Aminopyrrolecarboxamides, pharmaceutical compositions thereof and uses thereof

Abstract

The present application belongs to the field of medicine, and particularly relates to an aminopyrrole carboxamide compound, a pharmaceutical composition thereof and use thereof. The present application also relates to a pharmaceutical composition containing the compound as an active ingredient and use of the compound or the pharmaceutical composition for anti-HBV.

Description

Technical Field

[0001] This invention belongs to the pharmaceutical field, specifically relating to aminopyrrolamide compounds, their pharmaceutical compositions, and uses. Background Technology

[0002] Hepatitis B virus (HBV), which causes hepatitis B, remains one of the most concerning viruses globally. Currently, over 250 million people worldwide carry HBV, and these individuals have an increased risk of developing serious liver diseases, including cirrhosis, permanent liver scarring, liver failure, and cancer. The World Health Organization estimates that more than 780,000 people die from hepatitis B each year. According to data from the Hepatitis B Foundation, HBV accounts for 80% of factors inducing liver cancer, and the five-year survival rate for liver cancer patients is typically only 15%.

[0003] Despite the existence of effective vaccines, vaccine coverage remains unsatisfactory in HBV-endemic areas. Currently, two main classes of drugs are approved for the treatment of chronic hepatitis B. One class is interferon (IFN-α) and its pegylated form (Peg-IFN-α). IFN-α is an immunomodulator that nonspecifically induces the expression of interferon-stimulated genes (ISGs). These genes encode constitutive or secretory proteins with direct or indirect antiviral properties, thereby promoting the differentiation or activation of immune cells. After 48 weeks of subcutaneous IFN-α administration, the response rate in HBV patients is only 25%. Besides the low response rate, IFN-α also has other drawbacks, such as the need for injection, adverse reactions like severe flu-like symptoms, and contraindications in patients with compensated cirrhosis, severe hepatitis, autoimmune diseases, and mental illnesses. Another class of approved drugs are nucleoside analogs (NAs). Currently, five NAs have been approved: lamivudine (LMV), telbivudine (Ldt), adefovir dipivoxil (ADV), tenofovir (TFV), and entecavir (ETV). NAs reduce viral particles by directly inhibiting HBV polymerase activity, thereby disrupting the nucleocapsid-to-nucleus cycle of infected cells and theoretically reducing cccDNA expression. However, NA treatment fails to inhibit de novo cccDNA synthesis in newly infected cells. This indicates that residual viral particles during antiviral treatment can lead to new cell infection and cccDNA library reconstruction, potentially causing virological rebound if patients discontinue medication on their own. Therefore, functional cure of chronic hepatitis B virus infection is rare, and most patients require lifelong medication. However, long-term use of NAs can easily lead to viral mutations and drug resistance.

[0004] Given the limitations of current therapies, several drugs targeting the HBV replication cycle or enhancing the human immune response are under development and have entered clinical trials. Among these, the core protein in the HBV replication process is crucial for the packaging and reverse transcription of HBV pgRNA. Molecules developed targeting this protein are called core protein allosteric regulators or capsid assembly regulators (CpAMs). Based on their mechanisms of action, CpAMs can be divided into two categories: Class I CpAMs, represented by heteroaryl dihydropyrimidines (HAPs), enhance nucleocapsid formation kinetics, leading to misassembly of the nucleocapsid; and Class II CpAMs, represented by phenylacrylamide (PPAs) and sulfadiazine (SBAs) structures, accelerate nucleocapsid assembly and form morphologically normal nucleocapsids lacking viral pgRNA and HBV polymerase encapsulation.

[0005] Antiviral studies of nucleocapsid inhibitors BAY41-4109 (HAP) and JNJ-632 (SBA) with different mechanisms in human primary hepatocytes revealed that CpAMs not only inhibit HBV replication but also suppress HBV RNA transcription and antigen production, suggesting that CpAMs have a dual mechanism of action in inhibiting both early and late stages of viral infection. NVR3-778 (SBA), as an early-developed CpAM molecule, exhibited strong anti-HBV activity, showing positive results in EC50 in HepG2.2.15 cells. 50 The concentration was 0.4 μM. Similarly, it showed effective anti-HBV activity in a humanized liver mouse model infected with HBV. With advancements in science and technology, drug developers have now designed and screened molecular structures with in vitro anti-HBV activity at the nM level, such as GLP-26 and RG7907.

