Pyrazolecarbonylpiperazinone compounds, pharmaceutical compositions and their uses

Abstract

This application relates to the pharmaceutical technology, and more particularly to pyrazolecarbonylpiperazinone compounds, pharmaceutical compositions, and their uses. Pyrazolecarbonylpiperazinone compounds are those having the structure represented by formula I, or pharmaceutically acceptable salts, esters, amides, solvates, active metabolites, crystalline polymorphs, isotopically labeled isomers, or prodrugs of the structure represented by formula I. JPEG2026517508000029.jpg58170

Description

Technical Field

[0001] This application relates to the field of pharmaceutical technology, and particularly to pyrazole carbonyl piperazinone compounds, pharmaceutical compositions and their uses.

Background Art

[0002] Transforming growth factor-β (TGF-β) belongs to the TGF-β superfamily and plays an extremely important role in regulating cell growth and differentiation. Dysfunction of the TGF-β signaling pathway is associated with human diseases such as cancer, fibrosis, systemic sclerosis, and genetic diseases (Wakefield LM and Hill CS. Nat Rev Cancer 2013, 13:328-341). For example, in advanced cancer, TGF-β often promotes tumor progression and metastasis and acts as an oncogenic factor (Massague. J. Cell. 2008, 134:215~230).

[0003] There are mainly two types of TGF-β cell membrane receptors, namely TGF-β type I receptor and TGF-β type II receptor. In vertebrates, there are seven types of type I receptors (Activin-receptor like kinases), namely ALK1 (ACVRL1), ALK2 (ACVR1), ALK3 (BMPR1A), ALK4 (ACVR1B), ALK5 (TGFBR1), ALK6 (BMPR1B) and ALK7 (ACVR1C), and five types of type II receptors, namely TGFβRII, ACTRII, ACTRIIB, BMPRII, and AMHRII. Among these, the ALK6 gene belonging to the type I receptor, that is, the BMPR1B (bone morphogenetic protein receptor, type IB) gene, has been shown to be associated with various high-incidence diseases such as breast cancer, gastric cancer, and pulmonary hypertension. Therefore, the method of artificially inhibiting BMPR1B to treat related diseases is an urgent issue in the pharmaceutical field. [Overview of the project] [Problems that the invention aims to solve]

[0004] Based on the above, there is a need for a pyrazolecarbonylpiperazinone compound that selectively inhibits BMPR1B, possesses antitumor activity, and is expected to have applications in the clinical treatment of cancer. [Means for solving the problem]

[0005] A first aspect of this application provides a pyrazolecarbonylpiperazinone compound, which has the structure represented by formula I, or is a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, crystalline polymorph, isotope-labeled, isomer, or prodrug of the structure represented by formula I. [ka] (In the formula, ring A is selected from one of the following structures, [ka] Y is -(CH2) n - and n is 0, 1, 2 or 3, R 1 is -H, -D, unsubstituted or R 2 Substitutions of C1-C6 alkyl, C1-C6 alkoxy, and C6-C 10 Aryl or C3~C 10 Selected from heteroaryls, R 2 These are selected from -F, -Cl, -Br, -I, -OH, -COOH, C1-C6 alkyl, or C1-C6 alkoxy. The asterisk (*) indicates a connecting point.

[0006] A second aspect of this application provides a pharmaceutical composition comprising the pyrazolecarbonylpiperazinone compound and at least one pharmaceutically acceptable carrier.

[0007] A third aspect of this application provides uses for the pyrazolecarbonylpiperazinone compound, or the pharmaceutical composition, in the manufacture of a pharmaceutical for the treatment of one or more diseases, such as cancer and pulmonary hypertension.

[0008] A fourth aspect of this application provides the use of the pyrazolecarbonylpiperazinone compound as a BMPR1B inhibitor.

[0009] A fifth aspect of this application provides a method for inhibiting the expression of BMPR1B. The process includes contacting the pyrazolecarbonylpiperazinone compound with cells, biological tissue, or organoids containing BMPR1B.

[0010] A sixth aspect of this application provides a method for treating cancer or pulmonary hypertension, comprising the steps of administering to a patient a therapeutically effective amount of the pyrazolecarbonylpiperazinone compound, or administering to a patient a therapeutically effective amount of the pharmaceutical composition. [Brief explanation of the drawing]

[0011] [Figure 1] This shows the binding specificity of pyrazolecarbonylpiperazinone compounds to their targets based on kinase activity screening analysis. Figure 1A shows the effect of compound II-1 on the activity of 330 kinase enzymes, and Figures 1B and 1C show the results of kinase activity tests of compound II-1 against TGFBR1 and BMPR2 at different concentrations. [Figure 2] The PyMol software displays information on the interaction between the compound and BMPR1B in the pyrazolecarbonylpiperazinone compound-BMPR1B complex. The compound in Figure 2A is II-13, the compound in Figure 2B is II-11, the compound in Figure 2C is II-2, and the compound in Figure 2D is II-1. [Figure 3] The results of toxicity experiments on tumor cells using compounds II-1 and LY2157299 are shown. [Modes for carrying out the invention]

[0012] To facilitate understanding of this application, a more comprehensive description of this application will be provided below with reference to the relevant drawings. While the drawings illustrate preferred embodiments of this application, it can be implemented in a variety of forms and is not limited to the embodiments described herein. Rather, the purpose of presenting these embodiments is to provide a more complete and comprehensive understanding of the disclosures of this application.

[0013] Unless otherwise specified, all technical and scientific terms used herein have meanings that are generally understood by those skilled in the art. The terms used herein are for illustrative purposes of specific embodiments and are not intended to limit this application. The terms “and / or” used herein encompass any one or more of the listed items, or any combination thereof.

[0014] In this application, the disclosed technical features include confidential technical solutions consisting of the listed features, as well as open technical solutions that include the listed features.

[0015] In this application, unless otherwise specified, numerical ranges are understood as continuous ranges, including the minimum and maximum values ​​of the range, and all values ​​between the minimum and maximum values. Furthermore, if a range refers to an integer, it includes all integers between the minimum and maximum values ​​of that range. In addition, for features or characteristics to which multiple ranges are assigned, these ranges can be combined. In other words, unless otherwise specified, all ranges disclosed herein are understood to include any and all subranges contained therein.

[0016] In this application, unless otherwise specified, percentage content refers to mass percentage for solid-liquid mixtures and solid-phase-solid mixtures, and volume percentage for liquid-phase-liquid mixtures.

[0017] In this application, unless otherwise specified, percentage concentrations refer to the final concentration. The final concentration, as used herein, means the proportion of the component in the system after its addition.

[0018] Unless otherwise specified, the temperature parameters in this application may be constant temperature processing or processing within a specific temperature range. Constant temperature processing, as used herein, means that temperature fluctuations are permitted within a precision range controlled by the equipment.

