 200 μL of an acetonitrile solution of propranolol (50 ng / mL) as the internal standard was added, vortexed and mixed well, 100 μL of purified water was added, vortexed again and mixed well, centrifuged at 4000 rpm for 5 min, and the supernatant was taken for LC-MS / MS analysis. The detection conditions of LC-MS / MS are as follows. Chromatography column: Waters ACQUITYTM PREMIER HSS T3, 50 * 2.1 mm, 1.8 μm. Mobile phase: water (0.1% formic acid)-acetonitrile (gradient elution is performed according to the following table) TIFF2025524934000040.tif5094(3) Data processing After detecting the blood concentration by LC-MS / MS, pharmacokinetic parameters were calculated using WinNonlin 6.1 software and the non-compartmental model method. The results are shown in Table 3.

Table 3

[0063] Example 14 Pharmacokinetic study of the compound in rats (1) Experimental materials SD rats: male, 200 - 300 g, purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. Reagents: DMSO (dimethyl sulfoxide), PEG - 400 (polyethylene glycol 400), physiological saline, heparin, acetonitrile, formic acid, propranolol (internal standard) are all commercially available. Equipment: LC - MS / MS of Thermo Fisher (U300 UPLC, TSQ QUANTUMN ULTRA triple quadrupole mass spectrometer). (2) Experimental method The compound was weighed and dissolved in a DMSO-PEG-400-physiological saline (5:60:35, v / v / v) system, and after intragastric administration to rats, 200 μL of venous blood was collected into a heparin-added EP tube together with sodium fluoride at 15 min, 30 min, 1 h, 2 h, 5 h, 7 h, and 24 h, centrifuged at 12,000 rpm for 2 min, and the plasma was frozen and stored at -80 °C for the test. A certain amount of the test article was accurately weighed and dissolved in DMSO to a concentration of 2 mg / mL to obtain a stock solution. An appropriate amount of the compound stock solution was accurately taken, diluted by adding acetonitrile, and a standard series of solutions was prepared. 20 μL of each of the above standard series of solutions was accurately taken, 180 μL of blank plasma was added, vortexed and mixed, and plasma samples with plasma concentrations of 1, 3, 5, 10, 30, 100, 300, 1000, and 3000 ng / mL were prepared. Sample analysis was performed twice for each concentration, and a calibration curve was created. 30 μL of plasma was taken, 200 μL of an acetonitrile solution of propranolol (50 ng / mL) as the internal standard was added, vortexed and mixed well, 100 μL of purified water was added, vortexed and mixed well again, centrifuged at 4000 rpm for 5 min, and the supernatant was taken for LC-MS / MS analysis. The detection conditions for LC-MS / MS are as follows. Chromatography column: Waters ACQUITYTM PREMIER HSS T3, 50 * 2.1 mm, 1.8 μm. Mobile phase: water (0.1% formic acid)-acetonitrile (gradient elution is performed according to the following table) TIFF2025524934000042.tif4997(3) Data processing After detecting the blood concentration by LC-MS / MS, pharmacokinetic parameters were calculated using WinNonlin 6.1 software and the non-compartmental model method. The results are shown in Table 4.

Table 4

[0064] Example 15 Examples of Salt Screening Compound A was dissolved in various solvents such as isopropanol, acetone, and ethyl acetate, and an acid was added in the same molar ratio (a total of 17 acids as shown in the following table). After a certain reaction time, when the solution became clear, a poor solvent was added or the temperature was lowered to precipitate a solid. [Table 5] Compound A is an oil and can only precipitate as a solid after forming salts with some acids. As can be seen from the above table, the salt formation of Compound A is very difficult, and only several acids such as sulfuric acid, phosphoric acid, oxalic acid, and hydrochloric acid can form salts.