[0006] Currently, most anti-HBV drugs exhibit good in vitro anti-HBV activity. However, after entering clinical trials, some drugs have failed due to low efficacy and adverse reactions such as elevated ALT and relapse after drug withdrawal. Therefore, despite the availability of numerous drugs and methods for treating HBV, novel molecular structures and mechanisms of action for achieving a cure for hepatitis B remain urgently needed in the field of HBV treatment. Summary of the Invention

[0007] This invention provides aminopyrrolamide compounds or pharmaceutically acceptable salts thereof, the structures of which are shown below:

[0008] .

[0009] Pharmaceutical Composition

[0010] The present invention also provides a pharmaceutical composition comprising the aforementioned aminopyrrolamide compound or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable excipient.

[0011] Methods for preparing various pharmaceutical compositions containing a certain amount of active ingredient are known, or will be obvious to those skilled in the art according to the disclosure of the present invention. As described in REMINGTON'S PHARMACEUTICAL SCIENCES, Martin, EW, ed., Mack Publishing Company, 19th ed. (1995), the method for preparing the pharmaceutical composition includes incorporating appropriate pharmaceutical excipients, carriers, diluents, etc.

[0012] Medical Use

[0013] The present invention also provides the use of the aforementioned aminopyrrolamide compounds or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions thereof, in the preparation of medicaments for treating and / or preventing and / or reducing HBV infection.

[0014] The present invention also provides the use of the aforementioned aminopyrrolamide compounds or pharmaceutically acceptable salts thereof, or the pharmaceutical compositions thereof, in the preparation of medicaments for treating HBV infection.

[0015] In this invention, "treatment" generally refers to achieving the desired pharmacological and / or physiological effect. This effect may be preventative based on the complete or partial prevention of the disease or its symptoms; and / or therapeutic based on the partial or complete stabilization or cure of the disease and / or side effects resulting from the disease. As used herein, "treatment" encompasses any treatment of a patient's disease, including: (a) preventing the occurrence of a disease or symptoms in a patient who is susceptible to the disease or symptoms but has not yet been diagnosed with the disease; (b) suppressing the symptoms of the disease, i.e., preventing its progression; or (c) alleviating the symptoms of the disease, i.e., causing the disease or symptoms to regress.

[0016] The compounds of the present invention also include isomers of the compounds (e.g., stereoisomers, tautomers, etc.), solvates (e.g., hydrates), crystal forms (e.g., polymorphs, eutectics, etc.), prodrugs, metabolites, deuterated compounds, etc.

[0017] The term "stereoisomer" refers to isomers of the compounds of the present invention formed due to at least one asymmetric center. In compounds having one or more (e.g., 1, 2, 3, or 4) asymmetric centers, racemic mixtures, single enantiomers, diastereomer mixtures, and single diastereomers can be produced. The term "stereoisomer" includes conformational isomers and configurational isomers, wherein the configurational isomers primarily include cis-trans isomers and optical isomers. The compounds of the present invention can exist in stereoisomer form and therefore encompass all possible stereoisomeric forms, any combination thereof, or any mixture thereof. For example, single enantiomers, single diastereomers, or mixtures of more than one. When the compounds of the present invention contain an olefinic double bond, unless otherwise specified, they include cis and trans isomers, and any combination thereof.

[0018] The term "tautomer" refers to the functional group isomers of the compounds of the present invention that are produced by the rapid movement of an atom in the molecule between two positions, such as keto-enol tautomers, imine-enamine tautomers, etc. If the compounds of the present invention have tautomers, they can exist in the form of a single tautomer or a mixture thereof, preferably in the form of a more stable tautomer.