[0019] (Explanation of terms) The term "alkyl" refers to a monovalent residue produced by the loss of one hydrogen atom from a saturated hydrocarbon containing a primary (n) carbon atom, a secondary carbon atom, a tertiary carbon atom, a quaternary carbon atom, or a combination thereof. Expressions containing this term, such as "C1-C6 alkyl," refer to alkyl groups containing 1 to 6 carbon atoms, and each instance may independently be one of C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, or C6 alkyl. Suitable examples include methyl (Me, -CH3), ethyl (Et, -CH2CH3), 1-propyl (n-Pr, n-propyl, -CH2CH2CH3), 2-propyl (i-Pr, i-propyl, -CH(CH3)2), 1-butyl (n-Bu, n-butyl, -CH2CH2CH2CH3), 2-methyl-1-propyl (i-Bu, i-butyl, -CH2CH(CH3)2), and 2-butyl (s-Bu, s-butyl, -CH(CH3)). CH2CH3), 2-methyl-2-propyl(t-Bu, t-butyl, -C(CH3)3), 1-pentyl(n-pentyl, -CH2CH2CH2CH2CH3), 2-pentyl(-CH(CH3)CH2CH2CH3), 3-pentyl(-CH(CH2CH3)2), 2-methyl-2-butyl(-C(CH3)2CH2CH3), 3-methyl-2-butyl(-CH(CH3)CH(CH3)2), 3-methyl-1-butyl(-CH 2CH2CH(CH3)2), 2-methyl-1-butyl(-CH2CH(CH3)CH2CH3), 1-hexyl(-CH2CH2CH2CH2CH2CH3), 2-hexyl(-CH(CH3)CH2CH2CH2CH3), 3-hexyl(-CH(CH2CH3)(CH2CH2CH3)), 2-methyl-2-pentyl(-C(CH3)2CH2CH2CH3), 3-methyl-2-pentyl(-CH(CH3)CH(CH3)C Examples include, but are not limited to, H2CH3, 4-methyl-2-pentyl (-CH(CH3)CH2CH(CH3)2), 3-methyl-3-pentyl (-C(CH3)(CH2CH3)2), 2-methyl-3-pentyl (-CH(CH2CH3)CH(CH3)2), 2,3-dimethyl-2-butyl (-C(CH3)2CH(CH3)2), and 3,3-dimethyl-2-butyl (-CH(CH3)C(CH3)3).

[0020] The term "alkoxy" refers to a group having an -O-alkyl structure, that is, a group in which the alkyl group defined above is bonded to an adjacent group through an oxygen atom. Expressions containing this term, such as "C1-C6 alkoxy", refer to an alkyl moiety containing 1 to 6 carbon atoms, and each time it appears, it may independently be any of C1 alkoxy, C4 alkoxy, C5 alkoxy, or C6 alkoxy. Suitable examples include, but are not limited to, methoxy (-O-CH3 or -OMe), ethoxy (-O-CH2CH3 or -OEt), and tert-butoxy (-O-C(CH3)3 or -OtBu).

[0021] "Aryl" refers to an aromatic hydrocarbon group derived by removing one hydrogen atom from an aromatic ring compound, and may be any of monocyclic aryl, fused-ring aryl, or polycyclic aryl. In the case of polycyclic aryl, at least one is an aromatic ring system. For example, "C6-C 10 aryl" refers to an aryl group containing 6 to 10 carbon atoms, and each time it appears, it may independently be any of a C6 aryl group, a C7 aryl group, a C8 aryl group, a C9 aryl group, or a C 10 aryl group. Suitable examples include, but are not limited to, benzene, biphenyl, naphthalene, and their derivatives.

[0022] "Heteroaryl" refers to a group in which at least one carbon atom in an aryl group or a cyclopentadienyl group is replaced by a non-carbon atom such as an N atom, an O atom, or an S atom. For example, "C3-C 10"Heteroaryl" refers to a heteroaryl group containing 3 to 10 carbon atoms, and each instance may independently be a C3 heteroaryl, C4 heteroaryl, C5 heteroaryl, C6 heteroaryl, C7 heteroaryl, or C8 heteroaryl. Suitable examples include, but are not limited to, furan, benzofuran, thiophene, benzothiophene, pyrrole, pyrazole, triazole, imidazole, oxazole, oxadiazole, thiazole, tetrazole, indole, carbazole, pyrrole-imidazole, pyrrole-pyrrole, thiophene-pyrrole, thiophene-thiophene, furan-pyrrole, furan-furan, thiophene-furan, benzoisoxazole, benzoisothiazole, benzimidazole, pyridine, pyrazine, pyridazine, pyrimidine, triazine, quinoline, isoquinoline, cinnoline, quinoxaline, phenanthidine, primidine, quinazoline, and quinazolinone.

[0023] In this specification, the term “prodrug” refers to any compound that, upon administration to a living organism, generates a drug, i.e., an active ingredient, through spontaneous chemical reactions, enzymatic catalyzed chemical reactions, photolysis, and / or metabolic chemical reactions. Thus, a prodrug is a covalently modified analog or potential form of a therapeutically active compound. Suitable examples include, but are not limited to, carboxylic acid esters, carbonate esters, phosphate esters, nitrate esters, sulfate esters, sulfone esters, sulfinate esters, amino compounds, carbamates, azo compounds, phosphoramides, glucosides, ethers, and acetals.

[0024] "Medically acceptable" means a ligand, material, composition, and / or dosage form that is appropriate for administration to a patient within the bounds of reasonable medical judgment and that has a reasonable benefit-to-risk ratio.

[0025] "Medically acceptable carrier" refers to a pharmaceutically acceptable material, composition, or medium, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material. As used herein, the term "medically acceptable carrier" includes buffers suitable for drug administration, sterile water for injection, solvents, dispersions, coatings, antimicrobial and antifungal agents, isotonic agents, and absorption retarders. Each carrier must be "medically acceptable" in the sense of compatibility with other components in the formulation and non-harmful to the patient. Appropriate examples include: (1) sugars such as lactose, glucose, and sucrose; (2) starches such as corn starch, potato starch, and substituted or unsubstituted β-cyclodextrins; (3) cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and acetylcellulose; (4) powdered tragacanth gum; (5) malt; (6) gelatin; (7) talc; (8) excipients such as cocoa butter and suppository waxes; and (9) oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil. Examples of non-toxic, suitable substances used in pharmaceutical formulations include, but are not limited to, (10) glycols such as propylene glycol, (11) polyols such as glycerol, sorbitol, mannitol, and polyethylene glycol, (12) esters such as ethyl oleate and ethyl laurate, (13) agar, (14) buffers such as magnesium hydroxide and aluminum hydroxide, (15) alginic acid, (16) pyrogen-free water, (17) isotonic saline, (18) Ringer's solution, (19) ethanol, (20) phosphate buffer, and (21) other non-toxic, suitable substances used in pharmaceutical formulations.

[0026] A "pharmaceutically acceptable salt" refers to a salt formed by any compound having the given structure with an acid or base suitable for use as a pharmaceutical. pharmaceutically acceptable salts include both inorganic and organic salts. An example of a salt is a salt formed by the compound of this application with an acid. Acids suitable for salt formation include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, picric acid, benzoic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, and naphthalenesulfonic acid; and amino acids such as proline, phenylalanine, aspartic acid, and glutamic acid. Another example of a salt is a salt formed by the compound of this application with a base. Suitable bases for salt formation include, but are not limited to, alkali metal salts (e.g., sodium or potassium salts), alkaline earth metal salts (e.g., magnesium or calcium salts), ammonium salts (e.g., lower alkanolammonium salts and other pharmaceutically acceptable amine salts), such as methylamine salts, ethylamine salts, propylamine salts, dimethylamine salts, trimethylamine salts, diethylamine salts, triethylamine salts, tert-butylamine salts, ethylenediamine salts, hydroxyethylamine salts, dihydroxyethylamine salts, trihydroxyethylamine salts, and amine salts formed from morpholine, piperazine, and lysine, respectively.

[0027] "Medicinally acceptable esters and amides" refers to esters or amides suitable for medicinal use, formed by any of the compounds having the indicated structure with other compounds. Medicinally acceptable esters include organic esters and inorganic esters.

