WO2008154819A1 - Carbethoxy-substituted thiazolyl dihydropyrimidines - Google Patents

Abstract

Novel carbethoxy-substituted thiazolyl dihydropyrimidines and compositions thereof with other antiviral agents, suitable for combating hepatitis B virus (HBV) infections.

Description

 Ethoxycarbonyl-substituted thiazolyl dihydropyrimidine

 The present invention relates to a novel ethoxycarbonyl-substituted thiazolyl dihydropyrimidine, a process for the preparation thereof and its use as a medicament, especially as a medicament for the treatment and prevention of hepatitis B virus infection. The invention further relates to compositions of these dihydropyrimidines with other antiviral agents and, where appropriate, immunomodulators, and medicaments containing such compositions, especially compositions for the treatment and prevention of HBV infections such as hepatitis B. Background technique

 Hepatitis B virus belongs to the family of hepatic viruses. It can cause acute and/or persistent/progressive chronic conditions. Hepatitis B virus also causes many other clinical signs in pathological morphology - especially chronic inflammation of the liver, cirrhosis of the liver and carcinogenesis of hepatocytes. In addition, co-infection with hepatitis D can have an adverse effect in the development of the disease.

Interferon and lamivudine are conventional drugs approved for the treatment of chronic hepatitis. However, interferon is only moderately active and has harmful side effects; although lamivudine has good activity, its resistance develops rapidly during treatment and often has a rebound effect after stopping treatment. Lamivudine (3-butyl 0> has an IC ₅ Q value of 300 nM (Science, 299 (2003), 893-896).

 No. 7,074,784 discloses 6-aminoalkyldihydropyrimidines and their use as medicaments, especially for the treatment and prevention of hepatitis B virus infection.

 This patent discloses 54 different examples, and almost all of them (50) are compounds having a fluorine-substituted 2-pyridine residue at the position of R6. In all of the examples (54) of the patent, R3 is a methyl group.

 This patent teaches that compounds having longer alkyl linkages (e.g., ethyl) are weakly active or even inert.

In the patent, nine compounds (Examples 1, 5, 7, 9, 12, 24, 31, 34, 45) having the following structure have eight fluoro-substituted 2-pyridine residues having 8 positions. Only one example (Example 45) had R ⁶ = thiazol-2-yl.

 Summary of the invention

We have surprisingly found Example 45 of this patent having a thiosalt-2-yl substituent ((R ⁶ = thiazol-2-yl)ethyl ester (R ³ = ethyl) (R ^l = o-chloro and R ² = p-chloro), produced a 3-fold higher activity derivative, IC _{5 ,} lower than l nM (see Table 1)

Accordingly, the present invention relates to a compound represented by the formula (I) and an isomer thereof (la)

among them

R ¹ is o-chloro, R ² is p-chloro, R ³ is C ₂ -C ₃ alkyl, R ⁶ is thiazol-2-yl, X is methylene, and Z is morpholinyl.

 Preferably, in the compounds of the formula (I) and (la) of the present invention:

R ¹ is o-chloro, R ² is p-chloro, R ³ is ethyl, R ⁶ is thiazol-2-yl, X is methylene, and Z is morpholinyl.

 The invention also relates to enantiomers of the above compounds and to their respective mixtures. The racemic form can be separated by known means, which is essentially a homogeneous component of the stereoisomer.

 The compounds of the invention comprise isomers of the formulae (I) and (la) and mixtures thereof. The compounds of the invention may also be in the form of a salt. In the present invention, a physiologically acceptable salt is preferred.

The physiologically acceptable salt may be a mineral acid salt or an organic acid salt. Preferred are salts of inorganic acids such as, for example, hydrochloric acid, hydrobromic acid, phosphoric acid or sulfuric acid, or organic carboxylic acids or sulfonic acids such as acetic acid, maleic acid, fumaric acid, malic acid, citric acid, tartaric acid. , lactic acid, benzoic acid or methanesulfonic acid, ethanesulfonic acid, benzenesulfonic acid, toluenesulfonic acid or naphthalene-disulfuric acid The salt formed.