[0019] The term "solvate" refers to a substance formed by the association of the compound of the present invention with solvent molecules. The solvent may be an organic solvent (e.g., methanol, ethanol, propanol, acetonitrile, etc.), for example, the compound of the present invention can form an ethanolide with ethanol. The compound of the present invention can also form a hydrate with water. The amount of solvent may be stoichiometric or non-stoichiometric.

[0020] The term "crystal form" refers to the crystal structure of a substance. During crystallization, various factors alter the intramolecular or intermolecular bonding patterns, resulting in different arrangements of molecules or atoms in the crystal lattice, thus forming different crystal structures. The compounds of this invention can exist in one crystal structure or multiple crystal structures, i.e., they exhibit "polymorphism." The compounds of this invention can exist in different crystal forms.

[0021] The term "prodrug" refers to a derivative of the compound shown in this invention having specific substituents, which may have weak or no activity on its own, but after administration, is converted into the compound with specific substituents discovered in this invention under physiological conditions (e.g., through metabolism, solvation or other means) to produce the corresponding biological activity in vivo.

[0022] The term "metabolite" refers to the product obtained in vivo by the metabolism of the compound with specific substituents as discovered in this invention. A metabolite of a compound can be identified using techniques known in the art, and its activity can be characterized by experimental methods as described in this invention. Such products can be obtained by administering the compound through oxidation, reduction, hydrolysis, acylation, deacylation, esterification, defatting, enzymatic cleavage, etc. Accordingly, this invention includes metabolites of compounds, including metabolites produced by sufficiently exposing the compounds of this invention to mammals for a period of time.

[0023] The term "deuterated compound" refers to a compound of the present invention comprising at least one deuterium atom, specifically meaning that one or more hydrogen atoms in the compound of the present invention can be replaced or substituted by a deuterium atom. In some embodiments, the compound comprises two or more deuterium atoms. In some embodiments, the compound comprises 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or 12 deuterium atoms. Methods for synthesizing isotopes into organic compounds are known in the art.

[0024] The term "pharmaceutically acceptable salt" refers to the salt of the compounds of this invention, prepared by reacting a compound having specific substituents discovered in this invention with a relatively non-toxic acid or base. When the compounds of this invention contain relatively acidic functional groups, base addition salts can be obtained by contacting such compounds with a sufficient amount of base in a pure solution or a suitable inert solvent. When the compounds of this invention contain relatively basic functional groups, acid addition salts can be obtained by contacting such compounds with a sufficient amount of acid in a pure solution or a suitable inert solvent.

[0025] This invention also includes all pharmaceutically acceptable isotope labels that are identical to the compounds of this invention, except that one or more atoms are replaced by atoms having the same atomic number but with an atomic mass or mass number different from the dominant atomic mass or mass number in nature. Examples of isotopes suitable for inclusion in the compounds of this invention include, but are not limited to, isotopes of hydrogen (e.g., 2 H, 3 H, deuterium (D), tritium (T); carbon isotopes (e.g., H, deuterium (D), tritium (T)); 11 C 13 C and 14 C); isotopes of chlorine (e.g.) 37 Cl); isotopes of fluorine (e.g., Cl); 18 F); isotopes of iodine (e.g., F); 123 I and 125 I); nitrogen isotopes (e.g.) 13 N and 15 N); isotopes of oxygen (e.g., N); 15 O、 17 O and 18 O); isotopes of phosphorus (e.g., O); phosphorus isotopes ... 32 P); and isotopes of sulfur (e.g. 35 S).

[0026] The beneficial effects of this invention are:

[0027] The compounds of the present invention have excellent anti-HBV activity and can be used as drugs related to HBV prevention and / or improvement and / or treatment. Detailed Implementation

[0028] The present invention will be described in detail below with reference to examples, but this does not imply any adverse limitation on the invention. The compounds of the present invention can be prepared by various synthetic methods well known to those skilled in the art, including the specific embodiments listed below, embodiments formed by combining them with other chemical synthetic methods, and equivalent substitutions well known to those skilled in the art. Preferred embodiments include, but are not limited to, the embodiments of the present invention. It will be apparent to those skilled in the art that various changes and modifications can be made to the specific embodiments of the present invention without departing from the spirit and scope of the invention.

