WO2017202207A1 - Substituted naphthalene ring compound, pharmaceutical composition, and application thereof - Google Patents

Abstract

Provided are a substituted naphthalene ring compound, and a pharmaceutical composition and an application thereof, the naphthalene ring compound being the compound shown in formula (I), or a polymorph, a pharmaceutically acceptable salt, a prodrug, a stereoisomer, an isotopic variant, a hydrate, or a solvate thereof. The present naphthalene ring compound can act as an inhibitor of hepatitis C virus, having good hepatitis C virus protein NS5B inhibitory activity and good pharmacodynamic/pharmacokinetic properties, and is therefore suitable for preparing medicine for the treatment of hepatitis C virus infection.

Description

Substituted naphthalene ring compound and pharmaceutical composition and application thereof Technical field

The invention belongs to the technical field of medicine, and in particular relates to a hepatitis C virus inhibitor, a pharmaceutical composition and application thereof.

Background technique

HCV (Hepatitis C Virus) is an RNA virus belonging to the genus Hepacivirus genus in the Flaviviridae family. The encapsulated HCV virion contains a positive-stranded RNA genome that encodes all known virus-specific proteins in a single uninterrupted open reading frame. The open reading frame comprises approximately 9500 nucleotides and encodes a single large polyprotein of approximately 3000 amino acids. Polyproteins include core proteins, envelope proteins E1 and E2, membrane-bound protein P7, and non-structural proteins NS2, NS3, NS4A, NS4B, NS5A, and NS5B.

In recent years, the number of hepatitis C patients has increased year by year. From 2006 to 2010, there were 70,681 cases, 92,378 cases, 108,446 cases, 131,849 cases and 163,174 cases.

Prior to 2011, the therapeutic drugs for HCV were interferon and ribavirin. These two drugs, by modulating the immune system or opening various antiviral genes in the cell, create an environment that is not conducive to viral replication, actually targeting host cells. These drugs have many side effects because they act on every cell in the body. New direct-acting antiviral drugs can directly target viruses, which has great advantages. Among the direct acting antiviral drugs, protease inhibitors, NS5A inhibitors and NS5B polymerase inhibitors are the three major classes of anti-HCV drugs under investigation. There were two HCV drugs approved by the US FDA in 2014, namely Jelly's Harvoni and Abbott's Viekira Pak, all of which are compound preparations.

Among them, Aibowei's Viekira Pak is mainly composed of ombitasvir, paritaprevir and dasabuvir, and can be used for the treatment of patients with chronic genotype 1 (GT1) hepatitis C virus infection, and also for HCV/HIV-1 co-infection (hepatitis C virus combined with humans). Immunodeficiency virus type 1) patients and liver transplant patients. Dasabuvir is an indispensable NS5B polymerase inhibitor in this combination.

In addition, sofosbuvir is currently the first medicine in the world that can completely cure hepatitis C in the short term. It takes the oral route directly to the lesion, and the method has simple side effects and is highly sought after by patients. Sofibuvir is produced by Gilead, USA, and is marketed in the United States in 2013. It has been clinically proven to be effective in treating hepatitis C, type 1, 2, 3, and 4, including liver transplantation, liver cancer, and HCV/HIV-1 co-infection. Clinical Trials. This breakthrough is for patients with hepatitis C worldwide Come the gospel. The US Gilead company has extended the authorization of the new hepatitis C drug Sofibuvir to Egypt last year. Egypt is the country with the highest incidence of hepatitis C in the world. Subsequently agreed to produce sofibru generic drugs in India. Up to now, a total of eight Indian pharmaceutical companies have obtained Gilead's authorization to sell Sophie Buwei to 91 developing countries around the world (China is excluded). Sophie Buwei is currently in a blank space in China. On the one hand, there is a large demand, and on the other hand, it is drug-free. In the face of such awkward dilemma, some large hospitals are trying to solve this problem through video remote overseas medical treatment. Some patients who can't wait, have to consider going abroad to see a doctor.

Therefore, there is still a need in the art to develop compounds having inhibitory activity or better pharmacodynamic properties against the hepatitis C virus protein NS5B.

Summary of the invention

In view of the above technical problems, the present invention discloses a substituted naphthalene ring compound and a pharmaceutical composition thereof as a hepatitis C virus inhibitor which has better hepatitis C virus protein NS5B inhibitory activity and/or has better pharmacodynamics/ Pharmacokinetic properties.

