Papd5 And Papd7 Inhibitors For Treating A Hepatitis B Infection

Abstract

The present invention relates to a method for identifying a compound that prevents, ameliorates and/or inhibits hepatitis B virus (HBV) infections. The compound (i) reduces the expression and/or activity of PAP associated domain containing 5 (PAPD5) and/or PAP associated domain containing 7 (PAPD7); and/or (ii) binds to PAPD5 and/or PAPD7 and inhibits 5 propagation of HBV; and is identified as a compound that prevents, ameliorates and/or inhibits HBV infections. An inhibitor of PAPD5 and/or PAPD7 for use in treating and/or preventing HBV infections; as well as a combined preparation comprising an inhibitor of PAPD5 and an inhibitor of PAPD7 for simultaneous or sequential use in the treatment or prevention of HBV infections is also provided. The present invention includes a 10-pharmaceutical composition for use in treatment and/or prevention of HBV infections, and a method for monitoring therapeutic success during treatment of HBV infections.

Description

FIELD OF THE INVENTION

[0001] The present invention relates to a method for identifying a compound that prevents, ameliorates and/or inhibits a hepatitis B virus (HBV) infection, wherein a compound that (i) reduces the expression and/or activity of PAP associated domain containing 5 (PAPD5) and/or PAP associated domain containing 7 (PAPD7); and/or (ii) binds to PAPD5 and/or PAPD7 and inhibits propagation of HBV; is identified as a compound that prevents, ameliorates and/or inhibits a HBV infection. The invention also provides for an inhibitor of PAPD5 and/or PAPD7 for use in treating and/or preventing a HBV infection; as well as a combined preparation comprising an inhibitor of PAPD5 and an inhibitor of PAPD7 for simultaneous or sequential use in the treatment or prevention of a HBV infection. Also comprised in the present invention is a pharmaceutical composition for use in the treatment and/or prevention of a HBV infection, and a method for monitoring the therapeutic success during the treatment of a HBV infection.

BACKGROUND

[0002] The hepatitis B virus (HBV) is an enveloped, partially double-stranded DNA virus. The compact 3.2 kb HBV genome consists of four overlapping open reading frames (ORF), which encode for the core, polymerase (Pol), envelope and X-proteins. The Pol ORF is the longest and the envelope ORF is located within it, while the X and core ORFs overlap with the Pol ORF. The lifecycle of HBV has two main events: 1) generation of closed circular DNA (cccDNA) from relaxed circular (RC DNA), and 2) reverse transcription of pregenomic RNA (pgRNA) to produce RC DNA. Prior to the infection of host cells, the HBV genome exists within the virion as RC DNA. It has been determined that HBV virions are able to gain entry into host cells by non-specifically binding to the negatively charged proteoglycans present on the surface of human hepatocytes (Schulze, Hepatology, 46, (2007), 1759-68) and via the specific binding of HBV surface antigens (HBsAg) to the hepatocyte sodium-taurocholate cotransporting polypeptide (NTCP) receptor (Yan, J Virol, 87, (2013), 7977-91). The control of viral infection needs a tight surveillance of the host innate immune system which could respond within minutes to hours after infection to impact on the initial growth of the virus and limit the development of a chronic and persistent infection. Despite the available current treatments based on IFN and nucleos(t)ide analogues, the HBV infection remains a major health problem worldwide which concerns an estimated 350 million chronic carriers who have a higher risk of liver cirrhosis and hepatocellular carcinoma.

[0003] The secretion of antiviral cytokines in response to a HBV infection by the hepatocytes and/or the intra-hepatic immune cells plays a central role in the viral clearance of the infected liver. However, chronically infected patients only display a weak immune response due to various escape strategies adopted by the virus to counteract the host cell recognition systems and the subsequent antiviral responses.

[0004] Many observations showed that several HBV viral proteins could counteract the initial host cellular response by interfering with the viral recognition signaling system and subsequently the interferon (IFN) antiviral activity. Among these, the excessive secretion of HBV empty subviral particles (SVPs, HBsAg) are thought to participate to the maintenance of the immunological tolerant state observed in chronically infected patients (CHB). The persistent exposure to HBsAg and other viral antigens can lead to HBV-specific T-cell deletion or to progressive functional impairment (Kondo, Journal of Immunology (1993), 150, 4659-4671; Kondo, Journal of Medical Virology (2004), 74, 425-433; Fisicaro, Gastroenterology, (2010), 138, 682-93;). Moreover HBsAg has been reported to suppress the function of immune cells such as monocytes, dendritic cells (DCs) and natural killer (NK) cells by direct interaction (Op den Brouw, Immunology, (2009b), 126, 280-9; Woltman, PLoS One, (2011), 6, e15324; Shi, J Viral Hepat. (2012), 19, e26-33; Kondo. ISRN Gasteroenterology, (2013), Article ID 935295).

[0005] HBsAg quantification is a significant biomarker for prognosis and treatment response in chronic hepatitis B. However the achievement of HBsAg loss and seroconversion is rarely observed in chronically infected patients but remains one of the ultimate goals of therapy. Current therapy such as Nucleos(t)ide analogues are molecules that inhibit HBV DNA synthesis but are not directed at reducing HBsAg level. Nucleos(t)ide analogs, even with prolonged therapy, only show weak HBsAg clearance comparable to those observed naturally (between -1%-2%) (Janssen, Lancet, (2005), 365, 123-9; Marcellin, N. Engl. J. Med., (2004), 351, 1206-17; Buster, Hepatology, (2007), 46, 388-94).

[0006] Hepatitis B e-antigen (also called HBV envelope antigen or HBeAg) is a viral protein that is secreted by hepatitis B infected cells. HBeAg is associated with chronic hepatitis B infections and is used as a marker of active viral disease and a patient's degree of infectiousness.

[0007] The function of the hepatitis B virus precore or HBeAg is not completely known. However HBeAg is well known to play a key role in viral persistence. HBeAg is thought to promote HBV chronicity by functioning as an immunoregulatory protein. In particular, the HBeAg is a secreted accessory protein, which appears to attenuate the host immune response to the intracellular nucleocapsid protein (Walsh, Virology, 2011, 411(1):132-141). The HBeAg acts as an immune tolerogen contributing to HBV persistence, and possibly functions in utero considering that soluble HBeAg traverses the placenta (Walsh, Virology, 2011, 411(1):132-141). Furthermore, HBeAg downregulates: i) cellular genes controlling intracellular signaling; and ii) the Toll-like receptor 2 (TLR-2) to dampen the innate immune response to viral infection (Walsh, Virology, 2011, 411(1):132-141). In the absence of HBeAg, HBV replication is associated with upregulation of the TLR2 pathway (Walsh, Virology, 2011, 411(1):132-141). Accordingly, HBeAg has a significant role in modulating virus/host interactions to influence the host immune response (Walsh, Virology, 2011, 411(1):132-141). Thus, reducing HBeAg in HBeAg positive patient population may lead to reversal of HBV specific immunedysfunction (Milich, 1997, J. Viral. Hep. 4: 48-59; Milich, 1998, J. Immunol. 160: 2013-2021). In addition, the secreted HBeAg is significantly more efficient than the intracellular hepatitis core antigen (HBcAg) at eliciting T-cell tolerance, and the split T-cell tolerance between the HBeAg and the HBcAg and the clonal heterogeneity of HBc/HBeAg-specific T-cell tolerance may have significant implications for natural HBV infection and especially for precore-negative chronic hepatitis (Chen, 2005, Journal of Virology, 79: 3016-3027).

[0008] Accordingly, reducing secretion of HBeAg in addition to secretion of HBsAg would lead to an improved inhibition of development of a chronic HBV infection as compared to the inhibition of secretion of HBsAg alone. In addition, the highest rates of transmission of an acute infection to chronic (>80%) have been reported in cases of materno-fetal and neonatal HBV transmission from HBeAg-positive mothers (Liaw, Lancet, 2009, 373: 582-592; Liaw, Dig. Dis. Sci., 2010, 55: 2727-2734; and Hadziyannis, 2011, Journal of hepatology, 55: 183-191). Therefore, reducing HBeAg in an expected mother may not only reduce the patient's degree of infectiousness, but may also inhibit the development of a chronic HBV infection of her child.

[0009] Therefore, in the therapy of HBV there is an unmet medical need to inhibit viral expression, particularly to inhibit secretion of HBsAg and HBeAg (Wieland, S. F. & F. V. Chisari. J Virol, (2005), 79, 9369-80; Kumar et al. J Virol, (2011), 85, 987-95; Woltman et al. PLoS One, (2011), 6, e15324; Op den Brouw et al. Immunology, (2009b), 126, 280-9).

[0010] WO 03/022987 discloses for example in Table 7A 1298 genes that are upregulated in hepatitis C-positive tissue. One of the mentioned genes is topoisomerase-related function protein 4 (TRF4, AF089897). AF089897 is also called TRF4-2, which is quite similar to position 880 to 2340 of SEQ ID NO: 4 herein. The observation that a fragment of PAPD5 is upregulated slightly in hepatitis C positive cells does not provide any indication that inhibiting PAPD5 represents an effective therapy. WO 03/022987A2 does not disclose any hint that fragments of PAPD5 plays any critical role during hepatitis C infection at all. In addition, HCV and HBV are two completely different viruses leading to two completely different diseases with different etiologies, different progression and different medication. This is in line with the observation of the present inventors that the PAPD5 and PAPD7 inhibitors DHQ and THP are inactive against hepatitis C virus (HCV) or other viruses beside HBV (data not shown).

[0011] In WO 2010/040571 PAPD5 has been suggested in a long list of other genes as having a potential role in cell proliferation in metabolic and tumorous disease without the provision of any actual evidence.

[0012] In WO2013/166264 PAPD5 has been suggested in a long list of other genes as having a potential role in increasing viral replication without the provision of any actual evidence.

[0013] In WO 2017/066712 down regulation of PAPD5 in relation to the treatment and diagnosis of telomere diseases has been described. Five shRNA structures for this purpose have been described.

[0014] To our knowledge the expression of PAPD5 or PAPD7 has never been associated with HBV infection.

OBJECTIVE OF THE INVENTION

[0015] Thus, the technical problem underlying the present invention is the identification and provision of ameliorated means and methods for treating and/or preventing a HBV infection.

[0016] The technical problem is solved by the provision of the embodiments described herein and characterized in the claims.

SUMMARY OF INVENTION

[0017] One aspect of the present invention relates to a screening method, particularly to a method for identifying a compound that prevents, ameliorates and/or inhibits a HBV infection, comprising:

[0018] (a) contacting a test compound with

[0019] (a1) PAPD5 polypeptide and/or PAPD7 polypeptide; or

[0020] (a2) a cell expressing PAPD5 and/or PAPD7;

[0021] (b) measuring the expression and/or activity of PAPD5 and/or PAPD7 in the presence and absence of said test compound; and

[0022] (c) identifying a compound that reduces the expression and/or activity of PAPD5 and/or PAPD7 as a compound that prevents, ameliorates and/or inhibits a HBV infection.

[0023] A further aspect of the invention is a method for identifying a compound that prevents, ameliorates and/or inhibits a HBV infection, comprising:

[0024] (a) contacting a test compound with [0025] (i) PAPD5 and/or PAPD7 polypeptide; or [0026] (ii) a cell expressing PAPD5 and/or PAPD7;

[0027] (b) measuring whether the test compound binds to the PAPD5 and/or to PAPD7 polypeptide;

[0028] (c) measuring whether the test compound inhibits propagation of HBV; and

[0029] (d) identifying a compound that binds to PAPD5 and/or PAPD7 polypeptide and inhibits propagation of HBV as a compound that prevents, ameliorates and/or inhibits a HBV infection.

[0030] A further aspect of the present invention is an inhibitor of PAPD5 and/or PAPD7 for use in treating and/or preventing a HBV infection, wherein said inhibitor is

[0031] (a) a small molecule that binds to PAPD5 and/or PAPD7; or

[0032] (b) an antibody that specifically binds to PAPD5 and/or PAPD7.

[0033] The inhibitor for the use in treating or preventing HBV can be selected from compounds of Formula (I) or (II). In particular the inhibitors of Formula (III) and (IV) are useful in the invention.

BRIEF DESCRIPTION OF THE FIGURES

[0034] The Figures show:

[0035] FIG. 1: Pictures from 1-by-1 experiment with HBX129653/HBX129654 chemical probes and the 3 prey fragments.

[0036] FIG. 2: Pictures from 1-by-1 experiment with HBX129653/HBX129654 chemical probes and PAPD5/7 full length proteins.

[0037] FIG. 3: Pictures from competition assay using HBX129653 (DHQ) and MOL653/654 for competition

[0038] FIG. 4: Pictures from competition assay using HBX129654 (THP) and MOL653/654 for competition

[0039] FIG. 5: Pictures from competition assay using HBX129653 (DHQ) and INACT653/INACT654 for competition. MOL653 was included as positive control.

[0040] FIG. 6: Pictures from competition assay using HBX129654 (THP) and INACT653/INACT654 for competition. MOL653 was included as positive control.

[0041] FIG. 7: (A) SiRNA knock-down (KD) of PAPD5 and PAPD7 in HBV-infected dHepaRG leads to reduction in HBV expression. Differentiated HepaRG cells were infected with HBV and treated with siRNA against either PAPD5, PAPD7 or both (25 nM each) one day prior to HBV infection and on day 4 post infection. Supernatant were harvested on day 11, levels of HBsAg and HBeAg secreted in the supernatant were measured by ELISA and normalized to non-treated control. Cell toxicity and inhibition of gene expression was measured subsequently and also normalized to the non-treated control. (B) The same experiment as described in (A) was performed, with the exception that only the level of HBsAg secreted in the supernatant was measured.

DETAILED DESCRIPTION OF THE INVENTION

[0042] PAPD5 and PAPD7 are non-canonical poly(A)-polymerases that belong to the superfamily of polymerase .beta.-like nucleotidyl transferases. In context of the present invention it has surprisingly been shown that a compound that is useful for the therapeutic intervention of a HBV infection can successfully be identified by analyzing whether a test compound inhibits PAPD5 and/or PAPD7. Or, in other words, inhibition of PAPD5 and/or PAPD7 was identified in the appended examples as being an indicator for the efficacy of a compound to inhibit a HBV infection. The appended examples demonstrate that a dihydroquinolizinone compound having the formula (III) as shown herein below (herein called DHQ) and a tetrahydropyridopyrimidine compound having the formula (IV) as shown herein below (herein called THP) bind to PAPD5 and PAPD7 polypeptides. These compounds have the capacity to inhibit production of HBV surface antigen (HBsAg) and the expression of HBV RNA during HBV infection (WO 2015/113990 A1 and WO2016/177655). In addition, the appended examples show that inhibition of PAPD5 and/or PAPD7 by using siRNA leads to an inhibition of viral expression, particularly of the secretion of HBsAg and HBeAg as well as of the production of intracellular HBV mRNA. These results directly indicate that by reducing the amount and/or activity (e.g. the amount) of PAPD5 and/or PAPD7 an HBV infection (e.g. a chronic HBV infection) can be prevented or treated (i.e. ameliorated and/or inhibited). Thus, the present invention relates to a screening method, wherein a compound that reduces the expression and/or activity (e.g. the expression) of PAPD5 and/or PAPD7 (e.g. of PAPD5 and PAPD7) is identified as a compound that prevents and/or treats (i.e. ameliorates and/or inhibits) a HBV infection.

[0043] It has been found in context of the present invention that a compound antagonizes (i.e. inhibits) PAPD5 and/or PAPD7 leads to inhibition of HBV gene expression and replication; and thus, prevents, ameliorates and/or inhibits a HBV infection. Such a compound may lead to a reduction of the PAPD5 and/or PAPD7 expression and/or activity of 10-100%, preferably of 20-100%, more preferably of 30-100%, even more preferably of 40-100%, even more preferable of 50-100%, even more preferably of 60-100%, even more preferably of 70-100%, even more preferably of 80-100%, and most preferably of 90-100%.

[0044] In the herein provided screening method it is envisaged that the expression of PAPD5 and/or PAPD7 is measured (i.e. analyzed/determined) by using in step (a) a cell expressing PAPD5 and/or PAPD7, i.e. (ai). The activity of PAPD5 and/or PAPD7 may be measured (i.e. analyzed/determined) by using in step (a) either (ai) PAPD5 and/or PAPD7 polypeptide, e.g. in a cell-free preparation; or (aii) a cell expressing PAPD5 and/or PAPD7.

[0045] In one aspect of the invention, a compound that reduces the expression of PAPD5 and/or PAPD7 (e.g. of PAPD5, preferably of PAPD5 and PAPD7) is identified as a compound that prevents, ameliorates and/or inhibits (i.e. treats) HBV infection. In another aspect of the invention a compound that reduces the activity of PAPD5 and/or PAPD7 (e.g. of PAPD5, preferably of PAPD5 and PAPD7) is identified as a compound that prevents, ameliorates and/or inhibits (i.e. treats) a HBV infection. It is prioritized that a compound that reduces the expression and/or activity of PAPD5 or of both molecules, PAPD5 and PAPD7, is identified as a compound that prevents, ameliorates and/or inhibits a HBV infection. Most preferably, a compound that reduces the expression and/or activity of both molecules, PAPD5 and PAPD7, is identified as a compound that prevents, ameliorates and/or inhibits a HBV infection.

[0046] In accordance with the present invention a compound that prevents and/or treats (i.e. ameliorates and/or inhibits) a HBV infection can be identified (i.e. selected) by performing a first pre-selection step in order to identify a compound that binds to PAPD5 and/or PAPD7. Subsequently, in a second step, it may be evaluated whether a compound that has been identified as binding to PAPD5 and/or PAPD7 inhibits propagation of HBV. Thus, the present invention relates to a further screening method, wherein a compound that binds to PAPD5 and/or PAPD7 (e.g. to PAPD5 and PAPD7) and inhibits propagation of HBV is identified as a compound that prevents, ameliorates and/or inhibits (i.e. treats) a HBV infection.

[0047] Thus, the invention relates to a method for identifying a compound that prevents, ameliorates and/or inhibits a HBV infection, comprising:

[0048] (a) contacting a test compound with

[0049] (ai) PAPD5 polypeptide and/or PAPD7 polypeptide; or

[0050] (aii) a cell expressing PAPD5 and/or PAPD7;

[0051] (b) measuring whether the test compound binds to PAPD5 and/or to PAPD7;

[0052] (c) measuring whether the test compound inhibits propagation of HBV; and

[0053] (d) identifying a compound that binds to PAPD5 and/or PAPD7 and inhibits propagation of HBV as a compound that prevents, ameliorates and/or inhibits a HBV infection.

[0054] Thus, in accordance with the present invention a compound that binds to PAPD5 and/or PAPD7 (e.g. to PAPD5, preferably to PAPD5 and PAPD7) and inhibits propagation of HBV is identified as a compound that prevents, ameliorates and/or inhibits (i.e. treats) a HBV infection. It is prioritized that a compound that (i) binds to PAPD5, or that binds to both molecules, PAPD5 and PAPD7; and (ii) inhibits propagation of HBV, is identified as a compound that prevents, ameliorates and/or inhibits a HBV infection. Most preferably, a compound that binds to both molecules, PAPD5 and PAPD7, and inhibits propagation of HBV, is identified as a compound that prevents, ameliorates and/or inhibits a HBV infection.

[0055] The above described screening methods lead to the identification of a compound that prevents, ameliorates and/or inhibits a HBV infection. It is prioritized that said compounds ameliorates and/or inhibits (i.e. treats) a HBV infection. Thus, the herein provided screening methods are useful in the identification of a compound that treats a HBV infection.

[0056] In the context of the present invention, PAPD5 may be the PAPD5 polypeptide or the PAPD5 mRNA. It is prioritized in context of the screening methods provided herein that PAPD5 is the PAPD5 polypeptide. One aspect of the present invention relates to the herein provided screening methods, wherein the PAPD5 polypeptide is a polypeptide comprising or consisting of

[0057] (a) the amino acid sequence of SEQ ID NO: 1 or 2;

[0058] (b) an amino acid sequence having at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to an amino acid sequence of (a), wherein the polypeptide has poly-A polymerase function;

[0059] (c) the amino acid sequence of an enzymatically active fragment of SEQ ID NO: 1 or 2; or

[0060] (d) an amino acid sequence having at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to an amino acid sequence of (d), wherein the polypeptide has poly-A polymerase function.

[0061] Examples for enzymatically active fragments of SEQ ID NO: 1 or 2 (i.e. of PAPD5) are the nucleotidyltransferase domain at positions 145-256 of SEQ ID NO: 1 or 2, or the Cid1 poly A polymerase at positions 308-368 of SEQ ID NO: 1 or 2.

[0062] Another aspect of the present invention relates to the herein provided screening methods, wherein the cells expressing PAPD5 contain PAPD5 mRNA, a polynucleotide comprising or consisting of

[0063] (i) the nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 1 or 2;

[0064] (ii) a nucleotide sequence encoding an amino acid sequence having at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to SEQ ID NO: 1 or 2, wherein the polynucleotide encodes a polypeptide that has poly-A polymerase function;

[0065] (iii) the nucleotide sequence encoding an enzymatically active fragment of SEQ ID NO: 1 or 2;

[0066] (iv) a nucleotide sequence encoding an amino acid sequence having at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to an amino acid sequence of an enzymatically active fragment of SEQ ID NO: 1 or 2, wherein the polynucleotide encodes a polypeptide that has poly-A polymerase function;

[0067] (v) a nucleotide sequence comprising or consisting of SEQ ID NO: 4 or 5; or

[0068] (vi) a nucleotide sequence having at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to SEQ ID NO: 4 or 5, wherein the polypeptide expressed from the sequence has poly-A polymerase function; or

[0069] (vii) a pre-mRNA that when processed (i.e. spliced) leads to a polynucleotide of (v) or (vi).

[0070] In preferred embodiments, the PAPD5 mRNA may be a polynucleotide comprising or consisting of the nucleotide sequence of SEQ ID NO: 4 or 5. However, the PAPD5 mRNA may also be a polynucleotide comprising or consisting of a nucleotide sequence having at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to SEQ ID NO: 4 or 5, wherein the polynucleotide encodes a polypeptide that has poly-A polymerase function.

[0071] In context of the present invention PAPD7 may be the PAPD7 polypeptide or the PAPD7 mRNA. It is prioritized in context of the screening methods provided herein that PAPD7 is the PAPD7 polypeptide. One aspect of the present invention relates to the herein provided screening methods, wherein the PAPD7 polypeptide is a polypeptide comprising or consisting of

[0072] (a) the amino acid sequence of SEQ ID NO: 3;

[0073] (b) an amino acid sequence having at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to an amino acid sequence of (a), wherein the polypeptide has poly-A polymerase function;

[0074] (c) the amino acid sequence of an enzymatically active fragment of SEQ ID NO: 3; or

[0075] (d) an amino acid sequence having at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to an amino acid sequence of (c), wherein the polypeptide has poly-A polymerase function.

[0076] Examples for enzymatically active fragments of SEQ ID NO: 3 (i.e. of PAPD7) are the nucleotidyltransferase domain at positions 15-125 of SEQ ID NO: 3; or the Cid1 family poly A polymerase at positions 178-238 of SEQ ID NO: 3.

[0077] Another aspect of the present invention relates to the herein provided screening methods, wherein the cells expressing PAPD7 contain PAPD7 mRNA, a polynucleotide comprising or consisting of

[0078] (i) the nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 3;

[0079] (ii) a nucleotide sequence encoding an amino acid sequence having at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to SEQ ID NO: 3, wherein the polynucleotide encodes a polypeptide that has poly-A polymerase function;

[0080] (iii) the nucleotide sequence encoding an enzymatically active fragment of SEQ ID NO: 3; or (iv) a nucleotide sequence encoding an amino acid sequence having at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to an amino acid sequence of an enzymatically active fragment of SEQ ID NO: 3, wherein the polynucleotide encodes a polypeptide that has poly-A polymerase function; or

[0081] (v) a nucleotide sequence comprising or consisting of SEQ ID NO: 6; or

[0082] (vi) a nucleotide sequence having at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to SEQ ID NO: 6, wherein the polypeptide expressed from the sequence has poly-A polymerase function; or

[0083] (vii) a pre-mRNA that when processed (i.e. spliced) leads to a polynucleotide of (v) or (vi).

[0084] In preferred embodiments, the PAPD7 mRNA may be a polynucleotide comprising or consisting of the nucleotide sequence of SEQ ID NO: 6. However, the PAPD7 mRNA may also be a polynucleotide comprising or consisting of a nucleotide sequence having at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to SEQ ID NO: 6, wherein the polynucleotide encodes a polypeptide that has poly-A polymerase function.

[0085] In context of the present invention said cell may be a eukaryotic cell. For example, said cell may be a yeast cell or a vertebrate cell. Vertebrate cells include fish, avian, reptilian, amphibian, marsupial, and mammalian cells. Preferably, the cell is a mammalian cell, most preferably, a human cell. Mammalian cells also include feline, canine, bovine, equine, caprine, ovine, porcine murine, such as mice and rat, and rabbit cells. In the herein provided screening methods, the "cell" may endogenously express PAPD5 and/or PAPD7 or overexpress PAPD5 and/or PAPD7. For overexpressing PAPD5 and/or PAPD7 the cell may comprise the nucleotide sequence encoding the PAPD5 polypeptide and/or the PAPD7 polypeptide within an expression vector. In preferred embodiments the cell comprise a nucleotide sequence encoding the PAPD5 polypeptide and a nucleotide sequence encoding the PAPD7 polypeptide. The cell of the herein provided screening methods may be comprised in a non-human animal, e.g. a mouse, rat, rabbit or ferret.

[0086] In the above described screening method wherein the binding to PAPD5 and/or PAPD7 is measured, a compound may be identified as a compound that binds to PAPD5 polypeptide and/or PAPD7 polypeptide if it has a particular binding affinity to PAPD5 and/or PAPD7. For example, the compound that binds to PAPD5 and/or PAPD7 may have a dissociation constant (Kd) in the micromolar range; or, preferably, in the range of 100 nM to 1 pM.

[0087] In context of the present invention it may be measured (i.e. analyzed) whether the test compound specifically binds to PAPD5 polypeptide and/or PAPD7 polypeptide, i.e. whether the test compound exclusively or predominately binds to PAPAD5 and/or PAPD7. For example, it may be measured whether the test compound specifically binds to PAPD7. Preferably, it is measured whether the test compound specifically binds to PAPD5. More preferably it is measured whether the test compound binds to both, PAPD5 and PAPD7. For example, it may be measured whether the test compound specifically binds to PAPD5 and PAPD7.

[0088] For example, in the herein provided screening methods, binding of the test compound to PAPD5 and/or PAPD7 may be measured by conducting a yeast 3 hybrid screen. The Y3H system is a modified version of the yeast two-hybrid (Y2H) system adapted for the detection of drug-protein interactions. It requires coupling of the drug of interest with a ligand that can be anchored to a DNA-binding protein inside yeast cells. The interaction of the anchored drug with a target protein is then detected by linking their association to the transcriptional activation of a reporter gene; see, e.g. Johnsson, Nature Chem Bio, 2011, 7: 375-383; and Licitra, Proc Natl Acad Sci USA, 1996, 12; 93(23):12817-21. In such a yeast 3 hybrid screen an inactive free compound may be used for competition against the labeled test compound.

[0089] Binding of a test compound to PAPD5 and/or PAPD7 may also be measured by using Biacore, ChemoProteomics, or Microscale Thermophoresis.

[0090] A compound that inhibits the propagation of HBV may be a compound that reduces the expression of viral RNA, that reduces the production of viral DNA (HBV DNA) deriving from viral RNA (HBV RNA), that reduces the production of new viral particles (HBV particles), and/or that produces production and/or secretion of HBsAg and/or HBeAg. Thus, one aspect of the present invention relates to the herein provided screening methods, wherein the compound that inhibits propagation of HBV inhibits secretion of HBsAg, inhibits secretion of HBeAg, and/or inhibits production of intracellular HBV mRNA or HBV DNA. Preferably, a compound that inhibits the propagation of HBV is a compound that inhibits secretion of HBsAg, secretion of HBeAg and production of intracellular HBV mRNA.

[0091] For example, a compound that inhibits propagation of HBV may reduce the expression of viral RNA (HBV RNA), the production of viral DNA (HBV DNA) deriving from viral RNA, the production of new viral particles (HBV particles), the production and/or secretion of HBsAg and/or HBeAg by 10-100%, preferably by 20-100%, more preferably by 30-100%, even more preferably by 40-100%, even more preferable by 50-100%, even more preferably by 60-100%, even more preferably by 70-100%, even more preferably by 80-100%, and most preferably by 90-100%, when compared the untreated cells or animals or cell or animal treated with an appropriate control.

[0092] The herein provided screening methods may additionally comprise the step of comparing the test compound to a control. Said control may be an inactive test compound, wherein said inactive test compound is a compound that:

[0093] (i) does not reduce the expression and/or activity of PAPD5 and/or PAPD7; and/or

[0094] (ii) does not bind to PAPD5 and/or PAPD7 and does not inhibit propagation of HBV.

[0095] This inactive test compound has no activity against HBV, e.g. it does not lead to inhibition of secretion of HBsAg and HBeAg and to inhibition of production of intracellular HBV mRNA. For example, the inactive test compound may have an IC.sub.50 value in the inhibition of HBsAg of more than 3 In the herein provided screening method, the inactive test compound may be the compound "DHQ compound inactive" or the compound "THP compound inactive" as defined in the appended examples. In the screening method wherein expression and/or activity of PAPD5 and/or PAPD7 is measured, the test compound as defined above in (i) may be used. Alternatively, in the screening method wherein binding to PAPD5 and/or PAPD7 is measured, the test compound as defined above in (ii) may be used. An inactive compound can be designed from an active one, e.g., by chemical modification and/or chiral separation.

[0096] In the herein provided screening methods, the activity of PAPD5 and/or PAPD7 in the presence and absence of the test compound may be measured, e.g. by monitoring the in vitro polyadenylation of mRNA, e.g., as described in Rammelt, RNA, 2011, 17:1737-1746. In brief, a ribo-oligonucleotide A.sub.15 may be incubated with recombinant PAPD5 protein expressed in Escherichia coli in the presence of ATP(A), CTP (C), GTP(G), UTP(U), or a mixture of all four dNTPs, respectively.

