HK1152705B - Dihydropyrimidine compounds and use thereof for the treatment of viral diseases - Google Patents

Description

Technical Field

The present invention relates to a dihydropyrimidine compound of formula (I) and a process for preparing the same, a pharmaceutical composition comprising the same, as well as use of said compound or a pharmaceutically acceptable salt or a hydrate thereof as a medicament, in particular is a medicament for use in a method for the treatment and prevention of type B hepatitis (HB).

Background Art

Chronic type B hepatitis is a serious infectious disease caused by hepatitis B virus (HBV) and prevalent throughout the world, and is closely relevant to the occurrence of hepatic cirrhosis and liver cancer. China is a high-risk area of HB. The results of seroepidemiological survey of viral hepatitis in China from 1992 to 1995 indicated that the persons carrying the surface antigen (HBsAg) of hepatitis B virus in China accounted for 9.7% of the population, and it was estimated hat about 130 millions persons were HBV carriers. A study on the epidemiological situation of viral hepatitis in China demonstrated that the annual reported incidence of HB was increased from 21.9/100 thousands in 1990 to 53.3/100 thousands in 2003, showing an obvious ascending tendency (Wang Xiaojun, Zhang Rongzhen, Hu Yuansheng, et al, Monitoring of Diseases, 2004, 19(8): 290-292). Chronic HB not only seriously affects human health, but also imposes heavy economic burden on a family and society. Therefore, Chronic HB has become one of significant public health concerns in China.

The drugs useful for treating chronic HB mainly include two types - immunomodulator and nucleoside inhibitor of DNA polymerase (Loomba R., Liang T. J., Antivir. Ther. ,2006, 11(1): 1-15). The former includes: interferon-α2b (IFN-α2b, Intron A®) and PEGylated interferon-α2a (peg-IFN-α2a, Pegasys®); the latter includes: Lamivudine (Epivir-HBV®), Adefovir Dipivoxil (Hepsera®) and Entecavir(Baraclude®). Comparatively, the number and type of drugs available for treating HB in clinic are still limited. Thus, it is extremely important to continuously research and develop new safe and effective anti-virus drugs, in particular those having a completely new mechanism of action.

Deres et al. reported heteroaromatic ring substituted dihydropyrimidine (HAP) compounds represented by Bay41-4109 and Bay39-5493, which compounds could act to suppress the replication of HBV by preventing the normal formation of nucleocapsid. The preclinical data demonstrated that Bay41-4109 had relatively good drug metabolism parameters (Deres K., Schroder C. H., Paessens A., et al. Science, 2003, 299 (5608) :893-896). A study on its mechanism of action showed that HAP changed the angle between dimers forming nucleocapsid, resulting in the formation of unstable swelled nucleocapsid, and accelerating the degradation of core protein by interacting with 113-143 amino acid residues of core protein (Hacker H. J., Deres K., Mildenberger M., et al., Biochem. Pharmacol. 2003, 66(12):2273-2279). WO99/54326 and WO99/54329 disclosed 2-pyridyl substituted and 2-thiazolyl substituted dihydropyrimidine compounds, respectively.

Contents of the Invention

The present invention relates to a new dihydropyrimidine compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein

• R1 represents hydrogen or acetyl,
• R3 for occurrence once or more each represents fluoro, chloro, bromo, hydroxyl, nitro, methoxy, or methyl, and
• R4 represents a group represented by formula -XR12

wherein

• X represents oxygen, and
• R12 represents a straight or branched alkyl having up to 3 carbon atoms.

The compound of the present invention may include a compound of formula (I) and its isomer Ia and a mixture thereof. IfR¹ is hydrogen, isomers I and Ia exist in tautomeric equilibrium:

The compound of the present invention may exist in a stereomeric form, and said stereomeric forms are in enantiomeric or diastereoisomeric relationship. The present invention relates to these enantiomers or diastereoisomers or a mixture thereof. Like a diastereoisomer, a racemate may be resolved into a single stereomer by using known methods.

The compound of the present invention may also be in the form of a salt. Its pharmaceutically acceptable salt is preferred.