In this regard, the technical solution adopted by the present invention is:

A hepatitis C virus inhibitor, such as a naphthalene ring compound compound represented by formula (I), or a polymorphic form thereof, a pharmaceutically acceptable salt, a prodrug, a stereoisomer, an isotope variant, a hydrate or Solvent compound,

Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ , R ¹⁴ , R ¹⁵ , R ¹⁶ , R ¹⁷ , R ¹⁸ , R ¹⁹ , R ²⁰ , R ²¹ , R ²² , R ²³ , R ²⁴ , R ²⁵ are each independently hydrogen, deuterium, halogen or trifluoromethyl;

Additional conditions are R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ , R ¹⁴ , R ¹⁵ , R ¹⁶ And at least one of R ¹⁷ , R ¹⁸ , R ¹⁹ , R ²⁰ , R ²¹ , R ²² , R ²³ , R ²⁴ and R ²⁵ is deuterated or deuterated.

With this technical solution, the shape and volume of ruthenium in the drug molecule are substantially the same as hydrogen, if in the drug molecule Hydrogen is selectively replaced with hydrazine, and deuterated drugs generally retain their original biological activity and selectivity. At the same time, the inventors have confirmed through experiments that the binding of carbon-germanium bonds is more stable than the combination of carbon-hydrogen bonds, which can directly affect the absorption, distribution, metabolism and excretion of some drugs, thereby improving the efficacy, safety and tolerability of the drugs.

Preferably, the strontium isotope content of the cerium in the deuterated position is at least greater than the natural strontium isotope content (0.015%), preferably greater than 30%, more preferably greater than 50%, more preferably greater than 75%, and even more preferably greater than 95. %, more preferably greater than 99%.

Specifically, in the present invention, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ and R ¹⁴ , The cesium isotope content of each of the R ⁵ , R ¹⁶ , R ¹⁷ , R ¹⁸ , R ¹⁹ , R ²⁰ , R ²¹ , R ²² , R ²³ , R ²⁴ and R ²⁵ is at least 5%, preferably greater than 10 More preferably, more than 15%, more preferably more than 20%, more preferably more than 25%, more preferably more than 30%, more preferably more than 35%, more preferably more than 40%, more preferably more than 45% More preferably greater than 50%, more preferably greater than 55%, more preferably greater than 60%, more preferably greater than 65%, more preferably greater than 70%, more preferably greater than 75%, and even more preferably greater than 80%, More preferably greater than 85%, more preferably greater than 90%, more preferably greater than 95%, and even more preferably greater than 99%.

Preferably, R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ , R of the compound of formula (I) ^{And at least one} of R ¹⁵ , R ¹⁶ , R ¹⁷ , R ¹⁸ , R ¹⁹ , R ²⁰ , R ²¹ , R ²² , R ²³ , R ²⁴ and R ²⁵ contains at least one of R, more preferably two R氘, preferably three R 氘, more preferably four R 氘, more preferably five R 氘, more preferably six R 氘, more preferably seven R 氘, more preferably Eight R 氘, more preferably nine R 氘, more preferably ten R 氘, more preferably eleven R 氘, more preferably twelve R 氘, more preferably thirteen R contains 氘, more preferably fourteen R 氘, more preferably fifteen R 氘, more preferably sixteen R 氘, more preferably seventeen R 氘, more preferably eighteen R contains 氘, preferably 19 R contains 氘, more preferably 20 R contains 氘, more preferably twenty-one R contains 氘, more preferably twenty-two R contains 氘, more preferably two Thirteen R contain 氘, more preferably twenty-four R contains 氘, more preferably twenty-five R contains 氘.

As a further improvement of the present invention, R ¹ , R ² and R ³ are each independently hydrazine or hydrogen.

As a further improvement of the present invention, R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ and R ¹² are each independently hydrazine or hydrogen.

As a further improvement of the present invention, R ¹³ and R ¹⁴ are each independently hydrazine or hydrogen.

As a further improvement of the present invention, R ¹⁵ and R ¹⁶ are each independently hydrazine or hydrogen.

As a further improvement of the present invention, R ¹⁷ , R ¹⁸ , R ¹⁹ , R ²⁰ , R ²¹ and R ²² are each independently hydrazine or hydrogen.

As a further improvement of the present invention, R ²³ , R ²⁴ and R ²⁵ are each independently hydrazine or hydrogen.

In another preferred embodiment, R ¹ , R ² , and R ³ are deuterium.

In another preferred embodiment, R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ and R ¹² are 氘.

In another preferred embodiment, R ¹³ and R ¹⁴ are deuterium.

In another preferred embodiment, R ¹⁵ and R ¹⁶ are deuterium.

In another preferred embodiment, R ¹⁷ , R ¹⁸ , R ¹⁹ , R ²⁰ , R ²¹ and R ²² are 氘.

In another preferred embodiment, R ²³ , R ²⁴ , and R ²⁵ are 氘.

As a further improvement of the present invention, the compound is selected from the group consisting of the following compounds or a pharmaceutically acceptable salt thereof:

In another preferred embodiment, the compound does not include a non-deuterated compound.