[0028] If the compound represented by formula I has a carboxyl group, the compound represented by formula I or its active derivative (e.g., acylchloride, mixed acid anhydride, etc.) is reacted with the corresponding alcohol (e.g., C1-C6 alcohol, etc.) or its active derivative (e.g., (5-methyl-2-oxo-1,3-dioxolen-4-yl)methyl body, (pivaloyloxy)methyl body, benzofuranone body, (isopropoxycarbonyl)oxymethyl body, etc.), or ammonia or the corresponding amine (mono-C1-C6 alkylamine or di-C1-C6 alkylamine, etc.) according to a conventional method (condensation reaction of carboxylic acid with alcohol or amine), and the ester of the compound represented by formula I is reacted with the corresponding alcohol (e.g., C1-C6 alcohol, etc.) (transesterification reaction), or ammonia or the corresponding amine (mono-C1-C6 alkylamine or di-C1-C6 alkylamine, etc.) Medicinally acceptable esters or amides can be prepared by reacting (amidation reaction) or by reacting an alkali metal salt of the compound represented by formula I with the corresponding halide (e.g., C1-C6 alkyl chloride or bromide, (5-methyl-2-oxo-1,3-dioxacyclopenten-4-yl)methyl chloride or bromide, (pivaloyloxy)methyl chloride or bromide, benzofuranone chloride or bromide, [(isopropoxycarbonyl)oxy]methyl chloride or bromide, etc.). Similarly, if the compound represented by formula I has a hydroxyl group, medicamentally acceptable esters can be obtained by condensing the compound represented by formula I with carboxylic acids, acyl chlorides, acid anhydrides, etc., according to conventional methods.

[0029] The aforementioned esters include, for example, C1-C6 alkyl esters such as methyl esters, ethyl esters, propyl esters, isopropyl esters, butyl esters, sec-butyl esters, tert-butyl esters, pentyl esters, and hexyl esters; C3-C6 cycloalkyl esters such as cyclopentyl esters and cyclohexyl esters; and C6-C alkyl esters such as phenyl esters and naphthyl esters. 10 Aryl esters; such as benzyl esters, phenethyl esters, α-methylbenzyl esters, 3-phenylpropyl esters, 4-phenylbutyl esters, 6-phenylhexyl esters, diphenylmethyl esters, triphenylmethyl esters, etc. C6-C 10 Examples include aryl C1-C6 alkyl esters. Alternatively, they may be esters that can be hydrolyzed in vivo, such as (5-methyl-2-oxo-1,3-dioxacyclopenten-4-yl)methyl ester, (pivaloyloxy)methyl ester, benzofuranone ester, [(isopropoxycarbonyl)oxy]methyl ester, [(cyclohexyloxycarbonyl)oxy]methyl ester, and 1-[(cyclohexyloxycarbonyl)oxy]ethyl ester.

[0030] Examples of the amides include mono-C1-C6 alkylamides or mono-C3-C6 cycloalkylamides such as amide (-CONH2), N-formamide, N-acetamide, N-propionamide, N-isopropionamide, N-butylamide, N-sec-butylamide, N-tert-butylamide, N-pentanamide, N-hexaneamide, N-cyclopropionamide, N-cyclopentanamide, and N-cyclohexaneamide. Alternatively, amides such as N,N-diformamide, N,N-diacetamide, N,N-dipropionamide, N,N-diisopropionamide, N-methyl-N-acetamide, N-methyl-N-propionamide, N-methyl-N-butylamide, N-ethyl-N-propionamide, N-ethyl-N-butylamide, N-butyl-N-cyclopentanamide, N-ethyl-N-cyclopropionamide, and N,N-dicyclohexaneamide are di-C1-C6 alkylamides, N-C1-C6 alkyl-N-C3-C6 cycloalkylamides, or di-C3-C6 cycloalkylamides.

[0031] A "solvate" refers to a complex formed when a compound represented by general formula (I) coordinates with a solvent molecule to form a specific ratio. A "hydrate" refers to a complex formed when the compound of this application coordinates with water.

[0032] An "active metabolite" refers to a derivative of a compound that possesses its activity, which is produced during the metabolism of that compound.

[0033] "Crystal polymorphism" refers to the compound of this application that exists in different crystal lattice configurations.

[0034] "Isotope labeling" refers to the compound of this application that is labeled with an isotope. For example, the isotopes in the compound of this application may be various isotopes of elements such as H, C, N, O, P, F, and S, for example, 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 31 P,32 P, 35 S, 18 F, 36 This includes S, etc.

[0035] An "isomer" refers to an isomer resulting from a different spatial arrangement of atoms within a molecule. Because the compounds in this application contain structures such as chiral or asymmetric centers and double bonds, they may include various isomeric forms, including optical isomers, geometric isomers, tautomers, and atropisomers. These isomers, their single isomers, and racemates are all within the scope of this application. For example, optical isomers, specifically the optically active (R)-, (S)- isomers and D, L isomers, can be prepared by chiral separation, chiral synthesis, chiral reagents, or other conventional techniques. For instance, they can be converted to diastereomers by reacting with a suitable optically active substance (e.g., a chiral alcohol or Mosher's chloride), and then separated to convert them back to the corresponding single isomers (e.g., by hydrolysis). Furthermore, separation can also be performed by chromatography.

[0036] "Pharmaceutical compositions" can be prepared using methods well known in the pharmaceutical field and can be administered or applied via various routes depending on whether local or systemic treatment is required and the area to be treated.

[0037] (Method of administration) There are no particular restrictions on the dosage form or method of administration of the compound or pharmaceutical composition of this application.

[0038] Typical methods of administration include, but are not limited to, oral, intratumor, rectal, parenteral (intravenous, intramuscular, or subcutaneous) injection, and local administration.

[0039] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the active compound is mixed with at least one common inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with any of the following components: (a) fillers or bulking agents such as starch, lactose, sucrose, glucose, mannitol, or silicic acid; (b) binders such as hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, or gum arabic; (c) humectants such as glycerin; (d) disintegrants such as agar, calcium carbonate, potato starch or tapioca starch, alginic acid, certain complex silicates, or sodium carbonate; (e) absorption retarders such as paraffin wax; (f) absorption enhancers such as quaternary ammonium compounds; (g) wetting agents such as cetyl alcohol or glyceryl monostearate; (h) adsorbents such as kaolin; or (i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, or mixtures thereof. In the form of capsules, tablets, and pills, buffering agents may be included in the dosage form. Solid dosage forms such as tablets, sugar-coated tablets, capsules, pills, and granules can be prepared using coating agents and shells, such as enteric coatings or other materials well known in the art. These may contain opacifying agents, and the release of the active compound or compound in such compositions may be delayed in a portion of the gastrointestinal tract. Examples of usable embedding components include polymers and waxes. If necessary, the active compound may also be prepared in microcapsule form using one or more of the above excipients.

[0040] Liquid dosage forms for oral administration include pharmaceutically acceptable emulsions, solutions, suspensions, syrups, or tinctures. In addition to the active compound, the liquid dosage form may include inert diluents commonly used in the art, such as water or other solvents, solubilizers, emulsifiers, specifically, for example, ethanol, isopropanol, ethyl carbonate, ethyl acetate, propylene glycol, 1,3-butanediol, dimethylformamide, and oils, particularly cottonseed oil, peanut oil, corn germ oil, olive oil, castor oil, sesame oil, or mixtures thereof. In addition to these inert diluents, the composition may also include auxiliary agents such as wetting agents, emulsifiers and suspending agents, sweeteners, flavoring agents, and fragrances. For example, a suspension may include suspending agents, specifically, for example, ethoxylated isooctadecyl alcohol, polyoxyethylene sorbitol and dehydrated sorbitan esters, microcrystalline cellulose, aluminum methoxide, and agar, or mixtures thereof.

[0041] Parenteral injection compositions may include physiologically acceptable sterile aqueous or non-aqueous solutions, dispersions, suspensions, or emulsions, as well as sterile powders for redissolution in sterile injection solutions or dispersions. Suitable aqueous or non-aqueous carriers, diluents, solvents, or excipients include water, ethanol, polyols, and suitable mixtures thereof.

[0042] Topical dosage forms include ointments, powders, patches, sprays, and inhalants. These are prepared by mixing the active ingredient with a pharmaceutically acceptable carrier under sterile conditions, and any preservatives, buffers, or propellants as needed.