[0029] Unless otherwise specified, the raw materials and reagents used in the examples are all commercially available chemically pure materials.

[0030] Example 1: Preparation of Compound 2

[0031]

[0032] Step 1: Compound 2-1 (1.40 g, 9.14 mmol) (purchased from Jiangsu Aikon Biomedical R&D Co., Ltd.) was dissolved in tetrahydrofuran (50 mL). Lithium di(trimethylsilyl)amino (73.12 mL, 73.1 mmol) was added at room temperature and stirred for 2 hours. 3,4-Difluoroaniline (2.36 g, 18.28 mmol) (purchased from Jiangsu Aikon Biomedical R&D Co., Ltd.) was added at room temperature, and stirring continued for 18 hours. The reaction was quenched with water (50 mL). The mixture was extracted with ethyl acetate (50 mL), and the organic phase was washed with saturated brine (50 mL). The solution was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude compound. Compound 2-2 (760 mg) was obtained by column chromatography (PE / EA = 1:1), yielding 35.2%. MS (ESI): m / z 237.11 (M+H).

[0033] Step 2: Compound 2-2 (400 mg, 1.69 mmol) was dissolved in anhydrous dichloromethane (10 mL). Under ice bath conditions, (S)-2-oxo-2-((1,1,1-trifluoropropane-2-yl)amino)acetyl chloride (413.5 mg, 2.03 mmol) (as per Example 2 of CN116768776A for the synthesis of compound 2-e) and anhydrous aluminum trichloride (903.0 mg, 6.8 mmol) were added sequentially. The mixture was heated to room temperature and stirred for 5 hours. The reaction was quenched with ice. After adding water (30 mL), the mixture was extracted with dichloromethane (50 mL * 3). The organic phases were combined and washed with saturated brine (50 mL). The mixture was dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated. The residue was separated by column chromatography (PE / EA = 1:1) to obtain compound 2-3 (230 mg), with a yield of 33.7%. MS(ESI): m / z 404.06 (M+H).

[0034] 1H NMR (400 MHz, DMSO-d6) δ 12.26 (d, J = 3.9 Hz, 1H), 9.98 (s, 1H), 9.50 (d, J = 8.9 Hz, 1H), 7.94 – 7.84 (m, 2H), 7.51 – 7.33 (m, 2H), 4.79 –4.64 (m, 1H), 2.53 (s, 3H), 1.35 (d, J = 7.0 Hz, 3H).

[0035] Step 3: Compound 2-3 (150 mg, 0.37 mmol) was dissolved in ethyl acetate (5 mL). N-fluorobisbenzenesulfonamide (700.0 mg, 2.2 mmol) and 2,2,6,6-tetramethylpiperidine oxide (173.5 mg, 1.1 mmol) were added sequentially at room temperature. The mixture was heated to 50 °C and stirred for 18 hours. The reaction mixture was concentrated to obtain the crude compound. The residue was separated by column chromatography (PE / EA = 2:1) to give compound 2-4 (170 mg), yield 65.4%, as a yellow solid. MS (ESI): m / z 699.09 (M+H).

[0036] Step 4: Compounds 2-4 (170 mg, 0.24 mmol) were dissolved in 1,2-dichloroethane (7 mL), and trifluoromethanesulfonic acid (7 mL, 79.1 mmol) was added at room temperature. The mixture was heated to 100 °C and stirred for 15 minutes. After the reaction solution cooled to room temperature, it was poured into a saturated sodium bicarbonate solution (100 mL) to quench the reaction. The mixture was extracted with dichloromethane (50 mL), the organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated to obtain the crude compound. Column chromatography (PE / EA = 1:1) was used to separate compound 2 (25 mg), yield 24.6%. MS (ESI): m / z 419.22 (M+H).