The present invention also discloses a pharmaceutical composition comprising a pharmaceutically acceptable carrier and the hepatitis C virus inhibitor as described above, or a crystalline form, a pharmaceutically acceptable salt, a hydrate or a solvent thereof A pharmaceutical composition of a compound, stereoisomer, prodrug or isotopic variation.

As a further improvement of the present invention, the pharmaceutical composition further comprises other compounds such as interferon or interference a combination of ribavirin or other HCV inhibitors (eg, HCV polymerase inhibitors or HCV protease inhibitors).

The present invention also provides a method of preparing a pharmaceutical composition comprising the steps of: administering a pharmaceutically acceptable carrier to a hepatitis C virus inhibitor as described above, or a crystalline form thereof, a pharmaceutically acceptable salt, or a hydrate thereof Or the solvate is mixed to form a pharmaceutical composition.

The invention also discloses the use of a hepatitis C virus inhibitor as described above, characterized in that it is used for the preparation of a medicament for the treatment of hepatitis C virus infection.

The HCV includes a plurality of genotypes thereof and a plurality of gene subtypes, such as 1a, 1b, 2a, 2b, 3a, 3b, 4a, 5a, 6a.

As a further improvement of the present invention, the pharmaceutically acceptable carrier includes a glidant, a sweetener, a diluent, a preservative, a dye/colorant, a flavor enhancer, a surfactant, a wetting agent, a dispersant At least one of a disintegrant, a suspending agent, a stabilizer, an isotonic agent, a solvent or an emulsifier.

As a further improvement of the present invention, the pharmaceutical composition is a tablet, a pill, a capsule, a powder, a granule, an ointment, an emulsion, a suspension, a solution, a suppository, an injection, an inhalant, a gel, a microsphere or Aerosol.

Typical routes of administration of the pharmaceutical compositions of the invention include, but are not limited to, oral, rectal, transmucosal, enteral, or topical, transdermal, inhalation, parenteral, sublingual, intravaginal, intranasal, intraocular, intraperitoneal , intramuscular, subcutaneous, intravenous administration. Oral administration or injection administration is preferred.

The pharmaceutical composition of the present invention can be produced by a method known in the art, such as a conventional mixing method, a dissolution method, a granulation method, a sugar-coating method, a pulverization method, an emulsification method, a freeze-drying method, and the like.

It is to be understood that within the scope of the present invention, the various technical features of the present invention and the various technical features specifically described hereinafter (as in the embodiments) may be combined with each other to constitute a new or preferred technical solution. Due to space limitations, we will not repeat them here.

Herein, "halogen" means F, Cl, Br, and I unless otherwise specified. More preferably, the halogen atom is selected from the group consisting of F, Cl and Br.

As used herein, unless otherwise specified, "deuterated" means that one or more hydrogens in the compound or group are replaced by deuterium; deuteration may be monosubstituted, disubstituted, polysubstituted or fully substituted. The terms "one or more deuterated" are used interchangeably with "one or more deuterated".

As used herein, unless otherwise specified, "non-deuterated compound" means a compound containing a proportion of germanium atoms not higher than the natural helium isotope content (0.015%).

The invention also includes isotopically labeled compounds (also referred to as "isotopic variants"), equivalent to the original compounds disclosed herein. Examples of isotopes which may be listed as compounds of the present invention include hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine isotopes such as ² H, ³ H, ¹³ C, ¹⁴ C, ¹⁵ N, ¹⁷ O, ¹⁸ O, respectively. , ³¹ P, ³² P, ³⁵ S, ¹⁸ F and ³⁶ Cl. A compound of the formula (I) of the present invention containing the above isotope or other isotopic atom, or a polymorph, pharmaceutically acceptable salt, prodrug, stereoisomer, isotopic variation, hydrate or solvate thereof It is within the scope of the invention. Certain isotopically-labeled compounds of the present invention, such as the radioisotopes of ³ H and ¹⁴ C, are also among them, useful in tissue distribution experiments of drugs and substrates.氚, ie ³ H and carbon-14, ie ¹⁴ C, are easier to prepare and detect and are preferred in isotopes. In addition, heavier isotopic substitutions such as guanidine, or ² H, are preferred in certain therapies due to their good metabolic stability, such as increased half-life or reduced dosage in vivo, and therefore may be preferred in certain circumstances. Isotopically labeled compounds can be prepared in a conventional manner by substituting a readily available isotopically labeled reagent with a non-isotopic reagent using the protocol of the examples.