 The physiologically acceptable salt may also be a metal or ammonium salt of a compound of the invention. Particularly preferred examples are sodium, potassium, magnesium, or calcium salts, and from ammonia or organic amines such as ethylamine, di- or triethylamine, di- or triethanolamine, dicyclohexylamine, dimethylaminoethanol, An ammonium salt formed by arginine, lysine, ethylenediamine, or 2-phenylethylamine.

 The compound (I) of the present invention can be produced by the following method:

 [A] First, the benzaldehyde (II) and the β-ketoester represented by the formula (III) are converted into the general formula (IV) by adding or not adding a base or an acid, and if necessary, in the presence of an inert organic solvent. Shown benzylidene compounds:

Wherein Κλ R ² , R ³ , X and Z have the same meanings as described above.

Then, the latter and the hydrazine of the formula (V) or a salt thereof (such as, for example, a hydrochloride or an acetate), with or without the addition of a base or an acid, where appropriate, in the presence of an inert organic solvent React

Wherein R ⁶ means as described above, or

[B] combining a compound of the formula (III) with an aldehyde (Π) and a ruthenium (V) or a salt thereof For example, hydrochloride or acetate) with or without the addition of a base or acid, where appropriate, in the presence of an inert organic solvent; or

 [C] When X in the formula (I) is a methylene group, a compound represented by the formula (VI)

Wherein RR ² , R ³ and R ⁶ have the same meanings as defined above, and Y is a nucleophilic substituent such as chloride, bromide, iodide, methanesulfonyl or tosyl, and morpholine (VII)

 The reaction is carried out with or without the addition of a base, as appropriate, in the presence of an inert solvent. The compound (VI) can be produced, for example, by using the compound of the formula (VIII)

Wherein R ¹ , R ² , R ³ and R ⁶ have the same meanings as defined above, and a brominating agent such as N-bromosuccinate; imine, preferably in an inert solution, to give a compound of the formula (IX)

The latter having a nucleophilic substituent directly or in accordance with the conventional formula described in the literature After further conversion, the reaction with morpholine (VII).

[D] an aldehyde represented by the formula (II) and a compound represented by the formula (X), which are reacted in an inert solvent when appropriate,

Wherein R ³ , X and Z have the same meanings as defined above, and the oxime represented by the formula (V) has or does not carry a base adduct.

To prepare a compound of the formula (I) of the present invention, wherein X is a methylene group and z is a morpholinyl group, a chloroacetate ester represented by the formula (XI) can be reacted with morpholine (VII) to prepare Corresponding β-ketocarboxylate (ΠΙ)

Wherein R ^{3 has the} same meaning as described above.

 2-Chloro-4-fluoro-benzaldehyde (oxime) as a starting material is commercially available. The β-ketocarboxylate (III) used as a starting material is well known or can be analogized from known methods published in the literature [e.g., D. Borrmann, "Umsetzung von Diketen mit Alkoholen, Phenolen und Mercaptanen", in "Methoden der organischen Chemie" (Houben- Weyl), vol. VII/4, 230 ff (1968); Y. Oikawa, K. Sugano und O. Yonemitsu, J. Org. Chem. 43, 2087 ( 1978)].

 Compound (V) is known and can be prepared as described in WO-A-99/54326 and WO-A-99/54329.

 Morpholine (VII) is commercially available.

 Compounds (VIII) and (X) can be produced according to the procedure [A] or [B] described in WO-A-99/54326.

All inert organic solvents are suitable for the eight, B, C and D steps. Preferred among them are alcohols such as methanol, ethanol, isopropanol, ethers such as dioxins, diethyl ether, tetrahydrofuran, ethylene glycol monomethyl ether, ethylene glycol dimethyl ether, carboxylic acids such as glacial acetic acid, or dimethyl. Formamide, Dimethyl sulfoxide, acetonitrile, pyridine and hexamethylphosphoric triamide.