[0065] Example 16 Preparation of Oxalate The free base of Compound A (3.5 g) was dissolved in isopropanol (10 ml), oxalic acid dihydrate (689 mg) was added, and the mixture was stirred at room temperature for 1 h for reaction, concentrated to obtain an oil, isopropyl ether (50 ml) was added under an ice bath, stirred to precipitate a solid, stirred in the ice bath for 1 day, and filtered to obtain an oxalate solid. The free base of Compound A (2.4 g) and oxalic acid dihydrate (46 mg) were reacted in ethyl acetate (1 w / w) to form a salt, which was dropped into isopropyl ether (15 w / w) at -5°C, stirred for 1 day, filtered, and dried to obtain a solid. The free base of Compound A (2.2 g) and oxalic acid (70 mg) were reacted in isopropanol (2 ml), concentrated, isopropyl acetate (0.5 ml) was added, isopropyl ether (15 ml) was added at -15°C, stirred for 4 h, and filtered to obtain a solid. NMR data of oxalate: [Chemical formula] TIFF2025524934000046.tif159157 analysis: δ = 0.93 corresponds to the methyl hydrogens on the C atoms numbered 34 and 37. The number of protons is 6, and it is a doublet of triplets. δ = 1.28 corresponds to the methyl hydrogen on the C atom numbered 45. The number of protons is 3, and it is a broad doublet. δ = 1.34 corresponds to the methyl hydrogen on the C atom numbered 46, and the methylene hydrogens on the C atoms numbered 33 and 36. The number of protons is 7, and they are a triplet and two broad doublets respectively. δ = 1.52 corresponds to the methylene hydrogens on the C atoms numbered 32 and 35. The number of protons is 4, and it is a doublet of triplets. δ = 1.59 corresponds to the methyl hydrogen on the C atom numbered 39. The number of protons is 3, and it is a broad doublet. δ = 3.24, 3.29, 3.31, 3.35, 3.41, 3.59, and 3.68 correspond to the methylene hydrogens on the C atoms numbered 30, 31, 26, 11, 9, and 10 respectively. The number of protons is 12, and it is a triplet of multiplets. δ = 3.78 corresponds to the methyl hydrogen on the C atom numbered 25. The number of protons is 3, and it is a singlet. δ = 4.39 corresponds to the methine hydrogen on the C atom numbered 7. The number of protons is 1, and it is a doublet of broad doublets. δ = 5.08 corresponds to the methine hydrogen on the C atom numbered 44. The number of protons is 1, and it is a multiplet. δ = 5.93 corresponds to the methylene hydrogen on the C atom numbered 20. The number of protons is 2, and it is a triplet. δ = 6.63, 6.72, and 6.82 correspond to the methine hydrogens on the C atoms numbered 18, 15, and 14 respectively. The number of protons is 3, and they are a triplet of doublets. δ = 6.91 corresponds to the methine hydrogens on the benzene ring C atoms numbered 4 and [missing number]. The number of protons is 2, and it is a doublet of triplets. δ = 7.20 corresponds to the methine hydrogens on the benzene ring C atoms of No. 1 and No. 3. The number of protons is 2, and it is a doublet of triplets. δ = 7.51 corresponds to the methine hydrogen on the C atom of No. 38. The number of protons is 1, and it is a multiplet. δ = 11.05 corresponds to the carboxyl hydrogen on the C atom of oxalic acid. The number of protons is 2, and it is a singlet.

[0066] Example 17 Preparation of Phosphate The free base of Compound A (4.1 g) was dissolved in isopropanol (12 ml), phosphoric acid (800 mg) was added, and the reaction was carried out for 1 h. Isopropyl ether (80 ml) was added under an ice bath and stirred for 4 h to obtain a phosphate solid. NMR data of the phosphate:

Chemical Structure

[0067] Example 18 Preparation of hydrochloride The free base of compound A (202.5 mg) was weighed into a 5 mL vial, and MeOH (1.0 mL) and an equimolar ratio of hydrochloric acid methanol solution were added. It was suspended at -20 °C and stirred for 1 day. Methanol was removed by rotary evaporation, and the gel-like sample remained gel-like even after being subjected to vacuum drying. Isopropyl ether (2.0 mL) was added, and it was suspended and stirred at -20 °C for 1 day to precipitate a solid, which was centrifuged and subjected to vacuum drying to obtain a solid. The NMR spectrum is shown in Figure 2.

[0068] Example 19 Preparation of sulfate The free base of compound A (203.1 mg) was weighed into a 5 mL vial, an IPA (1.0 mL) solvent and an equimolar ratio of sulfuric acid were added, and it was suspended at -20 °C and stirred for 1 day. It was subjected to vacuum drying to remove IPA, and a gel-like sample was obtained. Isopropyl ether (2.0 mL) was added, and it was suspended and stirred at -20 °C for 1 day to precipitate a solid, which was centrifuged and dried under vacuum to obtain a solid. The NMR spectrum is shown in Figure 3.