Pharmaceutically acceptable salts include inorganic and organic salts. A preferred class of salts are the salts of the compounds of the invention with acids. Suitable acids for forming salts include, but are not limited to, mineral acids such as hydrochloric acid, hydrobromic acid, hydrofluoric acid, sulfuric acid, nitric acid, phosphoric acid; formic acid, acetic acid, trifluoroacetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, Organic acids such as 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, naphthalenesulfonic acid; Amino acids such as amino acid, phenylalanine, aspartic acid, and glutamic acid. Another preferred class of salts are the salts of the compounds of the invention with bases, such as alkali metal salts (for example sodium or potassium salts), alkaline earth metal salts (for example magnesium or calcium salts), ammonium salts (for example lower alkanolammonium). Salts and other pharmaceutically acceptable amine salts), such as methylamine, ethylamine, propylamine, dimethylamine, trimethylamine, diethylamine, triethylamine, tert-butyl A base amine salt, an ethylenediamine salt, a hydroxyethylamine salt, a dihydroxyethylamine salt, a trihydroxyethylamine salt, and an amine salt formed of morpholine, piperazine, and lysine, respectively.

The term "polymorph" refers to a different arrangement of chemical drug molecules, generally expressed as the presence of a pharmaceutical material in a solid state. A drug may exist in a plurality of crystalline forms, and different crystal forms of the same drug may have different dissolution and absorption in the body, thereby affecting the dissolution and release of the formulation.

The term "solvate" refers to a complex of a compound of the invention that is coordinated to a solvent molecule to form a specific ratio. "Hydrate" means a complex formed by the coordination of a compound of the invention with water.

The term "prodrug" refers to a compound that is converted in vivo to an active form having its medical effect by, for example, hydrolysis in blood. A prodrug is any covalently bonded carrier which, when administered to a patient, releases the compound of the invention in vivo. Prodrugs are typically prepared by modifying functional groups that cleave prodrugs in vivo to produce maternal complexes Things. Prodrugs include, for example, a compound of the invention wherein a hydroxy, amino or thiol group is bonded to any group which, when administered to a patient, can be cleaved to form a hydroxy, amino or thiol group. Thus, representative examples of prodrugs include, but are not limited to, covalent derivatives of the compounds of the invention formed by the hydroxyl, amino or thiol functional groups thereof with acetic acid, formic acid or benzoic acid. Further, in the case of a carboxylic acid (-COOH), an ester such as a methyl ester, an ethyl ester or the like can be used. The ester itself may be active and/or may hydrolyze under conditions in humans. Suitable pharmaceutically acceptable in vivo hydrolysable esters include those which readily decompose in the human body to release the parent acid or its salt.

The compounds of the invention may include one or more asymmetric centers, and thus may exist in a variety of "stereoisomer" forms, for example, enantiomeric and/or diastereomeric forms. For example, the compounds of the invention may be in the form of individual enantiomers, diastereomers or geometric isomers (e.g., cis and trans isomers), or may be in the form of a mixture of stereoisomers, A racemic mixture and a mixture rich in one or more stereoisomers are included. The isomers can be separated from the mixture by methods known to those skilled in the art, including: chiral high pressure liquid chromatography (HPLC) and the formation and crystallization of a chiral salt; or preferred isomers can be passed Prepared by asymmetric synthesis.

The beneficial effects of the present invention compared to the prior art are: First, the compound of the present invention has excellent inhibitory effect on the hepatitis C virus protein NS5A. Second, by deuteration this technique changes the metabolism of the compound in the organism, giving the compound better pharmacokinetic parameter characteristics. In this case, the dosage can be changed and a long-acting preparation can be formed to improve the applicability. Third, replacing the hydrogen atom in the compound with hydrazine increases the drug concentration of the compound in the animal due to its strontium isotope effect, thereby improving the drug efficacy. Fourth, replacing the hydrogen atom in the compound with hydrazine can inhibit certain metabolites and improve the safety of the compound.

detailed description

The preparation of the structural compound of the formula (I) of the present invention is more specifically described below, but these specific methods do not constitute any limitation to the present invention. The compounds of the present invention may also be conveniently prepared by combining various synthetic methods described in the specification or known in the art, and such combinations are readily made by those skilled in the art to which the present invention pertains.

Example 1 Preparation of N-(6-(3-tert-butyl-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-d3-methoxy benzene (naphthalen-2-yl)methanesulfonamide (compound D-1)

The specific synthesis steps are as follows:

Step 1. Synthesis of N-(2-cyanophenyl)pyridine amide (Compound 3).

Pyridine-2-carboxylic acid (1.23 g, 10 mmol), 2-cyanoaniline (1.42 g, 12 mmol), 2-(7-oxobenzotriazole)-N,N,N', N'-tetramethylurea hexafluorophosphate (HATU, 7.6 g, 20 mmol) and N,N-diisopropylethylamine (DIPEA, 5.17 g, 40 mmol), dissolved in 50 mL of DMF, stirred at room temperature 4- After 6 hours, after TLC detection reaction, it was diluted with ethyl acetate, washed with 5% aqueous citric acid solution, water, sodium hydrogen carbonate solution and saturated brine, concentrated, and then recrystallized with a small amount of isopropanol to obtain 1.23 g of product. Rate: 55.15%.

