RU2447074C2 - Pteridines effective as hcv inhibitors and methods for producing pteridines - Google Patents

Abstract

FIELD: medicine, pharmaceutics.

SUBSTANCE: present invention refers to the use of pteridines of formula (I), including new pteridine derivatives of formula (VII), their pharmaceutically acceptable salts or esters as HCV-replication inhibitors. The invention also refers to pharmaceutical compositions based on the compounds stated above and a method for HCV-replication inhibition. In general formula

R¹ independently means hydrogen; L means -NR⁸-, -NR⁸-C_1-6alkandiyl or

wherein a cycle indicated by a dashed line together with N and Z forms a cycle containing 5-8 members, and can be substituted by an aromatic monocyclic heterocycle having 6 members in a cycle which contains as cycle members one heteroatom specified in nitrogen, wherein said L cycle is attached to a pteridine cycle by a nitrogen atom; Z means N or CH; R² means hydroxyC_1-6alkyl, phenyl, Het², wherein said phenyl and Het² are independently optionally substituted by one or more substitutes specified in a group consisting of C_1-4alkyl, -COOR⁷, morpholin-4-yl, -CONR^4aR^4b, wherein R^4a and R^4b represent hydrogen, Het¹-C_1-4alkyl; or NR^4aR^4b, wherein R^4a and R^4b represent hydrogen, hydroxy C_1-4alkyl, or can optionally form together with a nitrogen atom whereto attached, a 5-6-members saturated cycle, optionally containing an oxygen atom; R³ means phenyl, Het² . The other radical values are specified in the patent claim.

EFFECT: preparing the compounds applicable as HCV-replication inhibitors.

16 cl, 1 dwg, 4 tbl, 26 ex

Description

The present invention relates to the use of pteridins as inhibitors of HCV replication, as well as the use of these compounds in pharmaceutical compositions intended for the treatment or control of HCV infections. In addition, the present invention relates to compounds, as such, and the use of these compounds in the form of drugs. The present invention also relates to methods for producing such compounds, pharmaceutical compositions comprising these compounds, and combinations of these compounds with other anti-HCV agents.

After discovering in 1989 as an agent involved in most non-A and B non-A and B viral hepatitis (Choo et al., Science 244, 359-362, 1989), hepatitis C virus (HCV) has been the focus of a variety of medical studies ( Lauer GM and Walker BD, New Eng. J Med. 345, 41-52, 2001). HCV is a member of the flaviviridae family of viruses of the genus hepacivirus and a closely related genus of flavivirus, including a number of viruses involved in human diseases, such as dengue virus and yellow fever virus, and the family of animal pestivirus, including cattle diarrhea virus (BVDV). HCV is a positively sensitive virus with single-stranded RNA and a genome of approximately 9600 bases. The genome includes both 5 'and 3' untranslated regions that borrow secondary RNA structures, and a central open reading frame that encodes a single polyprotein of approximately 3010-3030 amino acids. The polyprotein encodes ten gene products generated from the preceding polyprotein by organized series of co- and post-translational endoproteolytic cleavages mediated by proteases of both the host and the virus. Viral structural proteins include a core nucleocapsid protein and two envelope glycoproteins, E1 and E2. Non-structural (NS) proteins encode some essential viral enzymatic functions (helicase, polymerase, protease), as well as proteins of unknown function. Replication of the viral genome is mediated by an RNA-dependent DNA polymerase encoded by non-structural protein 5b (NS5B). Chimpanzee infection models have shown that in addition to polymerase, the functions of viral helicase and protease, which are both encoded by the bifunctional NS3 protein, are necessary for the replication of HCV RNA (Kolykhalov AA, Mihalik K., Feinstone, SM and Rice, CM J Virol. 74, 2046-2051, 2000). In addition to the NS3 serine protease, HCV also encodes a metalloproteinase in the NS2 region.

HCV replicates predominantly in hepatocytes, but is not completely cytopathic, leading to persistent infection. In particular, the lack of a powerful T-lymphocyte response and the high susceptibility of the virus to mutation seem to contribute to a high rate of chronic infection. There are 6 major HCV genotypes and over 50 subtypes that are geographically distributed. HCV type 1 is the dominant genotype in the United States and Europe. For example, type 1 HCV accounts for up to 70-75 percent of all HCV infections in the United States. The extensive genetic heterogeneity of HCV has important diagnostic and clinical manifestations, possibly explaining the difficulties in developing a vaccine and the lack of response to treatment. An estimated 170 million people worldwide are infected with the hepatitis C virus (HCV). Following the primary acute infection, most infected patients develop chronic hepatitis, which can progress to liver fibrosis leading to cirrhosis, end-stage liver disease, and HCC (hepatocellular carcinoma) (National Institutes of Health Consensus Development Conference Statement: Management of Hepatitis C. Hepatology 36, 5 Suppl. S3-S20, 2002). Hepatic cirrhosis due to HCV infection causes about 10,000 deaths per year in the United States alone and causes liver transplants. HCV transmission can occur by contact with contaminated blood or blood products, for example after blood transfusion or intravenous administration of a drug. The introduction of diagnostic tests used in blood screening has led to a decrease in the post-transfusion spread of HCV. However, given the slow progression to the final stage of liver disease, existing infections will continue to cause problems associated with severe drug burden and pose a heavy economic burden for decades (Kim WR Hepatology, 36, 5 Suppl. S30-S34, 2002 )

The treatment of this chronic disease does not satisfy clinical needs, as the current therapy is only partially effective and is limited by undesirable side effects.

Modern HCV therapy is based on (pegylated) alpha interferon (IFN-α) in combination with ribavirin. Such combination therapy causes a stable virological response in more than 40% of patients infected with genotype 1 virus and about 80% of patients infected with genotype 2 and 3 virus. In addition to limited efficacy against type 1 HCV, combination therapy has significant side effects and is poorly tolerated by many patients. For example, according to the registration protocols for testing pegylated interferon and ribavirin, significant side effects lead to discontinuation of treatment for approximately 10-14 percent of patients. The main side effects of combination therapy include flu-like symptoms, hematologic disorders and neuropsychiatric symptoms. The development of more effective, convenient and better tolerated treatments is a major public health concern.

So, there is a significant medical need for low molecular weight compounds that lead to inhibition of HCV replication.

Surprisingly, pteridine derivatives have antiviral activity against mammals infected with HCV, in particular such derivatives inhibit HCV replication. These compounds are useful, therefore, in the treatment or fight against HCV infections.

US 20040038856 describes methods for treating fibroproliferative disorders associated with TGF-β signaling by administering non-peptide, low molecular weight TGF-β inhibitors specifically binding to Type I TGF-β receptor (TGF-β-R1). Inhibitors are predominantly quinazoline derivatives.

WO 04/048930 further describes methods of counteracting the loss of β-adrenergic sensitivity in the β-adrenergic signal transduction pathway by administering an effective amount of a compound capable of inhibiting the transmission of the TGF-β signal through the TGF-β receptor.

WO 04/065392 relates to fused pyridines and pyrimidines and the use of these compounds as ALK-5 receptor ligands. In particular, the invention describes therapeutically active substituted quinoline and quinazoline compounds, the use of said compounds in therapy, especially in the treatment or prevention of disorders characterized by overproduction of transforming growth factor β (TGF-β), and pharmaceutical compositions for use in said therapy.

After the initial exposure to hepatitis C virus, HCV RNA can be detected in the blood after 1-3 weeks. On average, over 50 days, almost all patients develop impaired liver cells. Most patients are asymptomatic and anicteric. Only 25-35 percent show malaise, weakness or loss of appetite, and some become icteric. Antibodies to HCV (anti-HCV) can almost always be detected during illness. Anti-HCV can be detected in 50-70 percent of patients at the onset of symptoms and in approximately 90 percent of patients 3 months after infection. HCV infection goes away without treatment in only 15 percent of cases. A return to normal is characterized by the disappearance of HCV RNA in the blood and a return of liver enzymes to normal.

About 85 percent of HCV-infected patients cannot get rid of the virus within 6 months and chronic hepatitis develops with persistent, although sometimes intermittent, viremia. This ability to produce chronic hepatitis is one of the most striking features of HCV infection. Chronic hepatitis C is usually a progressively developing process, progressing, if it does, at a low rate without symptoms or physical signs in most patients during the first two decades after infection. Symptoms first appear in many patients with chronic hepatitis C during the development of advanced liver disease.

In chronic hepatitis, cells in the inflammatory zone infiltrate portal channels and can also collect in small clusters in the parenchyma. The latter case is usually accompanied by focal necrosis of liver cells. The marginal zone of the parenchyma and portal canals can become inflamed in the presence of liver cell necrosis in this area (border hepatitis). If and when the disease progresses, inflammation and death of liver cells can lead to fibrosis. A mild form of fibrosis does not extend beyond the portal canals and directly adjacent parenchyma. The more severe form of fibrosis leads to the formation of bridges between the portal canals and between the portal canals and the hepatic veins. Such fibrosis can progress to cirrhosis, defined as the state of diffuse fibrosis, in which the fibrous septa secretes clusters of liver cells into nodules. The size of fibrosis determines the stage of the disease and can easily be determined. Severe fibrosis and necrotic inflammatory changes precede progression to cirrhosis. When cirrhosis is established, complications that are secondary to liver failure and / or portal hypertension, such as spillage of bile, ascites, bleeding from varicose veins, and encephalopathy, can follow. The development of any of these complications means the transition from compensated to decompensated cirrhosis.

Chronic hepatitis C infection leads to cirrhosis in at least 20 percent of patients within 2 decades of infection. Cirrhosis and the final stage of liver disease can sometimes develop rapidly, especially in patients with concomitant alcoholism. Chronic HCV infection is associated with an increased risk of liver cancer. The underlying principle is that hepatocellular carcinoma (HCC) appears along with background inflammation, and the regeneration associated with chronic hepatitis lasts for about 3 or more decades. In most cases, HCV-induced HCC is observed in the presence of cirrhosis.

Liver fibrosis is one of the processes that occurs when the liver is affected. Such a lesion may result from the action of a virus (for example, chronic hepatitis type B or C) or other liver infections (for example, parasites, bacteria); chemicals (e.g. pharmaceuticals, recreational drugs, excessive amounts of alcohol, exposure to environmental pollutants); immune processes (e.g., autoimmune hepatitis); metabolic disorders (e.g., lipid, glycogen or metabolic disorders) or growth of carcinoma (primary or secondary hepatic carcinoma). Fibrosis is both a sign of liver damage and a potential factor contributing to liver failure through increasing cirrhosis of the liver.

Inhibition of the TGF kinase family has been found to be useful in the treatment of fibro-proliferative disorders, including liver fibrosis. However, as indicated above, liver fibrosis can be caused by various etiological agents, including hepatitis C. Most importantly, liver fibrosis is a specific condition in the progression of the disease in patients infected with HCV.

Unexpectedly, it was found that the compounds of the present invention inhibit HCV replication. HCV replication means the process of reproducing or creating copies of HCV RNA. For the purposes of this invention, replication of HCV means both replication of the HCV virus in general and replication of the HCV RNA genome.

It is important to treat HCV-infected patients in the early stages in order to avoid disease progression, thus avoiding the development of chronic hepatitis, liver fibrosis, cirrhosis, hepatocellular carcinoma (HCC) or death.