[0097] The expression of PAPD5 and/or PAPD7 in the presence and absence of the test compound may be measured, e.g. by using (q)PCR, western blot, or MassSpec.

[0098] Inhibition of propagation of HBV may be measured, e.g., by measuring whether the test compound has the activity to inhibit secretion of HBsAg and/or of HBeAg, and/or to inhibit production of intracellular HBV mRNA. Inhibition of secretion of HBsAg and/or HBeAg may be measured by ELISA, e.g. by using the CLIA ELISA Kit (Autobio Diagnostic) according to the manufacturers' instructions. Inhibition of production of intracellular HBV mRNA may be measured by real-time PCR, e.g. as described in the appended examples. Further methods for evaluating whether a test compound inhibits propagation of HBV are measuring secretion of HBV DNA by RT-qPCR e.g. as described in WO 2015/173208; Northern Blot; in-situ hybridization, or immuno-fluorescence.

[0099] For performing the herein provided screening methods publicly or commercially available molecule libraries may be used. Thus, in context of the invention the said test compound may be

[0100] (i) a small molecule of a screening library; or

[0101] (ii) a peptide of a phage display library, of an antibody fragment library, or derived from a cDNA library.

[0102] For example, the cDHA Human Liver (HLV) library or the cDNA Human Placenta (PLA) library of Hybrigenics Services SAS may be used.

[0103] In the herein provided screening method wherein the activity of PAPD5 polypeptide and/or PAPD7 polypeptide is measured, said activity of PAPD5 and PAPD7 is preferably the poly-A polymerase function (i.e. the poly-A polymerase activity). The poly-A polymerase function/activity of a polypeptide (e.g. of PAPD5 or PAPD7) may be measured, e.g. by monitoring the in vitro polyadenylation of mRNA, e.g. as described in Rammelt, RNA, 2011, 17:1737-1746. This method can also be used to measure the poly-A polymerase function of PAPD5 and/or PAPD7 in the presence and absence of a test compound.

[0104] The appended examples demonstrate that by inhibiting PAPD5 and/or PDPD7 polypeptide, the secretion of HBsAg and HBeAg as well as production of intracellular HBV mRNA can effectively be inhibited. These data demonstrate that an inhibitor of PAPD5 and/or PAPD7 can be used to prevent and/or treat a HBV infection.

[0105] Several compounds that have a certain efficacy in the treatment of a HBV infection have been described in the art (see, e.g. WO 2015/113990 A1 and WO 2016/177655). However, in context of the present invention it has surprisingly been found that anti-HBV agents that are completely different in structure (e.g. DHQ and THP) surprisingly and specifically bind to PAPD5 and PAPD7. In addition, the prior art also encompass agents less active in the inhibition of HBsAg production. Such agents have been shown in the appended examples to have less binding affinity to PAPD5 and PAPD7 (see, e.g., "DHQ inactive"). Indeed, the appended examples demonstrate a clear correlation between activity of the compound against a HBV infection and binding affinity towards PAPD5 and PAPD7. Thus, selectively using an anti-HBV agent that binds to PAPD5 and/or PAPD7 leads to particularly high anti-HBV efficacy. Furthermore, the present invention shows for the first time that a compound that inhibits PAPD5, PAPD7, or particularly PAPD5 and PAPD7 has an extraordinary high activity in terms of inhibition of secretion of HBsAg and HBeAg as well as of production of intracellular HBV mRNA. Reduction of secretion of HBsAg and HBeAg inhibits development of chronic HBV infection more effectively as compared to the reduction of secretion of HBsAg alone. In addition, inhibition of secresion of HBsAg and HBeAg reduces the infectiousness of a HBV infected person. Furthermore, reducing HBeAg in an expected mother may also inhibit the development of a chronic HBV infection of her child. Thus, the present invention unexpectedly demonstrates that selectively using compounds that inhibit PAPD5 and/or PAPD7 leads to an improved therapeutic success in the treatment of a HBV infection in terms of a considerably more effective reduction of HBsAg and HBeAg.

[0106] Accordingly, an aspect of the present invention is use of an inhibitor of PAPD5 and/or PAPD7 in the treatment of HBV infection, in particular a chronic HBV infection. In a further embodiment the invention relates to the use of an inhibitor of a PAPD5 and/or PAPD7 in reduction of the viral antigens HBsAg and HBeAg.

[0107] Thus, the present invention relates to an inhibitor of PAPD5 and/or PAPD7 for use in treating and/or preventing a HBV infection, wherein said inhibitor is

[0108] (i) a small molecule that binds to PAPD5 and/or PAPD7;

[0109] (ii) a RNA interference (RNAi) molecule against PAPD5 and/or PAPD7;

[0110] (iii) an antibody that specifically binds to PAPD5 and/or PAPD7; or

[0111] (iv) a genome editing machinery, comprising:

[0112] (a) a site-specific DNA nuclease or a polynucleotide encoding a site-specific DNA nuclease; and

[0113] (b) a guide RNA or a polynucleotide encoding a guide RNA.

[0114] The inhibitor of the present invention may also be a PAPD5 and/or PAPD7 specific locked nucleic acid (LNA) molecule.

[0115] It is envisaged that the inhibitor of the invention is used for treating (e.g. ameliorating) a HBV infection.

[0116] The inhibitor may be a molecule that specifically inhibits PAPD7. Preferably, the inhibitor is a molecule that specifically inhibits PAPD5. More preferably, the inhibitor inhibits both, PAPD5 and PAPD7. Thus, it is prioritized that the inhibitor of the present invention either inhibits PAPD5 or both, PAPD5 and PAPD7. Most preferably, the inhibitor of the present invention inhibits PAPD5 and PAPD7. In one aspect of the invention the inhibitor of the present invention inhibits both, PAPD5 and PAPD7 and leads to a reduction of secretion of HBsAg and/or HBeAg of at least 50% as compared to the no drug control (i.e. compared to cells or subjects to which no drug has been administrated).

[0117] The inhibitor of the present invention may have an IC.sub.50 value in the inhibition of HBsAg and HBeAg of below 3 .mu.M, preferably of below 2 .mu.M, more preferably belo 1 .mu.M, more preferably below 0.1 .mu.M, and most preferably below 0.01 .mu.M.

[0118] Genome editing by using a site-specific DNA nuclease (such as Cas9 or Cpf1) and a guide RNA is commonly known in the art and described, e.g., in "CRISPR-Cas: A Laboratory Manual", 2016, edited by Jennifer Doudna, ISBN 978-1-621821-31-1.

[0119] For example, if said site-specific DNA nuclease is a Cas9 nuclease, then the genome editing machinery preferably further comprises:

[0120] (i) at least one guide RNA consisting of at least one target sequence specific CRISPR RNA (crRNA) molecule and at least one trans-activating crRNA (tracrRNA) molecule;

[0121] (ii) a polynucleotide encoding the RNA molecules of (i);

[0122] (iii) at least one guide RNA, which is a chimeric RNA molecule comprising at least one target sequence specific crRNA and at least one tracrRNA; or

[0123] (iv) a polynucleotide encoding the chimeric RNA of (iii).

[0124] In an alternative example the site-specific DNA nuclease is a Cpf1 nuclease, and the genome editing machinery preferably further comprises:

[0125] (i) at least one guide RNA comprising a target sequence specific CRISPR RNA (crRNA) molecule; or

[0126] (ii) a polynucleotide encoding the RNA molecules of (i).

[0127] The herein provided inhibitor of PAPD5 and/or PAPD7 may also be a genome editing machinery that comprises at least one pre-assembled Cas9 protein-guide RNA ribonucleoprotein complex (RNP).

[0128] Herein, the guide RNA is designed to target the genomic PAPD5 or PAPD7 DNA. Alternatively, several guide RNAs are used, so that the genomic DNA of PAPD5 and of PAPD7 can be targeted. Inhibition of PAPD5 and/or PAPD7 may be achieved by introducing frame-shift knockout mutations into the genomic PAPD5 and/or PAPD7 DNA through non-homologous end-joining (NHEJ), or by modifying the genomic PAPD5 and/or PAPD7 DNA through homology-directed repair (HDR). How these mechanisms can be induced is commonly known in the art and described, e.g., in Heidenreich, 2016, Nat Rev Neurosci 17 36-44.

[0129] The inhibitor of the present invention may be a naturally occurring molecule, e.g. a naturally occurring antibody or a naturally occurring RNAi molecule. However, the inhibitor of the present invention may also be a non-naturally occurring molecule. For example, the inhibitor of the invention may be an antibody having an amino acid sequence that is not identical to naturally occurring antibodies or may be an antibody comprising at least one non-naturally occurring amino acid residue such as synthetic amino acids providing similar side chain functionality. For example, aromatic amino acids may be replaced with D- or L-naphthylalanine, D- or L-phenylglycine, D- or L-2-thienylalanine, D- or L-1-, 2-, 3-, or 4-pyrenylalanine, D- or L-3-thienylalanine, D- or L-(2-pyridinyl)-alanine, D- or L-(3-pyridinyl)-alanine, D- or L-(2-pyrazinyl)-alanine, D- or L-(4-isopropyl)-phenylglycine, D-(trifluoromethyl)-phenylglycine, D-(trifluoromethyl)-phenylalanine, D-p-fluorophenylalanine, D- or L-pbiphenylalanine D- or L-p-methoxybiphenylalanine, D- or L-2-indole(alkyl)alanines, and D- or Lalkylalanines wherein the alkyl group is selected from the group consisting of substituted or unsubstituted methyl, ethyl, propyl, hexyl, butyl, pentyl, isopropyl, iso-butyl, and iso-pentyl. Non-carboxylate amino acids can be made to possess a negative charge, as provided by phosphono- or sulfated amino acids, which are to be considered as non-limiting examples. Further non-natural amino acids are alkylated amino acids, made by combining an alkyl group with any natural amino acid. Basic natural amino acids such as lysine and arginine may be substituted with alkyl groups at the amine (NH.sub.2) functionality. Yet other substitutions on non-natural amino acids include nitrile derivatives (e.g., containing a CN-moiety in place of the CONH.sub.2 functionality) of asparagine or glutamine, and sulfoxide derivative of methionine.

[0130] Analogously, the inhibitor of the invention may be a RNAi molecule having a nucleotide sequence that is not identical to naturally occurring RNAi molecules or may be a RNAi molecule comprising at least one non-naturally occurring nucleotides, such as a oligonucleotide thiophosphate, a substituted ribo-oligonucleotide, a LNA molecule, a PNA molecule, a GNA (glycol nucleic acid) molecule, a TNA (threose nucleic acid) molecule, a morpholino polynucleotide, or a nucleic acid with a modified backbone such as polysiloxane, 2'-O-(2-methoxy) ethyl-phosphorothioate, or a nucleic acid with a substituent, such as methyl-, thio-, sulphate, benzoyl-, phenyl-, amino-, propyl-, chloro-, and methanocarbanucleoside, or a reporter molecule to facilitate its detection. The inhibitor of the invention may also be naturally occurring or a non-naturally occurring small molecule or genome editing machinery.

[0131] In context of the present invention, the herein provided inhibitor may

[0132] (i) bind to PAPD5 and/or PAPD7 polypeptide; and/or

[0133] (ii) inhibit expression and/or activity of PAPD5 and/or PAPD7.

[0134] For example, the inhibitor of the present invention may bind to PAPD5 polypeptide and inhibit activity of PAPD5 polypeptide. In another example, the inhibitor of the present invention binds to PAPD7 polypeptide and inhibits activity of PAPD7 polypeptide. It is prioritized herein that the inhibitor binds to both, PAPD5 and PAPD7 polypeptide and inhibits the activity of both, PAPD5 and PAPD7 polypeptide. The inhibitor of the present invention may inhibit the expression of PAPD5 or PAPD7; or may inhibit the expression of both, PAPD5 and PAPD7.

[0135] As described above, in context of the present invention it has been shown that a compound that inhibits PAPD5 and/or PAPD7 has a high activity in terms of inhibition of secretion of HBsAg and HBeAg as well as of production of intracellular HBV mRNA. Therefore, the inhibitor of the present invention reduces secretion of HBsAg and HBeAg. Due to the reduction of HBsAg secretion the inhibitor of the present invention inhibits development of chronic HBV infection. In particular, due to inhibition of HBeAg secretion, the inhibitor of the present invention more efficiently inhibits development of a chronic HBV infection as compared to a compound that only reduces secretion of HBsAg. In addition, reducing HBeAg in an expected mother may also inhibit the development of a chronic HBV infection of her child. Thus, due to the reduction of HBeAg secretion the inhibitor of the present invention inhibits development of a chronic HBV infection (such as development of a chronic HBV infection in the offspring of an HBV infected mother) and reduces the infectiousness of a HBV infected person. Accordingly, one aspect of the present invention related to the herein provided inhibitor, wherein the inhibitor reduces secretion of HBsAg and HBeAg. In line with this, a further aspect of the invention relates to the herein provided inhibitor, wherein the inhibitor inhibits development of chronic HBV infection and reduces the infectiousness of a HBV infected person. In a particular aspect of the invention, the herein provided inhibitor inhibits development of a chronic HBV infection in the offspring of a HBV infected mother. This mother is preferably HBeAg positive.

[0136] The subject to be treated with the inhibitor of the invention (or which prophylactically receives the inhibitor of the present invention) is preferably a human, more preferably a human patient who is HBsAg positive and/or HBeAg positive, even more preferably a human patient that is HBsAg positive and HBeAg positive. Said human patient may be an expected mother, e.g. an expected mother who is HBeAg positive and/or HBsAg positive, more preferably an expected mother who is HBeAg positive and HBsAg positive.

[0137] Compounds of the Invention

[0138] As described above, the inhibitor of the present invention may be a small molecule. For example, the inhibitor of the invention may be the compound of formula (I) or (II):

##STR00001##

[0139] wherein

[0140] R.sup.1 is hydrogen, halogen, C.sub.1-6alkyl, C.sub.1-6alkylamino or C.sub.1-6alkoxy;

[0141] R.sup.2 is hydrogen; halogen; C.sub.1-6alkyl, which is unsubstituted or once, twice or three times substituted by fluoro; C.sub.1-6alkoxy, which is unsubstituted or once, twice or three times substituted by fluoro; cyano; C.sub.3-7cycloalkyl; hydroxy or phenyl-C.sub.xH.sub.2x--O--;

[0142] R.sup.3 is hydrogen;

[0143] halogen;

[0144] C.sub.1-6alkyl, which is unsubstituted or once, twice or three times substituted by fluoro;

[0145] cyano;

[0146] pyrrolidinyl;

[0147] amino;

[0148] phenyl-C.sub.xH.sub.2x--N(C.sub.1-6alkyl)-;

[0149] C.sub.1-6alkoxycarbonylpiperazinyl;

[0150] or R.sup.7--O--, wherein R.sup.7 is hydrogen; C.sub.1-8alkyl, which is unsubstituted or substituted with one to three substituents independently selected from fluoro, hydroxy and C.sub.2-6alkenyl; C.sub.1-6alkoxyC.sub.1-6alkyl; C.sub.1-6alkoxyC.sub.1-6alkoxyC.sub.1-6alkyl; aminoC.sub.1-6alkyl; C.sub.1-6alkylcarbonylaminoC.sub.1-8alkyl; C.sub.1-6alkylsulfonylaminoC.sub.1-8alkyl; C.sub.1-6alkylsulfanylC.sub.1-6alkyl; C.sub.1-6alkylsulfonylC.sub.1-8alkyl; cyanoC.sub.1-6alkyl; C.sub.3-7cycloalkylC.sub.1-6alkyl; cyanoC.sub.3-7cycloalkylC.sub.1-6alkyl; phenylC.sub.1-6alkyl; pyrrolidinylcarbonylC.sub.1-6alkyl; C.sub.2-6alkynyl; hydroxyC.sub.1-6alkylC.sub.2-6alkynyl; aminoC.sub.1-6alkoxyC.sub.1-6alkyl; C.sub.1-6alkylaminoC.sub.1-6alkoxyC.sub.1-6alkyl; diC.sub.1-6alkylaminoC.sub.1-6alkoxyC.sub.1-6alkyl; carboxyC.sub.1-6alkyl; or C.sub.1-6alkoxycarbonylaminoC.sub.1-8alkyl; heteroarylC.sub.1-6alkyl, wherein heteroaryl is N-containing monocyclic heteroaryl; or heterocycloalkylC.sub.1-6alkyl, wherein heterocycloalkyl is monocyclic heterocycloalkyl;

[0151] R.sup.4 is hydrogen, halogen, C.sub.1-6alkyl, cyano or C.sub.1-6alkoxy;

[0152] provided that R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are not hydrogen simultaneously;

[0153] R.sup.5 is hydrogen or C.sub.1-6alkyl;

[0154] R.sup.6 is hydrogen; C.sub.1-6alkyl, which is unsubstituted or once, twice or three times substituted by fluoro; C.sub.3-7cycloalkyl, which is unsubstituted or once, twice or three times substituted by fluoro or C.sub.1-6alkyl; or phenyl-C.sub.xH.sub.2x--;

[0155] x is 1-6;

[0156] or a pharmaceutically acceptable salt, or an enantiomer thereof, or a diastereomer thereof;

##STR00002##

[0157] wherein

[0158] R.sup.1 is C.sub.1-6alkyl, C.sub.3-7cycloalkyl, haloC.sub.1-6alkyl, hydroxyC.sub.1-6alkyl, nitroC.sub.1-6alkyl, C.sub.1-6alkoxycarbonylC.sub.1-6alkyl, carboxyC.sub.1-6alkyl, di(C.sub.1-6alkoxycarbonyl)methylenyl, cyanoC.sub.1-6alkyl, C.sub.3-7cycloalkylC.sub.1-6alkyl, phenylC.sub.1-6alkyl, C.sub.1-6alkylsulfanylC.sub.1-6alkyl, C.sub.1-6alkylsufonylC.sub.1-6alkyl, aminoC.sub.1-6alkyl, C.sub.1-6alkylcarbonylaminoC.sub.1-6alkyl, C.sub.1-6alkylsufonylaminoC.sub.1-6alkyl, C.sub.1-6alkoxycarbonyl aminoC.sub.1-6alkyl, aminocarbonylC.sub.1-6alkyl, diC.sub.1-6alkylaminocarbonylC.sub.1-6alkyl, monocyclic heterocycloalkylC.sub.1-6alkyl or imidazolylC.sub.1-6alkyl;

[0159] R.sup.2 is aryl or heteroaryl, said aryl or heteroaryl being unsubstituted, or substituted by one, two, three or four substituents independently selected from C.sub.1-6alkyl, C.sub.3-7cycloalkyl, halogen, haloC.sub.1-6alkyl, cyano, nitro, hydroxy, haloC.sub.1-6alkoxy, --O--C.sub.xH.sub.2x--R.sup.3, --O--C.sub.yH.sub.2y--NHR.sup.6, --NR.sup.9R.sup.19, --SO.sub.2--R.sup.11, --SO.sub.2--NR.sup.12R.sup.13, carboxy, C.sub.1-6alkoxycarbonyl, --C(.dbd.O)--NR.sup.12R.sup.13, aryl, heteroaryl, monocyclic heterocycloalkyl and --O-monocyclic heterocycloalkyl; wherein monocyclic heterocycloalkyl is unsubstituted or substituted by C.sub.1-6alkyl, C.sub.3-7cycloalkyl, C.sub.1-6alkylcarbonyl, C.sub.1-6alkylsufonyl or C.sub.1-6alkoxycarbonyl;

[0160] R.sup.3 is hydrogen; C.sub.3-7cycloalkyl; haloC.sub.3-7cycloalkyl; hydroxy; hydroxyC.sub.1-6alkylC.sub.3-7cycloalkyl; C.sub.1-6alkoxy; monocyclic heterocycloalkyl; monocyclic heterocycloalkyl substituted by C.sub.1-6alkyl, C.sub.1-6alkylcarbonyl, C.sub.1-6alkylsufonyl, C.sub.3-7cycloalkyl or C.sub.1-6alkoxycarbonyl; --C(.dbd.O)--R.sup.4; C.sub.1-6alkylsulfinyl; --SO.sub.2--R.sup.5; --C(NHR.sup.7)--C(.dbd.O)--R.sup.8; carboxyC.sub.1-6alkoxy or aminocarbonylC.sub.1-6alkoxy; wherein

[0161] R.sup.4 is hydroxy, C.sub.1-6alkoxy, amino, C.sub.1-6alkylamino, tetrahydrofuranylamino, pyrrolidinyl or morpholinyl;

[0162] R.sup.5 is C.sub.1-6alkyl, C.sub.3-7cycloalkyl, hydroxy, amino, C.sub.1-6alkylamino or diC.sub.1-6alkylamino;

[0163] R.sup.7 is hydrogen or C.sub.1-6alkoxycarbonyl;

[0164] R.sup.8 is hydroxy or C.sub.1-6alkoxy;

[0165] R.sup.6 is hydrogen, C.sub.1-6alkylcarbonyl, haloC.sub.1-6alkylcarbonyl, C.sub.1-6alkoxycarbonyl,

[0166] C.sub.1-6alkylsulfonyl, C.sub.3-7cycloalkylsulfonyl or C.sub.1-6alkoxyC.sub.1-6alkylsulfonyl;

[0167] R.sup.9 and R.sup.19 are independently selected from hydrogen, C.sub.1-6alkyl, C.sub.3-7cycloalkyl, C.sub.1-6alkylcarbonyl, C.sub.1-6alkylsulfonyl, C.sub.3-7cycloalkylcarbonyl and C.sub.3-7cycloalkylsulfonyl; or

[0168] R.sup.9 and R.sup.19 together with the nitrogen to which they are attached form monocyclic heterocycloalkyl;

[0169] R.sup.11 is C.sub.1-6alkyl, haloC.sub.1-6alkyl, C.sub.3-7cycloalkyl, haloC.sub.3-7cycloalkyl, hydroxyC.sub.1-6alkyl, C.sub.1-6alkoxyC.sub.1-6alkyl, haloC.sub.1-6alkoxyC.sub.1-6alkyl, C.sub.3-7cycloalkylC.sub.1-6alkyl, aminoC.sub.1-6alkyl, C.sub.1-6alkylaminoC.sub.1-6alkyl, C.sub.1-6alkylcarbonylaminoC.sub.1-6alkyl, C.sub.1-6alkylsulfonylaminoC.sub.1-6alkyl, C.sub.1-6alkoxycarbonylaminoC.sub.1-6alkyl, C.sub.1-6alkylsulfenylC.sub.1-6alkyl, C.sub.1-6alkylsulfanylC.sub.1-6alkyl or C.sub.1-6alkylsulfonylC.sub.1-6alkyl;

[0170] R.sup.12 and R.sup.13 are independently selected from hydrogen, C.sub.1-6alkyl, C.sub.1-6alkoxyC.sub.1-6alkyl, haloC.sub.1-6alkyl, C.sub.3-7cycloalkyl and haloC.sub.3-7cycloalkyl; or

[0171] R.sup.12 and R.sup.13 together with the nitrogen to which they are attached form monocyclic heterocycloalkyl;

[0172] x is 1, 2, 3, 4, 5, 6, 7 or 8;

[0173] y is 1, 2, 3, 4, 5, 6, 7 or 8;

[0174] U, W and Z are independently selected from CH and N;

[0175] one of X and Y is N, and the other one is CH or N;

[0176] or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

[0177] In one aspect of the invention 6-methyl-2-oxo-9-pyrrolidin-1-yl-6,7-dihydrobenzo[a]quinolizine-3-carboxy- lic acid, 9-fluoro-6-methyl-2-oxo-6,7-dihydrobenzo[a]quinolizine-3-carboxy- lic acid, and 9,10-difluoro-6-methyl-2-oxo-6,7-dihydrobenzo[a]quinolizine-3-carboxylic acid are excluded from the compound of formula (I).

[0178] In one particular embodiment of the present invention the compounds of formulae (I) and (II) are excluded from the inhibitor of the present invention. Thus, one embodiment of the present invention relates to the inhibitor of the present invention, wherein said inhibitor is not a compound according to formula (I) or (II).

[0179] As described above, the appended examples demonstrate that the anti-HBV agents DHQ (i.e. a compound of formula (III)) and THP (i.e. a compound of formula (IV)) effectively bind to PAPD5 and PAPD7. Thus, it is prioritized in context of the invention that the inhibitor of the invention is the compound of formula (III) or (IV):

##STR00003##

[0180] In the appended examples also derivatives of the compounds of formulae (III) and (IV) having a linker and anchor ligand have been shown to have binding affinity to PAPD5 and PAPD7. These derivatives are shown below as formulae (V) and (VI), respectively. Thus, in one aspect of the present invention the inhibitor of the invention is the compound of formula (V) or (VI):

##STR00004##

[0181] In context of the present invention the inhibitor of the invention may be the compound according to formula (I), wherein the inhibitor is any one of the compounds as defined in items (1)-(19), below:

[0182] 1. A compound according to formula (I), wherein

[0183] R.sup.1 is hydrogen, fluoro, chloro, bromo, methyl, methylamino, methoxy or ethoxy;

[0184] R.sup.2 is hydrogen, fluoro, chloro, bromo, methyl, ethyl, trifluoromethyl, methoxy, ethoxy, propoxy, trifluoromethoxy, cyano, cyclopropyl, hydroxy or phenylmethyl-O--;

[0185] R.sup.3 is hydrogen, bromo, methyl, propyl, trifluoromethyl, cyano, phenylmethyl-N(methyl)-, tert-butoxycarbonylpiperazinyl, hydroxy, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, difluoromethylmethyl-O--, difluoromethylethyl-O--, trifluoromethoxy, trifluoromethylmethyl-O--, trifluoromethylethyl-O--, ethyldifluoromethyl-O--, vinyldifluoromethyl-O--, propargyl-O--, hydroxymethylpropargyl-O--, methoxyethyl-O--, methoxypropyl-O--, methoxybutyl-O--, ethoxyethyl-O--, methoxyethyl-O-ethyl-O--, aminoethyl-O--, aminopentyl-O--, aminohexyl-O--, aminooctyl-O--, tert-butoxycarbonylaminopentyl-O--, tert-butoxycarbonylaminohexyl-O--, tert-butoxycarbonylaminooctyl-O--, methylcarbonylaminoethyl-O--, methylcarbonylaminopentyl-O--, methylsulfonylaminoethyl-O--, methylsulfonylaminopentyl-O--, methylsulfonylethyl-O--, methylsulfonylpropyl-O--, methylsulfanylpropyl-O--, cyanopropyl-O--, cyanocyclopropylmethyl-O--, cyclopropylmethyl-O--, cyclohexylethyl-O--, hydroxyethyl-O--, hydroxypropyl-O--, hydroxy-dimethylpropyl-O--, hydroxy-difluoropropyl-O--, hydroxybutyl-O--, hydroxypentyl-O--, hydroxyhexyl-O--, aminoethyl-O-propyl-O--, ethylamino-ethyl-O-propyl-O--, imidazolylethyl-O--, pyrazolylpropyl-O--, triazolylpropyl-O--, morpholinylethyl-O--, morpholinylpropyl-O--, (2-oxo-pyrrolidinyl)ethyl-O--, (2-oxo-pyrrolidinyl)propyl-O--, phenylmethyl-O--, phenylethyl-O--, pyrrolidinylethyl-O--, pyrrolidinylpropyl-O--, pyrrolidinylcarbonylmethyl-O--, tetrahydropyranylmethyl-O-- or carboxypropyl-O--;

[0186] R.sup.4 is hydrogen, fluoro, chloro, bromo, methyl or cyano;

[0187] provided that R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are not hydrogen simultaneously;

[0188] R.sup.5 is hydrogen or methyl;

[0189] R.sup.6 is hydrogen, methyl, ethyl, propyl, isopropyl, isobutyl, tert-butyl, trifluoromethyl, trifluoromethylmethyl, cyclopropyl, cyclobutyl, methylcyclopropyl or phenylmethyl; or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

[0190] 2. A compound according to formula (I), wherein

[0191] R.sup.1 is hydrogen, halogen, C.sub.1-6alkylamino or C.sub.1-6alkoxy;

[0192] R.sup.2 is hydrogen, halogen, C.sub.1-6alkyl, C.sub.1-6alkoxy, C.sub.3-7cycloalkyl, hydroxy or phenyl-C.sub.xH.sub.2x--O--;

[0193] R.sup.3 is hydrogen;

[0194] halogen;

[0195] C.sub.1-6alkyl;

[0196] cyano;

[0197] phenyl-C.sub.xH.sub.2x--N(C.sub.1-6alkyl)-;

[0198] C.sub.1-6alkoxycarbonylpiperazinyl;

[0199] or R.sup.7--O--, wherein R.sup.7 is hydrogen; C.sub.1-6alkyl, which is unsubstituted or substituted with one to three substituents independently selected from fluoro, hydroxy and C.sub.2-6alkenyl; C.sub.1-6alkoxyC.sub.1-6alkyl; C.sub.1-6alkoxyC.sub.1-6alkoxyC.sub.1-6alkyl; aminoC.sub.1-8alkyl; C.sub.1-6alkylcarbonylaminoC.sub.1-8alkyl; C.sub.1-6alkylsulfonylaminoC.sub.1-8alkyl; C.sub.1-6alkylsulfanylC.sub.1-6alkyl; C.sub.1-6alkylsulfonylC.sub.1-6alkyl; cyanoC.sub.1-6alkyl; C.sub.3-7cycloalkylC.sub.1-6alkyl; cyanoC.sub.3-7cycloalkylC.sub.1-6alkyl; phenylC.sub.1-6alkyl; pyrrolidinylcarbonylC.sub.1-6alkyl; C.sub.2-6alkynyl; hydroxyC.sub.1-6alkylC.sub.2-6alkynyl; aminoC.sub.1-6alkoxyC.sub.1-6alkyl; C.sub.1-6alkylaminoC.sub.1-6alkoxyC.sub.1-6alkyl; carboxyC.sub.1-6alkyl; C.sub.1-6alkoxycarbonylaminoC.sub.1-8alkyl; heteroarylC.sub.1-6alkyl, wherein heteroaryl is N-containing monocyclic heteroaryl; or heterocycloalkylC.sub.1-6alkyl, wherein heterocycloalkyl is monocyclic heterocycloalkyl;

[0200] R.sup.4 is hydrogen, halogen, C.sub.1-6alkyl or cyano;

[0201] provided that R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are not hydrogen simultaneously;

[0202] R.sup.5 is hydrogen or C.sub.1-6alkyl;

[0203] R.sup.6 is hydrogen; C.sub.1-6alkyl, which is unsubstituted or once, twice or three times substituted by fluoro; C.sub.3-7cycloalkyl; C.sub.1-6alkylC.sub.3-7cycloalkyl; or phenyl-C.sub.xH.sub.2x--;

[0204] x is 1-6;

[0205] or a pharmaceutically acceptable salt, or enantiomer, or a diastereomer thereof.