Step 2. Synthesis of 6-hydroxy-2-naphthalene boronic acid (Compound 5).

6-bromo-2-naphthol (2.23 g, 10 mmol) was added to the reaction flask, and dissolved in 40 mL of anhydrous tetrahydrofuran. The temperature was lowered to -78 ° C, n-butyl lithium (10 ml, 25 mmol) was added dropwise under nitrogen atmosphere, and the reaction was carried out for 1 hour. Triisopropyl borate (2.82 g, 15 mmol) was added dropwise, and the reaction was carried out at room temperature for 0.5 hour, and then the reaction was continued to room temperature. After 0.5 hours, after the TLC reaction was completed, the pH was adjusted to be acidic by adding 1 M of dilute hydrochloric acid, and the organic phase was separated. The aqueous phase was extracted twice with diethyl ether. The organic phase was combined, washed with brine, concentrated, and purified by column chromatography. g product, yield 60.2%.

Step 3. Synthesis of 2-tert-butyl-4,6-diiodophenol (Compound 7).

To the reaction flask was added 2-tert-butylphenol (5.0 g, 33.3 mmol), sodium hydroxide (1.6 g, 40 mmol), dissolved in 65 mL of methanol, cooled to -2 ° C in an ice salt bath, and added sodium iodide (3.75 g). , 25mmol), then add 10% aqueous sodium hypochlorite solution (11.25mL), during which the internal temperature is controlled to not exceed 2 ° C, repeat this operation 3 times, and finally add a small amount of sodium hypochlorite to change the color of the reaction liquid from yellowish green to brown, and then rise to room temperature. The reaction was continued for 1 hour, and the reaction was completed by TLC. The mixture was cooled to 0 ° C, and a 16.7% aqueous sodium thiosulfate solution (25 mL) was added dropwise. After addition, concentrated hydrochloric acid was added dropwise to adjust the pH to 3.0, and the mixture was filtered, washed with water and dried in vacuo to give the product 12.1 g, yield: 91% .

Step 4. Synthesis of 1-tert-butyl-3,5-diiodo-2-d3-methoxybenzene (Compound 8).

Compound 7 (4.2 g, 10.44 mmol) was added to the reaction flask, dissolved in 25 mL of acetone, and then, then, then, and then, After the reaction was completed by TLC, the solvent was concentrated, and a small amount of n-heptane and water were added, and the organic phase was separated, washed with saturated brine, concentrated, and purified by silica gel column chromatography to give a bright yellow oily liquid 4.05 g, yield: 92.6%

Step 5. Synthesis of 1-(3-tert-butyl-5-iodo-4-d3-methoxyphenyl)pyrimidine-2,4(1H,3H)-dione (Compound 9).

Compound 8 (5.19 g, 14.33 mmol), uracil (1.93 g, 17.2 mmol), compound 3 (0.64 g, 2.87 mmol) and potassium phosphate (6.4 g, 30.1 mmol) were added to the reaction flask, and dissolved in 10 mL of DMSO. Nitrogen was bubbled for 0.5 hour, cuprous iodide (0.27 g, 1.43 mmol) was added, and bubbling was continued for 10 minutes. The reaction was heated to 60 ° C for 18 hours under nitrogen atmosphere. After the TLC reaction was completed, a small amount of ethyl acetate was added and diluted. The mixture was washed with saturated aqueous ammonium chloride and brine.

Step 6. 1-(3-tert-Butyl-5-(6-hydroxynaphthalen-2-yl)-4-d3-methoxyphenyl)pyrimidine-2,4(1H,3H)-dione (compound) 10) Synthesis.

Compound 9 (1.7 g, 4.26 mmol) and compound 5 (0.84 g, 4.47 mmol) were added to the reaction flask, dissolved in 30 mL of tetrahydrofuran, and nitrogen was bubbled for 0.5 hour, and potassium phosphate (1.81 g, 8.52 mmol) in 10 mL aqueous solution was added. Further, Pd ₂ (dba) ₃ (39 mg, 0.043 mmol) and ligand Xantphos (113.4 mg, 0.196 mmol) were added, and nitrogen gas was bubbled for 0.5 hour, and the temperature was raised to 65 ° C overnight. After the TLC reaction was completed, a small amount of saturated brine was added. The organic phase was separated, concentrated and purified by silica gel column chromatography to yield 1.15 g of product.

Step 7. 6-(3-tert-Butyl-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-d3-methoxyphenyl)naphthalene Synthesis of 2-yl-1,1,2,2,3,3,4,4,4-nonafluorobutane-1-sulfonate (Compound 11).