In addition, the compounds of the invention are valuable in that they can reduce the patient's HCV viral load to undetectable levels.

Description of drawings

Figure 1 illustrates the average concentration in plasma and tissues (n = 3) of compound No. 21 after a single oral administration at 20 mg basic equivalent / kg to male Swiss SPF mice (CD1).

Description of the invention

Thus, the present invention relates to the use of a compound of formula (I) for the manufacture of a medicament suitable for inhibiting HCV activity in mammals infected with HCV. The specified compound is a pteridine of the formula (I):

its N -oxide, salt, stereoisomer, racemic mixture, prodrug, ester or metabolite, where

R ¹ independently means hydrogen, amino, mono- or disubstituted amino, where amino substituent (s) may be selected from the group consisting of C _1-6 alkyl, C _2-6 alkenyl,

C _2-6 alkynyl, C _1-4 alkoxyC _1-4 alkyl, diC _1-4 alkilaminoC _1-4 alkyl, piperidin-1-ilC _1-4 alkyl arilC _1-6 alkyl, wherein the aryl group may further be substituted with C _1-4 alkyl or C _1-4 alkoxy;

L is —NR ⁸ -, —NR ⁸ —C _1-6 alkanediyl—, —NR ⁸ —CO — C _1-6 alkanediyl—, —NR ⁸ —SO ₂ —C _1-6 alkanediyl—, —O—, - OC _1-6 alkanediyl-, -O-CO-,

-O-CO-C _1-6 alkanediyl-, -S-, -SC _1-6 alkanediyl- or

where the cycle indicated by the dashed line, together with N and Z, forms a Het ¹ cycle containing 5-8 members, including the cyclic members of N and Z, and where the specified L cycle is attached to the pteridine cycle through a nitrogen atom;

Z is N or CH;

R ² is hydrogen, hydroxyC _1-6 alkyl, C _3-7 cycloalkyl, aryl, Het ¹ or Het ² , where each C _3-7 cycloalkyl, aryl, Het ¹ and Het ^{2 is} independently optionally substituted with one or more substituents selected from a group comprising C _1-4 alkyl,

C _2-4 alkenyl, C _2-4 alkynyl, polyhalogen C _1-4 alkyl, halogen, cyano, nitro, -COR ⁶ , -COOR ⁷ , -CONR ^4a R ^4b , -OR ⁷ , -OCOR ⁶ , -OCONR ^4a R ^4b , -NR ^4a R ^4b , -NR ^4a COR ⁶ , -NR ^4a CONR ^4a R ^4b , -NR ^4a SOR ⁵ , -NR ^4a SO ₂ R ⁵ , -SR ⁵ , -SOR ⁷ , -SO ₂ R ⁵ , -SO ₃ R ⁷ ,

—SO ₂ NR ^4a R ^4b , morpholin-4-yl, phenyl, aminophenyl and aminophenylcarbonyl, and wherein C _1-4 alkyl may be further substituted with —COOR ⁷ ;

R ³ means C _1-6 alkyl, C _3-7 cycloalkyl, aryl, aryl C _1-6 alkyl, Het ¹ , Het ² or Het ² -C _1-6 alkyl, each of which is independently optionally substituted with one or more substituents selected from the group consisting of C _1-4 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, polyhalogen C _1-4 alkyl, halogen, cyano, nitro, -COR ⁶ , -COOR ⁷ ,

-CONR ^4a R ^4b , -OR ⁷ , -OCOR ⁶ , -OCONR ^4a R ^4b , -NR ^4a R ^4b , -NR ^4a COR ⁶ , -NR ^4a COOR ⁷ , -NR ^4a CONR ^4a R ^4b , -NR ^4a SOR ⁵ , -NR ^4a SO ₂ R ⁵ , -SR ⁵ , -SOR ⁷ , -SO ₂ R ⁵ , -SO ₃ R ⁷ , -SO ₂ NR ^4a R ^4b ; and where R ^4a and R ^4b may optionally form, together with the nitrogen atom to which they are bonded, a 5-8 membered saturated, unsaturated or partially unsaturated ring, optionally containing one or two additional heteroatoms;

each of R ^4a and R ^4b independently means hydrogen, C _1-4 alkyl, hydroxyC _1-4 alkyl, Het ¹ C _1-4 alkyl, polyhaloC _1-4 alkyl, cyano or nitro;

each of R ⁵ independently means hydrogen or C _1-4 alkyl;

each of R ⁶ independently means hydrogen or C _1-4 alkyl;

each of R ⁷ independently means hydrogen or C _1-4 alkyl and

R ⁸ means hydrogen, C _1-10 alkyl, C _2-10 alkenyl, C _2-10 alkynyl,

C _1-10 alkylcarbonyl, amino C _1-10 alkyl, aryl, arylcarbonyl,

aryl C _1-10 alkyl, Het ¹ , Het ¹ C _1-6 alkyl or a protective group, where aryl is optionally substituted with 1-3 substituents selected from the group consisting of C _1-4 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, C _1-4 alkylcarbonyl, phenyl, C _1-4 alkylphenyl, phenylcarbonyl, aminophenyl,

amino C _1-4 alkylphenyl, aminophenylcarbonyl, halogen, —OR ⁶ , —NR ^4a R ^4b ,

-SR ⁵ , -SOR ⁵ , -NR ^4a SOR ⁵ , -NR ^4a SO ₂ R ⁵ , -SO ₂ R ⁵ , -OCOR ⁶ , -NR ^4a COR ⁶ ,

-NR ^4a CONR ^4a R ^4b , -NR ^4a COOR ⁶ , -OCONR ^4a R ^4b , -COOR ⁶ , -SO ₃ R ⁶ , -CONR ^4a R ^4b ,

-SO ₂ NR ^4a R ^4b , cyano, polyhaloC _1-4 alkyl and nitro;

Het ¹ as a group or part of a group means a saturated or unsaturated monocyclic, bicyclic or tricyclic heterocycle having preferably 3-12 members in a cycle, preferably 5-10 members in a cycle and more preferably 5-8 members in a cycle that contains as members one or more heteroatoms selected from the group consisting of nitrogen, oxygen or sulfur, and which is optionally substituted at one or more carbon atoms with substituents selected from the group consisting of C _1-6 alkyl,

C _1-6 alkoxy, halogen, hydroxy, oxo, optionally mono- or disubstituted amino, nitro, cyano, polyhaloC _1-4 alkyl, carboxyl, C _1-6 alkoxycarbonyl, C _3-7 cycloalkyl, optionally mono- or disubstituted aminocarbonyl, methylthio, methylsulfonyl, aryl and saturated or partially unsaturated monocyclic, bicyclic or tricyclic heterocycle having 3-12 members in the cycle, which contains as members of the cycle one or more heteroatoms selected from the group comprising nitrogen, oxygen or sulfur, and optional alternate E at any amino function are hydrogen, or C _1-4 alkyl;

Het ² as a group or part of a group means an aromatic monocyclic, bicyclic or tricyclic heterocycle having 3-14 members in a cycle, preferably 5-10 members in a cycle, and more preferably 5-6 members in a cycle that contains one or more heteroatoms each independently selected from the group consisting of nitrogen, oxygen or sulfur and which is optionally substituted on one or more carbon atoms with substituents selected from the group consisting of C _1-6 alkyl, optionally mono- or d substituted aminoC _1-6 alkyl, hydroxyC _1-6 alkyl, C _1-6 alkoxy, halogen, hydroxy, optionally mono- or disubstituted amino, nitro, cyano, polyhaloC _1-4 alkyl, carboxyl, C _1-6 alkoxycarbonyl, C _{3 -7} cycloalkyl, optionally mono- or disubstituted aminocarbonyl, methylthio, methylsulfonyl, aryl, Het ¹ and an aromatic monocyclic, bicyclic or tricyclic heterocycle having 3-12 members in the cycle, with hydrogen or C _1-4 being optional substituents on any amino functions alkyl; and

aryl as a group or part of a group means phenyl.

The present invention also relates to the use of a compound of formula (II) for the manufacture of a medicament suitable for inhibiting HCV activity in a mammal infected with HCV. The specified compound is a pteridine of the formula (II):

its N -oxide, salt, stereoisomer, racemic mixture, prodrug, ester or metabolite, where R ¹ , R ³ , R ^4a , R ^4b , R ⁵ , R ⁶ , R ⁷ , R ⁸ , Het ¹ and Het ² have the above values; Where

R ⁹ means C _1-4 alkyl, C _2-4 alkenyl, C _2-4 alkynyl, polyhalo C _1-4 alkyl, halogen, cyano, nitro, —COR ⁶ , —COOR ⁷ , —CONR ^4a R ^4b , —OR ⁷ , -OCOR ⁶ ,

-OCONR ^4a R ^4b , -NR ^4a R ^4b , -NR ^4a COR ⁶ , -NR ^4a CONR ^4a R ^4b , -NR ^4a SOR ⁵ , -NR ^4a SO ₂ R ⁵ , -SR ⁵ , -SOR ⁷ , -SO ₂ R ⁵ , —SO ₃ R ⁷ , —SO ₂ NR ^4a R ^4b , morpholin-4-yl, phenyl, aminophenyl or aminophenylcarbonyl and wherein C _1-4 alkyl may be further substituted with —COOR ⁷ ; and

n is 0, 1, 2, 3, or 4.

The present invention also relates to the use of a compound of formula (III) for the manufacture of a medicament suitable for inhibiting HCV activity in an HCV-infected mammal. The specified compound is a pteridine of the formula (III):

its N -oxide, salt, stereoisomer, racemic mixture, prodrug, ester or metabolite, where R ¹ , L, R ² , R ^4a , R ^4b , R ⁵ , R ⁶ , R ⁷ , R ⁸ , Het ¹ and Het ² have the above meanings; Where

R ¹⁰ means C _1-4 alkyl, C _2-4 alkenyl, C _2-4 alkynyl,

polyhalogen C _1-4 alkyl, halogen, cyano, nitro, -COR ⁶ , -COOR ⁷ ,

-CONR ^4a R ^4b , -OR ⁷ , -OCOR ⁶ , -OCONR ^4a R ^4b , -NR ^4a R ^4b , -NR ^4a COR ⁶ , -NR ^4a COOR ⁷ , -NR ^4a CONR ^4a R ^4b , -NR ^4a SOR ⁵ , -NR ^4a SO ₂ R ⁵ , -SR ⁵ , -SOR ⁷ , -SO ₂ R ⁵ , -SO ₃ R ⁷ and

-SO ₂ NR ^4a R ^4b and

m is 0, 1, 2, 3 or 4.