[0206] 3. A compound according to formula (I) or according to item 1 or 2, wherein R.sup.1 is hydrogen, fluoro, chloro, bromo, methylamino, methoxy or ethoxy;

[0207] R.sup.2 is hydrogen, fluoro, chloro, methyl, ethyl, methoxy, ethoxy, propoxy, cyclopropyl, hydroxy or phenylmethyl-O--;

[0208] R.sup.3 is hydrogen, bromo, methyl, propyl, cyano, phenylmethyl-N(methyl)-, tert-butoxycarbonylpiperazinyl, hydroxy, methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, difluoromethylmethyl-O--, difluoromethylethyl-O--, trifluoromethylmethyl-O--, ethyldifluoromethyl-O--, vinyldifluoromethyl-O--, propargyl-O--, hydroxymethylpropargyl-O--, methoxyethyl-O--, methoxypropyl-O--, methoxybutyl-O--, ethoxyethyl-O--, methoxyethyl-O-ethyl-O--, aminoethyl-O--, aminopentyl-O--, aminohexyl-O--, aminooctyl-O--, tert-butoxycarbonylaminopentyl-O--, tert-butoxycarbonylaminohexyl-O--, tert-butoxycarbonylaminooctyl-O--, methylcarbonylaminoethyl-O--, methylcarbonylaminopentyl-O--, methylsulfonylaminoethyl-O--, methylsulfonylaminopentyl-O--, methylsulfonylethyl-O--, methylsulfonylpropyl-O--, methylsulfanylpropyl-O--, cyanopropyl-O--, cyanocyclopropylmethyl-O--, cyclopropylmethyl-O--, cyclohexylethyl-O--, hydroxyethyl-O--, hydroxypropyl-O--, hydroxy-dimethylpropyl-O--, hydroxy-difluoropropyl-O--, hydroxybutyl-O--, hydroxypentyl-O--, hydroxyhexyl-O--, aminoethyl-O-propyl-O--, ethylamino-ethyl-O-propyl-O--, imidazolylethyl-O--, pyrazolylpropyl-O--, triazolylpropyl-O--, morpholinylethyl-O--, morpholinylpropyl-O--, (2-oxo-pyrrolidinyl)ethyl-O--, (2-oxo-pyrrolidinyl)propyl-O--, phenylmethyl-O--, phenylethyl-O--, pyrrolidinylethyl-O--, pyrrolidinylpropyl-O--, pyrrolidinylcarbonylmethyl-O--, tetrahydropyranylmethyl-O-- or carboxypropyl-O--;

[0209] R.sup.4 is hydrogen, chloro, bromo, methyl or cyano;

[0210] provided that R.sup.1, R.sup.2, R.sup.3 and R.sup.4 are not hydrogen simultaneously;

[0211] R.sup.5 is hydrogen or methyl;

[0212] R.sup.6 is hydrogen, methyl, ethyl, propyl, isopropyl, isobutyl, tert-butyl, trifluoromethyl, trifluoromethylmethyl, cyclopropyl, cyclobutyl, methylcyclopropyl or phenylmethyl; or a pharmaceutically acceptable salt, or enantiomer, or a diastereomer thereof.

[0213] 4. A compound according to formula (I) or item 2, wherein the compound is the compound of formula (IA):

##STR00005##

[0214] wherein

[0215] R.sup.1 is hydrogen, halogen or C.sub.1-6alkoxy;

[0216] R.sup.2 is hydrogen, halogen, C.sub.1-6alkyl, C.sub.1-6alkoxy, C.sub.3-7cycloalkyl, hydroxy or phenyl-C.sub.xH.sub.2x--O--;

[0217] R.sup.4 is hydrogen or halogen;

[0218] R.sup.5 is hydrogen or C.sub.1-6alkyl;

[0219] R.sup.5 is hydrogen; C.sub.1-6alkyl, which is unsubstituted or once, twice or three times substituted by fluoro; C.sub.3-7cycloalkyl; C.sub.1-6alkylC.sub.3-7cycloalkyl; or phenyl-C.sub.xH.sub.2x--;

[0220] R.sup.7 is hydrogen; C.sub.1-6alkyl, which is unsubstituted or substituted with one to three substituents independently selected from fluoro, hydroxy and ethenyl; C.sub.1-6alkoxyC.sub.1-6alkyl; C.sub.1-6alkoxyC.sub.1-6alkoxyC.sub.1-6alkyl; aminoC.sub.1-6alkyl; C.sub.1-6alkylcarbonylaminoC.sub.1-6alkyl; C.sub.1-6alkylsulfonylaminoC.sub.1-6alkyl; C.sub.1-6alkylsulfanylC.sub.1-6alkyl; C.sub.1-6alkylsulfonylC.sub.1-6alkyl; cyanoC.sub.1-6alkyl; C.sub.3-7cycloalkylC.sub.1-6alkyl; cyanoC.sub.3-7cycloalkylC.sub.1-6alkyl; phenylC.sub.1-6alkyl; pyrrolidinylcarbonylC.sub.1-6alkyl; C.sub.2-6alkynyl; hydroxyC.sub.1-6alkylC.sub.2-6alkynyl; aminoC.sub.1-6alkoxyC.sub.1-6alkyl; C.sub.1-6alkylaminoC.sub.1-6alkoxyC.sub.1-6alkyl; carboxyC.sub.1-6alkyl; C.sub.1-6alkoxycarbonylaminoC.sub.1-8alkyl; heteroarylC.sub.1-6alkyl, wherein heteroaryl is N-containing monocyclic heteroaryl; or heterocycloalkylC.sub.1-6alkyl, wherein heterocycloalkyl is monocyclic heterocycloalkyl;

[0221] x is 1-6;

[0222] or a pharmaceutically acceptable salt, or enantiomer, or a diastereomer thereof.

[0223] 5. A compound according to item 4, wherein

[0224] R.sup.1 is hydrogen, fluoro, chloro or methoxy;

[0225] R.sup.2 is hydrogen, fluoro, chloro, methyl, ethyl, methoxy, ethoxy, propoxy, cyclopropyl, hydroxy or phenylmethyl-O--;

[0226] R.sup.4 is hydrogen or chloro;

[0227] R.sup.5 is hydrogen or methyl;

[0228] R.sup.6 is hydrogen, methyl, ethyl, propyl, isopropyl, isobutyl, tert-butyl, trifluoromethyl, trifluoromethylmethyl, cyclopropyl, cyclobutyl, methylcyclopropyl or phenylmethyl;

[0229] R.sup.7 is hydrogen, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, difluoromethylmethyl, difluoromethylethyl, trifluoromethylmethyl, ethyldifluoromethyl, vinyldifluoromethyl, propargyl, hydroxymethylpropargyl, methoxyethyl, methoxypropyl, methoxybutyl, ethoxyethyl, methoxyethyl-O-ethyl, aminoethyl, aminopentyl, aminohexyl, aminooctyl, tert-butoxycarbonylaminopentyl, tert-butoxycarbonylaminohexyl, tert-butoxycarbonylaminooctyl, methylcarbonylaminoethyl, methylcarbonylaminopentyl, methylsulfonylaminoethyl, methylsulfonylaminopentyl, methylsulfonylethyl, methylsulfonylpropyl, methylsulfanylpropyl, cyanopropyl, cyanocyclopropylmethyl, cyclopropylmethyl, cyclohexylethyl, hydroxyethyl, hydroxypropyl, hydroxy-dimethylpropyl, hydroxy-difluoropropyl, hydroxybutyl, hydroxypentyl, hydroxyhexyl, aminoethyl-O-propyl, ethylamino-ethyl-O-propyl-, imidazolylethyl, pyrazolyipropyl, triazolylpropyl, morpholinylethyl, morpholinylpropyl, (2-oxo-pyrrolidinyl)ethyl, (2-oxo-pyrrolidinyl)propyl, phenylmethyl, phenylethyl, pyrrolidinylethyl, pyrrolidinylpropyl, pyrrolidinylcarbonylmethyl, tetrahydropyranylmethyl or carboxypropyl;

[0230] or a pharmaceutically acceptable salt, or enantiomer, or a diastereomer thereof.

[0231] 6. A compound according to item 4, wherein

[0232] R.sup.1 is hydrogen or halogen;

[0233] R.sup.2 is C.sub.1-6alkyl, halogen or C.sub.3-7cycloalkyl;

[0234] R.sup.4 is hydrogen;

[0235] R.sup.5 is hydrogen or C.sub.1-6alkyl;

[0236] R.sup.6 is C.sub.1-6alkyl or C.sub.1-6alkylC.sub.3-7cycloalkyl;

[0237] R.sup.7 is C.sub.1-6alkyl, C.sub.1-6alkoxyC.sub.1-6alkyl; or phenylC.sub.1-6alkyl;

[0238] or a pharmaceutically acceptable salt, or enantiomer, or a diastereomer thereof.

[0239] 7. A compound according to item 6, wherein

[0240] R.sup.1 is hydrogen, fluoro or chloro;

[0241] R.sup.2 is methyl, ethyl, fluoro, chloro or cyclopropyl;

[0242] R.sup.4 is hydrogen;

[0243] R.sup.5 is hydrogen or methyl;

[0244] R.sup.6 is methyl, ethyl, isopropyl, isobutyl, tert-butyl or methylcyclopropyl;

[0245] R.sup.7 is methyl, ethyl, methoxyethyl, methoxypropyl or phenylmethyl;

[0246] or a pharmaceutically acceptable salt, or enantiomer, or a diastereomer thereof.

[0247] 8. A compound according to item 4, wherein wherein

[0248] R.sup.1 is hydrogen;

[0249] R.sup.2 is C.sub.1-6alkoxy;

[0250] R.sup.4 is hydrogen or halogen;

[0251] R.sup.5 is hydrogen or C.sub.1-6alkyl;

[0252] R.sup.6 is hydrogen; C.sub.1-6alkyl, which is unsubstituted or once, twice or three times substituted by fluoro; C.sub.3-7cycloalkyl; C.sub.1-6alkylC.sub.3-7cycloalkyl; or phenyl-C.sub.xH.sub.2x--;

[0253] R.sup.7 is hydrogen; C.sub.1-6alkyl, which is unsubstituted or substituted with one to three substituents independently selected from fluoro, hydroxy and C.sub.2-6alkenyl; C.sub.1-6alkoxyC.sub.1-6alkyl; C.sub.1-6alkoxyC.sub.1-6alkoxyC.sub.1-6alkyl; aminoC.sub.1-6alkyl; C.sub.1-6alkylcarbonylaminoC.sub.1-8alkyl; C.sub.1-6alkylsulfonylaminoC.sub.1-8alkyl; C.sub.1-6alkylsulfanylC.sub.1-6alkyl; C.sub.1-6alkylsulfonylC.sub.1-6alkyl; cyanoC.sub.1-6alkyl; cyanoC.sub.3-7cycloalkylC.sub.1-6alkyl; C.sub.3-7cycloalkylC.sub.1-6alkyl; phenylC.sub.1-6alkyl; pyrrolidinylcarbonylC.sub.1-6alkyl; C.sub.2-6alkynyl; hydroxyC.sub.1-6alkylC.sub.2-6alkynyl; aminoC.sub.1-6alkoxyC.sub.1-6alkyl; C.sub.1-6alkylaminoC.sub.1-6alkoxyC.sub.1-6alkyl; carboxyC.sub.1-6alkyl; C.sub.1-6alkoxycarbonylaminoC.sub.1-8alkyl; imidazolylC.sub.1-6alkyl; pyrazolylC.sub.1-6alkyl; triazolylC.sub.1-6alkyl; morpholinylC.sub.1-6alkyl; (2-oxo-pyrrolidinyl)C.sub.1-6alkyl; pyrrolidinylC.sub.1-6alkyl; or tetrahydropyranylC.sub.1-6alkyl;

[0254] x is 1-6;

[0255] or a pharmaceutically acceptable salt, or enantiomer, or a diastereomer thereof.

[0256] 9. A compound according to item 8, wherein

[0257] R.sup.1 is hydrogen;

[0258] R.sup.2 is methoxy, ethoxy or propoxy;

[0259] R.sup.4 is hydrogen or chloro;

[0260] R.sup.5 is hydrogen or methyl;

[0261] R.sup.6 is hydrogen, methyl, ethyl, propyl, isopropyl, isobutyl, tert-butyl, trifluoromethyl, trifluoromethylmethyl, cyclopropyl, cyclobutyl, methylcyclopropyl or phenylmethyl;

[0262] R.sup.7 is hydrogen, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, difluoromethylmethyl, difluoromethylethyl, trifluoromethylmethyl, ethyldifluoromethyl, vinyldifluoromethyl, propargyl, hydroxymethylpropargyl, methoxyethyl, methoxypropyl, methoxybutyl, ethoxyethyl, methoxyethyl-O-ethyl, aminoethyl, aminopentyl, aminohexyl, aminooctyl, tert-butoxycarbonylaminopentyl, tert-butoxycarbonylaminohexyl, tert-butoxycarbonylaminooctyl, methylcarbonylaminoethyl, methylcarbonylaminopentyl, methylsulfonylaminoethyl, methylsulfonylaminopentyl, methylsulfonylethyl, methylsulfonylpropyl, methylsulfanylpropyl, cyanopropyl, cyanocyclopropylmethyl, cyclopropylmethyl, cyclohexylethyl, hydroxyethyl, hydroxypropyl, hydroxy-dimethylpropyl, hydroxy-difluoropropyl, hydroxybutyl, hydroxypentyl, hydroxyhexyl, aminoethyl-O-propyl, ethylamino-ethyl-O-propyl-, imidazolylethyl, pyrazolylpropyl, triazolylpropyl, morpholinylethyl, morpholinylpropyl, (2-oxo-pyrrolidinyl)ethyl, (2-oxo-pyrrolidinyl)propyl, phenylmethyl, phenylethyl, pyrrolidinylethyl, pyrrolidinylpropyl, pyrrolidinylcarbonylmethyl, tetrahydropyranylmethyl or carboxypropyl;

[0263] or a pharmaceutically acceptable salt, or enantiomer, or a diastereomer thereof.

[0264] 10. A compound according to item 4, wherein

[0265] R.sup.1 is hydrogen or halogen;

[0266] R.sup.2 is halogen, C.sub.1-6alkyl, C.sub.1-6alkoxy or C.sub.3-7cycloalkyl;

[0267] R.sup.4 is hydrogen;

[0268] R.sup.5 is hydrogen or C.sub.1-6alkyl;

[0269] R.sup.6 is C.sub.1-6alkyl, which is unsubstituted or once, twice or three times substituted by fluoro;

[0270] C.sub.3-7cycloalkyl or C.sub.1-6alkylC.sub.3-7cycloalkyl;

[0271] R.sup.7 is C.sub.1-6alkyl, which is unsubstituted or substituted with one to three substituents independently selected from fluoro and hydroxy; C.sub.1-6alkoxyC.sub.1-6alkyl; aminoC.sub.1-8alkyl; C.sub.1-6alkylcarbonylaminoC.sub.1-8alkyl; C.sub.1-6alkylsulfonylaminoC.sub.1-8alkyl; C.sub.1-6alkylsulfanylC.sub.1-6alkyl; C.sub.1-6alkylsulfonylC.sub.1-6alkyl; C.sub.3-7cycloalkylC.sub.1-6alkyl; phenylC.sub.1-6alkyl; C.sub.1-6alkylaminoC.sub.1-6alkoxyC.sub.1-6alkyl; C.sub.1-6alkoxycarbonylaminoC.sub.1-8alkyl; morpholinylC.sub.1-6alkyl or tetrahydropyranylC.sub.1-6alkyl; or a pharmaceutically acceptable salt, or enantiomer, or a diastereomer thereof.

[0272] 11. A compound according to item 10, wherein

[0273] R.sup.1 is hydrogen, fluoro, or chloro;

[0274] R.sup.2 is fluoro, chloro, methyl, ethyl, methoxy, ethoxy or cyclopropyl;

[0275] R.sup.4 is hydrogen;

[0276] R.sup.5 is hydrogen or methyl;

[0277] R.sup.6 is methyl, ethyl, isopropyl, isobutyl, tert-butyl, trifluoromethylmethyl, cyclobutyl or methylcyclopropyl;

[0278] R.sup.7 is methyl, ethyl, propyl, butyl, isobutyl, cyclopropylmethyl, difluoromethylmethyl, difluoroethylmethyl, difluoromethylethyl, trifluoromethylmethyl, ethyldifluoromethyl, methoxyethyl, methoxypropyl, ethoxyethyl, aminohexyl, aminooctyl, tert-butoxycarbonylaminopentyl, tert-butoxycarbonylaminooctyl, methylcarbonylaminopentyl, methylsulfonylaminopentyl, methylsulfonylpropyl, methylsulfanylpropyl, hydroxypropyl, hydroxy-dimethylpropyl, hydroxy-difluoropropyl, hydroxybutyl, hydroxypentyl, hydroxyhexyl, ethylamino-ethyl-O-propyl-, morpholinylethyl, morpholinylpropyl, phenylmethyl or tetrahydropyranylmethyl; or a pharmaceutically acceptable salt, or enantiomer, or a diastereomer thereof.

[0279] 12. A compound according to formula (I), item 1 or item 2, or a pharmaceutically acceptable salt, or enantiomer thereof, wherein R.sup.1 is hydrogen.

[0280] 13. A compound according to formula (I), item 1 or item 2, or a pharmaceutically acceptable salt, or enantiomer thereof, wherein R.sup.2 is halogen or C.sub.1-6alkoxy.

[0281] 14. A compound according to formula (I), item 1 or item 2, or a pharmaceutically acceptable salt, or enantiomer thereof, wherein R.sup.2 is chloro or methoxy.

[0282] 15. A compound according to formula (I), item 1 or item 2, or a pharmaceutically acceptable salt, or enantiomer thereof, wherein R.sup.5 is hydrogen.

[0283] 16. A compound according to formula (I), item 1 or item 2, or a pharmaceutically acceptable salt, or enantiomer thereof, wherein R.sup.6 is C.sub.1-6alkyl or C.sub.1-6alkylC.sub.3-7cycloalkyl.

[0284] 17. A compound according to formula (I), item 1 or item 2, or a pharmaceutically acceptable salt, or enantiomer thereof, wherein R.sup.6 is ethyl, isopropyl, tert-butyl or methylcyclopropyl.

[0285] 18. A compound according to formula (I), item 1 or item 2, or a pharmaceutically acceptable salt, or enantiomer thereof, wherein R.sup.7 is C.sub.1-6alkoxyC.sub.1-6alkyl, hydroxyC.sub.1-6alkyl or aminoC.sub.1-6alkyl.

[0286] 19. A compound according to formula (I), item 1 or item 2, or a pharmaceutically acceptable salt, or enantiomer thereof, wherein R.sup.7 is methoxyethyl, methoxypropyl, hydroxydimethylpropyl, hydroxybutyl, hydroxypentyl, hydroxyhexyl, aminobutyl, aminopentyl or aminohexyl.

[0287] In context of the present invention the inhibitor of the invention may also be the compound according to formula (II), wherein the inhibitor is any one of the compounds as defined in items (1)-(20), below:

[0288] 1. A compound according to formula (II), wherein

[0289] R.sup.1 is C.sub.1-6alkyl, C.sub.3-7cycloalkyl, hydroxyC.sub.1-6alkyl, C.sub.1-6alkoxycarbonylC.sub.1-6alkyl or carboxyC.sub.1-6alkyl;

[0290] R.sup.2 is phenyl substituted by one, two, three or four groups independently selected from C.sub.1-6alkyl, C.sub.3-7cycloalkyl, halogen, haloC.sub.1-6alkyl, cyano, nitro, hydroxy, haloC.sub.1-6alkoxy, tetrahydrofuranyloxy, --O--C.sub.xH.sub.2x--R.sup.3, --O--C.sub.yH.sub.2y--NHR.sup.6, --SO.sub.2--R.sup.11. --SO.sub.2--NR.sup.12R.sup.13, carboxy, C.sub.1-6alkoxycarbonyl and --C(.dbd.O)--NR.sup.12R.sup.13; pyridinyl substituted by one, two or three groups independently selected from halogen, C.sub.1-6alkyl, haloC.sub.1-6alkoxy, tetrahydropyranyloxy, --O--C.sub.xH.sub.2x--R.sup.3 and NR.sup.6R.sup.16; or pyrimidinyl substituted by C.sub.1-6alkyl and diC.sub.1-6alkylamino; wherein

[0291] R.sup.3 is hydrogen, C.sub.3-7cycloalkyl, haloC.sub.3-7cycloalkyl, hydroxy, hydroxyC.sub.1-6alkylC.sub.3-7cycloalkyl, C.sub.1-6alkoxy, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, thietanyl, 1,1-dioxothietanyl, 1,1-dioxothiolanyl, morpholinyl, oxopyrrolidinyl, oxomorpholinyl, oxopiperazinyl, C.sub.1-6alkoxycarbonyloxopiperazinyl, oxoimidazolidinyl, C.sub.1-6alkylpiperazinyl, C.sub.1-6alkylcarbonylpiperazinyl, C.sub.1-6alkylsulfonylpiperazinyl, C.sub.1-6alkoxycarbonylpiperazinyl, azetidinyl, C.sub.1-6alkylcarbonylazetidinyl, C.sub.1-6alkylsulfonylazetidinyl, C.sub.1-6alkoxycarbonylazetidinyl, --C(.dbd.O)--R.sup.4, C.sub.1-6alkylsulfinyl, --SO.sub.2--R.sup.5, --C(NHR.sup.7)--C(.dbd.O)--R.sup.8, carboxyC.sub.1-6alkoxy or aminocarbonylC.sub.1-6alkoxy;

[0292] wherein

[0293] R.sup.4 is hydroxy, C.sub.1-6alkoxy, amino, C.sub.1-6alkylamino, diC.sub.1-6alkylamino, tetrahydrofuranylamino, pyrrolidinyl or morpholinyl;

[0294] R.sup.5 is C.sub.1-6alkyl, hydroxy or amino;

[0295] R.sup.7 is hydrogen or C.sub.1-6alkoxycarbonyl;

[0296] R.sup.8 is hydroxy or C.sub.1-6alkoxy;

[0297] R.sup.6 is hydrogen, C.sub.1-6alkylcarbonyl, haloC.sub.1-6alkylcarbonyl, C.sub.1-6alkoxycarbonyl, C.sub.1-6alkylsulfonyl, C.sub.3-7cycloalkylsulfonyl or C.sub.1-6alkoxyC.sub.1-6alkylsulfonyl;

[0298] R.sup.9 and R.sup.10 are independently selected from hydrogen, C.sub.1-6alkyl and C.sub.1-6alkylsulfonyl; or

[0299] R.sup.9 and R.sup.10 together with the nitrogen to which they are attached form pyrrolidinyl, morpholinyl, piperidinyl, piperazinyl and oxopiperazinyl;

[0300] R.sup.11 is C.sub.1-6alkyl or C.sub.1-6alkoxyC.sub.1-6alkyl;

[0301] R.sup.12 and R.sup.13 are independently selected from hydrogen, C.sub.1-6alkyl and C.sub.1-6alkoxyC.sub.1-6alkyl;

[0302] x is 1, 2, 3, 4, 5, 6, 7 or 8;

[0303] y is 1, 2, 3, 4, 5, 6, 7 or 8;

[0304] U is CH;

[0305] W is CH;

[0306] Z is CH or N;

[0307] X is N;

[0308] Y is N;

[0309] or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

[0310] 2. A compound according to formula (II) or item 1, wherein

[0311] R.sup.1 is C.sub.1-6alkyl;

[0312] R.sup.2 is phenyl substituted by one, two, three or four groups independently selected from C.sub.1-6alkyl, C.sub.3-7cycloalkyl, halogen, haloC.sub.1-6alkyl, cyano, hydroxy, haloC.sub.1-6alkoxy, tetrahydrofuranyloxy, --O--C.sub.xH.sub.2x--R.sup.3, --O--C.sub.vH.sub.2y--NHR.sup.6, --SO.sub.2--NR.sup.12R.sup.13, carboxy, C.sub.1-6alkoxycarbonyl and --C(.dbd.O)--NR.sup.12R.sup.13; pyridinyl substituted by one, two or three groups independently selected from halogen, C.sub.1-6alkyl, haloC.sub.1-6alkoxy, tetrahydropyranyloxy, --O--C.sub.xH.sub.2x--R.sup.3 and NR.sup.9R.sup.10; or pyrimidinyl substituted by C.sub.1-6alkyl and diC.sub.1-6alkylamino; wherein

[0313] R.sup.3 is hydrogen, C.sub.3-7cycloalkyl, haloC.sub.3-7cycloalkyl, hydroxyC.sub.1-6alkylC.sub.3-7cycloalkyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, thietanyl, 1,1-dioxothietanyl, 1,1-dioxothiolanyl, oxopyrrolidinyl, oxomorpholinyl, oxopiperazinyl, C.sub.1-6alkoxycarbonyloxopiperazinyl, oxoimidazolidinyl, C.sub.1-6alkylpiperazinyl, C.sub.1-6alkylcarbonylpiperazinyl, C.sub.1-6alkylsulfonylpiperazinyl, C.sub.1-6alkoxycarbonylpiperazinyl, azetidinyl, C.sub.1-6alkylcarbonylazetidinyl. C.sub.1-6alkylsulfonylazetidinyl, C.sub.1-6alkoxycarbonylazetidinyl, --C(.dbd.O)--R.sup.4, C.sub.1-6alkylsulfinyl, --SO.sub.2--R.sup.5, --C(NHR.sup.7)--C(.dbd.O)--R.sup.8 carboxyC.sub.1-6alkoxy or aminocarbonylC.sub.1-6alkoxy; wherein

[0314] R.sup.4 is hydroxy, C.sub.1-6alkoxy, amino, C.sub.1-6alkylamino, tetrahydrofuranylamino, or morpholinyl;

[0315] R.sup.5 is C.sub.1-6alkyl, hydroxy or amino;

[0316] R.sup.7 is hydrogen or C.sub.1-6alkoxycarbonyl;

[0317] R.sup.8 is hydroxy or C.sub.1-6alkoxy;

[0318] R.sup.6 is hydrogen, C.sub.1-6alkylcarbonyl, haloC.sub.1-6alkylcarbonyl, C.sub.1-6alkoxycarbonyl,

[0319] C.sub.3-7cycloalkylsulfonyl or C.sub.1-6alkoxyC.sub.1-6alkylsulfonyl;

[0320] R.sup.9 and R.sup.13 are independently selected from hydrogen, C.sub.1-6alkyl and C.sub.1-6alkylsulfonyl; or

[0321] R.sup.9 and R.sup.10 together with the nitrogen to which they are attached form pyrrolidinyl, morpholinyl, piperazinyl and oxopiperazinyl;

[0322] R.sup.11 is C.sub.1-6alkoxyC.sub.1-6alkyl;

[0323] R.sup.12 and R.sup.13 are independently selected from hydrogen, C.sub.1-6alkyl and C.sub.1-6alkoxyC.sub.1-6alkyl;

[0324] x is 1, 2, 3, 4, 5, 6, 7 or 8;

[0325] y is 1, 2, 3, 4, 5, 6, 7 or 8;

[0326] U is CH;

[0327] W is CH;

[0328] Z is N;

[0329] X is N;

[0330] Y is N;

[0331] or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

[0332] 3. A compound according to formula (II), item 1 or item 2, or a pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R.sup.1 is methyl.