Compound 10 (1.15 g, 2.76 mmol) and potassium carbonate (0.57 g, 4.14 mmol) were added to the reaction flask, dissolved in 30 mL of DMF, and perfluorobutanesulfonyl fluoride (0.92 g, 3.04 mmol) was added under nitrogen and stirred. After the reaction was completed by TLC, the reaction was quenched with EtOAc (EtOAc) EtOAc (EtOAc) : 69.6%.

Step 8. N-(6-(3-tert-Butyl-5-(2,4-dioxo-3,4-dihydropyrimidin-1(2H)-yl)-2-d3-methoxybenzene Synthesis of naphthyl-2-yl)methanesulfonamide (Compound D-1).

Pd ₂ (dba) ₃ (1.65 mg, 0.0018 mmol), ligand Xantphos (2.54 mg, 0.0044 mmol) and potassium phosphate (108.26 mg, 0.51 mmol) were added to the reaction flask, and 5 mL of tert-amyl alcohol was added under nitrogen to heat. The reaction was carried out at 80 ° C for 0.5 hours. In a separate reaction flask, compound 11 (321 mg, 0.46 mmol) and methylsulfonamide (66.6 mg, 0.7 mmol) were added, 10 mL of tert-amyl alcohol was added, and the mixture was heated to 60 ° C under nitrogen for 0.5 hour, and the solution was transferred to the above. The reaction flask was heated to 85 ° C for 14 hours. After the reaction was completed by TLC, the solvent was evaporated, evaporated, evaporated, evaporated, evaporated. LC-MS (APCI): m/z =495. 1H NMR (300MHz, DMSO-d ₆ ) δ 11.42 (d, J = 1.9 Hz, 1H), 10.05 (s, 1H), 8.03 (s, 1H), 7.96 (t, J = 8.3 Hz, 2H), 7.80 (d, J = 7.9 Hz, 1H), 7.75 - 7.67 (m, 2H), 7.42 (dd, J = 8.9, 2.1 Hz, 1H), 7.37 (d, J = 2.6 Hz, 1H), 7.31 (d) , J = 2.7 Hz, 1H), 5.65 (dd, J = 7.9, 2.2 Hz, 1H), 3.08 (s, 3H), 1.42 (s, 9H).

Example 2

The compounds of the above examples were evaluated for biological activity.

To verify the effect of the compounds described herein on HCV, the inventors used the HCV Replicon System as an evaluation model. Since its first report in Science in 1999, the HCV replication system has become one of the most important tools for studying HCV RNA replication, pathogenicity and viral persistence, for example, the use of replicons has successfully demonstrated the 5' required for HCV RNA replication. - NCR minimum region, and the HCV replication subsystem has been successfully used as an evaluation model for antiviral drugs. The inventors of the present invention verified according to the methods described in Science, 1999, 285 (5424), 110-3, and J. Virol, 2003, 77(5), 3007-19.

(1) Detection of compound anti-HCV 1a and 1b genotype replicon activity

Detection of Compound Hepatitis C Virus Genotype 1a Using HCV-1a and HCV-1b Stably Transfected Replicon Cells And the inhibitory activity of the 1b replicon. This experiment will use Dasabuvir as a positive control compound.

Step 1: The compound was diluted 1:3 in 8 series points, double-replicated, and added to a 96-well plate. The DMSO was set to no compound control. The final concentration of DMSO in the cell culture was 0.5%.

Step 2: HCV-1a and 1b cells were separately suspended in a culture medium containing 10% FBS, and seeded into a 96-well plate containing the compound at a density of 8,000 cells per well. The cells were cultured for 3 days at 5% CO ₂ at 37 °C.

Step 3: The cytotoxicity of the compound against GT1b replicon was determined using CellTiter-Fluor (Promega).

Step 4: Detection of luciferase assay by Bright-Glo (Promega) for anti-hepatitis C virus activity.

Step Five: using GraphPad Prism data analysis software, the curve fitting and EC ₅₀ values _were calculated and the ₅₀ value CC.

Table 1 Comparison of anti-HCV genotype replicon activity of Example D-1 and control Dasabuvir

Numbering	HCV GT1a EC 50 (nM)	HCV GT1b EC 50 (nM)	HCV CC 50 (nM)
Dasabuvir	7.61	<2.32	>5000
D-1	<2.32	<2.32	>5000

The experimental results show that the compound of the present invention can inhibit multiple genotypes of HCV and exert superior anti-HCV effects by inhibiting the mechanism of HCV NS5B protein.

(2) Metabolic stability evaluation

Microsomal experiments: human liver microsomes: 0.5 mg/mL, Xenotech; rat liver microsomes: 0.5 mg/mL, Xenotech; coenzyme (NADPH/NADH): 1 mM, Sigma Life Science; magnesium chloride: 5 mM, 100 mM phosphate buffer Agent (pH 7.4).