 The reaction temperature can be varied within a relatively wide range. A temperature between 20 and 150 ° C is usually used, but is preferably at the boiling temperature of the selected solvent.

 The reaction can be carried out under atmospheric pressure or under high pressure. It is usually carried out under atmospheric pressure.

 The reaction can be carried out in the presence or absence of an acid or a base; however, it is preferred to carry out the reaction in the presence of a weak acid such as acetic acid or formic acid.

 One embodiment of the invention relates to the use of A) at least one of the above-described dihydropyrimidines,

B) At least one composition of another antiviral agent different from A).

 A detailed embodiment of the invention relates to a composition comprising A) a dihydropyrimidine, a B) HBV polymerase inhibitor and, if appropriate, a C) immunomodulator.

 Preferred immunomodulators C) include, for example, all interferons such as alpha-, beta- and gamma-interferons, especially a-2a- and a-2b-interferon, interleukins such as interleukin-2 Polypeptides such as thymosin-ct-1 and thymoctonan, imidazoquinoline derivatives such as 8 levamisole, immunoglobulins and therapeutic vaccines. .

 Accordingly, the present invention also relates to such compositions for the treatment and prevention of HBV infection and their use in the treatment of diseases caused by HBV.

 The use of the compositions of the invention is beneficial for the treatment of HBV-induced diseases, primarily synergistic antiviral activity, and Tox-50 relative to a single component (50% cell viability toxicity) relative to a single treatment of a single compound. Scope) The compositions of the invention are well tolerated.

 The HBV polymerase inhibitors used to achieve the objects of the present invention are in Ph. A. Furman et al., Antimicrobial Agents and Chemotherapy Vol. 36 (No. 12), 2688 (1992). Those materials disclosed in the endogenous polymerase experiments, as well as those described below, inhibit the formation of HBV DNA duplexes, resulting in a maximum of 50% of the activity values of zero.

HBV virions from culture suspensions integrate nucleoside 5'-triphosphate in vitro The DNA of HBV is in the positive strand. By using agarose gel electrophoresis, the product in which [α - ³² P]-deoxynucleoside 5'-triphosphate was incorporated into the virus 3.2 kb DNA was found, and there was no potential HBV polymerase-inhibiting property. substance. From the cell culture suspension of HepG2.2.15 cells, it was precipitated with polyethylene glycol and concentrated to obtain HBV virions. One part by volume of the clarified cell culture suspension was mixed with 1/4 part by volume of an aqueous solution containing 50% by weight of polyethylene glycol 8000 and 0.6 M of sodium chloride. The pellet was centrifuged at 2500 x g for 15 minutes, and the precipitate was resuspended in 2 ml of a buffer containing 0.05 M tris-HCl M (H 7.5), and dialyzed against the buffer containing 100 mM potassium chloride. The sample was frozen at -80 Torr. Each reaction mixture (100 μΐ) contains at least 10 ⁵ HBV virions, 50 mM tris-HCl (p.sub.H 7.5), 300 mM potassium chloride, 50 mM magnesium chloride, 0.1%® Nonident P-40 (non-ion Type of washing, Boehringer Mannheim) > ΙΟ μΜ άΑΤΡ, 10 μΜ dGTP , 10 μΜ dTTP; 10 μΟΊ [ ³² P]dCTP (3000 Ci/mmol; final concentration 33 nM) and 1 μΜ triphosphate form of polymerase potential inhibition Agent. The samples were incubated at 37 C for one hour and then stopped by the addition of 50 mM EDTA. Add 10% w/v SDS solution (10 g SDS per 90 ml water) to a final concentration of 1% by volume (based on the total volume of the solution) and add proteinase K to a final concentration of 1 mg/ml. It was then incubated at 37 ° C for 1 hour, extracted with an equal volume of phenol / chloroform / isoamyl alcohol (25:24:1 by volume) solution, and the DNA was precipitated from the aqueous phase containing ethanol. The DNA pellet was suspended in ΙΟμΙ gel buffer (10.8 g of tris, 5.5 g of boric acid, and 0.75 g of EDTA (= TBE buffer) in 1 liter of water) and separated by agarose gel electrophoresis. The gel therein is dried or transformed into a membrane using Southern transformation techniques. The amount of the resulting labeled DNA duplex is then detected by comparison to a negative control (=endo-pol reaction with a blank or inert control). The presence of a maximum 50% concentration of the control group indicates the presence of an HBV polymerase inhibitor.