[0043] In this specification, “pharmaceutical” includes any drug, compound, composition, or mixture that produces physiological and / or pharmacological effects in vivo or in vitro, and which often produce beneficial effects. The range of physiological and / or pharmacological effects that such “pharmaceutical” produces in vivo is not particularly limited and may include systemic effects or local effects only. The activity of such “pharmaceutical” is not particularly limited and may include active substances that interact with other substances or inactive substances that do not interact with other substances.

[0044] As used herein, “therapeutic dose” means the amount of the compound of this application that elicits a biological or medical response in an individual, for example, the amount of the compound of this application that produces a physiologically and / or pharmacologically favorable effect in an individual, including but not limited to reducing or inhibiting the activity of an enzyme or protein, improving or alleviating symptoms, reducing or delaying the progression of a disease, or preventing a disease.

[0045] The TGF-β superfamily is involved in many aspects of biology, including embryonic development, organogenesis, cell fate determination, immunomodulation, stress response, and stem cell function. Therefore, numerous studies have been conducted on TGF-β, and several TGF-β signaling pathway inhibitors, such as Galunisertib (LY2157299), are currently in clinical trials. LY2157299 is a small molecule inhibitor that selectively inhibits the kinase activity of TβRIs. It has been studied in various cancer patients (e.g., glioblastoma, pancreatic cancer, hepatocellular carcinoma) as monotherapy or in combination with standard antitumor regimens (Rodon J et al., Clin Cancer Res 2015, 21:553~560). In the initial Phase I trial, LY2157299 showed promising activity in glioblastoma patients. On the other hand, LY2157299 showed superior efficacy compared to gemcitabine in pancreatic cancer patients (Mellisi D et al., Cancer Res 2016, 76(14 Supplement):CT068-CT068). However, existing small molecule inhibitors have limitations in their ability to kill tumor cells and can selectively inhibit only a limited number of kinases. Therefore, the development of TGF-β signaling inhibitors that are more effective, target more kinases, and can be useful in treating a wider range of diseases, especially cancer, is an urgent issue in the medical field.

[0046] Based on the above background, a first aspect of this application provides a pyrazolecarbonylpiperazinone compound. The pyrazolecarbonylpiperazinone compound has the structure represented by formula I, or is a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, crystalline polymorph, isotope-labeled, isomer or prodrug of the structure represented by formula I.

[0047] [ka]

[0048] In the formula, ring A is selected from one of the following structures.

[0049] [ka]

[0050] Y is -(CH2) n - and n is 0, 1, 2 or 3, R 1 is --H, --D, unsubstituted or R 2 Substitutions of C1-C6 alkyl, C1-C6 alkoxy, and C6-C 10 Aryl or C3~C 10 Selected from heteroaryls, R 2 These are selected from -F, -Cl, -Br, -I, -OH, -COOH, C1-C6 alkyl, or C1-C6 alkoxy. The asterisk (*) indicates a connecting point.

[0051] The pyrazolecarbonylpiperazinone compounds provided in this application, through their special structural design, can form relatively stable complexes with BMPR1B via hydrogen bonding and hydrophobic interactions. This enables targeted binding to BMPR1B, allowing for highly selective inhibition of BMPR1B, effectively killing tumor cells and preventing tumor progression and metastasis. Compared with existing small molecule inhibitors such as LY2157299, the pyrazolecarbonylpiperazinone compounds of this application exhibit higher killing power against tumor cells.

[0052] In some embodiments, Y is -(CH2) n - where n is 0, 1, or 2, preferably Y is -(CH2) n - where n is 0 or 1. More preferably, Y is -(CH2) n - and n is either 0 or 1.

[0053] In some embodiments, R 1 is -H, -D, unsubstituted or R 2 Substitution of C1-C6 alkyl groups, C6-C 10 Aryl, or C3~C 10 Selected from heteroaryl groups. Preferably, R 1is -H, -D, unsubstituted or R 2 Selected from substituted C3-C6 heteroaryl groups. More preferably, R 1 is -H, -D, unsubstituted or R 2 A substitution is selected from furan, thiophene, pyrrole, pyrazole, imidazole, oxazole, isoxazole, thiazole, isothiazole, pyran, thiam, pyridine, pyridazine, or pyrimidine. More preferably, R 1 is selected from -H, -D, or one of the following substituents.

[0054] [ka]

[0055] Here, "*" indicates a connecting point.

[0056] R 1 When the heteroaryl group is selected, particularly a heteroaryl group substituted with a methyl or hydroxyl group, the resulting pyrazolecarbonylpiperazinone compound exhibits high binding energy to the BMPR1B docking target and low physiological toxicity. This is beneficial for improving the efficacy and biocompatibility of pyrazolecarbonylpiperazinone compounds as BMPR1B inhibitors, thereby increasing their potential for clinical application.

[0057] In some embodiments, R 2 is selected from -OH or C1-C6 alkyl. Preferably, R 2 is selected from -OH or C1-C4 alkyl. More preferably, R 2 The propyl group is selected from -OH, methyl, ethyl, n-propyl, or isopropyl.

[0058] In some embodiments, the pyrazolecarbonylpiperazinone compound has one of the structures shown in formulas I-1 to I-13, or is a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, crystalline polymorph, isotope-labeled, isomer, or prodrug of one of the structures shown in formulas I-1 to I-13.

[0059] [ka]

[0060] In some embodiments, the pyrazolecarbonylpiperazinone compound has the structure shown in Formula II, or is a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, crystalline polymorph, isotope-labeled, isomer, or prodrug of the structure shown in Formula II.

[0061] [ka]

[0062] In the formula, ring A, Y and R 1 This is the same as the definition described in any of the embodiments described above.

[0063] In some embodiments, the pyrazolecarbonylpiperazinone compound has one of the structures shown in formulas II-1 to II-13, or is a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, crystalline polymorph, isotope-labeled, isomer, or prodrug of one of the structures shown in formulas II-1 to II-13.

[0064] [ka]

[0065] A second aspect of this application provides a pharmaceutical composition comprising the pyrazolecarbonylpiperazinone compound and at least one pharmaceutically acceptable carrier.

[0066] A third aspect of this application provides uses for the pyrazolecarbonylpiperazinone compound, or the pharmaceutical composition, in the manufacture of a pharmaceutical for the treatment of one or more diseases, such as cancer and pulmonary hypertension.

[0067] In some embodiments, the cancer is one or more of ovarian cancer, breast cancer, glioma, and germ cell tumors.

[0068] A fourth aspect of this application provides the use of the pyrazolecarbonylpiperazinone compound as a BMPR1B inhibitor.

[0069] In some embodiments, the concentration of the pyrazolecarbonylpiperazinone compound is 10 μM to 100 μM. In any choice, the concentration of the pyrazolecarbonylpiperazinone compound may be, for example, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, or 90 μM.

[0070] A fifth aspect of this application provides a method for inhibiting BMPR1B expression, comprising the step of contacting cells, biological tissue, or organoids containing BMPR1B with a pyrazolecarbonylpiperazinone compound described in one or more embodiments.

[0071] In some embodiments, the concentration of the pyrazolecarbonylpiperazinone compound during contact treatment is 10 μM to 100 μM. In any choice, the concentration of the pyrazolecarbonylpiperazinone compound may be, for example, 20 μM, 30 μM, 40 μM, 50 μM, 60 μM, 70 μM, 80 μM, or 90 μM.

[0072] A sixth aspect of this application provides a method for treating cancer or pulmonary hypertension, comprising the steps of administering to a patient a therapeutically effective amount of the pyrazolecarbonylpiperazinone compound, or administering to a patient a therapeutically effective amount of the pharmaceutical composition.

[0073] In some embodiments, the cancer is one or more of ovarian cancer, breast cancer, and glioma.