Preparation of the stock solution: A powder of the compound of the compound example was accurately weighed and dissolved in DMSO to 5 mM.

Preparation of phosphate buffer (100 mM, pH 7.4): Mix 150 mL of pre-formed 0.5 M potassium dihydrogen phosphate and 700 mL of 0.5 M potassium dihydrogen phosphate solution, and adjust the mixture with 0.5 M potassium dihydrogen phosphate solution. The pH was adjusted to 7.4, diluted 5 times with ultrapure water before use, and magnesium chloride was added to obtain a phosphate buffer (100 mM) containing 100 mM potassium phosphate, 3.3 mM magnesium chloride, and a pH of 7.4.

A solution of NADPH regeneration system (containing 6.5 mM NADP, 16.5 mM G-6-P, 3 U/mL G-6-P D, 3.3 mM magnesium chloride) was prepared and placed on wet ice before use.

Formulation stop solution: acetonitrile solution containing 50 ng/mL propranolol hydrochloride and 200 ng/mL tolbutamide (internal standard). Take 25057.5 μL of phosphate buffer (pH 7.4) into a 50 mL centrifuge tube, add 812.5 μL of human liver microsomes, and mix to obtain a liver microsome dilution with a protein concentration of 0.625 mg/mL. Take 25057.5μL phosphate The buffer (pH 7.4) was added to a 50 mL centrifuge tube, and 812.5 μL of SD rat liver microsomes were added and mixed to obtain a liver microsome dilution having a protein concentration of 0.625 mg/mL.

Incubation of the sample: The stock solution of the corresponding compound was diluted to 0.25 mM with an aqueous solution containing 70% acetonitrile as a working solution, and was used. 398 μL of human liver microsomes or rat liver microsome dilutions were added to 96-well incubation plates (N=2), and 2 μL of 0.25 mM working solution was added and mixed.

Determination of metabolic stability: 300 μL of pre-cooled stop solution was added to each well of a 96-well deep well plate and placed on ice as a stop plate. The 96-well incubation plate and the NADPH regeneration system were placed in a 37 ° C water bath, shaken at 100 rpm, and pre-incubated for 5 min. 80 μL of the incubation solution was taken from each well of the incubation plate, added to the stopper plate, and mixed, and 20 μL of the NADPH regeneration system solution was added as a sample of 0 min. Then, 80 μL of the NADPH regeneration system solution was added to each well of the incubation plate to start the reaction and start timing. The corresponding compound had a reaction concentration of 1 μM and a protein concentration of 0.5 mg/mL. 100 μL of the reaction solution was taken at 10, 30, and 90 min, respectively, and added to the stopper, and the reaction was terminated by vortexing for 3 min. The plate was centrifuged at 5000 x g for 10 min at 4 °C. 100 μL of the supernatant was taken into a 96-well plate to which 100 μL of distilled water was previously added, mixed, and sample analysis was performed by LC-MS/MS.

Data analysis: The peak area of the corresponding compound and the internal standard was detected by LC-MS/MS system, and the ratio of the peak area of the compound to the internal standard was calculated. The slope is measured by the natural logarithm of the percentage of the remaining amount of the compound versus time, and t _1/2 and CL _{int are} calculated according to the following formula, where V/M is equal to 1/protein concentration.

The metabolic stability of human and rat liver microsomes was evaluated by simultaneously testing the compounds of the present invention and their compounds without deuteration. The half-life and liver intrinsic clearance as indicators of metabolic stability are shown in Table 2. The undeuterated compound Dasabuvir was used as a control sample in Table 2. As shown in Table 2, the compound of the present invention can significantly improve metabolic stability by comparison with the undeuterated compound Dasabuvir, and is thus more suitable as a hepatitis C virus inhibitor.

Table 2 Comparison of metabolic stability of Example D-1 and Dasabuvir control

(3) Rat pharmacokinetic experiments

OBJECTIVE: To investigate the pharmacokinetic behavior of the compounds of the present invention after administration of Dasabuvir and Compound D-1 to rats.

Experimental animals:

Type and strain: SD rat grade: SPF grade

Gender and quantity: male, 6

Weight range: 180 ~ 220g (actual weight range is 187 ~ 197g)

Source: Shanghai Xipuer Bikai Experimental Animal Co., Ltd.

Experimental and animal certificate number: SCXK (Shanghai) 2013-0016

experiment procedure:

Before the blood sample was collected, 20 L of 2M sodium fluoride solution (esterase inhibitor) was previously added to the EDTA-K2 anticoagulation tube, dried in an 80 degree oven, and stored in a 4 degree refrigerator.