The present invention also relates to the use of a compound of formula (IV) for the manufacture of a medicament suitable for inhibiting HCV activity in a mammal infected with HCV. The specified compound is a pteridine of the formula (IV):

its N -oxide, salt, stereoisomer, racemic mixture, prodrug, ester or metabolite, where R ¹ , R ^4a , R ^4b , R ⁵ , R ⁶ , R ⁷ , R ⁸ , Het ¹ and Het ² have the above meanings; Where

R ⁹ means C _1-4 alkyl, C _2-4 alkenyl, C _2-4 alkynyl,

polyhalogen C _1-4 alkyl, halogen, cyano, nitro, -COR ⁶ , -COOR ⁷ ,

-CONR ^4a R ^4b , -OR ⁷ , -OCOR ⁶ , -OCONR ^4a R ^4b , -NR ^4a R ^4b , -NR ^4a COR ⁶ ,

-NR ^4a CONR ^4a R ^4b , -NR ^4a SOR ⁵ , -NR ^4a SO ₂ R ⁵ , -SR ⁵ , -SOR ⁷ , -SO ₂ R ⁵ , -SO ₃ R ⁷ ,

—SO ₂ NR ^4a R ^4b , morpholin-4-yl, phenyl, aminophenyl or aminophenylcarbonyl, and wherein C _1-4 alkyl may be further substituted with —COOR ⁷ ;

R ¹⁰ means C _1-4 alkyl, C _2-4 alkenyl, C _2-4 alkynyl,

polyhalogen C _1-4 alkyl, halogen, cyano, nitro, -COR ⁶ , -COOR ⁷ ,

-CONR ^4a R ^4b , -OR ⁷ , -OCOR ⁶ , -OCONR ^4a R ^4b , -NR ^4a R ^4b , -NR ^4a COR ⁶ , -NR ^4a COOR ⁷ , -NR ^4a CONR ^4a R ^4b , -NR ^4a SOR ⁵ , -NR ^4a SO ₂ R ⁵ , -SR ⁵ , -SOR ⁷ , -SO ₂ R ⁵ , -SO ₃ R ⁷ and -SO ₂ NR ^4a R ^4b ;

n is 0, 1, 2, 3 or 4 and

m is 0, 1, 2, 3 or 4.

The present invention also relates to the use of a compound of formula (V) for the manufacture of a medicament suitable for inhibiting HCV activity in a mammal infected with HCV. These compounds are pteridines of the formula (V):

its salt, stereoisomer and racemic mixture, where

R ¹ means hydrogen or amino;

R ⁸ is hydrogen, C _1-6 alkyl, amino C _1-4 alkyl, phenyl C _1-4 alkyl, pyrrolidin-1-yl C _1-4 alkyl or C _1-6 alkoxycarbonyl;

each of R ⁹ is independently hydrogen, C _1-4 alkyl, —COR ⁶ , —COOR ^7, or —CONR ^4a R ^4b ;

n is 0, 1, 2, 3 or 4;

R ¹¹ means hydrogen, halogen or —NR ^4a R ^4b , where R ^4a and R ^4b may optionally form, with the nitrogen atom to which they are bonded, a 5-8 membered saturated, unsaturated or partially unsaturated ring, optionally containing one or two additional heteroatoms;

R ¹² means hydrogen, halogen, C _1-4 alkyl or

polyhaloC _1-4 alkyl;

R ⁶ means hydrogen or C _1-4 alkyl;

R ⁷ means hydrogen or C _1-4 alkyl and

R ^4a and R ^4b independently mean hydrogen, C _1-4 alkyl, 2-oxopyrrolidin-1-yl C _1-4 alkyl.

One of the embodiments relates to the use of compounds of the above formulas (II), (IV) or (V), where n is 1.

In another aspect, the invention relates to a method for inhibiting HCV replication in a mammal infected with HCV, said method comprising administering an HCV inhibitory effective amount of a compound of formula (I), (II), (III), (IV) or (V) as indicated above or defined hereinafter. In a noteworthy embodiment, a method of inhibiting HCV replication in a mammal infected with HCV comprises administering an HCV inhibitory effective amount of a compound of formula (II), (IV) or (V), where n is 1.

According to another aspect, the invention relates to a method for treating a HCV-infected mammal, said method comprising administering an HCV-inhibiting effective amount of a compound of formula (I), (II), (III), (IV) or (V) as described above or as defined herein in further. In a noteworthy embodiment, a method of treating a mammal infected with HCV comprises administering an HCV-inhibiting effective amount of a compound of formula (II), (IV) or (V), where n is 1.

In another embodiment, the present invention relates to the use of a compound of formula (VI) for the manufacture of a medicament for inhibiting HCV replication in a mammal infected with HCV. The specified compound is a pteridine of formula (VI):

its salt, stereoisomer and racemic mixture, where R ¹ , R ⁸ , R ⁹ , R ¹¹ , R ¹² , R ⁶ have the above meanings.

In yet another embodiment, the present invention relates to a method for inhibiting HCV replication in a mammal infected with HCV, said method comprising administering an HCV inhibitory effective amount of a compound of formula (VI) as described above or as defined hereinafter.

In another embodiment, the present invention relates to a method for treating a mammal infected with HCV, said method comprising administering an HCV-inhibiting effective amount of a compound of formula (VI) as described above or as defined hereinafter.

Further additional embodiments of the invention relate to the use of compounds of formulas (V) or (VI) for the manufacture of a medicament for inhibiting HCV replication in a mammal infected with HCV. These compounds are pteridines of the formula (V) or (VI), where an acceptable value of n is 1 and

R ¹ means hydrogen or amino;

R ⁸ is hydrogen, C _1-6 alkyl, phenyl C _1-4 alkyl;

R ⁹ is hydrogen, C _1-4 alkyl, —COR ⁶ , —COOR ^7, or —CONR ^4a R ^4b ;

R ¹¹ means hydrogen, fluorine or pyrrolidin-1-yl;

R ¹² is halogen, C _1-4 alkyl or polyhalo C _1-4 alkyl;

R ⁶ means hydrogen or C _1-4 alkyl;

R ⁷ means hydrogen or C _1-4 alkyl and

R ^4a and R ^4b independently mean hydrogen, C _1-4 alkyl, 2-oxopyrrolidin-1-yl C _1-4 alkyl.

Other embodiments of the invention relate to a method for inhibiting HCV replication in an HCV-infected mammal and a method for treating a HCV-infected mammal, the methods comprising administering an HCV-inhibiting effective amount of a compound of formula (V) or (VI), wherein an acceptable n value is 1 and R ¹ , R ⁸ , R ⁹ , R ¹¹ , R ¹² take the meanings indicated in the above paragraph.

Further embodiments of the invention relate to the use of compounds of formula (V) or (VI) for the manufacture of a medicament for inhibiting HCV replication in a mammal infected with HCV. These compounds are pteridines of the formula (V) or (VI), where an acceptable value of n is 1 and

R ¹ means hydrogen;

R ⁸ is hydrogen, C _1-6 alkyl, phenyl C _1-4 alkyl;

R ⁹ is hydrogen, C _1-4 alkyl or —COOR ⁷ ;

R ¹¹ means fluorine or pyrrolidin-1-yl;

R ¹² means halogen or C _1-4 alkyl and

R ⁷ means hydrogen or C _1-4 alkyl.

Thus, further embodiments of the invention relate to a method of inhibiting HCV replication in an organism of a mammal infected with HCV, and a method for treating a mammal infected with HCV, these methods include administering an HCV-inhibiting effective amount of a compound of formula (V) or (VI), where an acceptable value n is 1 and R ¹ , R ⁸ , R ⁹ , R ¹¹ , R ¹² are as defined above.

In another embodiment, the present invention relates to a pteridine of formula (VII):

its salt, stereoisomer and racemic mixture, where

R ¹ means hydrogen or amino;

R ⁸ is hydrogen, C _1-6 alkyl, phenyl C _1-4 alkyl;

R ⁹ is hydrogen, C _1-4 alkyl, —COR ⁶ , COOR ^7, or —CONR ^4a R ^4b ;

R ⁶ independently means hydrogen or C _1-4 alkyl;

each of R ⁷ independently means hydrogen or C _1-4 alkyl and

each of R ^4a and R ^4b independently means hydrogen, C _1-4 alkyl, 2-oxopyrrolidin-1-yl C _1-4 alkyl;

with the proviso that when R ⁸ is hydrogen, R ^{9 is} not hydrogen.

In another embodiment, the present invention relates to a pteridine of formula (VII), a salt thereof, a stereoisomer, and a racemic mixture, wherein

R ⁸ is C _1-6 alkyl, phenyl C _1-4 alkyl;

R ¹ , R ^4a , R ^4b R ⁶ , R ⁷ and R ⁹ take the values listed above.

In another embodiment, the present invention relates to a pteridine of formula (VII), a salt thereof, a stereoisomer, and a racemic mixture, wherein

R ⁹ is C _1-4 alkyl, —COR ⁶ , COOR ^7, or —CONR ^4a R ^4b ;

R ¹ , R ^4a , R ^4b R ⁶ , R ⁷ and R ⁸ are as defined above.

In yet another embodiment, the present invention relates to pteridine of formula (VIII):

its salt, stereoisomer and racemic mixture, where

R ¹ independently means hydrogen or amino;

R ⁸ is hydrogen, C _1-6 alkyl, phenyl C _1-4 alkyl;

R ⁹ is hydrogen, C _1-4 alkyl, —COR ⁶ , COOR ^7, or —CONR ^4a R ^4b ;

R ⁶ independently means hydrogen or C _1-4 alkyl;

each of R ⁷ independently means hydrogen or C _1-4 alkyl and

each of R ^4a and R ^4b independently means hydrogen, C _1-4 alkyl, 2-oxopyrrolidin-1-yl C _1-4 alkyl;

with the proviso that when R ⁸ is hydrogen, R ^{9 is} not hydrogen.

In another embodiment, the present invention relates to a pteridine of formula (VIII), a salt thereof, a stereoisomer, and a racemic mixture, wherein

R ⁹ is C _1-4 alkyl, —COR ⁶ , COOR ^7, or —CONR ^4a R ^4b ;

R ¹ , R ^4a , R ^4b , R ⁶ , R ⁷ and R ⁸ take the values listed in the preceding paragraph.

In another embodiment, the present invention relates to a pteridine of formula (VIII), a salt thereof, a stereoisomer, and a racemic mixture, wherein

R ⁸ is C _1-6 alkyl, phenyl C _1-4 alkyl;

R ¹ , R ^4a , R ^4b , R ⁶ , R ⁷ and R ⁹ take the values listed above.

In a further embodiment, the present invention relates to pteridine of the above formula (VII) or (VIII), where

R ¹ means hydrogen;

R ⁸ means hydrogen;

R ⁹ means C _1-4 alkyl.

In a further embodiment, the present invention relates to pteridine of the above formula (VII) or (VIII), where

R ¹ means hydrogen;

R ⁸ means C _1-6 alkyl;

R ⁹ means hydrogen.

A method for treating clinical conditions caused by HCV infection in a mammal, said method comprises administering an HCV inhibitory effective amount of a compound of formula (V), wherein R ¹ , R ⁸ , R ⁹ , R ¹¹ , R ¹² are as defined below.

The method of the preceding paragraph, wherein the clinical conditions are different from liver fibrosis.

The compounds of formulas (I), (II), (III), (IV), (V), (VI), (VII) and (VIII) are active against the HCV virus and are therefore useful as a medicine and for the manufacture of a medicine for the prevention, treatment or suppression of infection, clinical conditions or disease caused by HCV infection.

The compounds of formulas (I), (II), (III), (IV), (V), (VI), (VII) and (VIII) are active against the HCV virus and are therefore useful as a medicine and for the manufacture of a medicine for the prevention, treatment or suppression of clinical conditions caused by HCV infection, other than liver fibrosis.

The term "C _1-2 alkyl", as a group or part of a group, means linear and branched, saturated hydrocarbon radicals with 1-2 carbon atoms, such as, for example, methyl, ethyl and the like.