[0074] Synthesis of compounds (In the following cases, if there is a discrepancy between the compound name and structural formula, the structural formula takes precedence):

[0075] [ka]

[0076] Intermediate M1-n (0.12 mmol), (4aS,8aS)-octahydroquinoline-2(1H)-one (M2, 15 mg, 0.1 mmol), and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU, 380 mg, 1 mmol) were mixed, and then N,N-diisopropylethylamine (DIEA, 2 mL) and N,N-dimethylformamide (DMF, 20 mL) were added, and the mixture was stirred at room temperature for 18 hours. Ethyl acetate and water were added to the reaction mixture, the organic layer was separated, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:hexane = 3:7) to obtain the target product II-n.

[0077] Compounds II-1 and II-13 are obtained by synthetic route A. II-1: (4aS,8aS)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxalin-2(1H)-one (4aS,8aS)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxaline-2(1H)-one (12 mg, 41%), LC-MS m / z = 289.2 [M + H] + II-13: (4aS,8aS)-4-(1H-indazole-3-carbonyl)octahydroquinoxalin-2(1H)-one (4aS,8aS)-4-(1H-indazole-3-carbonyl)octahydroquinoxaline-2(1H)-one (17mg, 57%), LC-MS m / z=299.2[M+H] +

[0078] [ka]

[0079] Compound II-12 is obtained by synthetic route B. II-12: (4aS,8aS)-4-(4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine-3-carbonyl)octahydroquinoxalin-2(1H)-one (4aS,8aS)-4-(4,5,6,7-tetrahydro-1H-pyrazolo[4,3-c]pyridine-3-carbonyl)octahydroquinoxaline-2(1H)-one

[0080] (1) At 0°C, 1.5 mL of 1,4-dioxane solution containing 0.25 g (1.1 mmol) of di-tert-butyl dicarbonate (Boc2O) was added dropwise to 2 mL of 1 mol / L NaOH containing 4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylic acid (M3-1, 0.167 g, 1 mmol) and 0.6 mL of 1,4-dioxane solution. The reaction mixture was stirred at 0°C for 0.5 hours, and then allowed to react overnight at room temperature while maintaining the pH at 8-9. After the reaction, the mixture was diluted with 5 mL of H2O and extracted with n-hexane. The aqueous phase was acidified to pH 2-3 with citric acid and extracted with ethyl acetate. The organic phase was washed with saturated saline solution, dried over anhydrous magnesium sulfate, and concentrated under reduced pressure to obtain 5-(tert-butoxycarbonyl)-4,5,6,7-tetrahydro-2H-pyrazolo[4,3-c]pyridine-3-carboxylic acid (M3-2, 200 mg, 75%). LC-MS m / z = 268.1 [M + H] + That was the case. (2) M3-2 (20 mg, 0.075 mmol) was mixed with (4aS,8aS)-octahydroquinoline-2(1H)-one (M2, 15 mg, 0.1 mmol) and 2-(7-azabenzotriazole)-N,N,N',N'-tetramethylurea hexafluorophosphate (HATU, 38 mg, 0.1 mmol), and N,N-diisopropylethylamine (DIEA, 0.2 mL) and N,N-dimethylformamide (DMF, 2 mL) were added, and the mixture was stirred at room temperature for 18 hours. Ethyl acetate and water were added to the reaction mixture, the organic layer was separated, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:hexane = 3:7) to obtain tert-butyl 3-((4aS,8aS)-3-oxodeoxyquinoline-1-carbonyl)-1,4,6,7-tetrahydro-5H-pyrazole[4,3-c]pyridine-5-carboxylic acid ester (M3-3, 18 mg, 60%). LC-MS m / z = 404.2 [M+H] + That was the case. (3) M3-3 (18 mg, 0.04 mmol) and trifluoroacetic acid (TFA, 35 mg, 0.3 mmol) were added to 2 mL of dichloromethane (DCM), and the mixture was stirred at room temperature for 30 minutes. Ethyl acetate and water were then added to the reaction mixture, the organic layer was separated, dried over sodium sulfate, and concentrated under reduced pressure. II-12 (13 mg, 96%) was obtained. LC-MS m / z = 304.2 [M + H] + That was the case.

[0081] [ka]

[0082] (1) Sodium hydride (60% suspension, 1.07 g, 26.9 mmol) dissolved in tetrahydrofuran (5 mL) was cooled to 0°C, and a solution of M1-n (21.5 mmol) in tetrahydrofuran (50 mL) was added dropwise over 15 minutes, and the mixture was stirred at 0°C for 1 hour. A solution of (2-(chloromethoxy)ethyl)trimethylsilane (M4, 4.76 mL, 26.9 mmol) in tetrahydrofuran (50 mL) was added dropwise to the mixture, and the mixture was stirred at room temperature for 48 hours. Water and ethyl acetate were slowly added to the reaction mixture, the organic layer was separated, dried over sodium sulfate, and concentrated under reduced pressure to obtain M5-n. (2) In a 20 mL reactor, R 1 -YX(0.698 mmol, where X represents halogen, R 1 (and Y are as defined above), (4aS,8aS)-octahydroquinoxaline-2(1H)-one (M2, 108 mg, 0.698 mmol, 1.0 eq.), cesium carbonate (796 mg, 2.443 mmol, 2.5 eq.), 4,5-bisdiphenylphosphino-9,9-dimethylxanthene (20 mg, 0.035 mmol, 0.05 eq.), and palladium acetate (7.84 mg, 0.035 mmol, 0.05 eq.) were dissolved in tetrahydrofuran (3 mL) and blown with nitrogen for 10 minutes. The reaction mixture was heated to 70°C and maintained for 90 minutes. The mixture was filtered, washed with dichloromethane, and then concentrated. The residue was purified by silica gel column chromatography (0-10% ethyl acetate / dichloromethane) to obtain M6-n. (3) M6-n (1.0 mmol, 1.0 eq.) was dissolved in dichloromethane (10 mL), and M5-n (1.2 mmol, 1.2 eq.), triethylamine (200 mg, 2.0 mmol, 2.0 eq.), and hexafluorophosphate azabenzotriazole tetramethylurea (570 mg, 1.5 mmol, 1.5 eq.) were added to the mixture and stirred overnight. The reaction was observed using thin-layer chromatography, and when the reaction products were no longer visible, the mixture was diluted with dichloromethane, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1) to obtain M7-n. (4) At 5°C, triethylsilane (2.30 g, 19.8 mmol) was added to a solution of M7-n (3.95 mmol) in tetrahydrofuran (39.5 mL), and the mixture was stirred at room temperature for 3 hours. The reaction mixture was then concentrated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 20-100%) to obtain the target product II-n.

[0083] Compounds II-2 to II-10 are obtained via synthetic route C. II-2: (4aS,8aS)-1-(3-methylthiophen-2-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxalin-2(1H)-one (4aS,8aS)-1-(3-methylthiophen-2-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxaline-2(1H)-one (1504 mg, 99%), LC-MS m / z=384.2[M+H] + II-3: (4aS,8aS)-1-(2-methyloxazol-4-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxalin-2(1H)-one (4aS,8aS)-1-(2-methyloxazol-4-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxaline-2(1H)-one (1445 mg, 99%), LC-MS m / z=370.2[M+H] + II-4: (4aS,8aS)-1-(3-methylisothiazol-5-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxalin-2(1H)-one (4aS,8aS)-1-(3-methylisothiazol-5-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxaline-2(1H)-one (1507 mg, 99%), LC-MS m / z=386.2[M+H] + II-5: (4aS,8aS)-1-(2-methylpyridin-4-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxalin-2(1H)-one (4aS,8aS)-1-(2-methylpyridine-4-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxaline-2(1H)-one (1469 mg, 98%), LC-MS m / z=380.2[M+H] + II-6: (4aS,8aS)-1-(6-methylpyridin-2-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxalin-2(1H)-one (4aS,8aS)-1-(6-methylpyridine-2-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxaline-2(1H)-one (1484 mg, 99%), LC-MS m / z=380.2[M+H] + II-7: (4aS,8aS)-1-(6-hydroxypyridin-2-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxalin-2(1H)-one (4aS,8aS)-1-(6-hydroxypyridine-2-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxaline-2(1H)-one (1491 mg, 99%), LC-MS m / z=382.2[M+H] + II-8: (4aS,8aS)-1-(4-hydroxypyridin-2-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxalin-2(1H)-one (4aS,8aS)-1-(4-hydroxypyridine-2-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxaline-2(1H)-one (1491 mg, 99%), LC-MS m / z=382.2[M+H] + II-9: (4aS,8aS)-1-(2-methylpyrimidin-4-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxalin-2(1H)-one (4aS,8aS)-1-(2-methylpyrimidine-4-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxaline-2(1H)-one (1488 mg, 99%), LC-MS m / z=381.2[M+H] + II-10: (4aS,8aS)-1-(6-methylpyridazin-3-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxalin-2(1H)-one (4aS,8aS)-1-(6-methylpyridazin-3-yl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxaline-2(1H)-one (1413 mg, 94%), LC-MS m / z=381.2[M+H] +