[0333] 4. A compound according to formula (II), item 1 or item 2, wherein

[0334] R.sup.1 is C.sub.1-6alkyl;

[0335] R.sup.2 is phenyl substituted by one, two, three or four groups independently selected from C.sub.1-6alkyl, C.sub.3-7cycloalkyl, halogen, haloC.sub.1-6alkyl, cyano, hydroxy, haloC.sub.1-6alkoxy, tetrahydrofuranyloxy, --O--C.sub.xH.sub.2x--R.sup.3, --O--C.sub.vH.sub.2y--NHR.sup.6, --SO.sub.2--R.sup.11, --SO.sub.2--NR.sup.12R.sup.13, carboxy, C.sub.1-6alkoxycarbonyl and --C(.dbd.O)--NR.sup.12R.sup.12;

[0336] R.sup.3 is hydrogen, C.sub.3-7cycloalkyl, haloC.sub.3-7cycloalkyl, hydroxyC.sub.1-6alkylC.sub.3-7cycloalkyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, thietanyl, 1,1-dioxothietanyl, 1,1-dioxothiolanyl, oxopyrrolidinyl, oxomorpholinyl, oxopiperazinyl, C.sub.1-6alkoxycarbonyloxopiperazinyl, oxoimidazolidinyl, C.sub.1-6alkylpiperazinyl, C.sub.1-6alkylcarbonylpiperazinyl, C.sub.1-6alkylsulfonylpiperazinyl, C.sub.1-6alkoxycarbonylpiperazinyl, azetidinyl, C.sub.1-6alkylcarbonylazetidinyl, C.sub.1-6alkylsulfonylazetidinyl, C.sub.1-6alkoxycarbonylazetidinyl, --C(.dbd.O)--R.sup.4, C.sub.1-6alkylsulfinyl, --SO.sub.2--R.sup.5 or --C(NHR.sup.7)--C(.dbd.O)--R.sup.8;

[0337] wherein

[0338] R.sup.4 is hydroxy, C.sub.1-6alkoxy, amino, C.sub.1-6alkylamino, tetrahydrofuranylamino, or morpholinyl;

[0339] R.sup.5 is C.sub.1-6alkyl, hydroxy or amino;

[0340] R.sup.7 is hydrogen or C.sub.1-6alkoxycarbonyl;

[0341] R.sup.8 is hydroxy or C.sub.1-6alkoxy;

[0342] R.sup.6 is hydrogen, C.sub.1-6alkylcarbonyl, haloC.sub.1-6alkylcarbonyl, C.sub.1-6alkoxycarbonyl, C.sub.3-7cycloalkylsulfonyl or C.sub.1-6alkoxyC.sub.1-6alkylsulfonyl;

[0343] R.sup.11 is C.sub.1-6alkoxyC.sub.1-6alkyl;

[0344] R.sup.12 and R.sup.13 are independently selected from hydrogen, C.sub.1-6alkyl and C.sub.1-6alkoxyC.sub.1-6alkyl;

[0345] x is 1, 2, 3, 4, 5 or 6;

[0346] y is 1, 2, 3, 4, 5, 6, 7 or 8;

[0347] U is CH;

[0348] W is CH;

[0349] Z is N;

[0350] X is N;

[0351] Y is N;

[0352] or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

[0353] 5. A compound according to formula (II), or any one of items 1 to 4, wherein

[0354] R.sup.1 is methyl;

[0355] R.sup.2 is phenyl substituted by one, two, three or four groups independently selected from methyl, cyclopropyl, fluoro, chloro, iodo, trifluoromethyl, cyano, hydroxy, methoxy, difluoroethoxy, difluoromethoxy, trifluoroethoxy, trifluoromethoxy, cyclopropylmethoxy, difluorocyclopropylmethoxy, hydroxymethylcyclopropylmethoxy, oxetanylethoxy, oxetanylmethoxy, tetrahydrofuranylethoxy, tetrahydrofuranylmethoxy, tetrahydropyranylmethoxy, thietanylmethoxy, (1,1-dioxothietanyl)methoxy, (1,1-dioxothiolanyl)methoxy, oxopyrrolidinyipropoxy, oxomorpholinyipropoxy, oxopiperazinylpropoxy, (tert-butoxycarbonyloxopiperazinyl)propoxy, oxoimidazolidinylpropoxy, methylpiperazinylpropoxy, acetylpiperazinylpropoxy, methylsulfonylpiperazinylpropoxy, (tert-butoxycarbonylpiperazinyl)propoxy, azetidinylethoxy, acetylazetidinylethoxy, methylsulfonylazetidinylethoxy, (tert-butoxycarbonylazetidinyl)ethoxy, (tert-butoxycarbonylazetidinyl)methoxy, carboxybutoxy, carboxyethoxy, carboxyhexyloxy, carboxymethoxy, carboxypropoxy, methoxycarbonylbutoxy, ethoxycarbonylhexyloxy, aminocarbonylbutoxy, am inocarbonylhexyloxy, am inocarbonylmethoxy, aminocarbonylpropoxy, methylaminocarbonylpropoxy, tetrahydrofuranylaminocarbonylmethoxy, morpholinylcarbonylmethoxy, methylsulfinylpropoxy, methylsulfonylpropoxy, sulfopropoxy, aminosulfonylpropoxy, amino-carboxy-propoxy, (tert-butoxycarbonylamino)-carboxy-propoxy, (tert-butoxycarbonylamino)-(methoxycarbonyl)-propoxy, aminopropoxy, aminopentoxy, aminohexyloxy, aminooctyloxy, methylcarbonylaminopropoxy, chloropropylcarbonylaminopropoxy, (tert-butoxycarbonylamino)hexyloxy, (tert-butoxycarbonylamino)octyloxy, (tert-butoxycarbonylamino)pentoxy, (tert-butoxycarbonylamino)propoxy, cyclopropylsulfonylaminopropoxy, methoxyethylsulfonylaminopropoxy, methoxypropylsulfonyl, methoxypropylaminosulfonyl, N-methoxypropyl-N-methyl-aminosulfonyl, carboxy, methoxycarbonyl, methoxypropylaminocarbonyl, N-methoxypropyl-N-methyl-aminocarbonyl and tetrahydrofuranyloxy;

[0356] U is CH;

[0357] W is CH;

[0358] Z is N;

[0359] X is N;

[0360] Y is N;

[0361] or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

[0362] 6. A compound according to formula (II) or any one of items 1 to 4, or a pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R.sup.2 is phenyl substituted by one, two or three groups independently selected from halogen, C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, C.sub.3-7cycloalkylC.sub.1-6alkoxy and haloC.sub.3-7cycloalkylC.sub.1-6alkoxy.

[0363] 7. A compound according to formula (II), or any one of items 1 to 6, or a pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R.sup.2 is phenyl substituted by one, two or three groups independently selected from fluoro, chloro, methoxy, difluoroethoxy, trifluoroethoxy, cyclopropylmethoxy and difluorocyclopropylmethoxy.

[0364] 8. A compound according to formula (II), or any one of items 1, 2 and 4, wherein

[0365] R.sup.1 is C.sub.1-6alkyl;

[0366] R.sup.2 is phenyl substituted by two or three groups independently selected from halogen, cyano, haloC.sub.1-6alkoxy, --O--C.sub.xH.sub.2x--R.sup.3 and --O--C.sub.yH.sub.2y--NHR.sup.6;

[0367] R.sup.3 is hydrogen, C.sub.3-7cycloalkyl, haloC.sub.3-7cycloalkyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, C.sub.1-6alkylsulfonylazetidinyl, aminocarbonyl or C.sub.1-6alkylsulfonyl;

[0368] R.sup.6 is hydrogen or C.sub.1-6alkoxycarbonyl;

[0369] x is 1, 2, 3, 4, 5 or 6;

[0370] y is 1, 2, 3, 4, 5 or 6;

[0371] U is CH;

[0372] W is CH;

[0373] Z is N;

[0374] X is N;

[0375] Y is N;

[0376] or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

[0377] 9. A compound according to formula (II), or any one of items 1 to 5 and 8, wherein

[0378] R.sup.1 is methyl;

[0379] R.sup.2 is phenyl substituted by two or three groups independently selected from fluoro, chloro, cyano, methoxy, difluoroethoxy, trifluoroethoxy, cyclopropylmethoxy, difluorocyclopropylmethoxy, methylsulfonylpropoxy, aminocarbonylmethoxy, oxetanylmethoxy, oxetanylethoxy, tetrahydrofuranylmethoxy, tetrahydropyranylmethoxy, methylsulfonylazetidinylethoxy, aminohexyloxy and (tert-butoxycarbonylamino)propoxy;

[0380] U is CH;

[0381] W is CH;

[0382] Z is N;

[0383] X is N;

[0384] Y is N;

[0385] or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

[0386] 10. A compound according to formula (II), or item 1 or item 2, wherein

[0387] R.sup.1 is C.sub.1-6alkyl;

[0388] R.sup.2 is pyridinyl substituted by one, two or three groups independently selected from halogen,

[0389] C.sub.1-6alkyl, haloC.sub.1-6alkoxy, tetrahydropyranyloxy, --O--C.sub.xH.sub.2x--R.sup.3 and NR.sup.9R.sup.19;

[0390] R.sup.3 is hydrogen, C.sub.3-7cycloalkyl, thietanyl, tetrahydrofuranyl, tetrahydropyranyl, oxomorpholinyl, 1,1-dioxo-thietanyl, C.sub.1-6alkylcarbonylazetidinyl, C.sub.1-6alkylsulfonylazetidinyl, --C(.dbd.O)--R.sup.4, carboxyC.sub.1-6alkoxy or aminocarbonylC.sub.1-6alkoxy; wherein

[0391] R.sup.4 is hydroxy, C.sub.1-6alkoxy or amino;

[0392] R.sup.9 and R.sup.19 are independently selected from hydrogen, C.sub.1-6alkyl and C.sub.1-6alkylsulfonyl; or

[0393] R.sup.9 and R.sup.10 together with the nitrogen to which they are attached form pyrrolidinyl, morpholinyl, piperazinyl and oxopiperazinyl;

[0394] x is 1, 2, 3, 4, 5, 6, 7 or 8;

[0395] U is CH;

[0396] W is CH;

[0397] Z is N;

[0398] X is N;

[0399] Y is N;

[0400] or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

[0401] 11. A compound according to formula (II), or any one of items 1 to 3 and 10, wherein

[0402] R.sup.1 is methyl;

[0403] R.sup.2 is pyridinyl substituted by one, two or three groups independently selected from fluoro, chloro, iodo, methoxy, methyl, difluoroethoxy, tetrahydropyranyloxy, cyclopropylmethoxy, thietanylmethoxy, tetrahydrofuranylmethoxy, tetrahydropyranylmethoxy, oxomorpholinylpropoxy, (1,1-dioxo-thietanyl)methoxy, acetylazetidinylmethoxy, methylsulfonylazetidinylmethoxy, carboxybutoxy, carboxyheptyloxy, carboxyhexyloxy, carboxypentyloxy, carboxypropoxy, methoxycarbonylheptyloxy, aminocarbonylbutoxy, aminocarbonylheptyloxy, am inocarbonylhexyloxy, aminocarbonylmethoxy, aminocarbonylpentyloxy, am inocarbonylpropoxy, carboxymethoxypropoxy, aminocarbonylmethoxypropoxy, amino, methylamino, dimethylamino, methylsulfonylamino, pyrrolidinyl, morpholinyl, piperazinyl and oxopiperazinyl;

[0404] U is CH;

[0405] W is CH;

[0406] Z is N;

[0407] X is N;

[0408] Y is N;

[0409] or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

[0410] 12. A compound according to formula (II), or any one of items 1 to 3 and 10, or a pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R.sup.2 is pyridinyl substituted by one, two or three groups independently selected from halogen, C.sub.1-6alkoxy, haloC.sub.1-6alkoxy, C.sub.1-6alkylamino, diC.sub.1-6alkylamino, pyrrolidinyl and oxopiperazinyl.

[0411] 13. A compound according to formula (II), or any one of items 1 to 3, 10 and 11, or a pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R.sup.2 is pyridinyl substituted by one, two or three groups independently selected from fluoro, chloro, methoxy, difluoroethoxy, methylamino, dimethylamino, pyrrolidinyl and oxopiperazinyl.

[0412] 14. A compound according to formula (II), or any one of items 1, 2 and 10, wherein

[0413] R.sup.1 is C.sub.1-6alkyl;

[0414] R.sup.2 is pyridinyl substituted by two or three groups independently selected from halogen, haloC.sub.1-6alkoxy, --O--C.sub.xH.sub.2x--R.sup.3 and NR.sup.9R.sup.15;

[0415] R.sup.3 is hydrogen, tetrahydrofuranyl, tetrahydropyranyl, oxomorpholinyl or aminocarbonyl;

[0416] R.sup.9 and R.sup.19 are independently selected from hydrogen and C.sub.1-6alkyl; or

[0417] R.sup.9 and R.sup.15 together with the nitrogen to which they are attached form pyrrolidinyl and oxopiperazinyl;

[0418] x is 1, 2, 3, 4, 5 or 6;

[0419] U is CH;

[0420] W is CH;

[0421] Z is N;

[0422] X is N;

[0423] Y is N;

[0424] or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

[0425] 15. A compound according to formula (II), or item 1, or item 6, wherein

[0426] R.sup.1 is methyl;

[0427] R.sup.2 is pyridinyl substituted by two or three groups independently selected from fluoro, chloro, methoxy, difluoroethoxy, tetrahydrofuranylmethoxy, tetrahydropyranylmethoxy, oxomorpholinylpropoxy, aminocarbonylhexyloxy, methylamino, dimethylamino, pyrrolidinyl and oxopiperazinyl;

[0428] U is CH;

[0429] W is CH;

[0430] Z is N;

[0431] X is N;

[0432] Y is N;

[0433] or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

[0434] 16. A compound according to formula (II), or item 1, wherein

[0435] R.sup.1 is C.sub.1-6alkyl, C.sub.3-7cycloalkyl, hydroxyC.sub.1-6alkyl, C.sub.1-6alkoxycarbonylC.sub.1-6alkyl, or carboxyC.sub.1-6alkyl;

[0436] R.sup.2 is phenyl substituted by one, two or three groups independently selected from halogen, nitro, C.sub.1-6alkylsulfonyl, --O--C.sub.xH.sub.2x--R.sup.3 and --O--C.sub.yH.sub.2y--NHR.sup.6; or pyridinyl substituted by two groups independently selected from halogen, haloC.sub.1-6alkoxy, --O--C.sub.xH.sub.2x--R.sup.3 and NR.sup.9R.sup.18; wherein

[0437] R.sup.3 is hydrogen, C.sub.3-7cycloalkyl, hydroxy, C.sub.1-6alkoxy, tetrahydrofuranyl, tetrahydropyranyl, morpholinyl, --C(.dbd.O)--R.sup.4, --SO.sub.2--R.sup.5 or aminocarbonylC.sub.1-6alkoxy; wherein

[0438] R.sup.4 is hydroxy, C.sub.1-6alkoxy, amino, diC.sub.1-6alkylamino or pyrrolidinyl;

[0439] R.sup.5 is C.sub.1-6alkyl;

[0440] R.sup.6 is hydrogen or C.sub.1-6alkylsulfonyl;

[0441] R.sup.9 and R.sup.10 are C.sub.1-6alkyl; or

[0442] R.sup.9 and R.sup.10 together with the nitrogen to which they are attached form pyrrolidinyl, morpholinyl, piperidinyl and oxopiperazinyl;

[0443] x is 1, 2, 3, 4, 5 or 6;

[0444] y is 1, 2, 3, 4, 5 or 6;

[0445] U is CH;

[0446] W is CH;

[0447] Z is CH;

[0448] X is N;

[0449] Y is N;

[0450] or a pharmaceutically acceptable salt, or an enantiomer, or a diastereomer thereof.

[0451] 17. A compound according to formula (II), or item 1, or item 16, or a pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R.sup.1 is C.sub.1-6alkyl.

[0452] 18. A compound according to formula (II), or any one of items 1, 16 and 17, or a pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R.sup.1 is methyl.

[0453] 19. A compound according to formula (II), or any one of items 1 and 16 to 18, or a pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R.sup.2 is phenyl substituted by one, two or three groups independently selected from halogen and C.sub.1-6alkoxy; or pyridinyl substituted by two groups independently selected from halogen, diC.sub.1-6alkylamino, pyrrolidinyl, and oxopiperazinyl.

[0454] 20. A compound according to formula (II), or any one of items 1, and 16 to 19, or a pharmaceutically acceptable salt, or enantiomer, or diastereomer thereof, wherein R.sup.2 is phenyl substituted by one, two or three groups independently selected from fluoro and methoxy; or pyridinyl substituted by two groups independently selected from fluoro, dimethylamino, pyrrolidinyl and oxopiperazinyl.

[0455] As described above, the inhibitor of the present invention may also be a RNAi molecule against

[0456] PAPD5 and/or PAPD7. Said RNAi molecule may be a siRNA or a shRNA.

[0457] For example, the inhibitor of the present invention may be a siRNA that is directed against PAPD5, wherein said siRNA is any one of the following siRNAs:

[0458] PAPD5 siRNA Pool (L-010011-00-0010; ON-TARGETplus Human PAPD5):

[0459] siRNA 1 J-010011-05 Target Sequence: CAUCAAUGCUUUAUAUCGA (SEQ ID NO: 10)

[0460] siRNA 2 J-010011-06 Target Sequence: GGACGACACUUCAAUUAUU (SEQ ID NO: 11)

[0461] siRNA 3 J-010011-07 Target Sequence: GAUAAAGGAUGGUGGUUCA (SEQ ID NO: 12)

[0462] siRNA 4 J-010011-08 Target Sequence: GAAUAGACCUGAGCCUUCA (SEQ ID NO: 13)

[0463] The inhibitor of the present invention may also be a siRNA that is directed against PAPD7, wherein said siRNA is any one of the following siRNAs:

[0464] PAPD7 siRNA Pool (L-009807-00-0005; ON-TARGETplus Human PAPD7):

TABLE-US-00001 siRNA-1-J-009807-05-Target Sequence: (SEQ ID NO: 14) GGAGUGACGUUGAUUCAGA siRNA-2-J-009807-06-Target Sequence: (SEQ ID NO: 15) CGGAGUUCAUCAAGAAUUA siRNA-3-J-009807-07-Target Sequence: (SEQ ID NO: 16) CGGAGUUCAUCAAGAAUUA siRNA-4-J-009807-08-Target Sequence: (SEQ ID NO: 17) GCGAAUAGCCACAUGCAAU

[0465] Above, target sequences of suitable siRNAs are shown. The sequences of the corresponding siRNAs are directly complementary to these target sequences.

[0466] It is envisaged in context of the present invention that (a) siRNA(s) directed against PAPD5 is combined with (a) siRNA(s) directed against PAPD7, in order to inhibit expression of both, PAPD5 and PAPD7.

[0467] The appended examples surprisingly demonstrate that two anti-HBV agents that are completely different in structure (i.e. DHQ and THP) have a shared binding site for PAPD5 and PAPD7 or at least are binding in close proximity to each other. In particular, selected interaction domains (SIDs) within PAPD5 and PAPD7 have been identified. SIDs are the amino acid sequences that are shared by all prey fragments matching the same reference protein. Therefore, the SIDs correspond to the amino acid regions where the anti-HBV agents DHQ and THP bind to PAPD5 and PAPD7. Accordingly, binding to these regions leads to an inhibition of the activity of PAPD5 and PAPD7, which results in inhibition of propagation of HBV. Thus, the inhibitor of the invention may be an antibody that specifically binds to at least one SID of PAPD5 and/or of PAPD7. Accordingly, the inhibitor of the invention may be an antibody that specifically binds to the amino acid stretch of any one of SEQ ID NOs: 7-9. The inhibitor of the invention may also be an antibody that specifically binds to more than one of the amino acid stretches of SEQ ID NOs: 7-9.

[0468] Applications

[0469] In context of the present invention it has surprisingly been shown that the combined inhibition of PAPD5 and PAPD7 leads to a synergistic effect in the inhibition of HBV propagation. The appended examples show that reduction of the expression of PAPD5 alone leads to a reduction of the secretion of HBsAg and HBeAg of around 50%. Reduction of the expression of PAPD7 alone leads to a reduction of the secretion of HBsAg and HBeAg of not more than 15%. Simultaneous knock-down of PAPD5 and PAPD7 leads to a synergistic effect in the reduction of secretion of HBsAg and HBeAg that lies above the sum of the single knock-downs. Without being bound by theory, this synergistic effect may be due to a compensatory effect of PAPD5 and PAPD7 since both proteins have high sequence homology and same enzymatic functions.

[0470] Therefore, one embodiment of the present invention relates to a combined preparation comprising an inhibitor of PAPD5 and an inhibitor of PAPD7 for use in the treatment and/or prevention of a HBV infection. Thus, the present invention relates to a combined preparation comprising an inhibitor of PAPD5 and an inhibitor of PAPD7 for simultaneous or sequential use in the treatment and/or prevention of a HBV infection. It is envisaged in context of the invention that said combined preparation is used for treating (e.g. ameliorating) a HBV infection. The definitions disclosed herein in connection with the inhibitor of the present invention apply, mutatis mutandis, to the combined preparation of the present invention. The combined preparation may comprise a molecule that is a PAPD5 inhibitor and a separate molecule that is a PAPD7 inhibitor (e.g. two separate siRNA molecules or two separate small molecules). These two separate inhibitors may be formulated within one unit, e.g., within one pill or vial. Alternatively, these two separate inhibitors may be formulated separately, in separate units, e.g. separate pills or vials. The two separate inhibitors may be administered together, (i.e. simultaneously) or separately (i.e. sequentially) provided that the synergistic effect of the two inhibitors is achieved. In one aspect of the invention the combined preparation leads to a reduction of secretion of HBsAg and HBeAg of at least 50% as compared to the no drug control (i.e. compared to cells or subjects to which no drug is administrated).

[0471] The present invention also relates to a pharmaceutical composition for use in the treatment and/or prevention of a HBV infection, wherein the pharmaceutical composition comprises

[0472] (i) the inhibitor of the invention; or the combined preparation of the invention; and

[0473] (ii) optionally a pharmaceutically acceptable carrier.

[0474] Accordingly, the present invention relates to a method of treating and/or preventing a HBV infection, wherein the method comprises administering an effective amount of the inhibitor of the invention, the pharmaceutical composition of the invention, or of the combined preparation of the invention to a subject in need of such a treatment.

[0475] The inhibitor of the invention, the combined preparation of the invention, or the pharmaceutical composition of the invention may be used in a combination therapy. For example, the inhibitor of the invention, the combined preparation of the invention, or the pharmaceutical composition of the invention may be combined with other anti-HBV agents such as interferon alpha-2b, interferon alpha-2a, and interferon alphacon-1 (pegylated and unpegylated), ribavirin, lamivudine (3TC), entecavir, tenofovir, telbivudine (LdT), adefovir, or other emerging anti-HBV agents such as a HBV RNA replication inhibitor, a HBsAg secretion inhibitor, a HBV capsid inhibitor, an antisense oligomer (e.g. as described in WO2012/145697 and WO 2014/179629), a siRNA (e.g. described in WO 2005/014806, WO 2012/024170, WO 2012/2055362, WO 2013/003520, WO 2013/159109, WO 2017/027350 and WO2017/015175), a HBV therapeutic vaccine, a HBV prophylactic vaccine, a HBV antibody therapy (monoclonal or polyclonal), or TLR 2, 3, 7, 8 or 9 agonists for the treatment and/or prophylaxis of HBV.

[0476] The appended examples demonstrate that down regulation of PAPD5 and/or PAPD7 goes along with a reduction in the production of HBsAg and HBeAg as well as of intracellular HBV mRNA in HBV infected cells. These results indicate that the amount and/or activity of PAPD5 and/or PAPD7 can be used for monitoring therapeutic success during the treatment of a HBV infection, e.g. if treatment with an inhibitor of PAPD5 and/or PAPD7 is ongoing or has been performed. Thus, the present invention relates to a method for monitoring the therapeutic success during the treatment of a HBV infection, wherein the method comprises:

[0477] (a) analyzing in a sample obtained from a test subject the amount and/or activity of PAPD5 and/or PAPD7;

[0478] (b) comparing said amount and/or activity with reference data corresponding to the amount and/or activity of PAPD5 and/or PAPD7 of at least one reference subject; and

[0479] (c) predicting therapeutic success based on the comparison step (b).

[0480] In the monitoring method of the invention the test subject may be a human being who receives medication for a HBV infection or has received medication for a HBV infection. The medication may comprise anti-HBV agents as described above. The medication may also comprise an inhibitor of PAPD5 and/or PAPD.

[0481] In the monitoring method of the invention the reference data may correspond to the amount and/or activity of PAPD5 and/or PAPD7 in a sample of at least one reference subject. Said sample may be blood or a liver biopsy.

[0482] One aspect of the invention relates to the monitoring method of the invention, wherein the at least one reference subject has a HBV infection but did not receive medication for a HBV infection; and wherein in step (c) a decreased amount and/or activity of PAPD5 and/or PAPD7 of the test subject as compared to the reference data indicates therapeutic success in the treatment of a HBV infection. For example, said decreased amount and/or activity of PAPD5 and/or PAPD7 may mean that the amount and/or activity of PAPD5 and/or PAPD7 in the sample of the test subject is 0 to 90% of the amount and/or activity of PAPD5 and/or PAPD7 in the sample of the at least one reference subject. For example, said decreased amount and/or activity of PAPD5 and/or PAPD7 may be 0 to 80%, preferably 0 to 70%, more preferably 0 to 60%, even more preferably 0 to 50%, even more preferably 0 to 40%, even more preferably 0 to 30, even more preferably 0 to 20%, and most preferably 0 to 10% of the amount and/or activity of PAPD5 and/or PAPD7 in the sample of the at least one reference subject.

[0483] Another aspect of the invention relates to the monitoring method of the invention, wherein the at least one reference subject has a HBV infection and has received medication for a HBV infection; and wherein in step (c) an identical or similar amount and/or activity of PAPD5 and/or PAPD7 of the test subject as compared to the reference data indicates therapeutic success in the treatment of a HBV infection. A further aspect of the invention relates to the monitoring method of the invention, wherein the at least one reference subject does not have a HBV infection; and wherein in step (c) an identical or similar amount and/or activity of PAPD5 and/or PAPD7 of the test subject as compared to the reference data indicates therapeutic success in the treatment of a HBV infection. An identical or similar amount and/or activity of PAPD5 and/or PAPD7 may mean that the amount and/or activity of PAPD5 and/or PAPD7 in the sample of the test subject is 90-110% of the amount and/or activity of PAPD5 and/or PAPD7 in the sample of the at least one reference subject. For example, said identical or similar amount and/or activity of PAPD5 and/or PAPD7 may be 95-105% of the amount and/or activity of PAPD5 and/or PAPD7 in the sample of the at least one reference subject.

[0484] Also encompassed by the present invention is a cell or a non-human animal (e.g. a mouse, rat, ferret or rabbit) with increased, reduced or absent PAPD5 and/or PAPD7 expression that can be used for identifying and/or characterizing a compound that prevents and/or treats (e.g. ameliorates) a HBV infection. For example, said cell or non-human animal may comprise an exogenous nucleotide sequence encoding PAPD5 and/or PAPD7, e.g. cloned into an expression vector and operable linked to an exogenous promoter. Said cell or non-human animal may overexpress PAPD5 and/or PAPD7, preferably PAPD5 and PAPD7. Alternatively, said cell or non-human animal may have a knock-down of PAPD5 and/or PAPD7, preferably of PAPD5 and PAPD7.

Embodiments of the Invention

[0485] Thus, the present invention relates to the following items:

[0486] 1. A method for identifying a compound that prevents, ameliorates and/or inhibits a hepatitis B virus (HBV) infection, comprising:

[0487] (a) contacting a test compound with

[0488] (a1) PAP associated domain containing 5 (PAPD5) and/or PAP associated domain containing 7 (PAPD7); or

[0489] (a2) a cell expressing PAPD5 and/or PAPD7;

[0490] (b) measuring the expression and/or activity of PAPD5 and/or PAPD7 in the presence and absence of said test compound; and

[0491] (c) identifying a compound that reduces the expression and/or activity of PAPD5 and/or PAPD7 as a compound that prevents, ameliorates and/or inhibits a HBV infection.

[0492] 2. A method for identifying a compound that prevents, ameliorates and/or inhibits a HBV infection, comprising:

[0493] (a) contacting a test compound with

[0494] (a1) PAPD5 and/or PAPD7; or

[0495] (a2) a cell expressing PAPD5 and/or PAPD7;

[0496] (b) measuring whether the test compound binds to PAPD5 and/or to PAPD7;

[0497] (c) measuring whether the test compound inhibits propagation of HBV; and

[0498] (d) identifying a compound that binds to PAPD5 and/or PAPD7 and inhibits propagation of HBV as a compound that prevents, ameliorates and/or inhibits a HBV infection. 3. The method of item 1 or 2, wherein PAPD5 is the PAPD5 polypeptide or the PAPD5 mRNA.

[0499] 4. The method of item 3, wherein the PAPD5 polypeptide is a polypeptide comprising or consisting of

[0500] (i) the amino acid sequence of SEQ ID NO: 1 or 2;

[0501] (ii) an amino acid sequence having at least 80% identity to an amino acid sequence of (i), wherein the polypeptide has poly-A polymerase function;

[0502] (iii) the amino acid sequence of an enzymatically active fragment of SEQ ID NO: 1 or 2; or

[0503] (iv) an amino acid sequence having at least 80% identity to an amino acid sequence of (iii), wherein the polypeptide has poly-A polymerase function.

[0504] 5. The method of item 3, wherein the PAPD5 mRNA is a polynucleotide comprising or consisting of

[0505] (i) the nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 1 or 2;

[0506] (ii) a nucleotide sequence encoding an amino acid sequence having at least 80% identity to SEQ ID NO: 1 or 2, wherein the polynucleotide encodes a polypeptide that has poly-A polymerase function;

[0507] (iii) the nucleotide sequence encoding an enzymatically active fragment of SEQ ID NO: 1 or 2; or

[0508] (iv) a nucleotide sequence encoding an amino acid sequence having at least 80% identity to an amino acid sequence of an enzymatically active fragment of SEQ ID NO: 1 or 2, wherein the polynucleotide encodes a polypeptide that has poly-A polymerase function.

[0509] 6. The method of item 1 or 2, wherein PAPD7 is the PAPD7 polypeptide or the PAPD7 mRNA.

[0510] 7. The method of item 6, wherein the PAPD7 polypeptide is a polypeptide comprising or consisting of

[0511] (i) the amino acid sequence of SEQ ID NO: 3;

[0512] (ii) an amino acid sequence having at least 80% identity to an amino acid sequence of (i), wherein the polypeptide has poly-A polymerase function;

[0513] (iii) the amino acid sequence of an enzymatically active fragment of SEQ ID NO: 3; or

[0514] (iv) an amino acid sequence having at least 80% identity to an amino acid sequence of (iii), wherein the polypeptide has poly-A polymerase function.

[0515] 8. The method of item 6, wherein the PAPD7 mRNA is a polynucleotide comprising or consisting of

[0516] (i) the nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 3;

[0517] (ii) a nucleotide sequence encoding an amino acid sequence having at least 80% identity to SEQ ID NO: 3, wherein the polynucleotide encodes a polypeptide that has poly-A polymerase function;

[0518] (iii) the nucleotide sequence encoding an enzymatically active fragment of SEQ ID NO: 3; or

[0519] (iv) a nucleotide sequence encoding an amino acid sequence having at least 80% identity to an amino acid sequence of an enzymatically active fragment of SEQ ID NO: 3, wherein the polynucleotide encodes a polypeptide that has poly-A polymerase function.

[0520] 9. The method of any one of items 1 to 8, wherein said cell is a eukaryotic cell.

[0521] 10. The method of any one of items 2 to 9, wherein the compound that inhibits propagation of HBV inhibits secretion of HBV surface antigen (HBsAg), inhibits secretion of HBV envelope antigen (HBeAg), and/or inhibits production of intracellular HBV mRNA or HBV DNA.

[0522] 11. The method of any one of items 1 to 10, which additionally comprises the step of comparing the test compound to a control.