The term "C _1-4 alkyl", as a group or part of a group, means linear and branched, saturated hydrocarbon radicals with 1-4 carbon atoms, such as, for example, the groups indicated for the term C _1-2 alkyl, and propyl, butyl, 2-methylpropyl and the like.

The term “C _1-6 alkyl”, as a group or part of a group, means linear and branched, saturated hydrocarbon radicals with 1-6 carbon atoms, such as, for example, the groups indicated for the term C _1-4 alkyl, and pentyl, hexyl, 2-methylbutyl, 3-methylpentyl and the like.

The term "C _1-10 alkyl", as a group or part of a group, means linear and branched, saturated hydrocarbon radicals with 1-10 carbon atoms, such as, for example, the groups indicated for the term C _1-6 alkyl, and heptyl, octyl, nonyl, decyl and the like.

The term “C _2-4 alkenyl,” as a group or part of a group, means linear and branched, hydrocarbon radicals having saturated carbon-carbon bonds and at least one double bond and containing from 2 to 4 carbon atoms, such as for example, ethenyl, prop-1-enyl, but-1-enyl, but-2-enyl and the like. Preferred are C _2-4 alkenyls with one double bond.

The term “C _2-6 alkenyl,” as a group or part of a group, means linear and branched, hydrocarbon radicals having saturated carbon-carbon bonds and at least one double bond and containing from 2 to 6 carbon atoms, such as for example, the groups indicated for the term C _2-4 alkenyl and pent-1-enyl, pent-2-enyl, hex-1-enyl, hex-2-enyl, hex-3-enyl, 1-methylpent-2 -enyl and the like. Preferred are C _2-6 alkenyls with one double bond.

The term “C _2-10 alkenyl,” as a group or part of a group, means linear and branched, hydrocarbon radicals having saturated carbon-carbon bonds and at least one double bond and containing from 2 to 10 carbon atoms, such as for example, groups indicated for the term C _2-6 alkenyl, and hept-1-enyl, hept-2-enyl, 2-methylhept-1-enyl, oct-3-enyl, non-4-enyl, 1-methylnon -2-enyl and the like. Preferred are C _2-10 alkenyls with one double bond.

The term “C _2-4 alkynyl,” as a group or part of a group, means linear and branched, hydrocarbon radicals having saturated carbon-carbon bonds and at least one triple bond and containing from 2 to 4 carbon atoms, such as for example, ethynyl, prop-1-ynyl, but-1-ynyl, but-2-ynyl and the like. Preferred are C _2-4 alkynyls with one triple bond.

The term “C _2-6 alkynyl,” as a group or part of a group, means linear and branched, hydrocarbon radicals having saturated carbon-carbon bonds and at least one triple bond and containing from 2 to 6 carbon atoms, such as for example, the groups indicated for the term C _2-4 alkynyl and pent-1-ynyl, pent-2-ynyl, hex-1-ynyl, hex-2-ynyl, hex-3-ynyl, 1-methylpent-2 -inyl, pent-2-en-4-ynyl and the like. Preferred are C _2-6 alkynyls with one triple bond.

The term “C _2-10 alkynyl,” as a group or part of a group, means linear and branched, hydrocarbon radicals having saturated carbon-carbon bonds and at least one triple bond and containing from 2 to 10 carbon atoms, such as for example, the groups indicated for the term C _2-6 alkynyl and hept-1-ynyl, hept-2-ynyl, 2-methylhept-1-ynyl, oct-3-ynyl, non-4-ynyl, 1-methylnon -2-ynyl and the like. Preferred are C _2-10 alkynyls with one triple bond.

The term “C _1-6 alkanediyl”, as a group or part of a group, means divalent linear and branched hydrocarbons containing from 1 to 6 carbon atoms, such as, for example, methanediyl, 1,2-ethanediyl or 1,1-ethanediyl, 1,3-propanediyl, 1,3-butanediyl, 1,4-butanediyl, 1,3-pentanediyl, 1,5-pentanediyl, 1,4-hexanediyl, 1,6-hexanediyl and the like.

The term “C _3-7 cycloalkyl” refers to a group comprising cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl and cycloheptyl.

The term "aryl" is intended to include phenyl or naphthyl as a group or part of a group. In a preferred embodiment, the term “aryl”, as a group or part of a group, means phenyl.

The term “halogen” refers to a group comprising fluoro, chloro, bromo or iodo.

As used above and hereinafter, “polyhaloC _1-4 alkyl,” as a group or part of a group, means a mono- or polyhalo-substituted C _1-4 alkyl, for example 1,1,1-trifluoroethyl, 1,1-difluoroethyl, referred to below polyhalogenomethyl groups and the like. A preferred subgroup of polyhaloC _1-4 alkyl is polyhalogenomethyl, where the latter, as a group or part of a group, is defined as mono- or polyhalo-substituted methyl, in particular methyl with one or more fluorine atoms, for example difluoromethyl or trifluoromethyl. In the case where more than one halogen atom is attached to an alkyl group falling within the definition of an alkyl group in polyhalogenomethyl or polyhaloC _1-4 alkyl, said alkyl groups may be the same or different.

The term "protecting group" means an amine protecting group such as such as C _1-10 alkoxycarbonyl, arilC _1-10 alkoxycarbonyl, such as benzoyl, anizoil-, izobutiroil-, acetyl or tretbutilbenzoil (Breipohl et al. (1997) Tetrahedron 53, 14671-14686). The protecting group may also be an acid unstable protecting group, such as dimethoxytrityl.

It should also be noted that the positions of the radicals on the molecular moiety used in the definitions, unless otherwise indicated, can be any in such a moiety as long as the moiety is chemically stable. For example, pyridyl includes 2-pyridyl, 3-pyridyl and 4-pyridyl; pentyl includes 1-pentyl, 2-pentyl and 3-pentyl.

When any of the variable parameters (for example, halogen or C _1-4 alkyl) occurs more than once in any of these components, each value is independent.

It is understood that the N-oxide forms of the subject compounds include any one of the compounds of the present invention, where one or more nitrogen atoms are oxidized to the so-called N-oxide.

Salts of the compounds of the present invention intended for therapeutic use of the present invention are those where the counterion is pharmaceutically and physiologically acceptable. However, salts containing a pharmaceutically unacceptable counterion may also find use, for example, in the preparation or purification of a pharmaceutically acceptable compound of the present invention. All salts, whether pharmaceutically acceptable or not, are within the scope of the present invention.

Pharmaceutically acceptable or physiologically tolerable forms of the addition salts which the compounds of the present invention are capable of forming are usually prepared using suitable acids, such as, for example, inorganic acids, such as hydrohalogen acids, for example hydrochloric or hydrobromic acid, sulfuric, hemisulfuric, nitric, phosphoric and the like acids; or organic acids, e.g. , benzenesulfonic, p-toluenesulfonic, cyclamic, salicylic, p-aminosalicylic, pamic and the like acids.

Conversely, these forms of acid addition salts can be converted by treatment with a base into the free base form.

The compounds of the present invention containing an acidic proton can also be converted to the corresponding non-toxic metal or amino addition base salt form by treatment with suitable organic or inorganic bases. Suitable basic salt forms include, for example, ammonium salts, alkali and alkaline earth metal salts, for example, lithium, sodium, potassium, magnesium, calcium salts and the like, salts with organic bases, for example, such as benzathine, N-methyl- D-glucamine, hydrabamine, and salts with amino acids such as, for example, arginine, lysine and the like. Alternatively, when the carboxyl moiety is present on the compound of the present invention, the compound can also be prepared as a salt with a pharmaceutically acceptable cation.

Conversely, forms of base addition salts can be converted by treatment with a suitable acid into the free acid form.

The term “salts” also includes the hydrates and solvent addition forms that the compounds of the present invention are capable of forming. Examples of such forms are, for example, hydrates, alcoholates, and the like.

If any of the substituents of the present invention contains chiral centers, as is sometimes the case, the compounds of the present invention include all stereoisomers both as isolated stereoisomers and as mixtures of such stereoisomers.

The term stereochemically isomeric forms of the compounds of the present invention, as used above, means all possible compounds consisting of the same atoms linked by the same sequence of bonds, but having different three-dimensional structures that are not interchangeable, which can form the compounds of the present invention . Unless otherwise specified or indicated, the chemical name encompasses a mixture of all possible stereochemically isomeric forms that the compound may possess. The specified mixture may contain all diastereomers and / or enantiomers of the main molecular structure of the specified compounds. It is understood that all stereochemically isomeric forms of the compounds of the present invention, both in pure form or in admixture with each other, are included in the scope of the present invention.

Pure stereoisomeric forms of the compounds and intermediates, as mentioned here, are defined as isomers substantially free of other enantiomeric or diastereomeric forms of the same basic molecular structure of said compounds and intermediates. In particular, the term “stereoisomerically pure” means compounds or intermediates having a stereoisomeric excess of at least 80% (i.e., a minimum of 90% of one isomer and a maximum of 10% of other possible isomers) up to a stereoisomeric excess of 100% (i.e., 100% of one isomer and no other), more preferably compounds or intermediates having a stereoisomeric excess of 90% to 100%, even more preferably having a stereoisomeric excess of 94% to 100%, and most preferably having stereoisomeric and bytok from 97% to 100%. The terms “enantiomerically pure” and “diastereomerically pure” should be understood in a similar way, but, respectively, in relation to the enantiomeric excess considered, the diastereomeric excess of the mixture.

Pure stereoisomeric forms of the compounds and intermediates of the present invention can be obtained using methods known from the prior art. For example, enantiomers can be separated from each other by selective crystallization of the corresponding diastereomeric salts with optically active acids and bases. Examples of such optically active compounds are tartaric acid, dibenzoyl tartaric acid, ditoluoyl tartaric acid and camphorsulfonic acid. Alternatively, enantiomers can be separated by chromatographic methods using chiral stationary phases. Said pure stereochemically isomeric forms can also be obtained from the corresponding pure stereochemically isomeric forms of the appropriate starting materials, provided that the reaction proceeds stereospecifically. Advantageously, if a specific stereoisomer is required, said compound is synthesized by stereospecific methods of preparation. In such methods, enantiomerically pure starting materials are predominantly used.

The diastereomeric racemates of the compounds of the present invention can be obtained separately by conventional methods. Suitable physical separation methods that can be used are, for example, selective crystallization and chromatography, for example, column chromatography. The present compounds may also exist in tautomeric forms. Such forms, although not explicitly indicated in the above formula, are considered to be included within the scope of the present invention. For example, in the definition of Het ² , for example, 1,2,4-oxadiazole can be substituted with a hydroxy or mercapto group at the 5-position, thus being in equilibrium with the corresponding tautomeric form, as shown below.

The term “prodrug” is meant to mean pharmacologically acceptable derivatives such as esters, amides and phosphates, such that the in vivo derivative biotransformation product of the derivative is an active drug that meets the definition of the compounds of the present invention. Reference material provided in Goodman and Gilman (The Pharmacological Basis of Therapeutics, 8 ^{th ed, McGraw-Hill, Int} . Ed. 1992, "Biotransformation of Drugs", p . 13-15), describing prodrugs, incorporated herein as commonly . Prodrugs of a compound of the present invention are prepared by modifying the functional groups present in the compound so that the modifications are cleaved either by standard processing or in vivo to the parent compound. For example, a sulfhydryl-containing substituent must be bound to a carrier that makes the compound biologically inactive until it is recovered by the action of endogenous enzymes or, for example, enzymes directed at a particular receptor, or localized in the body.