[0084] [ka]

[0085] Compound II-11 is obtained via synthetic route D. II-11: (4aS,8aS)-1-((5-methyl-1H-pyrazol-4-yl)methyl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxalin-2(1H)-one (4aS,8aS)-1-((5-methyl-1H-pyrazole-4-yl)methyl)-4-(1,4,5,6-tetrahydrocyclopenta[c]pyrazole-3-carbonyl)octahydroquinoxaline-2(1H)-one

[0086] (1) Sodium hydride (60% suspension, 1.07 g, 26.9 mmol) dissolved in tetrahydrofuran (5 mL) was cooled to 0°C, and a solution of 5-methyl-1H-pyrazole-4-carbaldehyde (M8-1, 2.37 g, 21.5 mmol) in tetrahydrofuran (50 mL) was added dropwise over 15 minutes, and the mixture was stirred at 0°C for 1 hour. A solution of (2-(chloromethoxy)ethyl)trimethylsilane (M4, 4.76 mL, 26.9 mmol) in tetrahydrofuran (50 mL) was added dropwise to the mixture, and the mixture was stirred at room temperature for 48 hours. Water and ethyl acetate were slowly added to the reaction mixture, the organic layer was separated, dried over sodium sulfate, and concentrated under reduced pressure to obtain 5-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-carbaldehyde (M8-2, 4.96 g, 96%). LC-MS m / z = 240.1[M+H] + That was the case. (2) In a 20 mL reactor, M8-2 (168 mg, 0.698 mmol), (4aS,8aS)-octahydroquinoxaline-2(1H)-one (M2, 108 mg, 0.698 mmol, 1.0 eq.), cesium carbonate (796 mg, 2.443 mmol, 2.5 eq.), 4,5-bisdiphenylphosphino-9,9-dimethylxanthene (20 mg, 0.035 mmol, 0.05 eq.), and palladium acetate (7.84 mg, 0.035 mmol, 0.05 eq.) were dissolved in tetrahydrofuran (3 mL) and blown in with nitrogen for 10 minutes. The reaction was heated to 70°C and maintained for 90 minutes. The mixture was filtered, washed with dichloromethane, and then concentrated. The residue was purified by silica gel column chromatography (0-10% ethyl acetate / dichloromethane) to obtain (4aS,8aS)-1-((5-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-yl)methyl)octahydroquinoxaline-2(1H)-one (M8-3, 244 mg, 92.2%). LC-MS m / z = 378.3 [M+H] + That was the case. (3) M8-3 (379 mg, 1.0 mmol, 1.0 eq.) was dissolved in dichloromethane (10 mL), and 1-((2-(trimethylsilyl)ethoxy)methyl)-1,4,5,6-tetrahydrocyclopentapyrazole-3-carboxylic acid (M8-4, 339 mg, 1.2 mmol, 1.2 eq.), triethylamine (200 mg, 2.0 mmol, 2.0 eq.), and hexafluorophosphate azabenzotriazole tetramethylurea (570 mg, 1.5 mmol, 1.5 eq.) were added and the mixture was stirred overnight. The reaction was observed using thin-layer chromatography, and when the reactants were no longer visible, the mixture was diluted with dichloromethane, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate = 2:1) to obtain (4aS,8aS)-1-((5-methyl-1-((2-(trimethylsilyl)ethoxy)methyl)-1H-pyrazole-4-yl)methyl)-4-(1-((2-(trimethylsilyl)ethoxy)methyl)-1,4,5,6-tetrahydrocyclopentapyrazole-3-carbonyl)octahydroquinoxaline-2(1H)-one (M8-5, 424 mg, 66%). LC-MS m / z = 642.4 [M+H] + That was the case. (4) At 5°C, triethylsilane (2.30 g, 19.8 mmol) was added to a solution of M8-5 (2540 mg, 3.95 mmol) in tetrahydrofuran (39.5 mL), and the mixture was stirred at room temperature for 3 hours. The reaction mixture was then concentrated, washed with brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (ethyl acetate:petroleum ether = 20-100%) to obtain II-11 (1510 mg, 99%). LC-MS m / z = 383.2 [M + H] + That was the case.

[0087] Example 1 Using AutoDock Vina software, the pyrazolecarbonylpiperazinone compounds described in this application were molecularly docked with the BMPR1B target protein to create 13 docking structures. The binding energy of the optimal docking result between each pyrazolecarbonylpiperazinone compound and the BMPR1B target protein was calculated, and the results are shown in Table 1. The second column of Table 1 shows the binding free energy calculated by the AutoDock Vina molecular docking software; the more negative the value, the stronger the compound's binding ability to the target protein. According to the predicted values, the absolute values ​​of the binding energies of all the pyrazolecarbonylpiperazinone compounds significantly exceeded the threshold 3 set based on the target characteristics.

[0088] Cytotoxicity predictions were performed for the pyrazolecarbonylpiperazinone compounds related to this application using eToxPred software. The toxicity calculation results for each pyrazolecarbonylpiperazinone compound are shown in the third column of Table 1. According to the predictions, the toxicity of all pyrazolecarbonylpiperazinone compounds was below the threshold of 0.58 (a value above 0.58 indicates the possibility of toxicity).

[0089] [Table 1]

[0090] Example 2 330 kinase candidates (15–50 nM, 2.5 μL) were mixed with compound II-1 (2.5 mM, 25 nL, DMSO prepared) and incubated at 25°C for 10 minutes. Subsequently, a mixture of kinase peptide substrate (0.2 mg / mL, supplier: GenScript) and ATP (10–60 μM, supplier: Promega, product number: V915B) (total 2.5 μL, test buffer prepared) was added to the mixture and reacted at 25°C for 60–120 minutes. Finally, HTRF was performed. 1 or ADP-Glo 2The reaction products were quantitatively analyzed using a specific method. In the kinase activity tests, the consumption of each substrate was less than 10%. Two technical replicates were set up for each kinase activity measurement.

[0091] As a result, II-1 significantly inhibited the kinase activity of BMPR1B (Figure 1A; Note: All results are shown, but for readability, one kinase name is displayed on the x-axis for every 10 kinases). On the other hand, II-1 at the same concentration did not inhibit the kinase activity of TGFBR1 and BMPR2, and showed no inhibitory effect on TGFBR1 and BMPR2 at different concentrations (maximum concentration 12.5 μM, 3-fold serial dilution) (Figures 1B and 1C). This result indicates that II-1 has the ability to specifically bind to its target site.