[0523] 12. The method of item 11, wherein in said control an inactive test compound is used, wherein said inactive test compound is a compound that:

[0524] (i) does not reduce the expression and/or activity of PAPD5 and/or PAPD7; and/or

[0525] (ii) does not bind to PAPD5 and/or PAPD7 and does not inhibit propagation of HBV.

[0526] 13. The method of any one of items 1 to 12, wherein said test compound is a small molecule of a screening library; or

[0527] (ii) a peptide of a phage display library, of an antibody fragment library, or derived from a cDNA library.

[0528] 14. The method of any one of items 1 and 3 to 13, wherein the activity of PAPD5 and PAPD7 is the poly-A polymerase function.

[0529] 15. An inhibitor of PAPD5 and/or PAPD7 for use in treating and/or preventing a HBV infection, wherein said inhibitor is

[0530] (i) a small molecule that binds to PAPD5 and/or PAPD7;

[0531] (ii) a RNA interference (RNAi) molecule against PAPD5 and/or PAPD7;

[0532] (iii) an antibody that specifically binds to PAPD5 and/or PAPD7; or

[0533] (iv) a genome editing machinery, comprising:

[0534] (a) a site-specific DNA nuclease or a polynucleotide encoding a site-specific DNA nuclease; and

[0535] (b) a guide IRNA or a polynucleotide encoding a guide IRNA.

[0536] 16. The inhibitor for the use according to item 15, which

[0537] (i) binds to PAPD5 and/or PAPD7; and/or

[0538] (ii) inhibits expression and/or activity of PAPD5 and/or PAPD7.

[0539] 17. The inhibitor for the use according to item 15 or 16, wherein the inhibitor reduces secretion of HBsAg and HBeAg.

[0540] 18. The inhibitor for the use according to any one of items 15 to 17, wherein the inhibitor inhibits development of chronic HBV infection and/or reduces the infectiousness of a HBV infected person.

[0541] 19. The inhibitor for the use according to any one of items 15 to 18, wherein the inhibitor is the compound of formula (III) or (IV):

##STR00006##

[0542] 20. The inhibitor for the use according to any one of items 15 to 18, wherein the inhibitor is an RNAi molecule such as a siRNA or a shRNA.

[0543] 21. The inhibitor for the use according to any one of items 15 to 18, wherein the inhibitor is an antibody that specifically binds to the amino acid stretch of any one of SEQ ID NOs: 7-9.

[0544] 22. Combined preparation comprising an inhibitor of PAPD5 and an inhibitor of PAPD7 for simultaneous or sequential use in the treatment and/or prevention of a HBV infection.

[0545] 23. A pharmaceutical composition for use in the treatment and/or prevention of a HBV infection, wherein the pharmaceutical composition comprises

[0546] (i) the inhibitor for the use according to any one of items 15 to 21; or the combined preparation of item 22; and

[0547] (ii) optionally a pharmaceutically acceptable carrier.

[0548] 24. A method for monitoring the therapeutic success during the treatment of a HBV infection, wherein the method comprises:

[0549] (a) analyzing in a sample obtained from a test subject the amount and/or activity of PAPD5 and/or PAPD7;

[0550] (b) comparing said amount and/or activity with reference data corresponding to the amount and/or activity of PAPD5 and/or PAPD7 of at least one reference subject; and

[0551] (c) predicting therapeutic success based on the comparison step (b).

[0552] 25. The monitoring method of item 24, wherein the test subject is a human being who receives medication for a HBV infection or has received medication for a HBV infection.

[0553] 26. The monitoring method of item 24 or 25, wherein the reference data corresponds to the amount and/or activity of PAPD5 and/or PAPD7 in a sample of at least one reference subject.

[0554] 27. The monitoring method of any one of items 24 to 26, wherein the at least one reference subject has a HBV infection but did not receive medication for a HBV infection; and wherein in step (c) a decreased amount and/or activity of PAPD5 and/or PAPD7 of the test subject as compared to the reference data indicates therapeutic success in the treatment of a HBV infection.

[0555] 28. The monitoring method of item 27, wherein said decreased amount and/or activity of PAPD5 and/or PAPD7 means that the amount and/or activity of PAPD5 and/or PAPD7 in the sample of the test subject is 0 to 90% of the amount and/or activity of PAPD5 and/or PAPD7 in the sample of the at least one reference subject.

[0556] 29. The monitoring method of any one of items 24 to 26, wherein the at least one reference subject has a HBV infection and has received medication for a HBV infection; and wherein in step (c) an identical or similar amount and/or activity of PAPD5 and/or PAPD7 of the test subject as compared to the reference data indicates therapeutic success in the treatment of a HBV infection.

[0557] 30. The monitoring method of any one of items 24 to 26, wherein the at least one reference subject does not have a HBV infection; and wherein in step (c) an identical or similar amount and/or activity of PAPD5 and/or PAPD7 of the test subject as compared to the reference data indicates therapeutic success in the treatment of a HBV infection.

[0558] 31. The monitoring method of item 29 or 30, wherein said identical or similar amount and/or activity of PAPD5 and/or PAPD7 means that the amount and/or activity of PAPD5 and/or PAPD7 in the sample of the test subject is 90-110% of the amount and/or activity of PAPD5 and/or PAPD7 in the sample of the at least one reference subject.

[0559] Manufacture

[0560] A compound of formula (I) (i.e. a dihydroquinolizinone compound according to formula (I)) may be synthesized as described in WO 2015/113990 A1. In brief, a compound of formula (I) may be prepared by a method comprising the following steps:

[0561] (a) hydrolysis of a compound of formula (A)

##STR00007##

[0562] or

[0563] (b) hydrolysis of a compound of formula (B)

##STR00008##

[0564] wherein R.sup.1 to R.sup.7 and R.sup.9 are defined above with respect to formula (I) unless otherwise indicated.

[0565] In step (a) and step (b) a base such as lithium hydroxide or sodium hydroxide can for example be used.

[0566] A compound of formula (II) (i.e. a tetrahydropyridopyrimidine compound according to formula (II)) may be synthesized as described in WO2016/177655. In brief, a compound of formula (II) may be prepared by a method comprising one of the following steps:

[0567] (a) coupling of a compound of formula (A)

##STR00009##

[0568] with a compound of formula (B)

R.sup.1M (B)

[0569] in the presence of a Lewis acid;

[0570] (b) coupling of a compound of formula (C)

##STR00010##

[0571] with a compound of formula (D)

NHR.sup.9R.sup.10 (D)

[0572] in the presence of a base;

[0573] (c) coupling of a compound of formula (E)

##STR00011##

[0574] with a compound of formula (F)

R.sup.2-L.sup.2 (F);

[0575] wherein R.sup.1, R.sup.2, U, W, X, Y and Z are defined as above with respect to formula (II); M is H, Mg, Zn or Na; L.sup.1 is F, Cl or Br; and L.sup.2 is F, Cl or Br.

[0576] In step (a), the Lewis acid can for example be BF.sub.3.Et.sub.2O or Sc(OTf).sub.3;

[0577] In step (b), the base can for example be K.sub.2CO.sub.3 or DIEA;

[0578] In step (c), the reaction can be carried out in the presence of a base, and the base can for example be K.sub.2CO.sub.3 or DIEA. The reaction can also be carried out in the absence of a base.

[0579] Compositions

[0580] As described above, the invention relates to an inhibitor of PAPD5 and/or PAPD7 for use in treating and/or preventing a HBV infection; a combined preparation comprising an inhibitor of PAPD5 and an inhibitor of PAPD7 for use in the treatment and/or prevention of a HBV infection; and a pharmaceutical composition comprising said inhibitor or said combined preparation. Said pharmaceutical composition (i.e. medicament) optionally comprises a pharmaceutically acceptable carrier. Said pharmaceutical composition may further comprise a therapeutically acceptable diluent or excipient.

[0581] A typical pharmaceutical composition is prepared by mixing a PAPD5 inhibitor and/or a PAPD7 inhibitor and a carrier or excipient. Suitable carriers and excipients are well known to those skilled in the art and are described in detail in, e.g., Ansel, Ansel's Pharmaceutical Dosage Forms and Drug Delivery Systems, Philadelphia: Lippincott, Williams & Wilkins, 2004; Gennaro, Remington: The Science and Practice of Pharmacy, Philadelphia: Lippincott, Williams & Wilkins, 2000; and Rowe, Handbook of Pharmaceutical Excipients, Chicago, Pharmaceutical Press, 2005. The formulations may also include one or more buffers, stabilizing agents, surfactants, wetting agents, lubricating agents, emulsifiers, suspending agents, preservatives, antioxidants, opaquing agents, glidants, processing aids, colorants, sweeteners, perfuming agents, flavoring agents, diluents and other known additives to improve appearance of the drug or aid in the manufacturing of the pharmaceutical product (i.e., medicament). For example, the pharmaceutical composition of the invention may be formulated by mixing an inhibitor of PAPD5 and/or an inhibitor of PAPD7 at ambient temperature at an appropriate pH, and with the desired degree of purity, with physiologically acceptable carriers, i.e., carriers that are non-toxic to recipients at the dosages and concentrations employed into a suitable administration form. The pharmaceutical composition of the invention may be sterile.

[0582] The compounds according to the present invention may exist in the form of their pharmaceutically acceptable salts. The term "pharmaceutically acceptable salt" refers to conventional acid-addition salts or base-addition salts that retain the biological effectiveness and properties of the compounds of the present invention and are formed from suitable non-toxic organic or inorganic acids or organic or inorganic bases. Acid-addition salts include for example those derived from inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, sulfamic acid, phosphoric acid and nitric acid, and those derived from organic acids such as p-toluenesulfonic acid, salicylic acid, methanesulfonic acid, oxalic acid, succinic acid, citric acid, malic acid, lactic acid, fumaric acid, and the like. Base-addition salts include those derived from ammonium, potassium, sodium and, quaternary ammonium hydroxides, such as for example, tetramethyl ammonium hydroxide. The chemical modification of a pharmaceutical compound into a salt is a technique well known to pharmaceutical chemists in order to obtain improved physical and chemical stability, hygroscopicity, flowability and solubility of compounds. It is for example described in Bastin, Organic Process Research & Development 2000, 4, 427-435 or in Ansel, In: Pharmaceutical Dosage Forms and Drug Delivery Systems, 6th ed. (1995), pp. 196 and 1456-1457. For example, the pharmaceutically acceptable salt of the compounds provided herein may be a sodium salt.

[0583] Compounds contain one or several chiral centers can either be present as racemates, diastereomeric mixtures, or optically active single isomers. The racemates can be separated according to known methods into the enantiomers. Particularly, diastereomeric salts which can be separated by crystallization are formed from the racemic mixtures by reaction with an optically active acid such as e.g. D- or L-tartaric acid, mandelic acid, malic acid, lactic acid or camphorsulfonic acid.

[0584] The pharmaceutical composition of the invention is formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular mammal being treated, the clinical condition of the individual patient, the site of delivery of the agent, the method of administration, the scheduling of administration, the age and sex of the patients and other factors known to medical practitioners. Herein, an "effective amount" (also known as "(therapeutically) effective dose") means the amount of a compound that will elicit the biological or medical response of a subject that is being sought by a medical doctor or other clinician. The "effective amount" of the inhibitor of the invention, the combined preparation of the invention, or the pharmaceutical composition of the invention will be governed by such considerations, and is the minimum amount necessary to inhibit HBsAg and/or HBeAg. For example, such amount may be below the amount that is toxic to the cells of the recipient, or to the mammal as a whole.

[0585] For example, if the PAPD5 inhibitor and/or the PAPD7 inhibitor is/are (a) compound(s) according to formula (I) or (II), then the pharmaceutically effective amount administered parenterally per dose may be in the range of about 0.01 to 100 mg/kg, alternatively about 0.01 to 100 mg/kg of patient body weight per day, with the typical initial range of compound used being 0.3 to 15 mg/kg/day. In another example, if the PAPD5 inhibitor and/or the PAPD7 inhibitor is/are (a) compound(s) according to formula (I) or (II), oral unit dosage forms, such as tablets and capsules, preferably contain from about 0.1 to about 1000 mg.

[0586] The inhibitor of the invention, the combined preparation of the invention, or the pharmaceutical composition of the invention may be administered by any suitable means, including oral, topical (including buccal and sublingual), rectal, vaginal, transdermal, parenteral, subcutaneous, intraperitoneal, intrapulmonary, intradermal, intrathecal and epidural and intranasal, and, if desired for local treatment, intralesional administration. Parenteral infusions include intramuscular, intravenous, intraarterial, intraperitoneal, or subcutaneous administration.

[0587] The inhibitor of the invention, the combined preparation of the invention, or the pharmaceutical composition of the invention may be administered in any convenient administrative form, e.g., tablets, powders, capsules, solutions, dispersions, suspensions, syrups, sprays, suppositories, gels, emulsions, patches, etc. Such compositions may contain components conventional in pharmaceutical preparations, e.g., diluents, carriers, pH modifiers, sweeteners, bulking agents, and further active agents.

[0588] The inhibitor of the invention, the combined preparation of the invention, or the pharmaceutical composition of the invention are useful in the prevention and/or treatment of an HBV invention. They preferably inhibit secretion of HBsAg and/or HBeAg, most preferably of HBsAg and HBeAg.

Definitions

[0589] The terms "treatment", "treating", "treats" or the like are used herein to generally mean obtaining a desired pharmacological and/or physiological effect. This effect is therapeutic in terms of partially or completely curing a disease and/or adverse effect attributed to the disease. The term "treatment" as used herein covers any treatment of a disease in a subject and includes: (a) inhibiting the disease, i.e. arresting its development like the inhibition of increase of HBsAg and/or HBeAg; or (b) ameliorating (i.e. relieving) the disease, i.e. causing regression of the disease, like the repression of HBsAg and/or HBeAg production. Thus, a compound that ameliorates and/or inhibits a HBV infection is a compound that treats a HBV invention. Preferably, the term "treatment" as used herein relates to medical intervention of an already manifested disorder, like the treatment of an already defined and manifested HBV infection. Herein the term "preventing", "prevention" or "prevents" relates to a prophylactic treatment, i.e. to a measure or procedure the purpose of which is to prevent, rather than to cure a disease. Prevention means that a desired pharmacological and/or physiological effect is obtained that is prophylactic in terms of completely or partially preventing a disease or symptom thereof. Accordingly, herein "preventing a HBV infection" includes preventing a HBV infection from occurring in a subject, and preventing the occurrence of symptoms of a HBV infection.

[0590] For the purposes of the present invention the "subject" (or "patient") may be a vertebrate. In context of the present invention, the term "subject" includes both humans and other animals, particularly mammals, and other organisms. Thus, the herein provided means and methods are applicable to both human therapy and veterinary applications. Accordingly, herein the subject may be an animal such as a mouse, rat, hamster, rabbit, guinea pig, ferret, cat, dog, chicken, sheep, bovine species, horse, camel, or primate. Preferably, the subject is a mammal. More preferably the subject is human.

[0591] The term "hepatitis B virus infection" or "HBV infection" is commonly known in the art and refers to an infectious disease that is caused by the hepatitis B virus (HBV) and affects the liver. A HBV infection can be an acute or a chronic infection. Some infected persons have no symptoms during the initial infection and some develop a rapid onset of sickness with vomiting, yellowish skin, tiredness, dark urine and abdominal pain ("Hepatitis B Fact sheet N.sup.o 204". who.int. July 2014. Retrieved 4 Nov. 2014). Often these symptoms last a few weeks and can result in death. It may take 30 to 180 days for symptoms to begin. In those who get infected around the time of birth 90% develop a chronic hepatitis B infection while less than 10% of those infected after the age of five do ("Hepatitis B FAQs for the Public--Transmission", U.S. Centers for Disease Control and Prevention (CDC), retrieved 2011-11-29). Most of those with chronic disease have no symptoms; however, cirrhosis and liver cancer may eventually develop (Chang, 2007, Semin Fetal Neonatal Med, 12: 160-167). These complications result in the death of 15 to 25% of those with chronic disease ("Hepatitis B Fact sheet N.sup.o 204". who.int. July 2014, retrieved 4 Nov. 2014). Herein, the term "HBV infection" includes the acute and chronic hepatitis B infection. The term "HBV infection" also includes the asymptotic stage of the initial infection, the symptomatic stages, as well as the asymptotic chronic stage of the HBV infection.

[0592] Herein, an enzymatically active fragment of SEQ ID NO: 1 or 2 (i.e. of PAPD5) relates to those polypeptides that comprise a stretch of contiguous amino acid residues of SEQ ID NO: 1 or 2 (i.e. of PAPD5) and that retain a biological activity (i.e. functionality) of PAPD5, particularly the poly-A polymerase function. In line with this, herein, an enzymatically active fragment of SEQ ID NO: 3 (i.e. of PAPD7) relates to those polypeptides that comprise a stretch of contiguous amino acid residues of SEQ ID NO: 3 (i.e. of PAPD7) and that retain a biological activity (i.e. functionality) of PAPD7, particularly the poly-A polymerase function. Examples for enzymatically active fragments of PAPD5 and PAPD7 are the nucleotidyltransferase domain and the Cid1 poly A polymerase.

[0593] Herein, term "polypeptide" includes all molecules that comprise or consist of amino acid monomers linked by peptide (amide) bonds. Thus, the term "polypeptide" comprises all amino acid sequences, such as peptides, oliogopeptides, polypeptides and proteins. The "polypeptide" described herein may be a naturally occurring polypeptide or a non-naturally occurring polypeptide. The non-naturally occurring polypeptide may comprise at least one mutation (e.g. amino acid substitution, amino acid deletion or amino acid addition) as compared to the naturally occurring counterpart. The non-naturally occurring polypeptide may also be cloned in a vector and/or be operable linked to a promoter that is not the natural promoter of said polypeptide. Said promoter may be a constitutively active promoter. The term "amino acid" or "residue" as used herein includes both L- and D-isomers of the naturally occurring amino acids as well as of other amino acids (e.g., non-naturally-occurring amino acids, amino acids which are not encoded by nucleic acid sequences, synthetic amino acids etc.). Examples of naturally-occurring amino acids are alanine (Ala; A), arginine (Arg; R), asparagine (Asn; N), aspartic acid (Asp; D), cysteine (Cys; C), glutamine (Gin; Q), glutamic acid (Glu; E), glycine (Gly; G), histidine (His; H), isoleucine (Ile; I), leucine (Leu; L), lysine (Lys; K), methionine (Met; M), phenylalanine (Phe; F), proline (Pro; P), serine (Ser; 5), threonine (Thr; T), tryptophane (Trp; W), tyrosine (Tyr; Y), valine (Val; V). Post-translationally modified naturally-occurring amino acids are dehydrobutyrine (Dhb) and labionin (Lab). Examples for non-naturally occurring amino acids are described above. The non-naturally occurring polypeptide may comprise one or more non-amino acid substituents, or heterologous amino acid substituents, compared to the amino acid sequence of a naturally occurring form of the polypeptide, for example a reporter molecule or another ligand, covalently or non-covalently bound to the amino acid sequence.

[0594] The term "nucleotide sequence" or "polynucleotide" is commonly known in the art and comprises molecules comprising or consisting of naturally occurring molecules such as DNA and RNA as well as nucleic acid analogues such as, e.g., oligonucleotides thiophosphates, substituted ribo-oligonucleotides, LNA molecules, PNA molecules, GNA (glycol nucleic acid) molecules, TNA (threose nucleic acid) molecules, morpholino polynucleotides, or nucleic acids with modified backbones such as polysiloxane, and 2'-O-(2-methoxy) ethyl-phosphorothioate, or a nucleic acid with substituents, such as methyl-, thio-, sulphate, benzoyl-, phenyl-, amino-, propyl-, chloro-, and methanocarbanucleosides, or a reporter molecule to facilitate its detection. Furthermore, the term "nucleotide sequence" is to be construed equivalently with the term "nucleic acid molecule" in context of the present invention and may inter alia refer to DNA, RNA, PNA or LNA or hybrids thereof or any modification thereof that is known in the art (see, e.g., U.S. Pat. Nos. 5,525,711, 4,711,955, 5,792,608 or EP 302175 for examples of modifications). Nucleic acid residues comprised by the nucleic acid sequence described and provided herein may be naturally occurring nucleic acid residues or artificially produced nucleic acid residues. Examples for nucleic acid residues are adenine (A), guanine (G), cytosine (C), thymine (T), uracil (U), xanthine (X), and hypoxanthine (HX). As understood by the person of skill in the art, thymine (T) and uracil (U) may be used interchangeably depending on the respective type of polynucleotide. For example, as the skilled person is aware of, a thymine (T) as part of a DNA corresponds to an uracil (U) as part of the corresponding transcribed mRNA. The polynucleotides described and provided herein may be single- or double-stranded, linear or circular, natural or synthetic.

[0595] The nucleotide sequences provided herein may be cloned into a vector. The term "vector" as used herein includes plasmids, cosmids, viruses, bacteriophages and other vectors commonly used in genetic engineering. In a preferred embodiment, these vectors are suitable for the transformation of cells, like mammalian cells or yeast cells. Herein, the vector may be an expression vector. Generally, expression vectors have been widely described in the literature. They may comprise a selection marker gene and a replication-origin ensuring replication in the host, a promoter, and a termination signal for transcription. Between the promoter and the termination signal there may be at least one restriction site or a polylinker which enables the insertion of a nucleic acid sequence desired to be expressed. Non-limiting examples for the vector into which a nucleotide sequence provided herein may be cloned are adenoviral, adeno-associated viral (AAV), lentiviral, HIV-based lentiviral, nonviral minicircle-vectors, or other vectors for bacterial and eukaryotic expression systems.

[0596] Herein, the term "compound" means any molecule, including organic molecules such as small molecules, polynucleotides such as RNAi molecules, polypeptides such as antibodies, and inorganic compounds. The term "compound" also includes lipids, hormone analogs, polypeptide ligands, enzymes, receptors, channels, and antibody conjugates. For example, herein the compound may be an RNAi molecule against PAPD5 and/or PAPD7, an antibody that specifically binds to PAPD5 and/or PAPD7, or a small molecule binding to PAPD5 and/or PAPD7.

[0597] The term "inhibitor" is known in the art and relates to a compound/substance capable of fully or partially preventing or reducing the physiologic function (i.e. the activity) of (a) specific protein(s) (e.g. of PAPD5 and/or PAPD7). Inhibitors are also known as "antagonists". In the context of the present invention, the inhibitor of PAPD5 and/or PAPD7 may prevent or reduce or inhibit or inactivate the physiological activity of PAPD5 and/or PAPD7, respectively, e.g., upon binding of said compound/substance to PAPD5 and/or PAPD7, respectively. Binding of an inhibitor/antagonist to PAPD5 and/or PAPD7 may reduce the enzymatic function of PAPD5 and/or PAPD7 (i.e. the poly-A polymerase function) or may prevent the binding of an endogenous activating molecule to PAPD5 and/or PAPD7, and thereby inhibiting the activity (i.e. function) of these proteins. In the context of the present invention, an "inhibitor" of PAPD5 and/or PAPD7 may be capable of preventing the activity/function of PAPD5 and/or PAPD7, respectively, by preventing or reducing the expression of the PAPD5 and/or PAPD7 gene. Thus, an inhibitor of PAPD5 and/or PAPD7 may lead to a decreased expression level of PAPD5 and/or PAPD7 (e.g. decreased level of PAPD5 and/or PAPD7 mRNA, or of PAPD5 and/or PAPD7 protein) which is reflected in a decreased functionality (i.e. activity) of PAPD5 and/or PAPD7, wherein said function comprises the poly-A polymerase function. An inhibitor of PAPD5 and/or PAPD7, in the context of the present invention, accordingly, may also encompass transcriptional repressors of PAPD5 and/or PAPD7 expression that are capable of reducing the level of PAPD5 and/or PAPD7. Accordingly, all means and methods that result in a decrease in activity (which may be the result of a lower expression) or PAPD5 and/or PAPD7, are to be used as inhibitors of PAPD5 and/or PAPD7 in accordance with the present invention.

[0598] Herein, the term "RNA interference (RNAi) molecule" refers to any molecule inhibiting RNA expression or translation. A small interfering RNA (siRNA) is a double-stranded RNA molecule that, by binding complementary mRNA after transcription, leads to their degradation and loss in translation. A small hairpin RNA (shRNA) is an artificial RNA molecule with a hairpin structure which upon expression is able to reduce mRNA via the DICER and RNA reducing silencing complex (RISC). RNAi molecules can be designed on the base of the RNA sequence of the gene of interest. Corresponding RNAi can then be synthesized chemically or by in vitro transcription, or expressed from a vector or PCR product

[0599] The term "small molecule" refers to an organic compound with a low molecular weight (<900 daltons). Small molecules may help to regulate a biological process, and have generally a size on the order of 10.sup.-9 m. Many drugs are small molecules.

[0600] Herein the term "antibody" is used in the broadest sense and specifically encompasses intact monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g., bispecific antibodies) formed from at least two intact antibodies, and antibody fragments, so long as they exhibit the desired biological activity (i.e. specifically binding to PAPD5 and/or PAPD7). Also human, humanized, camelized or CDR-grafted antibodies are comprised. "Antibody fragments" comprise a portion of an intact antibody. The term "antibody fragments" includes antigen-binding portions, i.e., "antigen binding sites" (e.g., fragments, subsequences, complementarity determining regions (CDRs)) that retain capacity to bind an antigen (such as PAPD5 and/or PAPD7), comprising or alternatively consisting of, for example, (i) a Fab fragment, a monovalent fragment consisting of the VL, VH, CL and CH1 domains; (ii) a F(ab')2 fragment, a bivalent fragment comprising two Fab fragments linked by a disulfide bridge at the hinge region; (iii) a Fd fragment consisting of the VH and CH1 domains; (iv) a Fv fragment consisting of the VL and VH domains of a single arm of an antibody, (v) a dAb fragment (Ward; 1989; Nature 341; 544-546), which consists of a VH domain; and (vi) an isolated complementarity determining region (CDR). Antibody fragments or derivatives further comprise F(ab')2, Fv or scFv fragments or single chain antibodies.

[0601] The phrase "specifically bind(s)" or "bind(s) specifically" when referring to a binding molecule refers to a binding molecule (e.g. an antibody) which has intermediate or high binding affinity, exclusively or predominately, to a target molecule, preferably PAPD5 and/or PAPD7. The phrase "specifically binds to" refers to a binding reaction that is determinative of the presence of a target (preferably the PAPD5 and/or the PAPD7 protein) in the presence of a heterogeneous population of proteins and other biologics. Thus, under designated assay conditions, the specified binding molecules bind preferentially to a particular target (preferably the PAPD5 and/or PAPD7 protein) and do not bind in a significant amount to other components present in a test sample. Specific binding to a target protein under such conditions may require a binding molecule that is selected for its specificity for a particular target protein. A variety of assay formats may be used to select binding molecules that are specifically reactive with a particular target protein. For example, solid-phase ELISA immunoassays, immunoprecipitation, Biacore and Western blot may be used to identify binding molecules that specifically react with the PAPD5 and/or PAPD7 protein. The PAPD5 protein is most preferably a polypeptide that has the amino acid sequence as shown in SEQ ID NO: 1 or 2. However, the PAPD5 protein may also be a polypeptide having at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to the amino acid sequence of SEQ ID NO: 1 or 2 and being functional, wherein the function is poly-A polymerase function. The PAPD7 protein is most preferably a polypeptide that has the amino acid sequence as shown in SEQ ID NO: 3. However, the PAPD7 protein may also be a polypeptide having at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to the amino acid sequence of SEQ ID NO: 3 and being functional, wherein the function is poly-A polymerase function. Typically, a specific or selective reaction will be at least twice background signal or noise and more typically more than 10 times background. Or, in other words, the phrase "specifically binds to" refers to a binding reaction that is determinative of the presence of the target protein (preferably PAPD5 and/or PAPD7) in a heterogeneous population of proteins and other biologics. Typically, an antibody which specifically binds to a certain target (preferably PAPD5 and/or PAPD7) binds to said target with an association constant (K.sub.a) of at least about 1.times.10.sup.6 M.sup.-1 or 10.sup.7 M.sup.-1, or preferably about 10.sup.8 M.sup.-1 to 10.sup.9 M.sup.-1, or more preferably about 10.sup.10 M to 10.sup.11 M.sup.-1 or higher. Moreover, an antibody that specifically binds to a particular target (preferably PAPD5 and/or PAPD7) preferably binds to this target with an affinity that is at least two-fold greater than its affinity for binding to a non-specific target (e.g., BSA, casein) other than the predetermined target or a closely-related target.

[0602] In context of the present invention, the term "identity" or "percent identity" means that amino acid or nucleotide sequences have identities of at least 80%, preferably at least 90%, more preferably at least 95%, even more preferably at least 98%, and even more preferably at least 99% identity to the sequences shown herein, e.g. those of SEQ ID NO: 1, 2, or 3, wherein the higher identity values are preferred upon the lower ones. In accordance with the present invention, the term "identity/identities" or "percent identity/identities" in the context of two or more nucleic acid or amino acid sequences, refers to two or more sequences that are the same, or that have a specified percentage of amino acid residues or nucleotides that are the same (e.g., at least 80%, at least 90%, at least 95%, at least 98%, or at least 99% identity with the amino acid sequences of, e.g., SEQ ID NO: 1, 2 or 3, or with the nucleotide sequences of, e.g., SEQ ID NO: 4, 5 or 6), when compared and aligned for maximum correspondence over a window of comparison, or over a designated region as measured using a sequence comparison algorithm as known in the art, or by manual alignment and visual inspection.

[0603] Preferably the described identity to exists over a region that is at least about 50 amino acids, preferably at least 100 amino acids, more preferably at least 400 amino acids, more preferably at least 500 amino acids, more preferably at least 600 amino acids and most preferably all amino acids in length. In case of nucleotide sequences, the described identity most preferably exists over a region that is at least 100 nucleotides, preferably at least 1,000 nucleotides, more preferably at least 2,000 nucleotides and most preferably all nucleotides in length.