Prodrugs are characterized by excellent water solubility, increased bioavailability and are easily converted in the process of metabolism into active inhibitors in vivo.

It is also understood that the present invention includes all isotopes of atoms that make up the compounds under consideration. Isotopes include atoms having the same atomic number but differing in mass numbers. As a general example, and not by way of limitation, hydrogen isotopes include tritium and deuterium. Carbon isotopes include C-13 and C-14.

Whenever the term “compounds of the formula (I)”, “compounds of the formula (II)”, “compounds of the formula (III)”, “compounds of the formula (IV)”, “compounds of the formula (V)”, “compounds of formula (VI) "," compounds of formula (VII) "," compounds of formula (VIII) "," compounds of formulas (V) to (VIII) "or" compounds of the present invention "or a similar term, this term is intended to include compounds general formulas (I), (II), (III), (IV), (V), (VI), (VII), (VIII), N-oxides, salts, stereoisomeric forms, racemic mixtures, prodrugs, esters and metabolites indicated with of the connections, as well as quaternary nitrogen analogues. A noteworthy subgroup of compounds of formula (V) or any subgroup of such compounds are N-oxides, salts and all stereoisomeric forms of compounds of formula (V).

Embodiments of the present invention are those compounds of the present invention or any subgroup of such compounds wherein 4-pyridyl forms an N-oxide, for example, the N-oxide of compound No. 24.

N-oxide of compound No. 24

Other embodiments of the present invention are those compounds of the present invention or any subgroup of such compounds wherein the compound exists as an acid addition salt, wherein the salt is preferably selected from the group consisting of hydrochloride, hydrobromide, trifluoroacetate, fumarate, chloroacetate, methanesulfonate, oxalate, acetate and citrate.

Further embodiments of the present invention are those compounds of the present invention or any subgroup of such compounds wherein R ^{1 is} independently hydrogen, amino, mono- or disubstituted amino, where amino substituent (s) may be selected from the group consisting of C _1-6 alkyl , C _1-4 alkoxyC _1-4 alkyl, diC _1-4 alkylamino C _1-4 alkyl, piperidin-1-ylC _1-4 alkyl, aryl C _1-6 alkyl, wherein the aryl group may be further substituted with C _1-4 alkyl or C _1-4 alkoxy.

Other embodiments of the present invention are those compounds of the present invention or any subgroup of such compounds wherein R ^{1 is} independently hydrogen, amino, mono- or disubstituted amino, where amino substituent (s) may be selected from the group consisting of C _1-4 alkyl , C _1-4 alkoxyC _1-4 alkyl, diC _1-4 alkylamino C _1-4 alkyl, piperidin-1-ylC _1-4 alkyl, aryl C _1-6 alkyl, wherein the aryl group may be further substituted with C _1-4 alkoxy.

Other embodiments of the present invention are those compounds of the present invention or any subgroup of such compounds wherein R ^{1 is} independently hydrogen, amino, mono- or disubstituted amino, where amino substituent (s) may be selected from the group consisting of C _1-2 alkyl , C _1-2 alkoxyC _1-2 alkyl, diC _1-2 alkylamino C _1-2 alkyl, piperidin-1-ylC _1-2 alkyl, arylC _1-2 alkyl, where the aryl group may be further substituted with C _1-2 alkoxy.

Other embodiments of the present invention are those compounds of the present invention or any subgroup of such compounds wherein R ^{1 is} independently hydrogen, amino, mono- or disubstituted amino, where amino substituent (s) may be selected from the group consisting of methyl, methoxyethyl, dimethylaminoethyl , piperidin-1-yl-ethyl, benzyl, wherein the phenyl group may be further substituted with methoxy.

Other embodiments of the present invention are those compounds of the present invention or any subgroup of such compounds where R ^{1 is} independently hydrogen, amino or monosubstituted amino, where the amino substituent may be selected from the group consisting of methoxyethyl, dimethylaminoethyl, piperidin-1-yl-ethyl, benzyl where the phenyl group may be further substituted with methoxy.

Other embodiments of the present invention are those compounds of the present invention or any subgroup of such compounds wherein R ¹ independently is hydrogen or amino.

Further embodiments of the present invention are those compounds of the present invention or any subgroup thereof, wherein R ⁸ is hydrogen, C _1-10 alkyl, aminoC _1-10 alkyl, arilC _1-10 alkyl, Het ¹ C _1-6 alkyl or a protecting a group wherein aryl is optionally substituted with 1-3 substituents selected from the group consisting of C _1-4 alkyl, C _1-4 alkylcarbonyl, halogen, —OR ⁶ ,

-NR ^4a R ^4b , -SR ⁵ and polyhaloC _1-4 alkyl.

In other embodiments of the present invention are those compounds of the present invention or any subgroup thereof, wherein R ⁸ is hydrogen, C _1-6 alkyl, aminoC _1-6 alkyl, arilC _1-6 alkyl, Het ¹ C _1-6 alkyl or C _1-6 alkoxycarbonyl.

Other embodiments of the present invention are those compounds of the present invention or any subgroup of such compounds wherein R ⁸ is hydrogen, C _1-6 alkyl, amino C _1-4 alkyl, phenylC _1-4 alkyl, pyrrolidin-1-ylC _1-4 alkyl or C _1-6 alkoxycarbonyl.

Other embodiments of the present invention are those compounds of the present invention or any subgroup of such compounds wherein each of R ^{9 is} independently hydrogen, C _1-4 alkyl, polyhalo C _1-4 alkyl, halogen, —COR ⁶ , —COOR ⁷ ,

—CONR ^4a R ^4b , —OR ⁷ , —NR ^4a R ^4b , —NR ^4a COR ⁶ , —NR ^4a SO ₂ R ⁵ , —SR ⁵ or morpholin-4-yl, and wherein C _1-4 alkyl may be optionally substituted by -COOR ⁷ .

Other embodiments of the present invention are those compounds of the present invention or any subgroup of such compounds wherein each of R ^{9 is} independently hydrogen, C _1-4 alkyl, -COR ⁶ , -COOR ⁷ or -CONR ^4a R ^4b and where C _{1- 4} alkyl may be further substituted with —COOR ⁷ .

Further embodiments of the present invention are those compounds of the present invention or any subgroup of such compounds wherein each of R ^{9 is} independently hydrogen, C _1-4 alkyl, —COR ⁶ , —COOR ^7, or —CONR ^4a R ^4b .

Further embodiments of the present invention are those compounds of formulas (V), (VI) or any of a subgroup of such compounds wherein R ¹¹ is hydrogen, fluoro or pyrrolidin-1-yl.

Other embodiments of the present invention are those compounds of formulas (V), (VI) or any of a subgroup of such compounds wherein R ¹¹ is hydrogen or fluoro.

Other embodiments of the present invention are those compounds of formulas (V), (VI) or any of a subgroup of such compounds wherein R ¹² is halogen, C _1-4 alkyl or polyhalo C _1-4 alkyl.

Further embodiments of the present invention are those compounds of formulas (V), (VI) or any of a subgroup of such compounds wherein R ¹² is halogen or polyhalo C _1-4 alkyl.

Other embodiments of the present invention are those compounds of formulas (V), (VI) or any of a subgroup of such compounds wherein R ¹² is chloro, bromo, fluoro or trifluoromethyl.

Other embodiments of the present invention are those compounds of formulas (V), (VI) or any of a subgroup of such compounds wherein R ¹¹ is fluoro and R ¹² is chloro or bromo.

Other embodiments of the present invention are those compounds of formulas (V), (VI) or any of a subgroup of such compounds wherein R ¹¹ is hydrogen and R ¹² is chloro, bromo, fluoro or trifluoromethyl.

Compounds of particular interest are compounds of formula (V) listed in table 1 below, in particular compounds number 1, number 7, number 21, number 23, number 24 and number 25, and the corresponding N-oxides, salts and stereoisomers.

A number of synthetic methods can be used to prepare the compounds of the present invention. Typically, these compounds can be synthesized using interactions known from the prior art. Any synthesis method known in the art may be used. However, the following synthesis methods are convenient for preparing the compounds of the invention.

The compounds of formula (V) can be synthesized according to the following adapted procedure Wamhoff, H .; Kroth, E. Synthesis, 1994, 405-410, represented by Scheme 1.

Scheme 1

Essentially, methyl 3-amino-2-pyrazinecarboxylate (1a) is reacted with an acyl chloride in the presence of a suitable solvent such as chloroform or pyridine to give 3-acylaminopyrazine-2-carboxylates (1b). These 3-acylaminopyrazine-2-carboxylates (1b) are converted, for example, using ammonium hydroxide to 3-acylaminopyrazine-2-amides (1d). Optional 3-acylaminopyrazine-2-carboxamides (1d) can already be obtained by acylation of 3-amino-2-pyrazinecarboxamide (1c).

The 3-acylaminopyrazine-2-amides (1d) are then cyclized by addition of a base to obtain pteridin-4-ol derivatives of formula (1e). The alcohol can then be substituted with halogen using a halogenating agent such as thionyl chloride in a suitable solvent such as chloroform, dichloroethane or tetrahydrofuran (THF) in the presence of a catalytic amount of dimethylformamide (DMF). Then the nucleophilic substitution is carried out on the compound (1f) with an amine or alcohol of the formula HLR ² , together with a suitable base, such as TEA or DIPEA, in an organic solvent such as DCM, THF or DMF, to obtain the pteridine compounds of the formula (1g).

Alternatively, pteridin-4-ol can be converted by a single reactor to the pteridines of formula (V) by reacting compounds of formula (1e) with an amine or alcohol of formula HLR ² , together with a suitable base such as TEA or DIPEA, in the presence of benzotriazole-1 -yloxytrispyrrolidinophosphonium hexafluorophosphate (PyBOP). In the formula HLR ² H is hydrogen and L and R ² are as defined above in the definition of substituents of the compounds of formula (V).

Scheme 2

Alternatively, compounds of formula (V) can be prepared from the corresponding pteridinones as starting materials, followed by conversion of the compounds obtained to iminochlorides and subsequent substitution of a chlorine atom with a suitable amine, such as 4-aminopyridine, as shown below in Scheme 2.

(i) thionyl chloride, DMF; (ii) 4-aminonicotinic acid methyl ester, TEA; (iii) NaOH; (iv) PyBOP, TEA, HNR ^4a R ^4b .

Schemes 3 and 4 below provide alternative routes to pyridyl nuclei and further substitutions of these nuclei.

Scheme 3

Scheme 4

Compounds that are embodiments of the present invention are shown below in table 1:

Table 1

The route of administration and formulation of the compounds useful for the invention and related compounds depends on the nature of the condition of the particular patient being treated and the personal assessment of the practitioner; the formulation is determined by the route of administration. Since the compounds of the invention are low molecular weight, these compounds are usually administered orally, in a mixture with suitable pharmaceutical excipients, for the manufacture of tablets, capsules, syrups and the like. Suitable formulations for oral administration may also include trace components, such as buffers, flavoring agents and the like. Typically, the amount of active ingredient in the compositions varies between 5% -95% of the total composition, but the allowable deviation range depends on the carrier. Suitable carriers include substances such as sucrose, pectin, magnesium stearate, lactose, peanut butter, olive oil, water and the like.