Preferred HBV polymerase inhibitors B) include, for example, 3TCHi lamivudine = 4-amino-1-[(2R-cis)-2-(hydroxymethyl)-1.3-oxothio-5 -yl-]pyrimidine-2(1H)-one, cf. EP-B 382 526 (=US Pat. No. 5,047,407) and WO 91/11186 (=US Pat. No. 5,204,466); Adefovir dipivoxil =9-[2- [bis(pivaloyloxymethoxy)phosphonomethoxy]ethyl]adenine, cf. EP-B 481 214 (=US Pat. Nos. 5,663,159 and 5,792,756), US Pat. Nos. 4,724,233 and 4,808,716;

 BMS 200 475=[1S-(L. alpha., 3..alpha., 4..beta.)]-2-amino-1.9-dihydro-9-[4-hydroxy-3-(hydroxymethyl) -2-methylene-cyclopentyl]-6H-indol-6-one, cf. EP-B 481 754 (=US Pat. Nos. 5,206,244 and 5,340,816), WO 98/09964 and 99/41275;

Abacavir = (-)-(lS-cis)-4-[ ² -amino-6-(cyclopropylamine)-9H-indol-9-yl]-2-yl-cyclopentene- 1-methanol, cf. EP-B 349 242 (=US Pat. No. 5,049,671) and EP-B 434 450 (=US Pat. No. 5,034,394);

 FTC=(2R-cis)-4-amino-5-fluoro-l-[2-(hydroxymethyl)-1.3-oxothio-5-yl]-p-pyrimidine-2(1H)-one, Cf. WO 92/14743 (=US Pat. Nos. 5,204,466; 5,210,085; 5,539,116; 5,700,937; 5,728,575; 5,814,639; 5,827,727; 5,852,027; 5,892,025; 5,914,331; 5,914,400) and WO 92/18517;

PL-FDDC = 5-(6-amino-2-fluoro-9H-purin-9-yl)-tetrahydro-2-furanmethanol, cf. WO 94/27616 (=US Pat. Nos. 5,627,160; 5,561,120; 5,631,239 And 5, 830, 881); L-FMAU=l-(2-deoxy-2-fluoro-.beta.-L-arabinofuranosyl)-5-methyl-pyrimidine e-2.4(lH,3H)- Ketone, cf. WO 99/05157, WO 99/05158 and US Pat.

 A more preferred embodiment of the invention relates to a composition comprising the above dihydropyrimidines (I) and (la) and B) lamivudine.

Another preferred HBV antiviral agent B contains, for example, phenylacrylamide represented by the following formula

 among them

R ¹ and R ² , independently of each other, d- -fluorenyl or, together with the nitrogen atom at the position thereof, form a ring having 5 to 6 ring atoms including carbon and/or oxygen atoms. R ³ to R ¹² are independently hydrogen, cyano, C _r C ₄ -alkyl, optionally substituted for C _r C ₄ -nonyloxy, nitro, cyano or trifluoromethyl. R ¹³ is hydrogen, C _r C ₄ -alkyl, dC ₇ -acyl or aralkyl, and X is halogen or an optionally substituted C _r C ₄ -fluorenyl group.

 These phenyl acrylamides and their preparation are disclosed in WO 98/33501, the disclosure of which is hereby incorporated herein. AT-61 is a compound

Preferred immunomodulators C) include, for example, all interfering agents such as α-, β- and γ-interferons, in particular a-2a- and a-2b-interfering, interleukins such as interleukin-2 , polypeptides such as thymosin-ot-1 and thymoctonan, imidazoquinoline derivatives such as 8 levamisole, immunoglobulins and therapeutic vaccines.