[0604] Those having skills in the art will know how to determine percent identity between/among sequences using, for example, algorithms such as those based on CLUSTALW computer program (Thompson, 1994, Nucl Acids Res, 2: 4673-4680) or FASTDB (Brutlag, 1990, Comp App Biosci, 6: 237-245), as known in the art. Also available to those having skills in this art are the BLAST and BLAST 2.0 algorithms (Altschul, 1997, Nucl Acids Res 25: 3389-3402; Altschul, 1993, J Mol Evol, 36: 290-300; Altschul, 1990, J Mol Biol 215: 403-410). For example, BLAST 2.0, which stands for Basic Local Alignment Search Tool BLAST (Altschul, 1997, loc. cit.; Altschul, 1993, loc. cit.; Altschul, 1990, loc. cit.), can be used to search for local sequence alignments. BLAST, as discussed above, produces alignments of both nucleotide and amino acid sequences to determine sequence similarity. Because of the local nature of the alignments, BLAST is especially useful in determining exact matches or in identifying similar sequences.

[0605] Analogous computer techniques using BLAST (Altschul, 1997, loc. cit.; Altschul, 1993, loc. cit.; Altschul, 1990, loc. cit.) are used to search for identical or related molecules in nucleotide databases such as GenBank or EMBL.

[0606] Herein, the term "measuring" also means "analyzing" or "determining" (i.e. detecting and/or quantifying). For example, the term "measuring the expression and/or activity of PAPD5 and/or PAPD7" means determining the amount of PAPD5 and/or PAPD7 expression and/or activity, for example, determining the amount of the PAPD5 and/or PAPD7 polypeptide (i.e. protein). Methods for measuring (i.e. determining) the amount and/or activity of PAPD5 and/or PAPD7 protein are known in the art and described herein above. Analogously, the term "measuring whether a test compound binds to PAPD5 and/or PAPD7" means analyzing or determining (i.e. detecting) whether a test compound binds to PAPD5 and/or PAPD7, e.g. to the PAPD5 polypeptide (i.e. protein) and/or to the PAPD7 polypeptide (i.e. protein). In line with this, the term "measuring whether a test compound inhibits propagation of HBV" means analyzing or determining (i.e. detecting and/or quantifying) whether a test compound inhibits propagation of HBV.

[0607] As used herein, the term "C.sub.1-6alkyl" alone or in combination signifies a saturated, linear- or branched chain alkyl group containing 1 to 6, particularly 1 to 4 carbon atoms, for example methyl, ethyl, propyl, isopropyl, 1-butyl, 2-butyl, tert-butyl and the like. Particular "C.sub.1-6alkyl" groups are methyl, ethyl, isopropyl and tert-butyl.

[0608] The term "C.sub.3-7cycloalkyl", alone or in combination, refers to a saturated carbon ring containing from 3 to 7 carbon atoms, particularly from 3 to 6 carbon atoms, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like. Particular "C.sub.3-7cycloalkyl" groups are cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0609] The term "C.sub.2-6alkenyl" denotes an unsaturated, linear or branched chain alkenyl group containing 2 to 6, particularly 2 to 4 carbon atoms, for example vinyl, propenyl, allyl, butenyl and the like. Particular "C.sub.2-6alkenyl" group is allyl and vinyl.

[0610] The term "C.sub.2-6alkynyl" denotes an unsaturated, linear or branched chain alkynyl group containing 2 to 6, particularly 2 to 4 carbon atoms, for example ethynyl, 1-propynyl, propargyl, butynyl and the like. Particular "C.sub.2-6alkynyl" groups are ethynyl and 1-propynyl.

[0611] The term "C.sub.xH.sub.2x" alone or in combination signifies a saturated, linear- or branched chain alkyl group containing 1 to 6, particularly 1 to 4 carbon atoms. The term "C.sub.1-6alkoxy" alone or in combination signifies a group C.sub.1-6alkyl-O--, wherein the "C.sub.1-6alkyl" is as defined above; for example methoxy, ethoxy, propoxy, iso-propoxy, n-butoxy, iso-butoxy, 2-butoxy, tert-butoxy, pentoxy, hexyloxy and the like. Particular "C.sub.1-6alkoxy" groups are methoxy, ethoxy and propoxy.

[0612] The term "halogen" means fluorine, chlorine, bromine or iodine.

[0613] The term "haloC.sub.1-6alkyl" denotes a C.sub.1-6alkyl group wherein at least one of the hydrogen atoms of the C.sub.1-6alkyl group has been replaced by same or different halogen atoms, particularly fluoro atoms. Examples of haloC.sub.1-6alkyl include monofluoro-, difluoro- or trifluoro-methyl, -ethyl or -propyl, for example 3,3,3-trifluoropropyl, 3,3-difluoropropyl, 2-fluoroethyl, 2,2-difluoroethyl, 2,2,2-trifluoroethyl, fluoromethyl, difluoromethyl or trifluoromethyl. Particular "haloC.sub.1-6alkyl" group is difluoromethyl or trifluoromethyl.

[0614] The term "haloC.sub.1-6alkoxy" denotes a C.sub.1-6alkoxy group wherein at least one of the hydrogen atoms of the C.sub.1-6alkoxy group has been replaced by same or different halogen atoms, particularly fluoro atoms. Examples of haloC.sub.1-6alkoxyl include monofluoro-, difluoro- or trifluoro-methoxy, -ethoxy or -propoxy, for example fluoropropoxy, difluoropropoxy, trifluoropropoxy, fluoroethoxy, difluoroethoxy, trifluoroethoxy, fluoromethoxy, difluoromethoxy or trifluoromethoxy. Particular "haloC.sub.1-6alkoxy" group is 3-fluoropropoxy, 3,3-difluoropropoxy, 3,3,3-trifluoropropoxy, 2-fluoroethoxy, 2,2-difluoroethoxy, 2,2,2-trifluoroethoxy, fluoromethoxy, difluoromethoxy or trifluoromethoxy.

[0615] The term "C.sub.3-7cycloalkyl", alone or in combination, refers to a saturated carbon ring containing from 3 to 7 carbon atoms, particularly from 3 to 6 carbon atoms, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl and the like. Particular "C.sub.3-7cycloalkyl" groups are cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

[0616] The term "C.sub.1-6alkoxy" alone or in combination signifies a group C.sub.1-6alkyl-O--, wherein the "C.sub.1-6alkyl" is as defined above; for example methoxy, ethoxy, propoxy, iso-propoxy, n-butoxy, iso-butoxy, 2-butoxy, tert-butoxy and the like. Particular "C.sub.1-6alkoxy" groups are methoxy and ethoxy and more particularly methoxy.

[0617] The term "haloC.sub.3-7cycloalkyl" denotes a C.sub.3-7cycloalkyl group wherein at least one of the hydrogen atoms of the C.sub.3-7cycloalkyl group has been replaced by same or different halogen atoms, particularly fluoro atoms. Examples of haloC.sub.3-7cycloalkyl include monofluoro- or difluoro-cyclopropyl, -cyclobutyl, -cyclopentyl or -cyclohexyl, for example fluorocyclopropyl, difluorocyclopropyl, fluocyclobutyl, difluocyclobutyl, fluocyclopentyl, difluocyclopentyl, fluocyclohexyl or difluocyclohexyl. Particular "haloC.sub.1-6alkyl" group is difluorocyclopropyl.

[0618] With respect to formula (I) the term "amino", alone or in combination, refers to primary (--NH.sub.2), secondary (--NH--) or tertiary amino

##STR00012##

[0619] With respect to formula (II) the term "amino" denotes a group of the formula --NR'R'' wherein R' and R'' are independently hydrogen, C.sub.1-6alkyl, C.sub.1-6alkoxy, C.sub.3-7cycloalkyl, heteroC.sub.3-7cycloalkyl, aryl or heteroaryl. Alternatively, R' and R'', together with the nitrogen to which they are attached, can form a heteroC.sub.3-7cycloalkyl.

[0620] The term "carbonyl" alone or in combination refers to the group --C(O)--.

[0621] The term "cyano" alone or in combination refers to the group --CN.

[0622] The term "C.sub.1-6alkylsulfinyl" denotes a group --SO--C.sub.1-6alkyl, wherein C.sub.1-6alkyl group is defined above. Examples of C.sub.1-6alkylsulfinyl include methylsulfinyl and ethylsulfinyl.

[0623] The term "C.sub.1-6alkylsulfonyl" denotes a group --SO.sub.2--C.sub.1-6alkyl, wherein C.sub.1-6alkyl group is defined above. Examples of C.sub.1-6alkylsulfonyl include methylsulfonyl and ethylsulfonyl.

[0624] The term "monocyclic heteroaryl" denotes a monovalent aromatic heterocyclic mono-ring system of 5 to 8 ring atoms, comprising 1, 2, 3 or 4 heteroatoms selected from N, O and S, the remaining ring atoms being carbon. Examples of monocyclic heteroaryl moieties include pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, thiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl, triazinyl, azepinyl, diazepinyl, isoxazolyl, isothiazolyl and the like.

[0625] With regard to formula (I) the term "monocyclic heterocycloalkyl" refers to a monovalent saturated or partly unsaturated monocyclic ring system of 3 to 7 ring atoms, comprising 1, 2, or 3 ring heteroatoms selected from N, O and S, the remaining ring atoms being carbon. Examples for monocyclic heterocycloalkyl are aziridinyl, oxiranyl, azetidinyl, oxetanyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, tetrahydrofuranyl, tetrahydro-thienyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperazinyl, morpholinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholin-4-yl, azepanyl, diazepanyl, homopiperazinyl, or oxazepanyl. Particular "monocyclic heterocycloalkyl" groups are morpholinyl, 2-oxo-pyrrolidinyl, pyrrolidinyl, tetrahydropyranyl, and more particularly pyrrolidin-1-yl, 2-oxo-pyrrolidin-1-yl, tetrahydropyran-4-yl and morpholin-1-yl.

[0626] With regard to formula (II) the term "monocyclic heterocycloalkyl" is a monovalent saturated or partly unsaturated monocyclic ring system of 4 to 7 ring atoms, comprising 1, 2, or 3 ring heteroatoms selected from N, O and S, the remaining ring atoms being carbon. Examples for monocyclic heterocycloalkyl are aziridinyl, oxiranyl, azetidinyl, oxetanyl, pyrrolidinyl, 2-oxo-pyrrolidinyl, tetrahydrofuranyl, thietanyl, pyrazolidinyl, imidazolidinyl, oxazolidinyl, isoxazolidinyl, thiazolidinyl, piperidinyl, tetrahydropyranyl, tetrahydrothiopyranyl, piperazinyl, morpholinyl, 2-oxo-morpholinyl, 2-oxo-piperazinyl, thiomorpholinyl, 1,1-dioxo-thiomorpholin-4-yl, 1,1-dioxothiolanyl, 1,1-dioxothietanyl, oxoimidazolidinyl, azepanyl, diazepanyl, homopiperazinyl, or oxazepanyl. Particular "monocyclic heterocycloalkyl" groups are azetidinyl, oxetanyl, thietanyl, tetrahydrofuranyl, tetrahydropyranyl, 1,1-dioxothietanyl, 1,1-dioxothiolanyl, morpholinyl, pyrrolidinyl, piperidinyl, piperazinyl, oxoimidazolidinyl, 2-oxo-pyrrolidinyl, 2-oxo-morpholinyl and 2-oxo-piperazinyl. More particularly, "monocyclic heterocycloalkyl" groups are azetidinyl, pyrrolidinyl, morpholinyl, oxomorpholinyl, piperidinyl, piperazinyl and oxopiperazinyl.

[0627] The term "aryl" denotes a monovalent aromatic carbocyclic mono- or bicyclic ring system comprising 6 to 10 carbon ring atoms. Examples of aryl moieties include phenyl and naphthyl, Particular "aryl" is phenyl.

[0628] The term "heteroaryl" denotes a monovalent aromatic heterocyclic mono- or bicyclic ring system of 5 to 12 ring atoms, comprising 1, 2, 3 or 4 heteroatoms selected from N, O and S, the remaining ring atoms being carbon. Examples of heteroaryl moieties include pyrrolyl, furanyl, thienyl, imidazolyl, oxazolyl, thiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl, triazinyl, azepinyl, diazepinyl, isoxazolyl, benzofuranyl, isothiazolyl, benzothienyl, indolyl, isoindolyl, isobenzofuranyl, benzimidazolyl, benzoxazolyl, benzoisoxazolyl, benzothiazolyl, benzoisothiazolyl, benzooxadiazolyl, benzothiadiazolyl, benzotriazolyl, purinyl, quinolinyl, isoquinolinyl, quinazolinyl, or quinoxalinyl. Particular "heteroaryl" are pyridinyl and pyrimidinyl.

[0629] The term "N-containing monocyclic heteroaryl" refers to a monocyclic heteroaryl wherein at least one of the heteroatoms is N. Examples for N-containing monocyclic heteroaryl are pyrrolyl, imidazolyl, oxazolyl, thiazolyl, triazolyl, oxadiazolyl, thiadiazolyl, tetrazolyl, pyridinyl, pyrazinyl, pyrazolyl, pyridazinyl, pyrimidinyl, triazinyl, azepinyl, diazepinyl, isoxazolyl, isothiazolyl and the like. Particular "N-containing monocyclic heteroaryl" groups are imidazolyl, pyrazolyl and triazolyl, and more particularly imidazol-1-yl, pyrazol-1-yl and 1,2,4-triazol-1-yl.

[0630] The term "halogen" means fluorine, chlorine, bromine or iodine. Halogen is particularly fluorine, chlorine or bromine.

[0631] The term "hydroxy" alone or in combination refers to the group --OH.

##STR00013##

[0632] The term "2-oxo-pyrrolidinyl" alone or in combination refers to the group.

[0633] The term "sulfonyl" alone or in combination refers to the group --S(O).sub.2--.

[0634] The term "C.sub.1-6alkylamino" refers to amino group as defined above wherein at least one of the hydrogen atoms of the amino group is replaced by a C.sub.1-6alkyl group.

[0635] The term "C.sub.1-6alkylsulfonyl" refers to a group C.sub.1-6alkyl-S(O).sub.2--, wherein the "C.sub.1-6alkyl" is as defined above.

[0636] The term "aminocarbonyl" refers to a group amino-C(O)--, wherein the "amino" is as defined above.

[0637] The term "cyanoC.sub.3-7cycloalkyl" refers to C.sub.3-7cycloalkyl group as defined above wherein at least one of the hydrogen atoms of the C.sub.3-7cycloalkyl group is replaced by a cyano group.

[0638] The term "pyrrolidinylcarbonyl" refers to a group pyrrolidinyl-C(O)--.

[0639] The term "enantiomer" denotes two stereoisomers of a compound which are non-superimposable mirror images of one another.

[0640] The term "diastereomer" denotes a stereoisomer with two or more centers of chirality and whose molecules are not mirror images of one another. Diastereomers have different physical properties, e.g. melting points, boiling points, spectral properties, and reactivities.

[0641] The present invention is further described by reference to the non-limiting figures and examples.

[0642] Examples The Examples illustrate the invention.

[0643] Material and Methods

[0644] Compound Chemistry

[0645] Each one compound from the two chemical series DHQ and THP were synthesized to be suitable for the Y3H screening performed by HYBRIGENICS SERVICES SAS. Both compounds included PEG5 linker and were tagged with a Trimethoprim (TMP) anchor ligand (Table 1).

TABLE-US-00002 TABLE 1 TMP-tagged compound IDs Hybrigenics ID Structure DHQ com- pound- TMP HBX129653 ##STR00014## THP com- pound- TMP HBX129654 ##STR00015##

[0646] Y3H ULTImate YChemH.TM. Screen

[0647] The two compounds were provided by Roche to HYBRIGENICS SERVICES SAS and tested for permeability and toxicity. Compounds were then screened against HYBRIGENICS's cDNA Human placenta library (PLA). The screens were carried out according to the optimized cell-to-cell mating protocol developed for Hybrigenics ULTImate Y2H.TM. using at different compound concentration (Table 2).

TABLE-US-00003 TABLE 2 YChemH screens IDs Hybrigenics YChemH Probe ID Project YChemH screen Project ID concentration DHQ HBX129653 hgx4240 PLA_RP6_hgx4240v1_pB409_A 5 .mu.M compound THP HBX129654 hgx4241 PLA_RP6_hgx4241v1_pB409_A 10 .mu.M compound

[0648] Y3H ULTImate YChemH.TM. Dependency Assay

[0649] Clones obtained from the screen were picked in 96-well format and clones positive for growth under selective conditions (HIS+) were evaluated in a dependency assay using spot assays. Only clones that were able to grow on selective medium in the presence of the tagged compound were being picked up, processed (cell lysis, PCR, gene sequencing) and mapped for protein alignment using Blast analysis.

[0650] Y3H ULTImate YChemH.TM. 1-by-1 Validation Experiment Prey Fragments

[0651] In this validation step each one identified fragment prey and one chemical probe (HBX129653, HBX129654) is tested in a 1-by-1 experiment. The plasmids from 3 selected preys from the screening library were extracted from the yeast cells, amplified in E. coli and re-transformed into YHGX13 yeast cells. For each interaction, DO1, 1/10, 1/100 and 1/1000 of the diploid yeast culture expressing both hook and prey constructs were spotted on a selective medium without tryptophan, leucine and histidine and supplemented with the chemical probe and FK506. Interactions were tested in duplicate. One plate was used per chemical compound and concentration (DMSO, 5, 10 and 20 .mu.M of HBX129653, 5, 10 and 20 .mu.M of HBX129654, 5 .mu.M of HBX24786 Trimethoprim (TMP) and 5 .mu.M of HBX129634 (TMP-PEG5-OH)). Plates were incubated at 30.degree. C. for 3 days.

[0652] Y3H ULTImate YChemH.TM. 1-by-1 Validation Experiment Full Length Proteins

[0653] The coding sequence of full-length PAPD5var1 (NM 001040284.2) and PAPD7varX1 (XM_005248234.2) were reconstituted from an N-terminal codon-optimized gene fragment (to remove high GC content) and commercially available clones of the C-terminal regions of the proteins and cloned in frame with the Gal4 Activation Domain (AD) into plasmid pP7 (AD-Prey), derived from the original pGADGH (Bartel et al., 1993 in Cellular interactions in development: A practical approach. ed. Hartley, D. A., Oxford University Press, Oxford, pp. 153-179). The constructs were checked by sequencing the entire inserts. For each prey, a mini-mating was carried out between YHGX13 (Y187 ade2-101::loxP-kanMX-IoxP, mato) transformed with the prey plasmids and YPT6AT yeast cells (mata) transformed with the DHFR hook (Dihydrofolate reductase) to produce a diploid yeast culture. For each interaction, DO1, 1/10, 1/100 and 1/1000 of the diploid yeast culture expressing both hook and prey constructs were spotted on a selective medium without tryptophan, leucine and histidine and supplemented with the chemical probe and FK506. Interactions were tested in duplicate. One plate was used per chemical compound and concentration (DMSO, 5, 10 and 20 .mu.M of HBX129653, 5, 10 and 20 .mu.M of HBX129654, 5 .mu.M of HBX24786 Trimethoprim (TMP) and 5 .mu.M of HBX129634 (TMP-PEG5-OH)). Plates were incubated at 30.degree. C. for 3 days.

[0654] Y3H ULTImate YChemH.TM.--Competition with Free Compound

[0655] The competition assay is based on the previously described 1-by-1 validation with a constant concentration for the chemical probe (HBX129653, HBX 129654) and increasing concentrations of the parent compound of the chemical probe (MOL653, MOL654) or its inactive enantiomer (INACT653, INACT654) (Table 3). The competition assays were performed on selective medium at 8 concentrations of the free compound (0, 0.25, 0.5, 1, 2, 5, 10 and 20 .mu.M) and a consistent concentration for the tagged Y3H-compound (1 .mu.M).

TABLE-US-00004 TABLE 3 YChemH competition IDs Hybrigenics ID Structure DHQ compound- active MOL653 ##STR00016## DHQ compound- inactive INACT653 ##STR00017## THP compound- active MOL654 ##STR00018## THP compound- inactive INACT654 ##STR00019##

[0656] HepaRG Cell Culture

[0657] HepaRG cells (Biopredics International, Rennes, France, Cat #HPR101) were cultured at 37.degree. C. in a humidified atmosphere with 5% CO2 in complete HepaRG growth medium consisting of William's E Medium (GIBCO), Growth Medium Supplement (Biopredics, Cat #ADD710) and 1% (v/v) GlutaMAX-1 (Gibco #32551) and 1.times. Pen/Strep (Gibco, #15140) for 2 weeks. To initiate differentiation, 0.9% (v/v) DMSO (Sigma-Aldrich, D2650) was added to the growth medium on confluent cells. After one week, medium was replaced by complete differentiation medium (HepaRG growth medium supplemented with 1.8% (v/v) DMSO) in which cells were maintained for approximately 4 weeks with differentiation medium renewal every 7 days. Differentiated HepaRG cells (dHepaRG), displayed hepatocyte-like cell islands surrounded by monolayer of biliary-like cells. Prior to HBV infection and compound treatment, dHepaRG cells were seeded into collagen I coated 96-well plates (Gibco, Cat #A11428-03) at 60,000 cells per well in 100 .mu.L of complete differentiation medium. Cells were allowed to recover their differentiated phenotype in 96-well plates for approximately 1 week after plating prior to HBV infection.

[0658] HBV Infection of dHepaRG

[0659] dHepaRG cells were infected with HBV particles at an MOI of 30. The HBV particles were produced from HBV-producing HepG2.2.15 cells (Sells et al 1987 Proc Natl Acad Sci USA 84, 1005-1009). dHepaRG culture conditions, differentiation and HBV infection have been described previously (Hantz, 2009, J. Gen. Virol., 2009, 90: 127-135). In brief complete differentiation medium (120 .mu.L/well) containing 4% PEG-8000 and virus stock (20 to 30 GE/cell) was added. One day post-infection, the cells were washed three times with phosphate-buffered saline and medium (complete differentiation medium) was replaced every two days during the experiment.

[0660] siRNA Treatment of HBV-Infected HepaRG

[0661] A pool of four different siRNAs was acquired from GE Dharmacon (ON TARGETplus) (Table 4).

TABLE-US-00005 TABLE 4 Overview siRNAs siRNA ON TARGETplus (Cat.No.) Target Sequence SEQ ID NO siPAPD5 (Cat. No. # J-010011-05 CAUCAAUGCUUUAUAUCGA 10 L-010011-00-0010) J-010011-06 GGACGACACUUCAAUUAUU 11 J-010011-07 GAUAAAGGAUGGUGGUUCA 12 J-010011-08 GAAUAGACCUGAGCCUUCA 13 siPAPD7 (Cat. No. # J-009807-05 GGAGUGACGUUGAUUCAGA 14 L-009807-00-0005) J-009807-06 CGGAGUUCAUCAAGAAUUA 15 J-009807-07 CGGAGUUCAUCAAGAAUUA 16 J-009807-08 GCGAAUAGCCACAUGCAAU 17

[0662] One day before infection with HBV cells and 4 days after infection cells were treated with siRNA pool either against PAPD5, PAPD7, both or the non-targeting siRNA as control. The siRNAs were transfected using DharmaFect 4 (GE Dharmacon; Cat. No. T-2004-01) and OPTI-MEM (Thermo Scientific; Cat. No. 51985034) according to manufacturer's protocol. The cells were treated for 11 days.

[0663] HBV Antigen Measurements

[0664] To evaluate the impact on HBV antigen expression and secretion, supernatants were collected on Day 11. HBV HBsAg and HBeAg levels were measured using CLIA ELISA Kits (Autobio Diagnostic #CL0310-2, #CL0312-2), according to the manufacturer's protocol. Briefly, 25 .mu.L of supernatant per well were transferred to the respective antibody coated microtiter plate and 25 .mu.L of enzyme conjugate reagent were added. The plate was incubated for 60 min on a shaker at room temperature before the wells were washed five times with washing buffer using an automatic washer. 25 .mu.L of substrate A and B were added to each well. The plates were incubated on a shaker for 10 min at room temperature before luminescence was measured using an Envision luminescence reader (Perkin Elmer).

[0665] Cell Viability

[0666] After the removal of supernatant media from the HBV infected dHepaRG cells, cells were incubated with CellTiterGlo One Solution (Promega) to measure cell viability.

[0667] Real-Time PCR for Intracellular mRNA

[0668] For intracellular mRNA isolation, dHepaRG were washed once with PBS (Gibco) and lysed using the MagNA Pure "96 Cellular RNA Large Volume Kit" (Roche #05467535001). The lystates may be stored at at -80.degree. C. For the real-time qPCR reaction an AB7900 HT sequence detection system (Applied Biosystems), the TaqMan.RTM. Gene Expression Master Mix (ThermoFisher Scientific) were used. For detection of HBV mRNA HBV core-specific primer (Integrated DNA Technologies) (Table 5) and to measure reduction of PAPD5 and PAPD7, in the presence of siRNA, gene-specific TaqMan Expression Assay probes (ThermoFisher Scientific; PAPD5 Cat. No. 4331182; PAPD7 Cat. No. 4331182) were used. Samples were normalized using TaqMan Expression Assay probe against b-Actin (ThermoFisher Scientific; PAPD5 Cat. No. 4331182).

TABLE-US-00006 TABLE 5 HBV core specific TaqMan probes Name Dye Sequence SEQ ID NO HBV Forward CTG TGC CTT GGG TGG CTT T 18 core (F3_HBVcore) Primer Reverse AAG GAA AGA AGT CAG AAG GCA AAA 19 (R3_HBVcore) Probe FAM- AGC TCC AAA/ZEN/TTC TTT ATA AGG 20 (P3_HBVcore) MGB GTC GAT GTC CAT G

Example 1: DHQ and THP Binds to PAPD5 and PAPD7

[0669] PAPD5/7 were Identified in Y3H Ultimate YChemH Screen as Common Interaction Partner of DHQ and THP

[0670] Both proteins PAPD5 (variant 1: NP_001035374; variant 2: NP_001035375) and PAPD7 (XP_005248291) were identified by a numerous number of fragments in the Y3H screen for both compounds (DHQ and THP) as described in the Materials and Method section. The identified proteins were ranked with a confidence score of A (scale A-D) by HYBRIGENICS (Table 6).

TABLE-US-00007 TABLE 6 YChemH screen results for PAPD5/7 Hybrigenics Protein prey # of Confidence ID identified fragments score DHQ HBX129653 PAPD5 variant 1 28 A compound PAPD5 variant 2 1 N/A PAPD7 12 A THP HBX129654 PAPD5 variant 1 5 N/A compound PAPD5 variant 2 49 A PAPD7 24 A

[0671] PAPD5/7 Interaction with DHQ and THP could be Confirmed Using Y3H ULTImate YChemH 1-by-1 Validation of Identified Prey Fragments and Further with Full Length Proteins

[0672] In a first validation step three fragments identified in the first screen were selected for the 1-by-1 validation assay (as described in the Materials and Method section) and tested at three different concentrations (5, 10 and 20 .mu.M) (Table 7).

TABLE-US-00008 TABLE 7 interacting fragment selected for validation assay Interaction # Prey fragment ID Protein Prey A PLA_RP6_hgx4240v1_pB409_A-15 PAPD7 B PLA_RP6_hgx4241v1_pB409_A-112 PAPD5 variant 1 C PLA_RP6_hgx4240v1_pB409_A-24 PAPD5 variant 2

[0673] All three fragments could be validated as specific binders for DHQ and THP already at the lowest tested concentration (FIG. 1).

[0674] In a second validation step, full length proteins for PAPD5 and PAPD7 were synthesized and used for 1-by-1 validation (as described in the Materials and Method section) with DHQ and THP (Table 8).

TABLE-US-00009 TABLE 8 Reference ID for full length protein prey used in 1-by-1 validation assay Interaction # HYBRIGENICS Reference Protein Prey A hgx4386v1_pP7 PAPD5 var1 full length B hgx4388v2_pP7 PAPD7 var1 full length

[0675] The interaction between these full length proteins and the DHQ and THP compounds were confirmed at the lowest tested concentration and shown to be specific for the chemical probes (FIG. 2).

[0676] PAPD5/7 Interaction with DHQ and THP in Y3H can be Competed by Both Free Active Compound, but not the Inactive Enantiomer

[0677] After validation of binding of DHQ and THP to protein fragments and full length PAPD5 and PAPD7 the binding was confirmed in a Y3H ULTImate YChenH competition experiment (as described in the Materials and Method section) using either inactive or active free compound (Table 9). A decrease of loss of yeast growth in the presence of the parent active compound, but not in the presence of the inactive enantiomer, means that the parent compound competes with the chemical probe and interacts with the protein target.

[0678] For all tested compounds toxicity on non-selective medium at the highest concentration (20 .mu.M) was tested using CellTiter-Glo Luminescent Cell Viability Assay (Promega) according to the manufacturer's protocol. No toxicity was observed at this concentration for any compound as yeast growth was not affected (data not shown). For both active free parent compounds (DHQ and THP, MOL653 and MOL654, respectively) competition could be observed, with lower concentration needed for competing the binding to the full length protein than for the fragment interactions (FIG. 3+4). Successful cross competition suggests a shared binding side for DHQ and THP to PAPD5/7 or at least binding in close proximity to each other.

TABLE-US-00010 TABLE 9 Reference ID for protein prey used in competition assay Interaction # Prey fragment ID Protein Prey A PLA_RP6_hgx4241v1_pB409_A-112 PAPD5 var1 experimental fragment B hgx4386v1_pP7 PAPD5 var1 full length C PLA_RP6_hgx4240v1_pB409_A-15 PAPD7 varX1 experimental fragment D hgx4388v2_pP7 PAPD7 varX1 full length

Example 2: Inhibition of PAPD5 and/or PAPD7 with siRNA Results in Effective Treatment of HBV Infection

[0679] To correlate the binding of DHQ and THP to PAPD5/7 and the impact of these two proteins on HBV gene expression, we used RNAi technology to reduce these proteins in naturally HBV infected dHepaRG and to monitor the impact of this reduction on viral parameters. For that we used siRNA pools against PAPD5 and PAPD7 (see table 4) in HBV infected dHepaRG cells as described in the Materials and methods section.