The pharmaceutically acceptable salt includes, but not limited to, salts formed with various inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, phosphorous acid, hydrobromic acid and nitric acid, and salts formed with various organic acids such as maleic acid, fumaric acid, malic acid, succinic acid, tartaric acid, citric acid, acetic acid, lactic acid, benzoic acid, methamesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, palmic acid and etc.

The pharmaceutically acceptable salt further includes, but not limited to, metal salts of the compound of the present invention, such as sodium salt, potassium salt, magnesium salt or calcium salt, or ammonium salts formed with ammonia or an organic amine such as ethylamine, diethylamine, triethylamine, diethanolamine, triethanolamine, dicyclohexylamine, dimethylaminoethanol, arginine, lysine, ethylenediamine, or 2-phenylethylamine, and the like.

Some compounds in the present invention may be crystallized or recrystallized by using water or various organic solvents, and in this case, various solvates may be formed. The present invention includes those stoichiometric solvates, hydrates, and also compounds comprising variable amount of water formed when prepared using lyophilisation.

The present invention also relates to the compounds:

• Ethyl 2-(2,4,6-trifluorophenyl)-4-(2-chloro-4-fluorophenyl)-6-methyl-1,4-dihydro-pyrimidin-5-carboxylate;
• Methyl 2-(2,4,6-trifluorophenyl)-4-(2-chloro-4-fluorophenyl)-6-methyl-1,4-dihydro-pyrimidin-5-carboxylate;
• Ethyl 4-R-(2-chloro-4-fluorophcnyl)-2-(2,4,6-trifluorophenyl)-6-methyl-1,4-dihydro-pyrimidin-5-carboxylate,

or its pharmaceutically acceptable salt.

The compound in the present invention may be prepared by the following process:

1. A) under the condition of adding a base or acid or not, and in a suitable inert solvent, reacting an amidine of formula (II) or a salt thereof wherein R is defined as above, with an aldehyde of formula (III) wherein R³ is defined as above, and a compound of formula (IV)         CH₃CO-CH₂-CO-R⁴     (IV) wherein R⁴ is defined as above, or
2. B) under the condition of adding a base or acid or not, at a temperature of 20-150°C, and in a suitable inert solvent, reacting a compound of formula (V) or (VI) wherein R³, R⁴ are as defined above, with the compound of formula (II), or
3. C) reacting an aldehyde of formula (III) wherein R³ is defined as above, with a compound of formula (VII) wherein R⁴ is defined as above, and the amidine of formula (II), or
4. D) in the presence of an ammonium salt, reacting the aldehyde of formula (III) with the compound of formula (IV) and an iminoether of formula (VIII) wherein R is defined as above, and R' is C₁-C₄ alkyl.

The compound, wherein R¹ is acetyl, can be prepared by reacting the compound obtained according to the above process, with acetyl chloride or acetic anhydride in an inert solvent in the presence of an inorganic base or an organic base at a temperature of 20-150°C.

The process of the present invention is exemplified by using the following reaction schemes:

As to all of the reaction schemes A, B and C, suitable solvents are any inert organic solvents. These solvents preferably include alcohols such as ethanol, methanol, isopropanol, ethers such as dioxane, ethyl ether, THF, ethylene glycol monomethyl ether, ethylene glycol dimethyl ether or glacial acetic acid, dimethylformamide, dimethyl sulfoxide, acetonitrile, pyridine and hexamethylphosphamide.

The reaction temperature may vary within a relatively broad range. Usually, the reaction is carried out at a temperature of 20-150°C, but it is preferably carried out at the boiling point of the solvent.

The reaction may be carried out under normal pressure, or under an elevated pressure. Usually, the reaction is carried out under normal pressure.

The reaction may be carried out under the condition of adding a base or acid or not, but it is preferably carried out in the presence of a relatively weak acid such as acetic acid or formic acid.

The amidine of formula (II) as the starting substance is known in certain cases, or may be prepared from the corresponding cyano compound according to known methods described in literatures (cf. Diana, G. D., Yarinsky, A., Zalay, E. S., et al. J. Med. Chem. 1969, 12(9):791-793; Garigipati, R. S. Tetrahedron. Lett. 1990, 31(14):1969-1972; Boere, R. J., Oakley, R. T., Read, R. V. J Organometal. Chem. 1987, 331:161-167; Judkins, B. D., Allen, D. G., Cook, T. A. Synth. Commun. 1996, 26(23):4351-4367; Tommasi, R. A., Macchia, W. M., Parker, D. T. Tetrahedron. Lett. 1998, 39:5947-5950).