[0037] 1 H NMR (400 MHz, Chloroform-d) δ 10.90 (s, 1H), 7.85 (d, J = 9.9 Hz,1H), 7.74 – 7.60 (m, 1H), 7.42 (s, 1H), 7.19 – 7.05 (m, 2H), 5.95 (s, 1H), 4.71 – 4.54 (m, 1H), 2.85 (s, 3H), 1.42 (d, J = 7.0 Hz, 3H).

[0038] Example 2: In vitro anti-HBV activity assay of HepG2.2.15 cells

[0039] 1. Compound dilution: First, the 20 μM compound stock solution was serially diluted 3-fold to 8 concentration points using DMSO to obtain the compound at serially diluted levels of 20 μM, 6.67 μM, 2.22 μM, 0.741 μM, 0.247 μM, 0.0823 μM, 0.0274 μM, and 0.00914 μM. Then, the serially diluted compounds were further diluted 200-fold using culture medium to obtain final compound concentrations of 100 nM, 33.3 nM, 11.1 nM, 3.70 nM, 1.23 nM, 0.412 nM, 0.137 nM, and 0.0457 nM.

[0040] 2. On the first day, HepG2.2.15 cells (Wuhan Institute of Virology, Chinese Academy of Sciences) (6×10⁶ cells) were... 4 Cells (per well) were seeded into 96-well plates and incubated overnight at 37°C with 5% CO2. Wells containing only cell culture medium and no cells were used as a culture medium control. On the second day, culture medium containing different final concentrations of the compound obtained in step 1 was added to the cell plate, with two replicates for each concentration. The final concentration of DMSO in the cell culture medium was 0.5%, and the final volume in each well was 200 μl. Wells containing only 0.5% DMSO without the compound were used as a DMSO control. On the fifth day, the same fresh culture medium was added to both the compound-treated and control wells. On the eighth day, the cell supernatant from the culture wells was collected, and a portion of the sample was used to detect the HBV DNA content. A plasmid containing the full-length D-type HBV DNA sequence was used as a standard for qPCR, with a standard range of 10. 7 -10 1 The HBV DNA content in all samples was determined by qPCR using copies / μl. After collecting the supernatant, Cell Titer-Glo reagent was added to the cell culture wells and incubated at room temperature for 10 minutes. Then, the chemiluminescence values ​​in all wells were read using a microplate reader to calculate cell proliferation activity.

[0041] 3. Data Analysis: Calculate the inhibition percentage using the following formula:

[0042] HBV DNA inhibition rate % = (1 - DNA copy number in compound sample / DNA copy number in DMSO control) × 100%.

[0043] Cell Viability % = (luminescence value in compound sample – luminescence value in culture medium control) / (luminescence value in DMSO control – luminescence value in culture medium control)) × 100%.

[0044] The HBV DNA inhibition rate (%) and compound concentration were fitted using a four-parameter method [log(agonist) vs. response -- variable slope] using GraphPad Prism software to obtain the compound's anti-HBV replication activity EC50. 50 Accurate value.

[0045] Using the above experimental methods, the anti-HBV activity and corresponding cell proliferation inhibition activity of the compounds of this invention are shown in Table 1 below.

[0046] Table 1. HepG2.2.15 cell HBV DNA inhibition

[0047]

[0048] CN114466837A discloses a compound. Meanwhile, its anti-HBV activity against HepG2.2.15 cells was also disclosed. 50 The molecular weight is 17.4 nM. This indicates that the compound of the present invention exhibits superior anti-HBV activity.

[0049] As shown in Table 1, the compounds of the present invention have excellent anti-HBV activity and corresponding cell proliferation inhibitory activity.

[0050] Industrial applicability

[0051] The compounds of this invention have excellent anti-HBV activity and corresponding cell proliferation inhibitory activity, and can be used as drugs for the treatment or prevention of diseases related to this effect.

[0052] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. An aminopyrrolamide compound, characterized in that: The structure of the compound is shown below: 。 2. A pharmaceutical composition, characterized in that: It comprises the aminopyrrolidamide compound of claim 1, and pharmaceutically acceptable excipients.

3. Use of the aminopyrrolidone compound of claim 1 or the pharmaceutical composition of claim 2 in the preparation of a medicament against HBV infection.

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