[0680] Reduction of PAPD5 led to inhibition of viral expression measured by secreted HBsAg and HBeAg as well as intracellular HBV mRNA (measured using CLIA ELISA and real-time PCR as described in the Materials and Methods section). While the reduction of PAPD5 mRNA dramatically reduced HBV gene expression, inhibition of PAPD7 had a modest effect on HBV expression (FIG. 7). However, an enhanced synergistic anti-HBV activity was observed when siRNA against PAPD7 and PAPD5 were combined (FIG. 7), suggesting a compensative role for PAPD7 in the absence of PAPD5.

Example 3: DHQ and THP Effectively Reduces HBsAg and HBeAg

[0681] The potency of DHQ and THP and their variants against HBV infection were measured in HepG2.2.15 cells using HBsAg and HBeAg as read out.

[0682] HepG2.2.15 cells (Sells et al 1987 Proc Natl Acad Sci USA 84, 1005-1009) were cultured in 96 well plates (15.000 cells/well in 100 uL) in DMEM+GluTaMax-1 (GiBCO Cat. NO. 10569), 1% Pen Strep (Gibco Cat. No. 15140), 10% FBS (Clontech Cat. No. 631106), Geneticin 0.25 ug/ml (Invitrogen 10131035). The compounds were tested using three-fold serial dilutions in DMSO with a top concentration of 100 .mu.M and 9 serial dilutions. Each compound was tested in quadricate. The cells were incubated for 3 days, supernatants were collected and HBsAg and HBeAg were measured as described in the Materials and Methods section. The IC.sub.50 values of the tested compounds in the reduction of secretion of HBsAg and HBeAg are shown in the following:

[0683] HBX129653 (DHQ TMP): IC.sub.50 HBsAg 1.181 uM

[0684] HBX129654 (THP TMP): IC.sub.50 HBsAg 0.299 uM

[0685] MOL653 (DHQ free--active): IC.sub.50 HBsAg 0.003 uM; IC.sub.50 HBeAg 0.007 uM

[0686] MOL654 (THP free--active): IC.sub.50 HBsAg 0.003 uM

[0687] INACT653 (DHQ free--inactive): IC.sub.50 HBsAg 3.15 uM

[0688] INACT654 (THP free--inactive): IC.sub.50 HBsAg>25 uM

Sequence CWU 1

1

201698PRTHomo sapiens 1Met Arg Pro Arg Pro Arg Ser Ala Pro Gly Lys Pro Arg Arg Arg Ser1 5 10 15Arg Ala Arg Leu Arg Ser Ser Arg Thr Pro Ser Gly Gly Ala Ser Gly 20 25 30Gly Gly Gly Ser Ser Ser Ser Ser Ser Thr Ala Thr Gly Gly Ser Gly 35 40 45Ser Ser Thr Gly Ser Pro Gly Gly Ala Ala Ser Ala Pro Ala Pro Ala 50 55 60Pro Ala Gly Met Tyr Arg Ser Gly Glu Arg Leu Leu Gly Ser His Ala65 70 75 80Leu Pro Ala Glu Gln Arg Asp Phe Leu Pro Leu Glu Thr Thr Asn Asn 85 90 95Asn Asn Asn His His Gln Pro Gly Ala Trp Ala Arg Arg Ala Gly Ser 100 105 110Ser Ala Ser Ser Pro Pro Ser Ala Ser Ser Ser Pro His Pro Ser Ala 115 120 125Ala Val Pro Ala Ala Asp Pro Ala Asp Ser Ala Ser Gly Ser Ser Asn 130 135 140Lys Arg Lys Arg Asp Asn Lys Ala Ser Thr Tyr Gly Leu Asn Tyr Ser145 150 155 160Leu Leu Gln Pro Ser Gly Gly Arg Ala Ala Gly Gly Gly Arg Ala Asp 165 170 175Gly Gly Gly Val Val Tyr Ser Gly Thr Pro Trp Lys Arg Arg Asn Tyr 180 185 190Asn Gln Gly Val Val Gly Leu His Glu Glu Ile Ser Asp Phe Tyr Glu 195 200 205Tyr Met Ser Pro Arg Pro Glu Glu Glu Lys Met Arg Met Glu Val Val 210 215 220Asn Arg Ile Glu Ser Val Ile Lys Glu Leu Trp Pro Ser Ala Asp Val225 230 235 240Gln Ile Phe Gly Ser Phe Lys Thr Gly Leu Tyr Leu Pro Thr Ser Asp 245 250 255Ile Asp Leu Val Val Phe Gly Lys Trp Glu Asn Leu Pro Leu Trp Thr 260 265 270Leu Glu Glu Ala Leu Arg Lys His Lys Val Ala Asp Glu Asp Ser Val 275 280 285Lys Val Leu Asp Lys Ala Thr Val Pro Ile Ile Lys Leu Thr Asp Ser 290 295 300Phe Thr Glu Val Lys Val Asp Ile Ser Phe Asn Val Gln Asn Gly Val305 310 315 320Arg Ala Ala Asp Leu Ile Lys Asp Phe Thr Lys Lys Tyr Pro Val Leu 325 330 335Pro Tyr Leu Val Leu Val Leu Lys Gln Phe Leu Leu Gln Arg Asp Leu 340 345 350Asn Glu Val Phe Thr Gly Gly Ile Gly Ser Tyr Ser Leu Phe Leu Met 355 360 365Ala Val Ser Phe Leu Gln Leu His Pro Arg Glu Asp Ala Cys Ile Pro 370 375 380Asn Thr Asn Tyr Gly Val Leu Leu Ile Glu Phe Phe Glu Leu Tyr Gly385 390 395 400Arg His Phe Asn Tyr Leu Lys Thr Gly Ile Arg Ile Lys Asp Gly Gly 405 410 415Ser Tyr Val Ala Lys Asp Glu Val Gln Lys Asn Met Leu Asp Gly Tyr 420 425 430Arg Pro Ser Met Leu Tyr Ile Glu Asp Pro Leu Gln Pro Gly Asn Asp 435 440 445Val Gly Arg Ser Ser Tyr Gly Ala Met Gln Val Lys Gln Ala Phe Asp 450 455 460Tyr Ala Tyr Val Val Leu Ser His Ala Val Ser Pro Ile Ala Lys Tyr465 470 475 480Tyr Pro Asn Asn Glu Thr Glu Ser Ile Leu Gly Arg Ile Ile Arg Val 485 490 495Thr Asp Glu Val Ala Thr Tyr Arg Asp Trp Ile Ser Lys Gln Trp Gly 500 505 510Leu Lys Asn Arg Pro Glu Pro Ser Cys Asn Gly Asn Gly Val Thr Leu 515 520 525Ile Val Asp Thr Gln Gln Leu Asp Lys Cys Asn Asn Asn Leu Ser Glu 530 535 540Glu Asn Glu Ala Leu Gly Lys Cys Arg Ser Lys Thr Ser Glu Ser Leu545 550 555 560Ser Lys His Ser Ser Asn Ser Ser Ser Gly Pro Val Ser Ser Ser Ser 565 570 575Ala Thr Gln Ser Ser Ser Ser Asp Val Asp Ser Asp Ala Thr Pro Cys 580 585 590Lys Thr Pro Lys Gln Leu Leu Cys Arg Pro Ser Thr Gly Asn Arg Val 595 600 605Gly Ser Gln Asp Val Ser Leu Glu Ser Ser Gln Ala Val Gly Lys Met 610 615 620Gln Ser Thr Gln Thr Thr Asn Thr Ser Asn Ser Thr Asn Lys Ser Gln625 630 635 640His Gly Ser Ala Arg Leu Phe Arg Ser Ser Ser Lys Gly Phe Gln Gly 645 650 655Thr Thr Gln Thr Ser His Gly Ser Leu Met Thr Asn Lys Gln His Gln 660 665 670Gly Lys Ser Asn Asn Gln Tyr Tyr His Gly Lys Lys Arg Lys His Lys 675 680 685Arg Asp Ala Pro Leu Ser Asp Leu Cys Arg 690 6952698PRTHomo sapiens 2Met Arg Pro Arg Pro Arg Ser Ala Pro Gly Lys Pro Arg Arg Arg Ser1 5 10 15Arg Ala Arg Leu Arg Ser Ser Arg Thr Pro Ser Gly Gly Ala Ser Gly 20 25 30Gly Gly Gly Ser Ser Ser Ser Ser Ser Thr Ala Thr Gly Gly Ser Gly 35 40 45Ser Ser Thr Gly Ser Pro Gly Gly Ala Ala Ser Ala Pro Ala Pro Ala 50 55 60Pro Ala Gly Met Tyr Arg Ser Gly Glu Arg Leu Leu Gly Ser His Ala65 70 75 80Leu Pro Ala Glu Gln Arg Asp Phe Leu Pro Leu Glu Thr Thr Asn Asn 85 90 95Asn Asn Asn His His Gln Pro Gly Ala Trp Ala Arg Arg Ala Gly Ser 100 105 110Ser Ala Ser Ser Pro Pro Ser Ala Ser Ser Ser Pro His Pro Ser Ala 115 120 125Ala Val Pro Ala Ala Asp Pro Ala Asp Ser Ala Ser Gly Ser Ser Asn 130 135 140Lys Arg Lys Arg Asp Asn Lys Ala Ser Thr Tyr Gly Leu Asn Tyr Ser145 150 155 160Leu Leu Gln Pro Ser Gly Gly Arg Ala Ala Gly Gly Gly Arg Ala Asp 165 170 175Gly Gly Gly Val Val Tyr Ser Gly Thr Pro Trp Lys Arg Arg Asn Tyr 180 185 190Asn Gln Gly Val Val Gly Leu His Glu Glu Ile Ser Asp Phe Tyr Glu 195 200 205Tyr Met Ser Pro Arg Pro Glu Glu Glu Lys Met Arg Met Glu Val Val 210 215 220Asn Arg Ile Glu Ser Val Ile Lys Glu Leu Trp Pro Ser Ala Asp Val225 230 235 240Gln Ile Phe Gly Ser Phe Lys Thr Gly Leu Tyr Leu Pro Thr Ser Asp 245 250 255Ile Asp Leu Val Val Phe Gly Lys Trp Glu Asn Leu Pro Leu Trp Thr 260 265 270Leu Glu Glu Ala Leu Arg Lys His Lys Val Ala Asp Glu Asp Ser Val 275 280 285Lys Val Leu Asp Lys Ala Thr Val Pro Ile Ile Lys Leu Thr Asp Ser 290 295 300Phe Thr Glu Val Lys Val Asp Ile Ser Phe Asn Val Gln Asn Gly Val305 310 315 320Arg Ala Ala Asp Leu Ile Lys Asp Phe Thr Lys Lys Tyr Pro Val Leu 325 330 335Pro Tyr Leu Val Leu Val Leu Lys Gln Phe Leu Leu Gln Arg Asp Leu 340 345 350Asn Glu Val Phe Thr Gly Gly Ile Gly Ser Tyr Ser Leu Phe Leu Met 355 360 365Ala Val Ser Phe Leu Gln Leu His Pro Arg Glu Asp Ala Cys Ile Pro 370 375 380Asn Thr Asn Tyr Gly Val Leu Leu Ile Glu Phe Phe Glu Leu Tyr Gly385 390 395 400Arg His Phe Asn Tyr Leu Lys Thr Gly Ile Arg Ile Lys Asp Gly Gly 405 410 415Ser Tyr Val Ala Lys Asp Glu Val Gln Lys Asn Met Leu Asp Gly Tyr 420 425 430Arg Pro Ser Met Leu Tyr Ile Glu Asp Pro Leu Gln Pro Gly Asn Asp 435 440 445Val Gly Arg Ser Ser Tyr Gly Ala Met Gln Val Lys Gln Ala Phe Asp 450 455 460Tyr Ala Tyr Val Val Leu Ser His Ala Val Ser Pro Ile Ala Lys Tyr465 470 475 480Tyr Pro Asn Asn Glu Thr Glu Ser Ile Leu Gly Arg Ile Ile Arg Val 485 490 495Thr Asp Glu Val Ala Thr Tyr Arg Asp Trp Ile Ser Lys Gln Trp Gly 500 505 510Leu Lys Asn Arg Pro Glu Pro Ser Cys Asn Gly Asn Gly Val Thr Leu 515 520 525Ile Val Asp Thr Gln Gln Leu Asp Lys Cys Asn Asn Asn Leu Ser Glu 530 535 540Glu Asn Glu Ala Leu Gly Lys Cys Arg Ser Lys Thr Ser Glu Ser Leu545 550 555 560Ser Lys His Ser Ser Asn Ser Ser Ser Gly Pro Val Ser Ser Ser Ser 565 570 575Ala Thr Gln Ser Ser Ser Ser Asp Val Asp Ser Asp Ala Thr Pro Cys 580 585 590Lys Thr Pro Lys Gln Leu Leu Cys Arg Pro Ser Thr Gly Asn Arg Val 595 600 605Gly Ser Gln Asp Val Ser Leu Glu Ser Ser Gln Ala Val Gly Lys Met 610 615 620Gln Ser Thr Gln Thr Thr Asn Thr Ser Asn Ser Thr Asn Lys Ser Gln625 630 635 640His Gly Ser Ala Arg Leu Phe Arg Ser Ser Ser Lys Gly Phe Gln Gly 645 650 655Thr Thr Gln Thr Ser His Gly Ser Leu Met Thr Asn Lys Gln His Gln 660 665 670Gly Lys Ser Asn Asn Gln Tyr Tyr His Gly Lys Lys Arg Lys His Lys 675 680 685Arg Asp Ala Pro Leu Ser Asp Leu Cys Arg 690 6953792PRTHomo sapiens 3Met Asp Pro Arg Val Ala Trp Ile Gln Pro Glu Gln Lys Gly Pro Ala1 5 10 15Asn Ala Leu Trp Met Gln Ile Trp Glu Thr Ser Gln Gly Val Gly Arg 20 25 30Gly Gly Ser Gly Phe Ala Ser Tyr Phe Cys Leu Asn Ser Pro Ala Leu 35 40 45Asp Thr Ala Ala Ala Ala Gly Ala Ala Gly Arg Gly Ser Gly Gly Leu 50 55 60Gly Pro Ala Leu Pro Ala Ala Ser Pro Pro Pro Pro Gly Pro Thr Ala65 70 75 80Pro Ala Ala Leu Pro Pro Ala Leu Leu Thr Ala Leu Gly Pro Ala Ala 85 90 95Glu Gly Ala Arg Arg Leu His Lys Ser Pro Ser Leu Ser Ser Ser Ser 100 105 110Ser Ser Ser Ser Ser Asn Ala Glu Ser Gly Thr Glu Ser Pro Gly Cys 115 120 125Ser Ser Ser Ser Ser Ser Ser Ala Ser Leu Gly Arg Pro Gly Gly Gly 130 135 140Arg Gly Gly Ala Phe Phe Asn Phe Ala Asp Gly Ala Pro Ser Ala Pro145 150 155 160Gly Thr Ala Asn Gly His Pro Gly Pro Arg Gly Pro Ala Pro Ala Gly 165 170 175Ser Pro Ser Gln His Gln Phe His Pro Gly Arg Arg Lys Arg Glu Asn 180 185 190Lys Ala Ser Thr Tyr Gly Leu Asn Tyr Leu Leu Ser Gly Ser Arg Ala 195 200 205Ala Ala Leu Ser Gly Gly Gly Gly Pro Gly Ala Gln Ala Pro Arg Pro 210 215 220Gly Thr Pro Trp Lys Ser Arg Ala Tyr Ser Pro Gly Ile Gln Gly Leu225 230 235 240His Glu Glu Ile Ile Asp Phe Tyr Asn Phe Met Ser Pro Cys Pro Glu 245 250 255Glu Ala Ala Met Arg Arg Glu Val Val Lys Arg Ile Glu Thr Val Val 260 265 270Lys Asp Leu Trp Pro Thr Ala Asp Val Gln Ile Phe Gly Ser Phe Ser 275 280 285Thr Gly Leu Tyr Leu Pro Thr Ser Asp Ile Asp Leu Val Val Phe Gly 290 295 300Lys Trp Glu Arg Pro Pro Leu Gln Leu Leu Glu Gln Ala Leu Arg Lys305 310 315 320His Asn Val Ala Glu Pro Cys Ser Ile Lys Val Leu Asp Lys Ala Thr 325 330 335Val Pro Ile Ile Lys Leu Thr Asp Gln Glu Thr Glu Val Lys Val Asp 340 345 350Ile Ser Phe Asn Met Glu Thr Gly Val Arg Ala Ala Glu Phe Ile Lys 355 360 365Asn Tyr Met Lys Lys Tyr Ser Leu Leu Pro Tyr Leu Ile Leu Val Leu 370 375 380Lys Gln Phe Leu Leu Gln Arg Asp Leu Asn Glu Val Phe Thr Gly Gly385 390 395 400Ile Ser Ser Tyr Ser Leu Ile Leu Met Ala Ile Ser Phe Leu Gln Leu 405 410 415His Pro Arg Ile Asp Ala Arg Arg Ala Asp Glu Asn Leu Gly Met Leu 420 425 430Leu Val Glu Phe Phe Glu Leu Tyr Gly Arg Asn Phe Asn Tyr Leu Lys 435 440 445Thr Gly Ile Arg Ile Lys Glu Gly Gly Ala Tyr Ile Ala Lys Glu Glu 450 455 460Ile Met Lys Ala Met Thr Ser Gly Tyr Arg Pro Ser Met Leu Cys Ile465 470 475 480Glu Asp Pro Leu Leu Pro Gly Asn Asp Val Gly Arg Ser Ser Tyr Gly 485 490 495Ala Met Gln Val Lys Gln Val Phe Asp Tyr Ala Tyr Ile Val Leu Ser 500 505 510His Ala Val Ser Pro Leu Ala Arg Ser Tyr Pro Asn Arg Asp Ala Glu 515 520 525Ser Thr Leu Gly Arg Ile Ile Lys Val Thr Gln Glu Val Ile Asp Tyr 530 535 540Arg Arg Trp Ile Lys Glu Lys Trp Gly Ser Lys Ala His Pro Ser Pro545 550 555 560Gly Met Asp Ser Arg Ile Lys Ile Lys Glu Arg Ile Ala Thr Cys Asn 565 570 575Gly Glu Gln Thr Gln Asn Arg Glu Pro Glu Ser Pro Tyr Gly Gln Arg 580 585 590Leu Thr Leu Ser Leu Ser Ser Pro Gln Leu Leu Ser Ser Gly Ser Ser 595 600 605Ala Ser Ser Val Ser Ser Leu Ser Gly Ser Asp Val Asp Ser Asp Thr 610 615 620Pro Pro Cys Thr Thr Pro Ser Val Tyr Gln Phe Ser Leu Gln Ala Pro625 630 635 640Ala Pro Leu Met Ala Gly Leu Pro Thr Ala Leu Pro Met Pro Ser Gly 645 650 655Lys Pro Gln Pro Thr Thr Ser Arg Thr Leu Ile Met Thr Thr Asn Asn 660 665 670Gln Thr Arg Phe Thr Ile Pro Pro Pro Thr Leu Gly Val Ala Pro Val 675 680 685Pro Cys Arg Gln Ala Gly Val Glu Gly Thr Ala Ser Leu Lys Ala Val 690 695 700His His Met Ser Ser Pro Ala Ile Pro Ser Ala Ser Pro Asn Pro Leu705 710 715 720Ser Ser Pro His Leu Tyr His Lys Gln His Asn Gly Met Lys Leu Ser 725 730 735Met Lys Gly Ser His Gly His Thr Gln Gly Gly Gly Tyr Ser Ser Val 740 745 750Gly Ser Gly Gly Val Arg Pro Pro Val Gly Asn Arg Gly His His Gln 755 760 765Tyr Asn Arg Thr Gly Trp Arg Arg Lys Lys His Thr His Thr Arg Asp 770 775 780Ser Leu Pro Val Ser Leu Ser Arg785 79048114DNAHomo sapiens 4acaacgcgct ccctgcgggg cgggcggcaa cctccatgcg gcctcgtcca cgctcagcac 60cggggaagcc gaggcggaga agccgcgcgc gcctcagaag ctcccggacg cccagcggcg 120gcgcgagcgg cggcggcggc agcagcagca gcagcagcac ggccaccggc gggagcggca 180gcagcaccgg cagccccggc ggcgcggcct cggccccggc cccggccccg gccggcatgt 240atcgctccgg ggagcgcctg ctgggcagcc acgcgctgcc cgcggagcag cgggacttcc 300tgcccctaga gacgaccaac aacaacaaca accaccacca gcccggggcc tgggcccgcc 360gggcgggctc ctcggcgtcc tcgcctccct cggcgtcctc gtccccgcac ccttcggccg 420ccgtccccgc cgccgatcca gccgattcgg cctcgggcag cagcaacaag aggaagcgcg 480acaacaaggc cagcacgtat ggactcaact acagcctgct gcagcccagc ggagggcggg 540ccgcgggggg cggccgagca gacggcggcg gggtcgtgta cagcgggacc ccgtggaaac 600ggaggaacta caaccaggga gtcgtgggtc tgcatgaaga aatcagtgat ttttatgaat 660acatgtctcc aagacctgag gaggagaaga tgcggatgga ggtggtgaac aggatcgaga 720gtgtaattaa ggagctctgg cccagcgctg acgtccagat atttggaagt tttaaaactg 780gactttattt acctactagt gacatcgacc tagtggtgtt tgggaagtgg gagaacctac 840ccctctggac tctggaagaa gctcttcgga aacacaaagt cgcagatgag gattcggtga 900aagttttaga caaagcaact gtacctatta ttaaattaac agattctttt actgaagtga 960aagttgatat cagctttaat gtacagaatg gcgtgagagc agctgacctc atcaaagatt 1020ttaccaagaa atatcctgta ttgccatact tggttttagt attgaaacaa ttcctattgc 1080agagggacct taatgaagta tttacaggtg gaattggttc ttatagtctc tttttaatgg 1140cagtcagttt ccttcagtta catcccaggg aagatgcttg catccccaat acaaactatg 1200gtgttctctt aatagaattt tttgaattat atggacgaca cttcaattat ttaaagactg 1260gcatccggat aaaggatggt ggttcatatg tggccaaaga tgaagtacag aaaaatatgc 1320tagatggcta caggccatca atgctttata tcgaagatcc tttacaacca ggtaacgatg 1380ttggaaggag ttcatatggg gccatgcaag tgaagcaggc ctttgattat gcctacgttg 1440ttttgagtca tgctgtatca ccaatagcaa agtactatcc caacaatgaa acagaaagca 1500tactaggtag aataattaga gtaacagatg aagttgccac atatagagat tggatatcaa 1560agcagtgggg cttgaagaat agacctgagc cttcatgcaa tggaaatggt gttaccttga 1620tagtagatac tcagcagtta gataaatgta ataataatct atctgaagaa aatgaagccc 1680ttggaaaatg tagaagtaaa acctcggaat