The aldehyde of formula (III) as the starting substance is known, or may be prepared according to known methods described in literatures (cf. T. D. Harris and G. P. Roth, J. Org. Chem. 1979, 44, 146; DE 2165260, July 1972 ; DE 2401665, July 1974 ; Mijano et. al. CA 1963, 59, 13929c; E. Adler ,H. D. Bccker, Chem. Scand. 1961, 15, 849; E. P. Papadopoulos, M. Mardin, Ch. Issidoridis, J. Org. Chem. Soc. 1956, 78, 2543).

The β-keto carboxylate of formula (IV) as the starting substance is known, or may be prepared according to known methods described in literatures (cf. D. Berman: Reaction of Diketene with Alcohol, Phenol and Thiol, Houben-Weyl, Methods of Organic Chemistry, Vol. VII, pages 230-(1968); Y. Oikawa, K. sugano, O. Yonemitsu, J. Org. Chem. 1978, 43, 2087).

The stibylene-β-ketone ester of formula (V) or formula (VI) as the starting substance may be prepared according to known methods described in literatures (cf. G Jones, "Knoevenage Condensation", Organic Reaction, Vol. XV, pages 204- (1967)).

The enamino carboxylate of formula (VII) and iminoether of formula (VIII) as the starting substances are known, or may be prepared according to known methods described in literatures (cf. S. A. Glckman, A. C. Cope, J. Am. Chem. Soc. 1945, 67, 1017).

The compound of formula (I)or claim 2 in the present invention may be synthesized individually according to conventional method, or synthesized by libraries (each library includes at least two, or 5-1,000, optimally 10-100 compounds) according to mix-split method or parallel synthetic method of combinatorial chemistry, i.e., the compound may be synthesized according to a liquid phase method or a solid phase method.

As for more detailed information about the preparation of the compound of formula (1) or claim 2, please see the examples.

The antiviral effect of the compound of the present invention is determined according to the methods described by Sells, et al (M. A. Sells, M. L. Chen, G. Proc. Natl. Acad. Sci. 1987, 84, 1005-1009) and Korba, et al (B. E. Korba, J. L. Gerin Antiviral Research 1992, 19, 55-70).

The antiviral test was carried out in a 96-cell microtiter plate. The first column of the plate only contained a culture medium and HepG 2.2.15 cells, as a blank control.

First, a stock solution (50 mmol) of a test compound was dissolved in DMSO, and further diluted with a culture medium for growth of HepG 2.2.15 cells. Usually, the compound of the present invention was transferred by suction at a test concentration of 100 µg/ml (the first test concentration) to each cell of the second column of the microtiter plate, and then diluted with the culture medium for growth plus 2% fetal bovine serum (25 µL), by 2 times once, up to 2¹⁰ times at the maximum.

Then, 225 µL of a suspension of HepG 2.2.15 cells in the culture medium for growth plus 2% fetal bovine serum (5 x 10⁴ cells/ml) was added to each cell of the 96-cell microtiter plate.

The test mixture was incubated under the conditions of 37°C, 5% CO₂ for 4 days. Then, the supernatant was removed by suction, and to each cell 225 µL of a new-prepared culture medium for growth was added. Again, the compound of the present invention was added with a solution of 25 µL. The obtained mixture was further incubated for 4 days.

Prior to collection of the supernatant for determination of the antiviral effect, the change in cytotoxicity of HepG 2.2.15 cells was investigated by using optical microscopic technique or biochemical detection method (e.g., Alamar Blue staining or Trypan Blue staining).

Thereafter, the supernatant was collected, and sucked in vacuum to a 96-cell dot blot chamber covered by nylon film (used in accordance with the instructions for use provided by the manufacturer).