 Another preferred embodiment of the invention relates to a composition comprising A) the above-described dihydropyrimidines (I) and (la); B) lamivudine; and, where appropriate, C) interferon. Experimental description

Based on MA Sells Proc. Natl. Acad. Sci. 84, 1005 1009 (1987) and B. The antiviral effect of the compounds of the invention on hepatitis B virus is studied by the method described by E. Korba et al., Antiviral Research 19, 55 70 (1992).

 Antiviral testing was performed on 96-well microtiter plates. The first column only received medium and HepG2.2.15 cells as a virus control.

The mother liquor of the test compound (50 mM) was first dissolved in DMSO and then diluted with HepG2.2.15 medium. Typically, a 100 μΜ test concentration (1 st test concentration) of the compound of the invention is pipetted into the second test column of the microtiter plate each time, and then added to the medium in which 2% by weight fetal calf serum (25 μl in volume) is added. Diluted 2 ^1Q times in two steps.

Each well of the microtiter plate containing 2% by weight of fetal bovine serum contained 225 l of HepG 2.2.15 cell suspension (5 x 10 ⁴ cdls/ml). The test mixture was incubated for 4 days at 37 ° C, 5% C0 ₂ (v/v).

 The surface suspension was then aspirated and discarded, and 225 μM of freshly prepared medium was added to the wells. A 25 μΐ volume of a 1/10 concentration solution of the compound of the invention was added. The mixture was continued for 4 days.

 Changes in cytotoxicity of HepG2.2.15 cells were examined under light microscopy or by biochemical assays (eg, Alamar Blue stain or Trypan Blue stain) prior to collecting the antiviral effects of the suspension.

 Surface suspensions and/or cells were collected and vacuum coated to coat a 96-well spot chamber with a nylon membrane (according to the manufacturer's information) Determination of cytotoxin

 Detection of cytotoxic or cytotoxic changes induced by substances in HepG2.2.15 cells, for example, changes in cell morphology under a light microscope. Changes in these substances in HepG2.2.15 cells are evident compared to untreated cells, such as cytolysis, vacuoles, or changes in cell morphology. 50% toxicity (TOX.-50) means that 50% of the cells exhibit a morphology compared to the corresponding control cells.

The tolerance of some of the compounds of the invention is tested by other host cells such as, for example, HeLa cells, primary human peripheral hematopoietic cells or transformed cell lines such as H-9 cells. No change in cytotoxin was detected at concentrations of the compounds of the invention > 10 ^ 1 . Determination of antiviral effect

 After the surface suspension or dissolved cells were transferred to a nylon membrane of a spot device (as described above), the intracellular or extracellular suspension of HepG2.2.15 cells was denatured (1.5 M NaCl/0.5 N NaOH), neutralized ( 3 M NaCl/0.5 M Tris HCl, pH 7.5), then rinse (2xSSC) o The DNA was dried onto the membrane by incubating the filter at 120 °C for 2-4 hours.

DNA hybridization

 Viral DNA from treated HepG2.2.15 cells on nylon filters is typically detected by a non-radioactive, digoxin-labeled hepatitis B-specific DNA probe, labeled with digoxin, and according to the manufacturer Instructions for purification and hybridization.

Prehybridization and hybridization in 5xSSC, lx blocking reagent, 0.1% (by weight) N- lauroyl sarcosine, 0.02% (wt.) SDS and ₁₀₀ μ β for herring sperm DNA. Pre-hybridization is at 60. C was run for 30 minutes and then specifically hybridized with 20 to 40 ng/ml of digoxin-labeled, denatured HBV-specific DNA (14 hours, 60 °C). Wash the filter. Detection of HBV-DNA with digoxin antibody