Rats, males, weighing 187-197 g, were randomly divided into 2 groups. They were fasted overnight in the afternoon before the experiment but were free to drink water. Food was given 4 h after administration. Group A was given 3 mg/kg of Dasabuvir, and group B was given 3 mg/kg of compound of formula T-1. Blood was taken from the orbital vein of rats 100-200 L at 15 min, 30 min, 1, 2, 3, 5, 8 and 10 h after administration. Left and right, placed in an EDTA-K2 anticoagulated 0.5mL Eppendorf tube, mix immediately, after anticoagulation, gently invert the tube as soon as possible 5-6 times, the blood is taken and placed in the ice box, Blood samples were centrifuged at 4000 rpm, 10 min, and 4 ° C for 30 min, and all plasma was collected and stored at -20 ° C immediately. Plasma concentrations in plasma at each time point were determined after sample collection at all time points.

According to the average blood drug concentration-time data obtained after the above, the male SD rats were treated with Winnonin software according to the non-compartmental statistical moment theory to give Dasabuvir (3 mg/kg) and the compound of the example (3 mg/kg). The relevant pharmacokinetic parameters are shown in Table 3.

Table 3 Comparison of rat pharmacokinetic experiments with Example D-1 and Dasabuvir control

The experimental results show that the present inventors have found that the compound of the present invention has superior activity to Dasabuvir and has excellent pharmacokinetic properties as compared with Dasabuvir, and thus is more suitable as a compound for inhibiting the hepatitis C virus protein NS5B, and is suitable for Preparation of a medicament for treating hepatitis C virus infection.

It is to be understood that the examples are merely illustrative of the invention and are not intended to limit the scope of the invention, and the experimental methods in which the specific conditions are not indicated in the examples, usually in accordance with conventional conditions or in accordance with the conditions suggested by the manufacturer. Parts and percentages are parts by weight and percentage by weight unless otherwise stated.

The above is a further detailed description of the present invention in connection with the specific preferred embodiments, and the specific embodiments of the present invention are not limited to the description. It will be apparent to those skilled in the art that the present invention may be made without departing from the spirit and scope of the invention.

Claims (10)

1. A hepatitis C virus inhibitor characterized by a naphthalene ring compound represented by formula (I), or a polymorph thereof, a pharmaceutically acceptable salt, a prodrug, a stereoisomer, an isotope variant , hydrate or solvent compound,

   

   Wherein R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ , R ¹⁴ , R ¹⁵ , R ¹⁶ , R ¹⁷ , R ¹⁸ , R ¹⁹ , R ²⁰ , R ²¹ , R ²² , R ²³ , R ²⁴ , R ²⁵ are each independently hydrogen, deuterium, halogen or trifluoromethyl;

   Additional conditions are R ¹ , R ² , R ³ , R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ , R ¹² , R ¹³ , R ¹⁴ , R ¹⁵ , R ¹⁶ And at least one of R ¹⁷ , R ¹⁸ , R ¹⁹ , R ²⁰ , R ²¹ , R ²² , R ²³ , R ²⁴ and R ²⁵ is deuterated or deuterated.
2. The hepatitis C virus inhibitor according to claim 1, wherein each of R ¹ , R ² and R ^{3 is} independently hydrazine or hydrogen.
3. The hepatitis C virus inhibitor according to claim 1, wherein R ⁴ , R ⁵ , R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ and R ¹² are each independently anthracene or hydrogen.
4. The hepatitis C virus inhibitor according to claim 1, wherein each of R ¹³ and R ^{14 is} independently hydrazine or hydrogen.
5. The hepatitis C virus inhibitor according to claim 1, wherein each of R ¹⁵ and R ^{16 is} independently hydrazine or hydrogen.
6. The hepatitis C virus inhibitor according to claim 1, wherein each of R ¹⁷ , R ¹⁸ , R ¹⁹ , R ²⁰ , R ²¹ and R ^{22 is} independently hydrazine or hydrogen.
7. The hepatitis C virus inhibitor according to claim 1, wherein each of R ²³ , R ²⁴ and R ^{25 is} independently hydrazine or hydrogen.
8. The hepatitis C virus inhibitor according to claim 1, wherein the compound is selected from the group consisting of the following compounds or a pharmaceutically acceptable salt thereof:

   

   
9. A pharmaceutical composition comprising a pharmaceutically acceptable carrier and a hepatitis C virus inhibitor according to any one of claims 1 to 7, or a crystalline form thereof, a pharmaceutically acceptable salt, A pharmaceutical composition of a hydrate or solvate, stereoisomer, prodrug or isotopic variation.
10. Use of a hepatitis C virus inhibitor according to any one of claims 1 to 7, which is for use in the preparation of a medicament for treating hepatitis C virus infection.