Compounds useful in the invention may also be administered via suppositories or other transmucosal carriers. Typically, such compositions include excipients that facilitate the passage of the compound through mucose, such as pharmaceutically acceptable detergents.

The compounds may also be applied topically or in a composition intended for penetration through the skin. Such formulations include lotions, creams, ointments and the like, formulated by known methods.

The compounds may also be administered by injection, including intravenous, intramuscular, subcutaneous or intraperitoneal injection. Typical formulations for this application are liquid formulations in isotonic solvents such as Hank's solution and Ringer's solution.

Alternative formulations include nasal sprays, liposome formulations, sustained release formulations, and the like, known in the art.

Any suitable formulations may be used. See a manual on prior art formulation methods of Remington's Pharmaceutical Sciences, latest edition, Mack Publishing Company, Easton, PA. Reference to this manual is standard in the art.

Dosages of the compounds of the invention depend on a number of factors that vary from patient to patient. However, it is generally believed that a daily oral dose of 0.001-100 mg / kg total body weight, preferably 0.01-50 mg / kg, and more preferably 0.01 mg / kg-10 mg / kg, should be used. The dosage regimen changes, again, depending on the conditions being treated and at the discretion of the practitioner.

It should be noted that the compounds of the invention can be administered either as individual active ingredients or as mixtures of several possible embodiments of the above formula. In addition, the compounds of the invention can be used as separate therapeutic agents or in combination with other therapeutic agents.

Due to the favorable antiviral properties, as follows from the examples, the compounds of the present invention are useful for the treatment of patients infected with HCV and for the prevention of these patients. In general, the compounds of the present invention may be useful in the treatment of warm-blooded animals infected with flaviviruses. Conditions that can be prevented or treated by the compounds of the present invention include, in particular, conditions caused by HCV and other pathogenic flaviviruses, such as yellow fever virus, Dengue fever (types 1-4), St. Louis encephalitis, Japanese encephalitis, Murray Valley encephalitis, West Nile virus and Kunjin virus. Conditions caused by HCV include progressive liver fibrosis, inflammation and necrosis leading to cirrhosis, end-stage liver disease and HCC; and in the case of other pathogenic flaviviruses, conditions include yellow fever, dengue fever, hemorrhagic fever and encephalitis.

The compounds of the present invention or any subgroup of these compounds may be useful as a medicine against the above conditions. Said use as a medicine or method of treatment includes the systemic administration of HCV-infected patients in an amount effective to suppress conditions caused by HCV and other pathogenic flaviviruses. Therefore, the compounds of the present invention can be used to make a medicament suitable for treating conditions caused by HCV and other pathogenic flaviviruses.

According to an embodiment, the invention relates to the use of a compound of formula (V) or any subgroup of said compounds as defined above for the manufacture of a medicament for the treatment or control of an infection or a condition caused by HCV infection in a mammal. The invention also relates to a method for treating a flavivirus infection or a disease caused by a flavivirus infection, comprising administering to a mammal in need of such treatment an effective amount of a compound of formula (V) or any subgroup of said compounds as defined above.

In another embodiment, the present invention relates to the use of a compound of formula (V) or any subgroup of said compounds as defined above for the manufacture of a medicament useful for inhibiting HCV activity in a mammal infected with flaviviruses, in particular HCV. In yet another embodiment, the present invention relates to the use of a compound of formula (V) or any subgroup of said compounds as defined above for the manufacture of a medicament useful for inhibiting HCV activity in a mammal infected with flaviviruses, wherein said HCV is inhibited in the replication step.

Also, a combination of a previously known anti-HCV compound, such as, for example, interferon-α (IFN-α), pegylated interferon-α and / or ribavirin, and the compounds of the present invention can be used as a medicine in combination therapy. The term "combination therapy" refers to a product which optionally contains (a) a compound of the present invention and (b) optionally another anti-HCV compound, as a combined preparation for simultaneous, separate or sequential use for the treatment of HCV infections, in particular the treatment of infections HCV type 1. Thus, for combating HCV infections or for appropriate treatment, the compounds of this invention can be administered together, in combination, for example, with substances such as interferon-α (IFN-α), peg interferon-α and / or ribavirin, as well as therapeutic agents based on antibodies directed against HCV epitopes, small interfering RNA (Si RNA), ribozymes, DNA enzymes, antisense RNA, antagonists, for example, NS3 protease, NS3 helicase and NS5B polymerase.

Therefore, the present invention relates to the use of a compound of formula (V) or any subgroup of said compounds as defined above for the manufacture of a medicament suitable for inhibiting HCV activity in a mammal infected with HCV viruses, wherein said medicament is used in combination therapy, said combination therapy preferably includes a compound of formula (V) and (pegylated) IFN-α and / or ribavirin.

One skilled in the art will recognize that the compounds of formula (V) can be investigated in the HCV replication cell system based on Lohmann et al. (1999) Science 285: 110-113, with additional modifications described in Krieger et al. (2001) Journal of Virology 75: 4614-4624 (incorporated herein by reference), further illustrated in the examples section. Such a model, although not a complete HCV infectious model, is widely accepted as the most reasonable and effective model of independent HCV RNA replication currently available. Compounds exhibiting anti-HCV activity in this cell model are considered suitable for further development for use in the treatment of HCV infections in mammals. It should be understood that it is important to distinguish compounds that specifically interfere with HCV functions from compounds that exhibit a cytotoxic or cytostatic effect in the HCV replication model and, as a result, cause a decrease in the concentration of HCV RNA or the concentration of the bound reporter enzyme. Studies known in the field of cellular cytotoxicity assessment are based, for example, on the activity of mitochondrial enzymes using fluorogenic redox dyes such as resazurin. In addition, cell counting tests exist to evaluate non-selective inhibition of the activity of a linked reporter enzyme, such as yellow-red luciferase. Appropriate cell types can be endowed with stable transfection using a luciferase reporter gene, the expression of which depends on the constitutively active promoter of the gene, and such cells can be used for screening counts to evaluate non-selective inhibitors.

All patents, patent applications, and articles mentioned earlier or below are incorporated herein by reference.

Examples

The following examples are considered illustrative but not limiting.

Example 1

Synthesis of 2- (5-bromo-2-fluorophenyl) -4- (4-pyridylamino) pteridine, compound No. 1

3- (5-Bromo-2-fluorobenzoylamino) pyrazine-2-carboxylic acid methyl ester 102

Pyridine (7.75 g, 98.0 mmol) was added at 0 ° C under N ₂ to a solution of 3-aminopyrazine-2-carboxylic acid methyl ester 101 (1.5 g, 9.80 mmol) and 5-bromo- 2-fluorobenzoyl chloride (9.45 g, 49.0 mmol) in CH ₂ Cl ₂ . The reaction mixture was heated to 40 ° C. for 4 hours, then cooled to room temperature. The reaction mixture was quenched with 20 ml of ethanol, evaporated, partitioned between CH ₂ Cl ₂ and 1N NaHCO ₃ , dried (Na ₂ SO ₄ ) and evaporated. The crude product is pulverized with EtOH, filtered, washed with EtOH and diethyl ether to give 2.6 g of 3- (5-bromo-2-fluorobenzoylamino) pyrazine-2-carboxylic acid methyl ester 102 as a white powder (LC-MS- analysis).

3- (5-Bromo-2-fluorobenzoylamino) pyrazine-2-carboxamide 103

A mixture of 3- (5-bromo-2-fluorobenzoylamino) pyrazine-2-carboxylic acid methyl ester 102 (2.6 g, 7.34 mmol) and NH ₄ OH (15 ml) in ethanol (50 ml) is heated to boiling point reflux for 10 min. Then the reaction mixture was cooled to room temperature and the precipitate was filtered off, washed with ethanol and diethyl ether to obtain 2.1 g of the title product 103 as a white powder (LC-MS analysis).

2- (5-Bromo-2-fluorophenyl) pteridin-4-one 104

A mixture of 3- (5-bromo-2-fluorobenzoylamino) pyrazine-2-carboxamide 103 (2.3 g, 6.78 mmol) and KOH (3.81 g, 67.8 mmol) in H ₂ O (60 ml) and DMSO (20 ml) was stirred at room temperature for 45 minutes. Acidification to pH 5 (control with pH indicator paper) with AcOH followed by 50 ml of ice-cold water gives a precipitate which is separated by filtration, washed with H ₂ O, acetonitrile and diethyl ether to give 1.83 g of the title Product 104 as a white powder (LC-MS analysis).

2- (5-Bromo-2-fluorophenyl) -4- (4-pyridylamino) pteridine, compound No. 1

Triethylamine (1.04 ml, 7.17 mmol) is added to a solution of 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 (800 mg, 2.49 mmol), 4-aminopyridine (469 mg, 4 98 mmol) and PyBOP (2.59 g, 4.98 mmol) in CH ₂ Cl ₂ . After 12 hours, the reaction mixture was partitioned between CH ₂ Cl ₂ / petroleum ether (2: 1, 300 ml) and ice-cold 1N HCl (300 ml). The pH of the aqueous phase is adjusted to 12 with conc. NaOH and extracted with AcOEt, dried (Na ₂ SO ₄ ) and evaporated. The residue was pulverized in a mixture of CH ₂ Cl ₂ / petroleum ether (2: 1, 15 ml), separated by filtration and washed with diethyl ether. The product was purified by column chromatography (AcOEt / CH ₂ Cl ₂ / MeOH 5: 4: 1 mixture) to give 325 mg of the title product 1 as a yellow powder (LC-MS analysis).

Example 2

Synthesis of 4 - [[2- (5-bromo-2-fluorophenyl) pteridin-4-yl] amino] nicotinic acid, compound No. 16, and 4 - [[2- (5-bromo-2-fluorophenyl) pteridin-4 -yl] amino] -N- [3- (2-oxopyrrolidin-1-yl) propyl] nicotinamide, compound No. 21

2- (5-Bromo-2-fluorophenyl) 4-chloropteridine 106

Thionyl chloride (371 mg, 3.11 mmol) is added to a stirred suspension of 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 (200 mg, 0.623 mmol) in chloroform (5 ml) and dry DMF (100 μl ) The reaction mixture is heated to boiling point under reflux, under nitrogen atmosphere, for 1 h (starting material passes HPLC). The solvent is removed in vacuo. The residue was then pulverized into Et ₂ O and separated by filtration to obtain 210 mg of the title product 106 as a yellow solid (LC-MS analysis).

4 - [[2- (5-Bromo-2-fluorophenyl) pteridin-4-yl] amino] nicotinic acid methyl ester 107

To a solution of 2- (5-bromo-2-fluorophenyl) -4-chloropteridin 106 (200 mg, 0.589 mmol), 4-aminonicotinic acid methyl ester (224 mg, 1.47 mmol) in dichloroethane (5 ml) was added dropwise triethylamine (300 μl, 2.07 mmol). The resulting mixture was heated to 70 ° C for 15 minutes, then quenched with silica and purified by column chromatography (AcOEt / CH ₂ Cl ₂ / triethylamine mixture, 70/19/1). Crystallization from AcOEt / Et ₂ O gave 200 mg of the title product 107 as yellow prismatic crystals (LC-MS analysis).