Determination of cytotoxicity

The substance-induced change in cytotoxicity in HepG 2.2.15 cells or in inhibition of the cells could be determined by using, e.g., optical microscopic technique, and expressed by the change of cell morphology. Such substance-induced changes, e.g., cell lysis, cavity formation or change in cell morphology, in HepG 2.2.15 cells were apparent as compared to the untreated cells. Taking the observed pathological change of cells as index, the pathological change of cells was observed under microscope after 8 days, a complete destroy being indicated as 4; 75% being indicated as 3; 50% being indicated as 2; 25% being indicated as 1; and no pathological change being indicated as 0. The average extents of pathological change of cells and the inhibition percentages at various concentrations were calculated. According to Reed&Muench method, a half toxic concentration (TC₅₀) and a maximum non-toxic concentration (TC₀) were calculated.

TC₅₀ refers to the concentration of the compound of the present invention when 50% of cells have a similar morphology to the corresponding cells as a control.

Determination of antiviral activity

After the supernatant was transferred onto the nylon film of the dot blot device (see the contents hereinabove), the supernatant of HepG 2.2.15 cells was denaturated (1.5 M NaCl/0.5 M NaOH), neutralized (3 M NaCl/0.5 M Tris. HCl, pH 7.5) and washed (2 x SSC). Then, the filter film was kept at 120°C for 2-4 hours, whereby DNA was baked on the filter film.

DNA hybridization

Usually, viral DNA of HepG 2.2.15 cells treated on the nylon film was detected by using a non-radioactive digoxigenin labeled HB-specific DNA probe. Wherein, each time the probe was labeled with digoxigenin, purified and hybridized according to the directions for use given by the manufacturer.

Briefly, pre-hybridization and hybridization were conducted with 5 x SSC, 1 x blocking agent, 0.1% N-lauroyl sarcosine, 0.02% SDS and 100 µg sperm DNA of black carp. After pre-hybridization at 60°C for 30 min, a specific hybridization (at 60°C for 14 hr) was conducted with 20-40 ng/ml digoxigenin labeled denaturalized HBV specific DNA. Then, the film was washed, followed by HBV DNA detection with digoxigenin antibody.

The immunology detection of digoxigenin labeled DNA was conducted according to the directions for use given by the manufacturer.

Briefly speaking, the film was washed and pre-hybridized with a blocking agent (in accordance with the directions for use given by the manufacturer), and then hybridized with an anti-DIG antibody that had been previously coupled onto an alkaline phosphatase for 30 min. After washing, an alkaline phosphatase substrate CSPD was added, and cultured together with the filter for 5 min, and then wrapped in a plastic film, followed by further culturing at 37°C for 15 min. The filter was exposed to X-ray, and the chemical luminous signal (culturing for 10 min to 2 hr according to the signal strength) of HB-specific DNA on the film was detected, whereby a half inhibitory concentration (IC₅₀) was calculated.

The half inhibitory concentration (IC₅₀) refers to the concentration of the compound of the present invention which reduces the HB-specific band by 50% as compared with the untreated sample.

The compound of the present invention exhibits a relatively strong antiviral effect. This kind of compound has unexpected antiviral activity to HBV, and thus is adapted to be used for treating various virus-caused diseases, in particular acute and chronic permanent diseases caused by HBV viral infection. Chronic viral diseases caused by HBV may lead to various syndromes having different extents of severity. As well known, chronic HBV infection may lead to hepatic cirrhosis and (or) liver cell carcinoma.

Examples of indications capable of being treated by the compound of the present invention include: acute and chronic viral infections capable of leading to infectious hepatitis, such as HB viral infection, and particularly preferred chronic HB viral infection and acute HB viral infection.

The pharmaceutical composition comprising the compound of the present invention may be administered in any of the following routes: orally, inhaled by spray, rectally, nasally, vaginally, topically, parenterally such as subcutaneous, intravenous, intramuscular, intraperitoneal, intrathecal, intraventricular, intrasternal or intracranial infection or infusion, or administered with the aid of an explanted reservoir, wherein the administration routes by orally, intramuscular, intraperitoneal or intravenous injection are preferred.

The compound of the present invention or the pharmaceutical composition comprising the compound of the present invention may be administered in a unit dosage form. The dosage form may be in a liquid form, or a solid form. The liquid form includes true solution, colloids, particulates, emulsions, suspensions. Other dosage forms include tablets, capsules, dropping pills, aerosols, pills, powder, solutions, suspensions, emulsions, granules, suppository, lyophilized powder for injection, clathrates, implants, patches, liniment, and the like.