Immunodetection of digoxin-labeled DNA was performed according to the manufacturer's instructions: The wash filter was hybridized in a blocking reagent (according to the manufacturer's instructions). Hybridization was then carried out using an anti-DIG antibody coupled to alkaline phosphatase for 30 minutes. After the washing step, the substrate of alkaline phosphatase, CSPD, was incubated with the filter for 5 minutes, then coated with a plastic film, and for another 15 minutes at 37 ° C. The filter was exposed to the X-ray layer. HBV DNA was seen emission signal (incubation depending on signal strength: 10 to 2 minutes) at half maximum inhibitory concentration (IC ₅ o, 50% inhibitory concentration) of a 50% reduction compared to the compounds of the present invention and an untreated sample. The concentration of intracellular or extracellular hepatitis B virus.

The compounds of the present invention exhibited an effective antiviral effect of IC _{5 ()} below 1 nM, which was unexpected. Thus, the compounds of the invention are useful in the treatment of viral-induced diseases, particularly acute and chronic persistent HBV viral infections. Chronic viral disease caused by HBV It can lead to serious morbidity, and chronic hepatitis B virus infection can lead to cirrhosis and/or hepatocellular carcinogenesis in many cases.

 For the compounds of the present invention, the indicated regions that may be mentioned are, for example, the treatment of acute and chronic viral infections that may result in infectious hepatitis, for example, hepatitis B virus infection. The compounds of the invention are especially suitable for the treatment of chronic hepatitis B infection and acute and chronic hepatitis B virus infection.

 The invention includes a pharmaceutical preparation comprising, in addition to a non-toxic, inert, pharmaceutically suitable carrier, one or more compounds (I) or (la) or compositions of the invention or one or more activities Ingredient (I) or (la) or a composition of the invention.

 The active ingredients (I) and (Ia) used in the above pharmaceutical preparations have a concentration of from about 0.1 to 99.5% by weight, preferably from about 0.5 to 95% by weight, based on the whole of the mixture.

 The above pharmaceutical preparations may also contain other active pharmaceutical ingredients other than the compounds (I) and (la).

 The content ratio of components A, B and suitable C in the composition of the invention may vary within wide limits, preferably from 5 to 500 mg of A/10 to 1000 mg of B, especially from 10 to 200 mg. A/20-400 mg of B.

 Component C, if appropriate, can also be used, preferably in an amount of from 1 to 10 million, more preferably from 2 to 7 million, I.U. (international unit), three times a week for a period of more than one year.

 The compound or composition of the present invention for use in the above pharmaceutical preparations is usually present in a concentration of from about 0.1% to about 99.5%, preferably from about 0.5% to about 95%, by weight based on the total of the mixture.

 The above pharmaceutical preparations can be prepared by a known conventional method, for example, by mixing the active ingredient with a carrier.

It is generally advantageous to take a total dose of from about 0.05 to about 500, preferably from 0.1 to 100 mg/kg body weight per 24 hours, whether in humans or veterinary, for a single dose of multiple doses. The ideal effect. The active ingredient contained in a single dose is preferably in a total amount of from about 0.1 to about 80, preferably from 0.1 to 30 mg/kg of body weight. No It is necessary to vary the above dosage, especially depending on the type and weight of the subject to be treated, the nature and severity of the condition, the type of preparation, the manner in which the medicine is administered, and the time or interval at which the medicine is administered.

 Accordingly, the present invention also relates to the above compounds and compositions for use in the control of diseases.

 The invention further relates to a medicament comprising at least one of the above compounds or compositions and, where appropriate, one or more other active pharmaceutical ingredients.

 The invention further relates to the use of the above compounds and compositions for the preparation of a medicament for the treatment and prevention of the above mentioned diseases, in particular viral diseases, in particular hepatitis B.

 Unless otherwise indicated, the percentages in the following examples are by weight. The proportion of the solvent in the mixed solution refers to the volume ratio.