4 - [[2- (5-Bromo-2-fluorophenyl) pteridin-4-yl] amino] nicotinic acid 16

Solution of methyl ester 4 - [[2- (5-bromo-2-fluorophenyl) pteridin-4-yl] amino] nicotinic acid 107 (200 mg, 0.439 mmol) and NaOH (44 mg, 1.10 mmol) in a mixture of THF / MeOH / H ₂ O (3: 2: 1, 5 ml), stirred at room temperature for 3 hours. The solvent was evaporated and the residue was dissolved in H ₂ O, neutralized with AcOH, filtered and successively washed with H ₂ O, MeOH and diethyl ether to give 160 mg of the title product 16 as a yellow powder (LC-MS analysis).

4 - [[2- (5-bromo-2-fluorophenyl) pteridin-4-yl] amino] -N- [3- (2-oxopyrrolidin-1-yl) propyl] nicotinamide 21

Triethylamine (140 μl, 1.00 mmol) is slowly added to the solution of 4 - [[2- (5-bromo-2-fluorophenyl) pteridin-4-yl] amino] nicotinic acid 16 (150 mg, 0.340 mmol), PyBOP ( 0.350 mg, 0.68 mmol) and 1- (3-aminopropyl) pyrrolidinone (97 mg, 0.68 mmol) in CH ₂ Cl ₂ (10 ml). After 15 minutes at room temperature, the reaction mixture was evaporated and the residue was purified by column chromatography (from AcOEt / CH ₂ Cl ₂ , 2: 1 + 1% triethylamine to AcOEt / CH ₂ Cl ₂ / MeOH, 7: 2: 1 + 1% triethylamine). The yellow powder was recrystallized from EtOH to give 58 mg of the title product 21 as yellow prismatic crystals (LC-MS analysis).

Example 3

Synthesis of 2- (5-Bromo-2-pyrrolidin-1-ylphenyl) -4- (3-methyl-4-pyridylamino) pteridine, compound No. 6

2- (5-Bromo-2-pyrrolidin-1-ylphenyl) pteridin-4-one 110

A solution of 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 in pyrrolidine is heated in a microwave cavity (Power = 270 W, Temp. = 110 ° C) for 12 min. Pyrrolidine was evaporated, then the residue was partitioned between 0.5 N NaHCO ₃ and CH ₂ Cl ₂ , dried (Na ₂ SO ₄ ) and evaporated. Trituration in Et ₂ O gave the title product 110 as a yellow powder (LC-MS analysis).

2- (5-Bromo-2-pyrrolidin-1-yl-phenyl) -4- (3-methyl-4-pyridylamino) pteridine 6

The title product was synthesized by reacting 2- (5-bromo-2-pyrrolidin-1-ylphenyl) pteridin-4-one 110 and 4-amino-3-methylpyridine, following the procedure described for 4- (4-pyridylamino) - 2- (5-bromo-2-fluorophenyl) pteridine 1 (LC-MS analysis).

Example 4

Synthesis of 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine, compound No. 3

4-Butylaminopyridine 113

A solution of 4-chloropyridine (2.0 g, 17.6 mmol), 50% butylamine in water (30 ml) was heated to 150 ° C in a sealed ampoule for 24 hours. The reaction mixture was evaporated, partitioned between 0.1 N NaOH and CH ₂ Cl ₂ , dried (Na ₂ SO ₄ ) and evaporated. Crystallization in 4: 1 diethyl ether / petroleum ether gave 2.4 g of the title product 113 as a white powder (LC-MS analysis).

4 - [(Butyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine 3

The title product was synthesized by reacting 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 with 4-butylaminopyridine 113 , following the procedure described for 4- (4-pyridylamino) -2- (5-bromo- 2-fluorophenyl) pteridine 1 (LC-MS analysis).

Example 5

Synthesis of 2- (3-fluorophenyl) -4- (4-pyridylamino) pteridine, compound No. 27

The title product was synthesized by reacting 2- (3-fluorophenyl) pteridin-4-one with 4-aminopyridine, following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo- 2-fluorophenyl) pteridine 3 .

Example 6

Synthesis of 2- (5-bromo-2-fluorophenyl) -4 - [(methyl) (4-pyridyl) amino] pteridine, compound No. 2

The title product was synthesized by reacting 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 with 4- (methylamino) pyridine, following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 7

Synthesis of 2- (5-bromo-2-fluorophenyl) -4 - [(3,3-dimethylbutyl) (4-pyridyl) amino] pteridine, compound No. 4

The title product was synthesized by reacting 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 with 4- (3,3-dimethylbutylamino) pyridine, following the procedure described for 4 - [(butyl) (4- pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 8

Synthesis of 4 - [(benzyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine, compound No. 5

The title product was synthesized by reacting 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 with 4- (benzylamino) pyridine, following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 9

Synthesis of 2- (5-bromo-2-fluorophenyl) -4- (3-methyl-4-pyridylamino) pteridine, compound No. 7

The title product was synthesized by reacting 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 with 4-amino-3-methylpyridine, following the procedure described for 4 - [(butyl) (4-pyridyl) amino ] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 10

Synthesis of 2- (5-bromo-2-fluorophenyl) -4- (phenylamino) pteridine, compound No. 8

The title product was synthesized by reacting 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 with aniline, following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5 β-bromo-2-fluorophenyl) pteridine 3.

Example 11

Synthesis of 2- (5-bromo-2-fluorophenyl) -4- (2-tolylamino) pteridine, compound No. 9

The title product was synthesized by reacting 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 with 2-methylaniline, following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 12

Synthesis of 2- (5-bromo-2-fluorophenyl) -4- [4- (2-pyridyl) piperazin-1-yl] pteridine, compound No. 12

The title product was synthesized by reacting 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 with 1- (2-pyridyl) piperazine following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 13

Synthesis of 2- (5-bromo-2-fluorophenyl) -4 - [(methyl) (phenyl) amino] pteridine, compound No. 10

The title product was synthesized by reacting 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 with N-methylaniline, following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 14

Synthesis of 2- (5-bromo-2-fluorophenyl) -4- (2-hydroxyethylamino) pteridine, compound No. 11

The title product was synthesized by reacting 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 with 2-hydroxyethylamine, following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 15

Synthesis of 2- (5-bromo-2-fluorophenyl) -4- (4-morpholinophenylamino) pteridine, compound No. 14

The title product was synthesized by reacting 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 with 4- (4-morpholino) aniline, following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 16

Synthesis of 2- (5-bromo-2-fluorophenyl) -4- (2-methyl-4-pyridylamino) pteridine, compound No. 15

The title product was synthesized by reacting 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 with 4-amino-2-methylpyridine, following the procedure described for 4 - [(butyl) (4-pyridyl) amino ] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 17

Synthesis of 2- (5-bromo-2-fluorophenyl) -4 - [[2- (pyrrolidin-1-yl) ethyl] - (4-pyridyl) amino] pteridine, compound No. 17

The title product was synthesized by reacting 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 with 4- [2- (pyrrolidin-1-yl) ethylamino] pyridine, following the procedure described for 4 - [( butyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 18

Synthesis of 2- (5-bromo-2-fluorophenyl) -4 - [(phenethyl) (4-pyridyl) amino] - pteridine, compound No. 22

The title product was synthesized by reacting 2- (5-bromo-2-fluorophenyl) pteridin-4-one 104 with 4- (phenethylamino) pyridine, following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 19

Synthesis of 2- (2-methyl-6-pyridyl) -4 - [(3-methyl-4-pyridyl) amino] pteridine, compound No. 19

The title product was synthesized by reacting 2- (2-methyl-6-pyridyl) pteridin-4-one with 4-amino-3-methylpyridine following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 20

Synthesis of 2- (3-chlorophenyl) -4- (4-pyridylamino) pteridine, compound No. 18

The title product was synthesized by reacting 2- (3-chlorophenyl) pteridin-4-one with 4-aminopyridine, following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo- 2-fluorophenyl) pteridine 3.

Example 21

Synthesis of 2- (5-chloro-2-fluorophenyl) -4- (3-ethyl-4-pyridylamino) pteridine, compound No. 23

The title product was synthesized by reacting 2- (5-chloro-2-fluorophenyl) pteridin-4-one with 4-amino-3-ethylpyridine following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 22

Synthesis of 2- (5-chloro-2-fluorophenyl) -4- (3-methyl-4-pyridylamino) pteridine, compound No. 25

The title product was synthesized by reacting 2- (5-chloro-2-fluorophenyl) pteridin-4-one with 4-amino-3-methylpyridine, following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo-2-fluorophenyl) pteridine 3.

Example 23

Synthesis of 2- (5-chloro-2-fluorophenyl) -4- (4-pyridylamino) pteridine, compound No. 24

The title product was synthesized by reacting 2- (5-chloro-2-fluorophenyl) pteridin-4-one with 4-aminopyridine, following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- ( 5-bromo-2-fluorophenyl) pteridine 3.

Example 24

Synthesis of 2- (3-trifluoromethylphenyl) -4- (4-pyridylamino) pteridine, compound No. 26

The title product was synthesized by reacting 2- (3-trifluoromethylphenyl) pteridin-4-one with 4-aminopyridine, following the procedure described for 4 - [(butyl) (4-pyridyl) amino] -2- (5-bromo- 2-fluorophenyl) pteridine 3.

Table 2 below shows the LC-MS data for the synthesized compounds:

m / z means mass to charge ratio

RT means retention time

Example 25

The activity of the compounds of formula (V) in tests for HCV replication

Stable Replication Cell Reporter Test:

The compounds of the present invention are tested for activity in inhibiting HCV RNA replication by testing on cells. The test shows that the considered compounds have activity against HCV replicons that are functional in cell culture. The cell test is based on a bicistronic expression construct as described in Lohmann et al. (1999) Science vol. 285 pp. 110-113, with the modifications described in Krieger et al. (2001) Journal of Virology 75: 4614-4624, using a multi-target screening strategy. In essence, the method is as follows.

A stably transfected Huh-7 luc / neo cell line (hereinafter referred to as Huh-Luc) was used in the assay. The specified cell line has an encoded RNA bicistronic expression construct comprising non-mutant NS3-NS5B regions of HCV type 1b translated from the source site of the internal ribosome (IRES) of the encephalomyocarditis virus (EMCV), with the previous part of the reporter (FfL luciferase) and part of the selectable marker neo ^R , neomycin phosphotransferase). The construct is surrounded by 5'- and 3'-NTRs (untranslated regions) of HCV type 1b. Continuous cultivation of replication cells in the presence of G418 (neo ^R ) depends on the replication of HCV RNA. Stably transfected replication cells expressing HCV RNA, which replicates independently and at high levels, in particular coding for luciferase, are used for screening antiviral compounds.

Experimental cell test procedure:

Replication cells are plated on 384-well plates in the presence of test and control compounds added at various concentrations. After incubation for three days, HCV replication is evaluated by assaying for luciferase activity (using standard luciferase assay substrates and reagents and a Perkin Elmer ViewLux ^Tm ultraHTS microplate imaging device). Replication cells in control cultures are characterized by high expression of luciferase in the absence of any inhibitor. The inhibitory activity of the compound against Huh-Luc cells is monitored for luciferase activity, giving a dose-response curve for each test compound. Then, the EC50 values are calculated, the indicated value corresponds to the amount of compound required to reduce by 50% the level of measured luciferase activity, or more precisely means the ability of genetically linked replication HCV RNA to reproduce by cell division.