The pharmaceutical composition of the present invention may further comprise a commonly used carrier that includes, but not limited to, ion exchanger, alumina, aluminum stearate, lecithin, serum protein such as human serum protein, buffer substance such as phosphate, glycerin, sorbic acid, potassium sorbate, a mixture of partial glycerine esters of saturated vegetable fatty acids, water, salt or electrolyte, such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salt, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose, polyethylene glycol, sodium carboxymethylcellose, polyacrylate, beeswax, lanolin, and the like. The amount of the carrier in the pharmaceutical composition may be 1% to 98% by weight, usually about 80% by weight. For the convenience, topical anesthetic, antiseptic, buffer and etc. may be directly dissolved in the carrier.

Oral tablets and capsules may comprise excipients e.g., binders such as syrup, Arabic gum, sorbitol, tragacanth, or polyvinylpyrrolidone, fillers such as lactose, sucrose, corn starch, calcium phosphate, sorbitol, aminoacetic acid, lubricant such as magnesium stearate, saponite, polyethylene glycol, silica, disintegrating agent such as potato starch, or acceptable moisturizing agent such as sodium lauryl sulfate. Tablets may be coated by using known methods in pharmaceutics.

Oral solution may be made as a suspension of water and oil, a solution, an emulsion, a syrup or an elixir, or made as a dried product to which water or other medium is added before use. This liquid preparation may comprise conventional additives, e.g., suspending agent such as sorbitol, cellulose methyl ether, glucose syrup, gelatin, hydroxyethyl cellulose, carboxymethyl cellulose, aluminum stearate gel, hydrogenated edible grease; emulsifying agent such as lecithin, sorbitan monooleate, Arabic gum; or non-aqueous carrier (possibly including edible oil), such as almond oil, grease such as glycerin, ethylene glycol, or ethanol; antiseptic such as methyl or propyl p-hydroxybenzoate, sorbic acid. If desired, a flavoring agent or a colorant may be added.

Suppository may comprise a conventional suppository substrate, such as cocoa butter or other glyceride.

For non-gastric administration, the liquid dosage form is usually made of the compound and a sterilized carrier. The preferred carrier is water. According to the carrier selected and the drug concentration, the compound can be dissolved in the carrier or made into a suspension. When making an injection solution, the compound is firstly dissolved in water, and then filtered and sterilized before being packaged into an enclosed bottle or ampoule.

For topical application on skin, the compound of the present invention may be made into a suitable form of ointment, lotion or cream, wherein the active ingredient is suspended or dissolved in one or more carrier(s). The carrier used for an ointment includes, but not limited to, mineral oil, liquid vaseline, white vaseline, propylene glycol, polyoxyethylene, polyoxypropylene, emulsified wax and water; the carrier used for a lotion and a cream includes, but not limited to, mineral oil, sorbitan monostearic ester, Tween 60, cetyl esters wax, hexadecylene aromatic alcohol, 2-octyl dodecanol, benzanol and water.

In the above preparations, the active compound exists in a concentration of about 0.1 to 99.5% by weight, preferably about 0.5 to 95% by weight, based on the total weight of the mixture.

The above preparations may further comprise other pharmaceutically active compounds, in addition to the compound of formula (I) or claim 2.

In general, it has been proved that, advantageously, whether in human medicine or in veterinary medicine, the total dose of the active compound of the present invention is about 0.5 to 500 mg every 24 hr, preferably 1 to 100 mg per kg body weight. If appropriate, the drug is administrated by single dose for multiple times, to thereby achieve the desired effect. The amount of the active compound in a single dose is preferably about 1 to 80 mg, more preferably 1 to 50 mg per kg weight body. Nevertheless, the dose may also be varied according to the type and body weight of the object to be treated, the kind and extent of severity of diseases, the type of the preparation and the administration manner of the drug, and the administration period or the time interval.

Concrete Modes for Carrying Out the Invention

The following examples are preferred embodiments of the present invention, and shall not be understood to limit the present invention in any manner.

The melting point of the compound was determined by using RY-1 melting point apparatus, and the thermometer was not calibrated. Mass spectrum was determined by using Micromass ZabSpec high resolution mass spectrograph (resolution 1000). ¹H NMR was determined by using JNM-ECA-400 superconductive NMR spectrometer, the working frequency being ¹H NMR 400MHz, ¹³C NMR 100MHz.