[0092] To further verify the above experiments from a structural perspective, we performed structural analysis on the complexes formed by compounds II-1, II-2, II-11, and II-13 with BMPR1B using PyMol software (supplier: Schrodinger). As shown in Figure 2, the structures of the complexes were obtained by molecular docking. The N- and C-terminal regions of BMPR1B are shown in cartoon models of different shapes. The loop structure of BMPR1B and the activation loop structure that stimulates catalytic activity are shown in cartoon models of different grayscale values. The pyrazolecarbonylpiperazinone compounds are shown in stick-ring models. The interacting amino acid backbone / side chains are shown in stick models. Hydrogen bonding interactions are shown by solid lines. Hydrophobic interactions are shown by dashed lines. This result also indicates that the pyrazolecarbonylpiperazinone compounds have structural properties that allow them to bind with BMPR1B.

[0093] Example 3 OVCAR3 (ovarian cancer), Hela (breast cancer), U251 (glioblastoma) tumor cells, H9 cells, and HEK293 non-tumor cells were cultured in DMEM / F12 + 10% FBS + 1% P / S medium. After the cells reached 70% confluence, they were digested with 0.25% Trypsin digestate at 37°C for 2-3 minutes, and digestion was stopped with DMEM / F12. The cell suspension was collected in a 15 mL centrifuge tube, centrifuged at 1000 rpm for 5 minutes, the supernatant was removed, and the cells were resuspended in DMEM / F12 + 10% FBS + 1% P / S medium and measured on a hemocytometer. Based on the measurement results, the cells were resuspended to the corresponding concentration using DMEM / F12 + 10% FBS + 1% P / S medium, and tumor cells were cultured at 1 × 10⁶ per well. 4 Cells were seeded in 96-well plates at a seeding rate of 100 μL / well. From day 1 to day 7 after cell seeding, the cells were treated with compound II-1 (pyrazolecarbonylpiperazinone compound diluted to 10 μM, 30 μM, and 50 μM using DMEM + 10% FBS medium) at concentrations of 0 μM, 10 μM, 30 μM, and 50 μM, and each concentration gradient was repeated three times.

[0094] On days 1, 3, 5, and 7 after treatment, absorbance at OD450nm (instrument) was measured using the CCK-8 cell proliferation and toxicity test kit (Vazyme, A311-02). Absorbance from wells containing only culture medium or only CCK-8 Solution was used as a blank control, and the test was repeated three times for each concentration. Based on the absorbance results, cell viability at different time points was calculated, and the results are shown in Table 2.

[0095] The cell viability rate is calculated as follows: At the same time point in the same cell, the absorbance detected from cells without compound II-1 treatment is set as the baseline value (100%). The absorbance detected from cells treated with different concentrations is compared to the value for the CK group, and the ratio of absorbance to the CK group is calculated. For example, the viability rate of H9 cells treated with 30 μM low molecular weight compound on day 4 is calculated as follows. [OD(D4)30μM iPS] / [OD(D4)0μM H9]×100%

[0096] [Table 2]

[0097] The results showed that when three types of tumor cells—Hela, U251, and OVCAR3—were treated with the same concentration of the low-molecular-weight compound II-1, cell viability tended to gradually decrease over time, and this trend continued as the concentration of compound II-1 increased. Among these, U251 cells showed a more sensitive response to the low-molecular-weight compound, exhibiting even lower cell viability compared to Hela and OVCAR3 cells.

[0098] Even for H9 cells, an atypical representative of embryonic tumors (e.g., germ cell tumors), the survival rate of H9 cells similarly tended to gradually decrease with the passage of time and increasing concentration of compound II-1. On the other hand, in non-tumor cells, HEK293, no clear change in cell survival rate was observed with the passage of time and increasing concentration of compound II-1. This result indicates that compound II-1 has a more specific killing effect on tumor cells.

[0099] Example 3 In this example, glioblastoma cells U251 were treated with compound II-1 and the control compound LY2157299. Samples were collected at 12 and 72 hours of induction, and total RNA was extracted using Rneasy Mini or Micro Kit (QIAGEN) with the same target compound used during treatment. cDNA was synthesized from 1 mg of RNA using the SuperScript III First-Strand Synthesis System (Invitrogen). Quantitative PCR labeling and reaction were performed using SYBR Premix Ex Taq (TaKaRa) and Thermal Cycler Dice Real Time System (TaKaRa), with beta-Actin used as the internal standard. All data were analyzed using the delta-Ct method. Each test was repeated three times, and analysis of variance was performed. Primer sequences for identifying genes encoding different cell markers are shown in Table 3. The detection results are shown in Figure 3. The experimental results showed that, at the same concentration, compound II-1 exhibited a higher apoptotic activity than the antitumor molecule LY2157299, and after 72 hours of treatment, fewer residual cells were observed in the wells treated with compound II-1. Therefore, compared to the control compound LY2157299, the pyrazolecarbonylpiperazinone compound of this application was shown to have a stronger killing effect on tumor cells.

[0100] [Table 3]

[0101] The technical features of the embodiments described above can be combined in any way, and for the sake of brevity, not all possible combinations of the technical features in the embodiments described above have been explained. However, as long as these combinations of technical features are inconsistent, they should all be considered to fall within the scope described herein.

[0102] The embodiments described above represent only a few embodiments of this application, and although the descriptions are relatively specific and detailed, they should not be understood as limiting the scope of the patent for the invention. Those skilled in the art will note that several modifications and improvements can be made without departing from the concept of this application, and all of these fall within the scope of protection. Therefore, the scope of protection sought by the patent for this application should be determined based on the attached claims, and the specification and drawings are used to interpret the content of the claims.

[0103] (Note) (Note 1) A pyrazolecarbonylpiperazinone compound having the structure represented by formula I, or a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, crystalline polymorph, isotope-labeled, isomer, or prodrug of the structure represented by formula I. [ka] (In the formula, ring A is selected from one of the following structures, [ka] Y is -(CH2) n - and n is 0, 1, 2 or 3, R 1 is -H, -D, unsubstituted or R 2 Substitutions of C1-C6 alkyl, C1-C6 alkoxy, and C6-C 10 Aryl or C3~C 10 Selected from heteroaryls, R 2 These are selected from -F, -Cl, -Br, -I, -OH, -COOH, C1-C6 alkyl, or C1-C6 alkoxy. The asterisk (*) indicates a connecting point.

[0104] (Note 2) (1) Y is -(CH2) n - and n is 0, 1 or 2, (2)R 1 is -H, -D, unsubstituted or R 2 substituted C1-C6 alkyl, C6-C 10 aryl or C3-C 10 heteroaryl, (3)R 2 is selected from -OH or C1-C6 alkyl, A pyrazolecarbonylpiperazinone compound according to Supplementary Note 1, characterized by satisfying one or more of the conditions.

[0105] (Supplementary Note 3) (1) Y is -(CH2) n -, n is 0 or 1, (2)R 1 is -H, -D, unsubstituted or R 2 substituted C3-C6 heteroaryl, (3)R 2 is selected from -OH or C1-C4 alkyl, A pyrazolecarbonylpiperazinone compound according to Supplementary Note 2, characterized by satisfying one or more of the conditions.

[0106] (Supplementary Note 4) (1) Y is -(CH2) n -, n is 0 or 1, (2)R 1 is, -H, -D, unsubstituted or R 2 substituted furan, thiophene, pyrrole, pyrazole, imidazole, oxazole, isoxazole, thiazole, isothiazole, pyran, thiopyran, pyridine, pyridazine or pyrimidine, (3)R 2 is selected from -OH, methyl, ethyl, n-propyl or isopropyl, A pyrazolecarbonylpiperazinone compound according to Supplementary Note 3, characterized by satisfying one or more of the conditions.

[0107] (Supplementary Note 5) R 1A pyrazolecarbonylpiperazinone compound according to any one of the appendices 1 to 4, characterized in that -H, -D, or any of the following substituents are selected. [ka] (The asterisk (*) indicates a connecting point.)