It was found that the studied compounds have the following activity:

Table 3 Connection number HCV replication activity (μM) one 0.352 13 4.9 21 0.058 6 eighteen 3 2.2 27 3.0 2 3.65 four 0.48 5 0.99 7 0.95 8 > 32 9 eleven 12 > 32 10 eleven eleven > 32 fourteen 8.7 fifteen 3.56 22 1.96 19 12 eighteen 1,5 23 0.52 25 0.48 24 0.78 26 2.0

Example 26

Pharmacokinetic profile of compound No. 21 in the body of male Swiss SPF (CD1) mice

Compound No. 21 was dissolved in a 10% solution of hydroxypropyl-b-cyclodextrin (HP-in-CD) to a final concentration of 1 mg basic equivalent / ml, pH 4.36.

A solution of compound No. 21 was orally administered to three animals, giving a dose of 20 mg basic equivalent / kg. Blood samples are taken at the time of 30 minutes, 1, 2, 4, 8, and 24 hours after an oral dose of the drug. Plasma is obtained after centrifugation at 4 ° C for 10 minutes, at approximately 1900 × g.

The heart and liver of each animal received an oral dose are prepared, receiving individual samples, and weighed. Tissue samples are homogenized in demineralized water.

Plasma and tissue samples were analyzed for Compound No. 21 using an appropriate LC-MS / MS assay.

Limited pharmacokinetic analysis is performed using WinNonlin (TM) Professional (Version 4.0.1). For all data, analysis without compartmentalization is used, applying the lin / log trapezoid rule with lin / log interpolation. The variability of the results between animals is indicated by the standard deviation (st dev).

Information on the average concentrations in plasma and tissues and some basic pharmacokinetic parameters are shown in table 4 and figure 1.

Conclusion: the test concentration of the oral composition, equal to 1.0 mg basic equivalent / ml, leads to the required dose of 20 mg basic equivalent / kg when administered orally. There are no stability problems on the day of administration.

Plasma

After a single oral administration of compound No. 21 at 20 mg basic equivalent / kg, the levels are quantifiable over a period of up to 8 hours after administration of the dose (table 4 and figure 1). The average maximum plasma concentration (C _max ), equal to 220 ng / ml, is achieved for a period of 0.5 h after administration of the dose (T _max ), which indicates the rapid absorption of the compound. The average half-life (t _{1/2 (2-8 hours)} ) is 2.9 hours. The exposure found from AUC _0-inf is 332 ng.h / ml.

the cloth

As can be seen from figure 1, the studied tissue together with the plasma have exactly the same concentration-time profiles, which indicates the equilibrium distribution between the plasma and the tested tissues. The average maximum concentration in tissues (C _max ) is reached at the same time moment as in plasma, for a period of 0.5 hours after the dose, which indicates the rapid establishment of equilibrium. The highest concentration is observed in the liver (4057 ng / g), then in the heart (678 ng / g), with ratios of tissue to plasma concentrations equal to 24 and 3.8, respectively (table 4 and figure 1). The average half-life (t _{1/2 (2-8 h)} ) established for the liver is 3.5 and 3.4 hours for the heart, which is comparable with the corresponding parameter for plasma (2.9 hours). Tissue levels are reduced, this decrease is similar to plasma levels, and for a period of 8 hours after dosing, only low levels of the compound are still determined, which is evidence of the absence of significant retention.

Table 4
Average plasma and tissue levels (n = 3) along with some basic pharmacokinetic parameters of compound No. 21 after a single oral administration at 20 mg basic equivalent / kg to male Swiss SPF mice (CD1) Compound No. 21 (ng / ml or g / ml) Time Plasma stdev Heart stdev Liver stdev 0.5
one
2
four
8
24 220
166
21.7
20.3
5.52
BQL ¹⁾ ± 100
± 160
± 6.7
± 5.1
± 1.20 678
557
92.9
83.6
29.1
BQL ¹⁾ ± 258
± 403
± 49.9
± 35.9
± 6.0 4057
2937
611
627
201
BQL ¹⁾ ± 1730
± 1852
± 131
± 189
± 59 C _max (ng / ml)
T _max (h)
t _{1/2 (2-8 h)} (h)
AUC _(0-8h) (ng.h / ml)
AUC _0-inf (ng.h / ml)
Tissue / plasma ratio 220
0.5
2.9
309
332
- 678
0.5
3.4
1119
1262
3.8 ²⁾ 4057
0.5
3,5
6965
7973
24 ^{2) 1)} BQL = below the limit of quantification. The LLOQ is 0.500 ng / ml for plasma and varies from 5.00 to 13.16 ng / g for tissue.
²⁾ value based on AUC _inf

Claims (16)

1. The use of the compounds of formula (I) for the manufacture of a medicament for inhibiting HCV replication, said compound has the formula (I)

a pharmaceutically acceptable salt or ester, wherein R ¹ independently is hydrogen;
L is —NR ⁸ -, —NR ⁸ —C _1-6 alkanediyl, or

where the cycle indicated by the dashed line, together with N and Z forms a cycle containing 5-8 members, and can be replaced by an aromatic monocyclic heterocycle having 6 members in the cycle, which contains as the members of the cycle one heteroatom selected from nitrogen, where L cycle attached to the pteridine cycle through a nitrogen atom;
Z is N or CH;
R ² is hydroxyC _1-6 alkyl, phenyl, Het ² , wherein said phenyl and Het ^{2 are} independently, optionally substituted with one or more substituents selected from the group consisting of C _1-4 alkyl, —COOR ⁷ , morpholin-4-yl , -CONR ^4a R ^4b , where R ^4a and R ^4b are hydrogen, Het ¹ -C _1-4 alkyl; or NR ^4a R ^4b , where R ^4a and R ^4b are hydrogen, hydroxyC _1-4 alkyl, or may optionally form, with the nitrogen atom to which they are bonded, a 5-6 membered saturated ring, optionally containing an oxygen atom;
R ³ means phenyl, Het ² , each of which is independently, optionally substituted with one or more substituents selected from the group consisting of halogen, polyhaloC _1-4 alkyl, NR ^4a R ^4b , and where R ^4a and R ^4b form together with the atom the nitrogen to which they are attached is a 5-8 membered saturated ring;
each of R ⁷ independently means hydrogen or C _1-4 alkyl, and
R ⁸ is hydrogen, C _1-10 alkyl, phenyl C _1-10 alkyl, amino-C _1-10 alkyl; Het ¹ -C _1-4 alkyl Het ¹ ;
Het ¹ means a saturated or partially unsaturated monocyclic heterocycle having 5-6 members in the cycle, which contains as members of the cycle one or two heteroatoms selected from the group consisting of nitrogen, oxygen, and which is optionally substituted at one or more carbon atoms substituents selected from the group consisting of C _1-6 alkyl, oxo;
Het ² as a group or part of a group means an aromatic monocyclic heterocycle having 6 members in a cycle, which contains one heteroatom selected from nitrogen as cycle members. 2. The use according to claim 1, where the compound has the formula (II)

a pharmaceutically acceptable salt or ester, wherein R ¹ , R ³ , R ^4a , R ^4b R ⁷ , R ⁸ , Het ¹ and Het ² are as defined in claim 1; Where
R ⁹ is C _1-4 alkyl, —COOR ⁷ , —CONR ^4a R ^4b , morpholin-4-yl; and
n is 0, 1, 2, 3, or 4. 3. The use according to claim 1, where the compound has the formula (III)

a pharmaceutically acceptable salt or ester, wherein R ¹ , L, R ² , R ^4a , R ^4b R ⁷ , R ⁸ , Het ¹ and Het ² are as defined in claim 1; Where
R ¹⁰ means halogen, polyhaloC _1-4 alkyl, —NR ^4a R ^4b ; and
m is 0, 1, 2, 3 or 4. 4. The use according to claim 1, where the compound has the formula (IV)

a pharmaceutically acceptable salt or ester thereof, wherein R ¹ , R ^4a , R ^4b R ⁷ , R ⁸ , Het ¹ and Het ² are as defined in claim 1; Where
R ⁹ is C _1-4 alkyl, —COOR ⁷ , —CONR ^4a R ^4b , morpholin-4-yl;
R ¹⁰ means halogen, polyhaloC _1-4 alkyl, —NR ^4a R ^4b ;
n is 0, 1, 2, 3 or 4 and
m is 0, 1, 2, 3 or 4. 5. The use according to claim 1, where the compound has the formula (V)

its pharmaceutically acceptable salt, where
R ¹ means hydrogen;
R ⁸ is hydrogen, C _1-6 alkyl, amino C _1-4 alkyl, phenyl C _1-4 alkyl;
each of R ⁹ is independently C _1-4 alkyl —COOR ⁷ or —CONR ^4a R ^4b , and R ^4a and R ^4b independently are hydrogen, 2-oxopyrrolidin-1-yl-C _1-4 alkyl;
n is 0, 1, 2, 3 or 4;
R ¹¹ is halogen or —NR ^4a R ^4b , where R ^4a and R ^4b form, together with the nitrogen atom to which they are bonded, a 5–8 membered saturated ring;
R ¹² means halogen or polyhaloC _1-4 alkyl;
R ⁷ means hydrogen or C _1-4 alkyl. 6. The use according to claim 5, where the compound has the formula (VI)

its pharmaceutically acceptable salt, where R ¹ , R ⁸ , R ⁹ , R ¹¹ , R ¹² take the values defined in accordance with claim 5. 7. A method for inhibiting HCV replication, wherein said method comprises administering an HCV inhibitory effective amount of a compound according to any one of claims 1 to 6. 8. The compound of formula (VII)

and a pharmaceutically acceptable salt thereof, wherein R ¹ is hydrogen;
R ⁸ is hydrogen, C _1-6 alkyl, phenyl C _1-4 alkyl;
R ⁹ means hydrogen, C _1-6 alkyl;
with the proviso that when R ⁸ is hydrogen, R ^{9 is} not hydrogen. 9. The compound of claim 8, where R ⁹ means C _1-4 alkyl;
R ¹ and R ⁸ take the values defined in clause 8. 10. The compound of formula (VIII)

its pharmaceutically acceptable salt, where
R ¹ independently means hydrogen;
R ⁸ is hydrogen, C _1-6 alkyl, phenyl C _1-4 alkyl;
R ⁹ means hydrogen, C _1-6 alkyl;
with the proviso that when R ⁸ is hydrogen, R ^{9 is} not hydrogen. 11. The compound of claim 10, where R ⁹ means C _1-4 alkyl;
R ¹ and R ⁸ take the values defined in clause 10. 12. The compound of claim 10, where
R ⁸ means C _1-6 alkyl, phenylC _1-4 alkyl;
R ¹ and R ⁹ take the values defined in clause 10. 13. The compound according to any one of claims 8 to 12, where R ¹ means hydrogen;
R ⁸ means hydrogen;
R ⁹ means C _1-4 alkyl. 14. The compound according to any one of claims 8 to 13, where
R ¹ means hydrogen;
R ⁸ means C _1-6 alkyl;
R ⁹ means hydrogen. 15. A compound according to any one of claims 8-14 for use as a medicament for inhibiting HCV replication. 16. A pharmaceutical composition having the properties of an inhibitor of HCV replication, comprising one or more compounds according to any one of claims 8-14.

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