Examples Example 58 Preparation of ethyl 2-(2,4,6-trifluorophenyl)-4-(2-chloro-4-fluorophenyl)-6-methy-1,4-dihydro-pyrimidin-5-carboxylate

2 mmol 2,4,6-Trifluorobenzatnidine acetate (Judkins, B. D., Allen, D. G., Cook, T. A. Synth. Commun. 1996, 26(23):4351-4367), 2 mmol 2-chloro-4-fluorobenzaldehyde, 2 mmol methyl acetoacetate and 2.2 mmol sodium acetate were reacted under reflux in 10 ml anhydrous ethanol for 20 hr, concentrated, and then ethyl acetate and water were added to separate the layers. The ethyl acetate layer was dried over anhydrous sodium sulfate, and separated by a column chromatography to obtain 0.29 g of a colorless fine needle crystal with mp 184-186°C; ¹H-NMR (400MHz, DMSO-d₆) δ 1.02-1.05(3H, t,J=7.3Hz); 2.32(3H,s); 3.92-3.95 (2H,m, J=7.3Hz); 5.97(1H,s); 7.21-7.45(5H,m); MS(EI) 426(M⁺).

Example 59 Preparation of methyl 2-(2,4,6-trifluorophenyl)-4-(2-chloro-4-fluorophenyl)-6-methyl-1,4-dihydro-pyrimidin-5-carboxylate

Using the method of Example 58, while using methyl acetoacetate in place of ethyl acetoacetate, 0.27 g of a colorless crystal was obtained, with mp 112-114°C; ¹H-NMR (400MHz, DMSO-d₆) δ 2.33(3H,s); 3.51 (3H,s); 5.96(1H,s); 7.20-7.43(5H,m); 9.9(1H,s); MS(EI) 412(M⁺).

Example 62 Preparation of ethyl 4-R-(2-chloro-4-fluorophenyl)-2-(2,4,6-trifluorophenyl)-6-methyl-1,4-dihydro-pyrimidin-5-carboxylate

(-)-enantiomer of the compound of Example 58, and obtained by chiral separation as a colorless needle crystal. ¹H-NMR (400MHz,DMSO-d₆) δ 1.02-1.05(3H,t, J=7.0 Hz); 2.32(3H,s); 3.92-3.96(2H,m,J=7.0Hz); 5.97(1H,s); 7.21-7.45(5H,m); 9.86(1H,s); MS(EI) 426(M⁺) ; [α]_D=-92.38(methanol).

Example 63 Determination of cytotoxicity and antiviral activity of the selected compounds

The cytotoxicity and antiviral activity of the compounds according to this invention were determined in accordance with the methods described above, and the results were showed in Table 1. Table 1. Inhibitory effects of the selected compounds on HBV DNA

Example No.
58	2.60	32.50
59	3.39	>30.30
62	2.01	117.23

Claims (4)

1. A compound of formula (I), or a pharmaceutical acceptable salt thereof, wherein

   R¹ represents hydrogen or acetyl,

   R³ for occurrence once or more each represents fluoro, chloro, bromo, hydroxyl, nitro, methoxy, or methyl, and

   R⁴ represents a group represented by formula -XR¹², wherein X represents oxygen, and R¹² represents a straight or branched alkyl having up to 3 carbon atoms.
2. A compound which is selected from the group consisting of:

   ethyl2-(2,4,6-trifluorophenyl)-4-(2-chloro-4-fluorophenyl)-6-methyl-1,4-dihydro-pirimidin-5-carboxylate,

   methyl 2-(2,4,6-trifluorophenyl)-4-(2-chloro-4-fluorophenyl)-6-methyl-1,4-dihydro-p pirimidin-5-carboxylate, and

   ethyl 4-R-(2-chloro-4-fluorophenyl)-2-(2,4,6-trifluorophenyl)-6-methyl-1,4-dihydro-pirimidin-5-carboxylate,

   or a pharmaceutically acceptable salt thereof.
3. A pharmaceutical composition comprising a compound according to claim 1 or 2, and at least one pharmaceutically acceptable carrier.
4. The compound according to claim 1 or 2 for use in a method for the treatment of an acute or chronic disease caused by hepatitis B virus infection.