[0108] (Note 6) A pyrazolecarbonylpiperazinone compound according to any one of the appendices 1 to 4, characterized by having a structure represented by any one of formulas I-1 to I-13, or being a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, crystalline polymorph, isotope-labeled, isomer, or prodrug of a structure represented by any one of formulas I-1 to I-13. [ka]

[0109] (Note 7) A pyrazolecarbonylpiperazinone compound according to any one of the appendices 1 to 4, characterized by having the structure shown in formula II, or being a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, crystalline polymorph, isotope-labeled, isomer, or prodrug of the structure shown in formula II. [ka] (A ring, Y and R ring) 1 This is the same as being defined in any one of the appendices 1-4.

[0110] (Note 8) A pyrazolecarbonylpiperazinone compound according to any one of the appendices 1 to 4, characterized by having a structure represented by any one of formulas II-1 to II-13, or being a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, crystalline polymorph, isotope-labeled, isomer, or prodrug of a structure represented by any one of formulas II-1 to II-13. [ka]

[0111] (Note 9) A pharmaceutical composition comprising a pyrazolecarbonylpiperazinone compound described in any one of the appendices 1 to 8, and at least one pharmaceutically acceptable carrier.

[0112] (Note 10) Uses of a pyrazolecarbonylpiperazinone compound described in any one of Appendix 1 to 8, or a pharmaceutical composition described in Appendix 9, in the manufacture of a pharmaceutical for the treatment of one or more diseases of cancer, pulmonary hypertension, etc.

[0113] (Note 11) The use described in Appendix 10, characterized in that the cancer is one or more of ovarian cancer, breast cancer, glioma, and germ cell tumor.

[0114] (Note 12) Uses of any pyrazolecarbonylpiperazinone compound listed in any one of the appendices 1 to 8 as a BMPR1B inhibitor.

[0115] (Note 13) The use described in Appendix 12, characterized in that the concentration of the pyrazolecarbonylpiperazinone compound is 10 μM to 100 μM.

[0116] (Note 14) A method for inhibiting BMPR1B expression, comprising the step of contacting cells, biological tissues, or organoids containing BMPR1B with a pyrazolecarbonylpiperazinone compound described in any one of Appendix 1 to 8.

[0117] (Note 15) The method according to Appendix 14, characterized in that the concentration of the pyrazolecarbonylpiperazinone compound is 10 μM to 100 μM when performing the contact treatment.

[0118] (Note 16) A method for treating cancer or pulmonary hypertension, comprising the steps of administering to a patient a therapeutically effective amount of a pyrazolecarbonylpiperazinone compound described in any one of appendices 1 to 8, or administering to a patient a therapeutically effective amount of a pharmaceutical composition described in appendice 9.

[0119] (Note 17) The method according to Appendix 16, characterized in that the cancer is one or more of ovarian cancer, breast cancer, and glioma.

Claims

1. A pyrazolecarbonylpiperazinone compound having the structure represented by formula I, or a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, crystalline polymorph, isotope-labeled, isomer, or prodrug of the structure represented by formula I. 【Chemistry 1】 (In the formula, ring A is selected from one of the following structures, 【Chemistry 2】 Y is - (CH ２ ) ｎ - and n is 0, 1, 2 or 3, R １ is -H, -D, unsubstituted or R ２ substituted C １ to C ６ alkyl, C １ to C ６ alkoxy, C ６ to C １０ aryl or C ３ to C １０ selected from heteroaryl, R ２ -F, -Cl, -Br, -I, -OH, -COOH, C １ ~C ６ Alkyl or C １ ~C ６ Selected from alkoxy, (* indicates a connecting point.)

2. (1) Y is - (CH ２ ) ｎ - and n is 0, 1 or 2, (2) R １ is -H, -D, unsubstituted or R ２ Substitution C １ ~C ６ Alkyl, C ６ ~C １０ Aryl or C ３ ~C １０ Selected from heteroaryls, (3) Caution ２ is -OH or C １ ~C ６ Selected from alkyl groups, The pyrazolecarbonylpiperazinone compound according to claim 1, characterized in that it satisfies one or more conditions.

3. (1) Y is - (CH ２ ) ｎ - and n is 0 or 1, (2) R １ is -H, -D, unsubstituted or R ２ Substitution C ３ ~C ６ Selected from heteroaryls, (3) Caution ２ is -OH or C １ ~C ４ Selected from alkyl groups, The pyrazolecarbonylpiperazinone compound according to claim 2, characterized in that it satisfies one or more conditions.

4. (1) Y is - (CH ２ ) ｎ - and n is 0 or 1, (2) R １ is -H, -D, unsubstituted or R ２ Substitutions are selected from furan, thiophene, pyrrole, pyrazole, imidazole, oxazole, isoxazole, thiazole, isothiazole, pyran, thiopyran, pyridine, pyridazine, or pyrimidine. (3) Caution ２ is selected from -OH, methyl, ethyl, n-propyl or isopropyl. The pyrazolecarbonylpiperazinone compound according to claim 3, characterized in that it satisfies one or more conditions.

5. R １ A pyrazolecarbonylpiperazinone compound according to any one of claims 1 to 4, characterized in that -H, -D, or any of the following substituents are selected. 【Transformation 3】 (The asterisk "*" indicates a connecting point.)

6. A pyrazolecarbonylpiperazinone compound according to any one of claims 1 to 4, characterized in that it has a structure represented by any one of formulas I-1 to I-13, or is a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, crystalline polymorph, isotope-labeled, isomer, or prodrug of a structure represented by any one of formulas I-1 to I-13. 【Chemistry 4】

7. A pyrazolecarbonylpiperazinone compound according to any one of claims 1 to 4, characterized in that it has the structure represented by formula II, or is a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, crystalline polymorph, isotope-labeled, isomer, or prodrug of the structure represented by formula II. 【Transformation 5】 (A ring, Y and R １ (This is the same as defined in any one of claims 1 to 4.)

8. A pyrazolecarbonylpiperazinone compound according to any one of claims 1 to 4, characterized in that it has a structure represented by any one of formulas II-1 to II-13, or is a pharmaceutically acceptable salt, ester, amide, solvate, active metabolite, crystalline polymorph, isotope-labeled, isomer, or prodrug of a structure represented by any one of formulas II-1 to II-13. 【Transformation 6】

9. A pharmaceutical composition comprising a pyrazolecarbonylpiperazinone compound according to any one of claims 1 to 8 and at least one pharmaceutically acceptable carrier.

10. Uses of a pyrazolecarbonylpiperazinone compound according to any one of claims 1 to 8, or a pharmaceutical composition according to claim 9, in the manufacture of a pharmaceutical for treating one or more diseases of cancer, pulmonary hypertension.

11. The use according to claim 10, characterized in that the cancer is one or more of ovarian cancer, breast cancer, glioma, and germ cell tumor.

12. Use of the pyrazolecarbonylpiperazinone compound according to any one of claims 1 to 8 as a BMPR1B inhibitor.

13. The use according to claim 12, characterized in that the concentration of the pyrazolecarbonylpiperazinone compound is 10 μM to 100 μM.

14. A method for inhibiting the expression of BMPR1B, comprising the step of contacting cells, biological tissues, or organoids containing BMPR1B with a pyrazolecarbonylpiperazinone compound according to any one of claims 1 to 8.

15. The method according to claim 14, characterized in that the concentration of the pyrazolecarbonylpiperazinone compound is 10 μM to 100 μM when performing the contact treatment.

16. A method for treating cancer or pulmonary hypertension, comprising the steps of administering to a patient a therapeutically effective amount of a pyrazolecarbonylpiperazinone compound according to any one of claims 1 to 8, or administering to a patient a therapeutically effective amount of a pharmaceutical composition according to claim 9.

17. The method according to claim 16, characterized in that the cancer is one or more of ovarian cancer, breast cancer, and glioma.

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