 Example

 A. Intermediates Example 1

Ethyl 4-(2,4-dichlorophenyl)-2-(thiazol-2-yl)-6-methyl-1,4-dihydropyrimidine-5-carboxylate

 10.0 g (57.1 mmol) of 2,4-dichlorobenzaldehyde, 7.4 g (57.1 mmol) ethyl acetoacetate, 9.3 g (57.1 mmol) 2-mercapto-thiazole hydrochloride and 5.7 g (69.5 mmol) Sodium acetate was dissolved or suspended in 500 ml of ethanol and refluxed for 16 hours. Cool to room temperature, filter with air, and wash the residue to remove inorganic salts. Product obtained 11.8 g (52.5%)

Melting point: 164-166. C. Example 2 Ethyl 6-bromomethyl 4-(2,4-dichlorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5-carboxylate

 8.0 g (20.2 mmol) of ethyl 4-(2,4-dichlorophenyl)-2-(thiazol-2-yl)-6-methyl-1,4-dihydropyrimidine prepared in Example 1 The -5-carboxylate was added to 140 ml of carbon tetrachloride and heated to 50 under an argon atmosphere. C, a clear solution is obtained. At this temperature, 4.36 g (24.2 mmol) of N-bromosuccinimide was added and kept at this temperature for 10 minutes. Cool immediately, filter at room temperature, and concentrate under reduced pressure. The purity of the product was determined by HPLC to be higher than 90% and used as a starting material for the next step.

RiN) ⁶⁸ (petroleum ether / ethyl acetate = ^{8: 2)}

B. Preparation Examples Example 3

Ethyl 4-(2,4-dichlorophenyl)-2-(thiazol-2-yl)-6-(4-morpholinomethyl)-1,4-dihydropyrimidine-

5.0 g of the freshly prepared ethyl-6-bromomethyl-4-(2-chloro-4-chlorophenyl)-2-(thiazol-2-yl)-1,4-dihydropyrimidine-5 -carboxylate added to 60 ml of methanol to form a solution The solution was mixed with 5 times the amount of morpholine and stirred at room temperature for 30 minutes. The solution was diluted with water and extracted with ethyl acetate.

Production: 4.1g

Melting point: 155-157 ° C.

R _f = 0.48 (petroleum ether / ethyl acetate = 8 : 2)

 The enantiomer prepared in Example 3 was separated on a chiral column (Daicel Chiralpak AS-H, mobile phase: n-hexane / ethanol = 99/1).

 The anti-HBV active compounds of Example 3 were all enantiomers with a longer retention time. The activity data for the compounds of the invention are listed below:

Example No. IC ₅₀ (nM)

 3 0.6

(-)-3 0.3 Treatment of HepG2.2.15 cells produced by hepatitis B virus using the compounds of the present invention results in a decrease in intracellular and/or extracellular viral DNA. Industrial applicability

Experiments show that the compounds of the present invention exhibit IC _5. An effective antiviral effect of less than 1 nM, whereby the compounds of the invention can be used in virus-induced diseases, especially acute and chronic persistent HBV viruses, according to the methods described herein or by methods known to those skilled in the art. Treatment of infection.

Claims

Claim

1. A compound represented by the formula (I) and its isomer (la), and an enantiomer thereof and a salt thereof:

 among them:

R ¹ is o-chloro, R ² is p-chloro, R ³ is dC ₄ alkyl, R ⁶ is thiazolyl-2-yl, X is methylene, and Z is morpholinyl.

2. A compound according to claim 1 and an enantiomer thereof and a salt thereof, wherein: R ¹ is o-chloro, R ² is p-chloro, R ³ is methyl or ethyl, and R ⁶ is Thiazolyl-2-yl, X is methylene, and Z is morpholinyl.