[0069] Example 20: Study on Stability Sulfate, phosphate, hydrochloride, and oxalate were each left standing for one week under the conditions of 25°C / 60%RH and 40°C / 75%RH, and the chemical stability of the samples was tested by HPLC. From the above table, it was found that the oxalate and phosphate obtained according to the present invention have good stability and are superior to hydrochloride and sulfate.

[0070] Example 21: Study on Hygroscopicity The hygroscopicity of sulfate, phosphate, hydrochloride, and oxalate was evaluated by a dynamic vapor sorption (DVS) apparatus. In this test, the change rate of the mass of the sample when the humidity changed under a constant temperature condition of 25°C was collected. The results of the DVS test are as follows. From the above table, it was found that the oxalate and phosphate obtained according to the present invention have low hygroscopicity and are superior to hydrochloride and sulfate.

[0071] The above examples are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above examples, and other changes, modifications, substitutions, combinations, and simplifications made without departing from the spirit and principle of the present invention are all equivalent substitution forms and are all included in the protection scope of the present invention.

Claims

1. A salt of a compound represented by formula (I), characterized by the following: 【Chemical 1】 (M is an inorganic acid or an organic acid, where n:x = 1:1 to 2.)

2. The inorganic acid is selected from sulfuric acid, hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, carbonic acid, or nitric acid, The organic acid is selected from benzoic acid, 2,5-dihydroxybenzoic acid, 4-acetamidobenzoic acid, 4-aminobenzoic acid, oxalic acid, acetic acid, dichloroacetic acid, trichloroacetic acid, hexanoic acid, adipic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, octanoic acid, decanoic acid, cinnamic acid, citric acid, aspartic acid, gluconic acid, glutamic acid, lactic acid, malic acid, mandelic acid, pyroglutamic acid, tartaric acid, succinic acid, formic acid, fumaric acid, gentisic acid, glutaric acid, valeric acid, aspartic acid, lauric acid, camphoric acid, maleic acid, malonic acid, nicotinic acid, propionic acid, salicylic acid, 4-aminosalicylic acid, sebacic acid, stearic acid, succinic acid, palmitic acid, pamoic acid, trifluoroacetic acid, thiocyanic acid, p-toluenesulfonic acid, and L-malic acid, A salt of the compound represented by formula (I) according to claim 1, characterized in that it is selected.

3. A salt of the compound represented by formula (I) according to claim 1, characterized in that the inorganic acid is selected from phosphoric acid.

4. A salt of the compound represented by formula (I) according to claim 1, characterized in that the inorganic acid is selected from phosphoric acid, where n = 3 and x = 3 to 6.

5. A salt of the compound represented by formula (I) according to claim 1, characterized in that the inorganic acid is selected from phosphoric acid, where n = 3, x = 3, n = 3, x = 4, or n = 3, x = 5, or n = 3, x = 6.

6. A salt of the compound represented by formula (I) according to claim 1, characterized in that the organic acid is selected from oxalic acid.

7. A salt of the compound represented by formula (I) according to claim 1, characterized in that the organic acid is selected from oxalic acid, where n = 1 and x = 1.

8. A method for producing a salt of the compound according to any one of claims 1 to 7, characterized by mixing compound A and an acid. 【Chemical 2】

9. The method for production according to claim 8, characterized in that the molar ratio of the compound to the acid molecule is 1:1 to 2.

10. Step 1) weighing an appropriate amount of free alkali and dissolving it in a good solvent; Step 2) weighing out an appropriate amount of ionic acid and adding it to Step 1 to react and form a salt; Step 3) back-titrating the reaction solution of step 2 into a poor solvent and stirring to precipitate; and 4) subjecting the mixture to high speed centrifugation or settling to obtain a salt of the compound.

11. A pharmaceutical composition comprising a salt of the compound of formula (I) according to any one of claims 1 to 7 and one or more pharmaceutically acceptable carriers.

12. Use of the salt of the compound of formula (I) according to any one of claims 1 to 7 in the manufacture of a medicament for treating and / or preventing endothelin A (ETA) receptor antagonism-related diseases.

13. The use according to claim 12, characterized in that the diseases include chronic kidney disease, IgA, FSGS, Alport and hypertension diseases.

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