 3. A compound having the following structure, an enantiomer thereof, a tautomer thereof (la) and a salt thereof

 4. A method of preparing a compound of claim 1 comprising the steps of:

[A] First, the benzaldehyde represented by the formula (Π) is reacted with the β-ketoester represented by the formula (III). Forming a benzylidene compound represented by the formula (IV):

Wherein: I ¹ , R ² , R ³ , X and Z have the meanings as defined in claim 1, and then the benzylidene compound represented by the formula (IV) and the oxime represented by the formula (V) or Its salt, or other compound reacts,

Wherein R ⁶ has the meanings recited in claim 1;

 [B] reacting a compound of the formula (III) with an aldehyde (II) and hydrazine (V) or a salt thereof, or other compound by a one-step method,

 [C] When X in the formula (I) is a methylene group, the compound of the formula (VI) is reacted with morpholine (VII) or a salt thereof:

Wherein R ¹ , R ² , R ³ and R ⁶ have the above meanings, and Y is a nucleophilic substituent,

 [D] The aldehyde represented by the formula (II) is reacted with a compound of the formula (X) and an anthracene of the formula (V) or a salt thereof:

NH ₂ (X)

Wherein R ³ , X and Z have the meanings recited in claim 1.

 5. A compound of the formula (XII):

Wherein R ¹ , R ² , R ³ and R ⁶ have the meanings as described in claims 1-3 and the compounds are used as intermediates for the synthesis of compound (I) or (la).

 6. A compound for use in the control of a disease according to any one of claims 1-3.

 A pharmaceutical composition comprising at least one compound according to any one of claims 1 to 3, further containing other active pharmaceutical ingredient, as appropriate.

 8. Use of a compound according to claims 1 to 3 for the manufacture of a medicament for the treatment and prevention of a viral disease.

 9. Use of a compound according to any of claims 1 to 3 for the manufacture of a medicament for the treatment and prophylaxis of hepatitis B infection.

 10. A method of treating hepatitis B comprising administering to a mammal an effective amount of a compound of claim 1.

 A method of treating a disease caused by a hepatitis B infection, which comprises administering to a mammal an effective amount of the compound of claim 1.

 12. The method of claim 11 wherein the disease is hepatitis.

 13. The method of claim 11 wherein said disease is cirrhosis of the liver.

14. The method of claim 11 wherein said disease is hepatocellular carcinoma.

A pharmaceutical preparation comprising one or more compounds of claim 1 and a pharmaceutically acceptable carrier.

 16. A composition consisting of the following components:

 A) at least one dihydropyrimidine as claimed in any one of claims 1 to 3,

B) at least one HBV antiviral agent different from A and, where appropriate,

C) at least one immunomodulator.

 17. The composition of claim 16 wherein component B is a HBV polymerase inhibitor.

 18. The composition of claim 16 wherein component B is lamivudine.

 The composition according to claim 16, wherein component B is selected from the group consisting of a compound represented by the following formula and a salt thereof

 among them

R ¹ and R ² , independently of each other, dC ₄ -alkyl or, together with the gas atom at the position thereof, form a ring having 5 to 6 ring atoms including carbon and/or oxygen atoms,

R ³ to R ¹² are each independently hydrogen, halogen, alkyl, optionally substituted dC ₄ -alkoxy, nitro, cyano or trifluoromethyl,

R ¹³ is hydrogen, C _r C ₄ -fluorenyl, CrC ₇ -acyl or aralkyl, and X is halogen or an optionally substituted dC alkyl group.

20. The composition of claim 19, wherein the compound has the structure:

The composition according to any one of claims 16 to 20, wherein the immunomodulator C comprises an interferon.

 22. A composition comprising:

 A) the dihydropyrimidine of claims 1 to 3,

 B) lamivudine and, when appropriate,

 C) Interferon.

 23. Process for the preparation of a composition according to any one of claims 16 to 22, characterized in that the components A, B and, where appropriate, the components are combined or mixed in a suitable manner.

 24. A composition for controlling disease according to any one of claims 16 to 22.

 25. A medicament comprising at least one composition according to any one of claims 16 to 22, and, where appropriate, other active pharmaceutical ingredients.

 Use of a composition according to any one of claims 16 to 22 for the manufacture of a medicament for the treatment or prevention of a viral disease.

 27. Use of a composition according to any of claims 16 to 22 for the manufacture of a medicament for the treatment and prevention of hepatitis B infection.