ctcttagtaa acactcttca aactcttcat 1740caggtccagt gtcgtcctct tctgccacac agtccagctc tagtgatgta gattccgatg 1800caacaccatg caaaaccccg aaacagctgc tttgccgtcc gtccactggg aaccgagtag 1860ggtcgcaaga tgtatccttg gagtcctctc aggcagttgg gaaaatgcaa agcacccaaa 1920ccactaacac atccaacagc accaacaaat ctcagcatgg atcagcaagg ctctttcgtt 1980cttccagcaa aggcttccaa ggtacaactc aaacaagcca tggttccttg atgacaaaca 2040aacaacatca aggcaaatcc aataatcagt attaccatgg caaaaagagg aaacacaaga 2100gggacgcgcc cctctcagac ctctgtagat agtcagcgct gcgcggtgga ctgtcttctc 2160tgtgcaatga tctcatgctc aggacagttg cgcagggact cctgggagat attcaggagc 2220ctcacactgt tcagacgttg acttagcaac tgcgtttttt cccagctcgc cacagaatgg 2280atcatgaaga ctgacaactg caaaaaaaac aaaacaaaac aaaaaaaaaa gcaagcaaaa 2340aagagggaaa aaaaaggctg cttatttgat aagtcatatg ctacaacagg gtcattttaa 2400gatttaaagc ttgaatgtaa aataaatata tttctcattg gctttatgca gagttatagg 2460gaatagtatt cagtgttggt agggtgatag aaacaaaaaa cagtatcaga ggatgaggtg 2520gggaaggaaa acaaaggtat ctgataggaa gtccagattc caaaggggaa agtgatctgt 2580gcatgttttt tttttaaata tttttgcata tatttaccat tttattgtgt gtatatatag 2640aagaccatat aggagattga tatttgtaat agtggatttg ttaataatac tttttacata 2700acattactgt ttaaattgta aacagatttt ttctcaggat tagtttgaaa aataatctaa 2760attgtcatct taacatccat atatagggaa gtgattagtt ctattactca atttgttttt 2820ctcagcattg aaatgactta atagaaccct tgtgtcctgc tgcaaaaatt tttcctctct 2880aaagaaaagg tttatggtgg caaatgatgt ttattttatt ttgtaaaaaa aaaaaaatgt 2940actatgtact tttgtgtaaa cactgaaaaa tctctggtca tctccgagaa ttaacttgca 3000actgttttct atagtgctgt cgtcttgggc aatgggcaat tacatgactt tgtgtttgct 3060tcctttgcag tctttttttt ttccccccat ttcttcctaa taggaaaaaa aaaaaaaaaa 3120aggtcaccca tgtctggtct cattcctgtt gcagtgaaac ttcgagttcc acagactttg 3180catgctggct tctctaaccc tgtgtgctgc gtgtgcctgt ttctcatctc ttattctttt 3240taaaattcat gcttaactac tgtgggagaa taactgtaaa cagctttaat taaatcatac 3300ttataaaaaa ctattttctt atattccact ctatgctttt ggtattgttg atctttacaa 3360attaaatggt ctttgataat ggatctattt tgtattgcct tattaagacc aaatacttct 3420tgtcatccca ttctttatcc tcttctttca tggaattgtt atcgttaatt aaaacttttt 3480taaacattgg cttgtttcaa tcatactgta aattttggtt gtagtcagct ttgagtgcaa 3540tgagatgtat aattctgtta tcattacctg ttgagtttga aactcagttg ggaatattta 3600atataataga atgtaagtga catttctgaa aatgctttct ttcagggtga aagctcttat 3660gtttagcatc aatgtgtatg gctctgttaa atgcagccat ttctgagacg agattctttt 3720atatatatat acatataaag tactattggc ttttaggagt ttcttttata tacatttatg 3780aaatactgaa gaccaatcag accattaatg gacacttagt gtaacttttt ataaagaaaa 3840taatgctaaa gtaagaccaa aactgatgtc atcactgaaa ttaacaattt tcaatatgtt 3900catattttaa ttcacaatgg aaaaatgtgt tccaaaactg gaaactcata gtactcgtgt 3960aaactgtgga agatttcaaa tgtgatgtta ttttgacaat gttttaaatt ttagagtcac 4020attttattct gatcagaatt tttattgaga tgttgagctt ttgtttttga aactagtttg 4080tcataacatt gtgcataatc acagtattta ttttctagga caattgtgaa tgtgtagact 4140tatgtttact gctaagggaa caattattta taaaataata ttaaatccag tattagctgc 4200ctatttcaga cacttaatac ttgcagagat ctatgttaca tttaccacac tgaagttttt 4260tttgttgttt tttgtttgtt tttaaagaat caccctcatt gttgaaagta aatgtactct 4320tagggtgcga atattagtgt tccaataagc atgtgattat attaaggtgg tggtagcggg 4380aagataattc tgattccatt gggaatctta ggttttcgta aatttattgg gaaaatagtt 4440tttcctgtac tgctgaagtt tctttttggt aaacagtatc tttctaaaag aaaaaagcat 4500gaaggagaaa ttgaggtgtg tatacatttc ctcaaatgac cagcattgta ttcgtgaata 4560ctgtgtatct tgcagtgaac agtgtggaag ctgttcattt ttcaatctga agtaaaatac 4620tttcaagaac ttttagtttg cctgctcatt tgttttatac atttcatcta tttgactcct 4680atcttatttc ttttttgagt tttaatactt cctatatttt gtgaatatat cagaaatgtg 4740tcatttatat attagagtcc attcatatcc atgaatcata accttccttt gctaatactt 4800gttgaatggg attttacaaa ttctccctca ctctggtgac atttctcagg cagtcatgta 4860tgtgtacctg gccattagaa atattaatat ttaaagactg ttttttagag gagctgatgg 4920gttggtgagg tgtcagcaca aaatcttact ggttatgttt tgatgataaa agtatatcca 4980ttttttccct ccagctttaa ggtgactgtg aaggtgcctg gttttgaatg tctttgtttg 5040gtttggagat gtcgcactca gttttcaaat ctagcttgga tctgtaggac ctatgttttt 5100tacaagtaat tgccctccag tcttcaacag ttgattctgt tttattttta tcctgttttg 5160agtgtacttt acctttactt gcattttgag cctcattaat atttaggtta tttgatttgg 5220ctccagatat tcctagatct gcacagggca aaacatgggc tatagggtga gcatttttaa 5280ttgtcttttt ctgctggaac cttatatctc tccatgtgtt ttctgctcct tccctccccc 5340atgaaatggt aagtgtgact tgtgtttgcc tgaacctgtg gactagtgtt tggggtttct 5400ggaaacacta gagggtcaga aaagagtaat gaccaccgtg acgtgcagga ttctcttgct 5460gtgacatgtt cattgcaaag ccctctccag tgactaggag gtgtagttat taaggttgat 5520ctgttagaaa tcaccattat taggtattag tggtagatgt tgctgatact tttattggtc 5580atgactacat ctcagtttta ctttaatatt gatctatagt ttgatcagtt ccttgaattc 5640taatatgttg atttctcagt gtttctgtca ctaaccaaga atgtttctag gcagttggtt 5700gcttcacagt caaaactaaa tggtaaacta tcaaaaatac attcccaatt ttgctgtgat 5760aaatattgaa atgttaaaat taatgaacag aagaatttat tcttacccat ctattcttgt 5820tctcctagtt cattaaactt tcagttattg gaaaggcaca ttctcaaagt attttatgag 5880caaaatattc tataaatgcg tctaacaaac ctaattgaat ataaaagtta tatttagtag 5940ttactgttga tagtaatttt catcagggtc atagttcatc tagtaaaata tttagagaat 6000gatgttaaca ttccagcatt aaagtgggaa caaagattta tatatgaaat tccttaaaag 6060agttcatctt gccttggttt ctgaccctca agactctagc tacctgccat cttgtcaaaa 6120catttgtggg tagaataagt gttaaagatc aaattttaat atgcttctcg atatttaaca 6180tagctaagaa gccagatttt actgtagaag ttatttacat gatttgaaaa cttgacctaa 6240ctggaagcct ttttctcagt catcttgttc taagccatct tgacttcaca cccttagcga 6300cttttctttt ttttttggtc aaagataatg agctaaatat atatagacgt tgaatgttga 6360caaaattatt aaccagaaaa attgcttata aaggctgctg atctatttga tacctagaat 6420taaatatttg aggacagttt ttagttaata aactgctaat gtttatttta ctgtctctca 6480ggtttttggt ttttttaaaa aaaatgtgtt tggcctttac attttctact taagtgtgta 6540ctttattgag tttaaccttg tctgtagcct agtagcctga aagaaaagga gacagaacca 6600gagagatgga tgtagtgcat tccctttggt tattacacat ttgtggtagc tcctggattt 6660actgagagat attttagcta tgtcaataag aacagctaat gatgtggaaa tcaggtgttc 6720tcttgtgtat ttcagtgaac atttttatta gtagttgcat atcatctcta gttccacatt 6780ttaacttaac gtctttgtgg cttcaccact gagctacctt tcactacacc agcttctgtg 6840tggcctggta acatggaagg tctctcctaa ggacagtctg gacgtatttt gggggaatgt 6900tatttatctt aaagatgcct agaaacaaaa cgcatatagt accagtgaga aactatgaag 6960taaacaagtt gctcaggccg ggcatggtgg ctcacgcctg taatcccagc actttgggag 7020gccgaagcgg gaggatggct tgaggctggg agtttgagac cttcatctct taaaaaaaca 7080aacaaaaacc tgaatggtga ggtgtggtgg aattgggtag gggagggaaa ggaggacttg 7140gaaaagcatt ctccaaagcc agcaacttgg tgaagttcag tacttgcctc ttagaggtta 7200ggccatgcct ttcaaagaga gtgaaatgat gggttatcag ccacattctt ggagttaata 7260tttttcttca tctttcagtt tgggttctgt gctattcata gttcttccct aagaccattt 7320cattattacc ttttatattt agttgcaatt tattataata tgttgttttg tccctgaact 7380taatctccta attttaagat cctctctgat ttttgcatat tgaaacttac agaagtcact 7440ttaaaaaagt cttttgaaag tcctacaatc ctaaaataaa tcacaagctt gtttgttaga 7500cgtgtcaaga gtctccagtc tttactacta aaaagcagca ctgccttaac acacattgtt 7560atgggtgaaa agtgagggac gaccagtgta gtttctggat ataaagtgtg aaggactgtt 7620gagttaaaca tttttagtgg aatatacata gataacgtgt atttagaaac tttggtgaag 7680ccagtatttg tttttagtaa cctttttatg tatttccttc tttgattagc attgtcttca 7740gtgttaagaa atgtggactc ctgtgaggtg ctggaggttt gaatcatctt gaaaactttc 7800caatcttgtc tagttaccac tgcagagaca ctaaggaatt taccagaaaa agatatttga 7860tacaagtgat ttaagaaatc tcaacatttc ctgaggccgt atcactgggc aaccagtgat 7920gaaaactatg aatgaattgc acacctggaa gattttttaa gctaatgaca gtttcttcaa 7980agatgtcaat tatttgcctt ggaaatttta taaattgcat ttctatgcac atcggcctct 8040agtgcttacc actcggttta ttattcataa tctgcaattc aataaaggct ttgtgttttc 8100atttatcttc aaaa 811457973DNAHomo sapiens 5acaacgcgct ccctgcgggg cgggcggcaa cctccatgcg gcctcgtcca cgctcagcac 60cggggaagcc gaggcggaga agccgcgcgc gcctcagaag ctcccggacg cccagcggcg 120gcgcgagcgg cggcggcggc agcagcagca gcagcagcac ggccaccggc gggagcggca 180gcagcaccgg cagccccggc ggcgcggcct cggccccggc cccggccccg gccggcatgt 240atcgctccgg ggagcgcctg ctgggcagcc acgcgctgcc cgcggagcag cgggacttcc 300tgcccctaga gacgaccaac aacaacaaca accaccacca gcccggggcc tgggcccgcc 360gggcgggctc ctcggcgtcc tcgcctccct cggcgtcctc gtccccgcac ccttcggccg 420ccgtccccgc cgccgatcca gccgattcgg cctcgggcag cagcaacaag aggaagcgcg 480acaacaaggc cagcacgtat ggactcaact acagcctgct gcagcccagc ggagggcggg 540ccgcgggggg cggccgagca gacggcggcg gggtcgtgta cagcgggacc ccgtggaaac 600ggaggaacta caaccaggga gtcgtgggtc tgcatgaaga aatcagtgat ttttatgaat 660acatgtctcc aagacctgag gaggagaaga tgcggatgga ggtggtgaac aggatcgaga 720gtgtaattaa ggagctctgg cccagcgctg acgtccagat atttggaagt tttaaaactg 780gactttattt acctactagt gacatcgacc tagtggtgtt tgggaagtgg gagaacctac 840ccctctggac tctggaagaa gctcttcgga aacacaaagt cgcagatgag gattcggtga 900aagttttaga caaagcaact gtacctatta ttaaattaac agattctttt actgaagtga 960aagttgatat cagctttaat gtacagaatg gcgtgagagc agctgacctc atcaaagatt 1020ttaccaagaa atatcctgta ttgccatact tggttttagt attgaaacaa ttcctattgc 1080agagggacct taatgaagta tttacaggtg gaattggttc ttatagtctc tttttaatgg 1140cagtcagttt ccttcagtta catcccaggg aagatgcttg catccccaat acaaactatg 1200gtgttctctt aatagaattt tttgaattat atggacgaca cttcaattat ttaaagactg 1260gcatccggat aaaggatggt ggttcatatg tggccaaaga tgaagtacag aaaaatatgc 1320tagatggcta caggccatca atgctttata tcgaagatcc tttacaacca ggtaacgatg 1380ttggaaggag ttcatatggg gccatgcaag tgaagcaggc ctttgattat gcctacgttg 1440ttttgagtca tgctgtatca ccaatagcaa agtactatcc caacaatgaa acagaaagca 1500tactaggtag aataattaga gtaacagatg aagttgccac atatagagat tggatatcaa 1560agcagtgggg cttgaagaat agacctgagc cttcatgcaa tggtccagtg tcgtcctctt 1620ctgccacaca gtccagctct agtgatgtag attccgatgc aacaccatgc aaaaccccga 1680aacagctgct ttgccgtccg tccactggga accgagtagg gtcgcaagat gtatccttgg 1740agtcctctca ggcagttggg aaaatgcaaa gcacccaaac cactaacaca tccaacagca 1800ccaacaaatc tcagcatgga tcagcaaggc tctttcgttc ttccagcaaa ggcttccaag 1860gtacaactca aacaagccat ggttccttga tgacaaacaa acaacatcaa ggcaaatcca 1920ataatcagta ttaccatggc aaaaagagga aacacaagag ggacgcgccc ctctcagacc 1980tctgtagata gtcagcgctg cgcggtggac tgtcttctct gtgcaatgat ctcatgctca 2040ggacagttgc gcagggactc ctgggagata ttcaggagcc tcacactgtt cagacgttga 2100cttagcaact gcgttttttc ccagctcgcc acagaatgga tcatgaagac tgacaactgc 2160aaaaaaaaca aaacaaaaca aaaaaaaaag caagcaaaaa agagggaaaa aaaaggctgc 2220ttatttgata agtcatatgc tacaacaggg tcattttaag atttaaagct tgaatgtaaa 2280ataaatatat ttctcattgg ctttatgcag agttataggg aatagtattc agtgttggta 2340gggtgataga aacaaaaaac agtatcagag gatgaggtgg ggaaggaaaa caaaggtatc 2400tgataggaag tccagattcc aaaggggaaa gtgatctgtg catgtttttt ttttaaatat 2460ttttgcatat atttaccatt ttattgtgtg tatatataga agaccatata ggagattgat 2520atttgtaata gtggatttgt taataatact ttttacataa cattactgtt taaattgtaa 2580acagattttt tctcaggatt agtttgaaaa ataatctaaa ttgtcatctt aacatccata 2640tatagggaag tgattagttc tattactcaa tttgtttttc tcagcattga aatgacttaa 2700tagaaccctt gtgtcctgct gcaaaaattt ttcctctcta aagaaaaggt ttatggtggc 2760aaatgatgtt tattttattt tgtaaaaaaa aaaaaatgta ctatgtactt ttgtgtaaac 2820actgaaaaat ctctggtcat ctccgagaat taacttgcaa ctgttttcta tagtgctgtc 2880gtcttgggca atgggcaatt acatgacttt gtgtttgctt cctttgcagt cttttttttt 2940tccccccatt tcttcctaat aggaaaaaaa aaaaaaaaaa ggtcacccat gtctggtctc 3000attcctgttg cagtgaaact tcgagttcca cagactttgc atgctggctt ctctaaccct 3060gtgtgctgcg tgtgcctgtt tctcatctct tattcttttt aaaattcatg cttaactact 3120gtgggagaat aactgtaaac agctttaatt aaatcatact tataaaaaac tattttctta 3180tattccactc tatgcttttg gtattgttga tctttacaaa ttaaatggtc tttgataatg 3240gatctatttt gtattgcctt attaagacca aatacttctt gtcatcccat tctttatcct 3300cttctttcat ggaattgtta tcgttaatta aaactttttt aaacattggc ttgtttcaat 3360catactgtaa attttggttg tagtcagctt tgagtgcaat gagatgtata attctgttat 3420cattacctgt tgagtttgaa actcagttgg gaatatttaa tataatagaa tgtaagtgac 3480atttctgaaa atgctttctt tcagggtgaa agctcttatg tttagcatca atgtgtatgg 3540ctctgttaaa tgcagccatt tctgagacga gattctttta tatatatata catataaagt 3600actattggct tttaggagtt tcttttatat acatttatga aatactgaag accaatcaga 3660ccattaatgg acacttagtg taacttttta taaagaaaat aatgctaaag taagaccaaa 3720actgatgtca tcactgaaat taacaatttt caatatgttc atattttaat tcacaatgga 3780aaaatgtgtt ccaaaactgg aaactcatag tactcgtgta aactgtggaa gatttcaaat 3840gtgatgttat tttgacaatg ttttaaattt tagagtcaca ttttattctg atcagaattt 3900ttattgagat gttgagcttt tgtttttgaa actagtttgt cataacattg tgcataatca 3960cagtatttat tttctaggac aattgtgaat gtgtagactt atgtttactg ctaagggaac 4020aattatttat aaaataatat taaatccagt attagctgcc tatttcagac acttaatact 4080tgcagagatc tatgttacat ttaccacact gaagtttttt ttgttgtttt ttgtttgttt 4140ttaaagaatc accctcattg ttgaaagtaa atgtactctt agggtgcgaa tattagtgtt 4200ccaataagca tgtgattata ttaaggtggt ggtagcggga agataattct gattccattg 4260ggaatcttag gttttcgtaa atttattggg aaaatagttt ttcctgtact gctgaagttt 4320ctttttggta aacagtatct ttctaaaaga aaaaagcatg aaggagaaat tgaggtgtgt 4380atacatttcc tcaaatgacc agcattgtat tcgtgaatac tgtgtatctt gcagtgaaca 4440gtgtggaagc tgttcatttt tcaatctgaa gtaaaatact ttcaagaact tttagtttgc 4500ctgctcattt gttttataca tttcatctat ttgactccta tcttatttct tttttgagtt 4560ttaatacttc ctatattttg tgaatatatc agaaatgtgt catttatata ttagagtcca 4620ttcatatcca tgaatcataa ccttcctttg ctaatacttg ttgaatggga ttttacaaat 4680tctccctcac tctggtgaca tttctcaggc agtcatgtat gtgtacctgg ccattagaaa 4740tattaatatt taaagactgt tttttagagg agctgatggg ttggtgaggt gtcagcacaa 4800aatcttactg gttatgtttt gatgataaaa gtatatccat tttttccctc cagctttaag 4860gtgactgtga aggtgcctgg ttttgaatgt ctttgtttgg tttggagatg tcgcactcag 4920ttttcaaatc tagcttggat ctgtaggacc tatgtttttt acaagtaatt gccctccagt 4980cttcaacagt tgattctgtt ttatttttat cctgttttga gtgtacttta cctttacttg 5040cattttgagc ctcattaata tttaggttat ttgatttggc tccagatatt cctagatctg 5100cacagggcaa aacatgggct atagggtgag catttttaat tgtctttttc tgctggaacc 5160ttatatctct ccatgtgttt tctgctcctt ccctccccca tgaaatggta agtgtgactt 5220gtgtttgcct gaacctgtgg actagtgttt ggggtttctg gaaacactag agggtcagaa 5280aagagtaatg accaccgtga cgtgcaggat tctcttgctg tgacatgttc attgcaaagc 5340cctctccagt gactaggagg tgtagttatt aaggttgatc tgttagaaat caccattatt 5400aggtattagt ggtagatgtt gctgatactt ttattggtca tgactacatc tcagttttac 5460tttaatattg atctatagtt tgatcagttc cttgaattct aatatgttga tttctcagtg 5520tttctgtcac taaccaagaa tgtttctagg cagttggttg cttcacagtc aaaactaaat 5580ggtaaactat caaaaataca ttcccaattt tgctgtgata aatattgaaa tgttaaaatt 5640aatgaacaga agaatttatt cttacccatc tattcttgtt ctcctagttc attaaacttt 5700cagttattgg aaaggcacat tctcaaagta ttttatgagc aaaatattct ataaatgcgt 5760ctaacaaacc taattgaata taaaagttat atttagtagt tactgttgat agtaattttc 5820atcagggtca tagttcatct agtaaaatat ttagagaatg atgttaacat tccagcatta 5880aagtgggaac aaagatttat atatgaaatt ccttaaaaga gttcatcttg ccttggtttc 5940tgaccctcaa gactctagct acctgccatc ttgtcaaaac atttgtgggt agaataagtg 6000ttaaagatca aattttaata tgcttctcga tatttaacat agctaagaag ccagatttta 6060ctgtagaagt tatttacatg atttgaaaac ttgacctaac tggaagcctt tttctcagtc 6120atcttgttct aagccatctt gacttcacac ccttagcgac ttttcttttt tttttggtca 6180aagataatga gctaaatata tatagacgtt gaatgttgac aaaattatta accagaaaaa 6240ttgcttataa aggctgctga tctatttgat acctagaatt aaatatttga ggacagtttt 6300tagttaataa actgctaatg tttattttac tgtctctcag gtttttggtt tttttaaaaa 6360aaatgtgttt ggcctttaca ttttctactt aagtgtgtac tttattgagt ttaaccttgt 6420ctgtagccta gtagcctgaa agaaaaggag acagaaccag agagatggat gtagtgcatt 6480ccctttggtt attacacatt tgtggtagct cctggattta ctgagagata ttttagctat 6540gtcaataaga acagctaatg atgtggaaat caggtgttct cttgtgtatt tcagtgaaca 6600tttttattag tagttgcata tcatctctag ttccacattt taacttaacg tctttgtggc 6660ttcaccactg agctaccttt cactacacca gcttctgtgt ggcctggtaa catggaaggt 6720ctctcctaag gacagtctgg acgtattttg ggggaatgtt atttatctta aagatgccta 6780gaaacaaaac gcatatagta ccagtgagaa actatgaagt aaacaagttg ctcaggccgg 6840gcatggtggc tcacgcctgt aatcccagca ctttgggagg ccgaagcggg aggatggctt 6900gaggctggga gtttgagacc ttcatctctt aaaaaaacaa acaaaaacct gaatggtgag 6960gtgtggtgga attgggtagg ggagggaaag gaggacttgg aaaagcattc tccaaagcca 7020gcaacttggt gaagttcagt acttgcctct tagaggttag gccatgcctt tcaaagagag 7080tgaaatgatg ggttatcagc cacattcttg gagttaatat ttttcttcat ctttcagttt 7140gggttctgtg ctattcatag ttcttcccta agaccatttc attattacct tttatattta 7200gttgcaattt attataatat gttgttttgt ccctgaactt aatctcctaa ttttaagatc 7260ctctctgatt tttgcatatt gaaacttaca gaagtcactt taaaaaagtc ttttgaaagt 7320cctacaatcc taaaataaat cacaagcttg tttgttagac gtgtcaagag tctccagtct 7380ttactactaa aaagcagcac tgccttaaca cacattgtta tgggtgaaaa gtgagggacg 7440accagtgtag tttctggata taaagtgtga aggactgttg agttaaacat ttttagtgga 7500atatacatag ataacgtgta tttagaaact ttggtgaagc cagtatttgt ttttagtaac 7560ctttttatgt atttccttct ttgattagca ttgtcttcag tgttaagaaa tgtggactcc 7620tgtgaggtgc tggaggtttg aatcatcttg aaaactttcc aatcttgtct agttaccact 7680gcagagacac taaggaattt accagaaaaa gatatttgat acaagtgatt taagaaatct 7740caacatttcc tgaggccgta tcactgggca accagtgatg aaaactatga atgaattgca 7800cacctggaag attttttaag ctaatgacag tttcttcaaa gatgtcaatt atttgccttg 7860gaaattttat aaattgcatt tctatgcaca tcggcctcta gtgcttacca ctcggtttat 7920tattcataat ctgcaattca ataaaggctt tgtgttttca tttatcttca aaa 797364511DNAHomo sapiens 6gggcgcgcgg gccccgcggg ggcggcgcgt ggatggatcc gcgcgtggcc tggatccagc 60ccgagcagaa ggggccggcc aatgccctgt ggatgcagat ctgggagacc tcgcagggcg 120tgggccgcgg cggctcgggc ttcgcgtcct atttctgcct caactcgccg gcgctggaca 180cggcggccgc ggcgggggcg gccgggcggg gcagtggcgg cctgggcccc gcgctgcccg 240ccgcgtcgcc cccgccgccc ggccccaccg cgcccgccgc gctgcccccc gcgctgctga 300cggcgctggg gcccgcggcc gagggcgcgc ggcgcttgca caagtcgccg tcgctgtcgt 360cctcgtcgtc gtcctcctcg tccaacgcgg agtcgggcac cgagagcccc ggctgctcgt 420cgtcgtcctc cagcagcgcc tcgctgggcc ggccgggcgg cggccgcggc ggcgccttct 480tcaacttcgc cgacggcgcg cccagcgccc ctggcacagc caacgggcac cccgggccgc 540gcggccccgc gcccgccggc tccccgtcgc agcaccagtt ccacccgggt

cgccggaaac 600gcgagaacaa ggccagcacc tacggcctca actacctgct gtccggcagc cgcgcggccg 660ctctcagcgg agggggcggc cccggggccc aggcgccgcg gcccggcacc ccgtggaaga 720gccgcgcgta cagcccgggc atccagggac tacatgagga aataattgac ttttataact 780tcatgtcccc ttgtcctgaa gaagcagcta tgagaagaga ggtggtgaaa cggatcgaaa 840ctgtggtgaa agacctttgg ccgacggctg atgtacagat atttggcagc tttagtacag 900gtctttatct tccaactagc gacatagacc tggtggtctt cgggaaatgg gagcgtcctc 960ctttacagct gctggagcaa gccctgcgga agcacaacgt ggctgagccg tgttccatca 1020aagtccttga caaggctacg gtaccaataa taaagctcac agatcaggag actgaagtga 1080aagttgacat cagctttaac atggagacgg gcgtccgggc agcggagttc atcaagaatt 1140acatgaagaa atattcattg ctgccttact tgattttagt attgaaacag ttccttctgc 1200agagggacct gaatgaagtt tttacaggtg gaattagctc atacagccta attttaatgg 1260ccattagctt tctacagttg catccaagaa ttgatgcccg gagagctgat gaaaaccttg 1320gaatgcttct tgtagaattt tttgaactct atgggagaaa ttttaattac ttgaaaaccg 1380gtattagaat caaagaagga ggtgcctata tcgccaaaga ggagatcatg aaagccatga 1440ccagcgggta cagaccgtcg atgctgtgca ttgaggaccc cctgctgcca gggaatgacg 1500ttggccggag ctcctatggc gccatgcagg tgaagcaggt cttcgattat gcctacatag 1560tgctcagcca tgctgtgtca ccgctggcca ggtcctatcc aaacagagac gccgaaagta 1620ctttaggaag aatcatcaaa gtaactcagg aggtgattga ctaccggagg tggatcaaag 1680agaagtgggg cagcaaagcc cacccgtcgc caggcatgga cagcaggatc aagatcaaag 1740agcgaatagc cacatgcaat ggggagcaga cgcagaaccg agagcccgag tctccctatg 1800gccagcgctt gactttgtcg ctgtccagcc cccagctcct gtcttcaggc tcctcggcct 1860cttctgtgtc ttcactttct gggagtgacg ttgattcaga cacaccgccc tgcacaacgc 1920ccagtgttta ccagttcagt ctgcaagcgc cagctcctct catggccggc ttacccaccg 1980ccttgccaat gcccagtggc aaacctcagc ccaccacttc cagaacactg atcatgacaa 2040ccaacaatca gaccaggttt actatacctc caccgaccct aggggttgct cctgttcctt 2100gcagacaagc tggtgtagaa ggaactgcgt ctttgaaagc cgtccaccac atgtcttccc 2160cggccattcc ctcagcgtcc cccaacccgc tctcgagccc tcatctgtat cataagcagc 2220acaacggcat gaaactgtcc atgaagggct ctcacggcca cacccaaggc ggcggctaca 2280gctctgtggg tagcggaggt gtgcggcccc ctgtgggcaa caggggacac caccagtata 2340accgcaccgg ctggaggagg aaaaaacaca cacacacacg ggacagtctg cccgtgagcc 2400tcagcagata atggctcctg gctgcgtcag cctcccccac ccctctgcag actgccccgc 2460ggcctcggcc accggcaggg gaaccgagac cagcaccccg cacgtcagcc gggctcgcgg 2520cacgcccgcc gctgatcact ctgcatgttt cttcgtgtgg tggtcgcgtc catcttcaag 2580aacagctcgt tgtgctcatc tgtgaagcct tattaaacgt ggacgttgtt ttctgccttc 2640ccaggattct tccttcagtg ctgaggcagg tcgggctcag gaactgcagg gacgtgaaca 2700tgcgcttgcg gtttgaggta gccgtgtctg ttccttcgcg gtttgctatt ttcatttcct 2760gttcgtcaaa gcagcagagg agatcaaacc ccgttcgtgt gtctttcctc cacggataag 2820cttgggaggt cattgtttta ctgccctcac attttgtttg aaatttcaga actgtttttc 2880tatgtaaata ttgaaaactt atgatttgtg caataactca gatatttttt atttaatttc 2940ctattttcac ataagttata tttaagggag gagggaattt tttttaaaca agcttaggtc 3000ctttcccgag ctgcattttc taagttgggt catcgtgtcg gctggttgtc tgacgagcat 3060cgttacaaac accatgatga ggggtttggg gttttatttt gatgtctttt cttttggtcg 3120gaagtgagtg aaggagccag gtcgccctga aggttttcca aagggcttgg ctccagagcc 3180acctggcaga ctgcccgtgg ccctgctgtc gggccccagg ccgttgtcct gctctgacca 3240cagagtttta atgttttggt tttcacttct tttaaactgg acaacaaatc cagcatttca 3300agtgccagaa gtataacttt ctaaggagag aagggttgtc acattataaa atctttagga 3360aaatgtgaac tggaaaacgc ttcggtcagt tttagtgaca tagcctgtga tgatgggtct 3420ggtgactatt attgcggacc gtggtaccca gttttaggaa tgtggagaaa ggaattctgt 3480tgattccgtt gaggaatctg tagcgtatgc attcgttctg ttaagagcaa atctaggaga 3540agtgcttcag ctgcccagtg cgccgtgggg agtgttttaa cggatcgtgt cgcaggagag 3600cacagcccag cgttggggcc gggaccgctg gcgcccgacg tcggaagcat acaggtatac 3660tatgcaagtg tattctgcca caacaaccac tgtctttgtt accttttttt gaacaagaat 3720atatccatcc tgcctaaccc tgagtttttg gagcaccaca gttgtcctgg gagttggttg 3780catcttgtag gccatctgac ttcctgtttt taaaacgggg gtctggtctt gctaaacact 3840acaggtaggt tggtctttga agtccactag tggagaatgt caagacaaga tacttattac 3900catgacatct gatgcatgtg cagcagtggg gagttctaga ttgatctctg aatgtgatcg 3960acgcccagca aggacaagct ttaaaatgtc tgcggtctgc ccttttgaag caggactggc 4020tcactctgtc attgggagct gtcagctgcg actgcaggtt ctctaggagg cattccagaa 4080tagagtagca cactgtgtct gcagttctcg atgaccgaaa gttatcaaaa atatttaaaa 4140tatttaaatt gtgaacctat tgataaagaa tatttataaa aactgatctg taggcctgta 4200ctaatctcta cgcattagca atattgactg taaacccaca ttaaggaaac cactacgggt 4260ctggcagtgc gtgtcccgtg gggtgtgcat tttaaaactc gattcataga cacaggtacc 4320atgttccatt tccgtcatgg tgaagcaaat gaattggcct ggctaccact gtggtcgcgt 4380gctacaggtt tgacaaaaag atatcatgtt tcgatttttt tgtgtgtgga caacaatatg 4440gaagctaaaa ttgacatatt tttatgtaaa gtttttctat tctttgattt ttaataaact 4500ttggaaacca g 45117474PRTHomo sapiens 7Thr Tyr Gly Leu Asn Tyr Ser Leu Leu Gln Pro Ser Gly Gly Arg Ala1 5 10 15Ala Gly Gly Gly Arg Ala Asp Gly Gly Gly Val Val Tyr Ser Gly Thr 20 25 30Pro Trp Lys Arg Arg Asn Tyr Asn Gln Gly Val Val Gly Leu His Glu 35 40 45Glu Ile Ser Asp Phe Tyr Glu Tyr Met Ser Pro Arg Pro Glu Glu Glu 50 55 60Lys Met Arg Met Glu Val Val Asn Arg Ile Glu Ser Val Ile Lys Glu65 70 75 80Leu Trp Pro Ser Ala Asp Val Gln Ile Phe Gly Ser Phe Lys Thr Gly 85 90 95Leu Tyr Leu Pro Thr Ser Asp Ile Asp Leu Val Val Phe Gly Lys Trp 100 105 110Glu Asn Leu Pro Leu Trp Thr Leu Glu Glu Ala Leu Arg Lys His Lys 115 120 125Val Ala Asp Glu Asp Ser Val Lys Val Leu Asp Lys Ala Thr Val Pro 130 135 140Ile Ile Lys Leu Thr Asp Ser Phe Thr Glu Val Lys Val Asp Ile Ser145 150 155 160Phe Asn Val Gln Asn Gly Val Arg Ala Ala Asp Leu Ile Lys Asp Phe 165 170 175Thr Lys Lys Tyr Pro Val Leu Pro Tyr Leu Val Leu Val Leu Lys Gln 180 185 190Phe Leu Leu Gln Arg Asp Leu Asn Glu Val Phe Thr Gly Gly Ile Gly 195 200 205Ser Tyr Ser Leu Phe Leu Met Ala Val Ser Phe Leu Gln Leu His Pro 210 215 220Arg Glu Asp Ala Cys Ile Pro Asn Thr Asn Tyr Gly Val Leu Leu Ile225 230 235 240Glu Phe Phe Glu Leu Tyr Gly Arg His Phe Asn Tyr Leu Lys Thr Gly 245 250 255Ile Arg Ile Lys Asp Gly Gly Ser Tyr Val Ala Lys Asp Glu Val Gln 260 265 270Lys Asn Met Leu Asp Gly Tyr Arg Pro Ser Met Leu Tyr Ile Glu Asp 275 280 285Pro Leu Gln Pro Gly Asn Asp Val Gly Arg Ser Ser Tyr Gly Ala Met 290 295 300Gln Val Lys Gln Ala Phe Asp Tyr Ala Tyr Val Val Leu Ser His Ala305 310 315 320Val Ser Pro Ile Ala Lys Tyr Tyr Pro Asn Asn Glu Thr Glu Ser Ile 325 330 335Leu Gly Arg Ile Ile Arg Val Thr Asp Glu Val Ala Thr Tyr Arg Asp 340 345 350Trp Ile Ser Lys Gln Trp Gly Leu Lys Asn Arg Pro Glu Pro Ser Cys 355 360 365Asn Gly Asn Gly Val Thr Leu Ile Val Asp Thr Gln Gln Leu Asp Lys 370 375 380Cys Asn Asn Asn Leu Ser Glu Glu Asn Glu Ala Leu Gly Lys Cys Arg385 390 395 400Ser Lys Thr Ser Glu Ser Leu Ser Lys His Ser Ser Asn Ser Ser Ser 405 410 415Gly Pro Val Ser Ser Ser Ser Ala Thr Gln Ser Ser Ser Ser Asp Val 420 425 430Asp Ser Asp Ala Thr Pro Cys Lys Thr Pro Lys Gln Leu Leu Cys Arg 435 440 445Pro Ser Thr Gly Asn Arg Val Gly Ser Gln Asp Val Ser Leu Glu Ser 450 455 460Ser Gln Ala Val Gly Lys Met Gln Ser Thr465 4708419PRTHomo sapiens 8Leu Leu Gln Pro Ser Gly Gly Arg Ala Ala Gly Gly Gly Arg Ala Asp1 5 10 15Gly Gly Gly Val Val Tyr Ser Gly Thr Pro Trp Lys Arg Arg Asn Tyr 20 25 30Asn Gln Gly Val Val Gly Leu His Glu Glu Ile Ser Asp Phe Tyr Glu 35 40 45Tyr Met Ser Pro Arg Pro Glu Glu Glu Lys Met Arg Met Glu Val Val 50 55 60Asn Arg Ile Glu Ser Val Ile Lys Glu Leu Trp Pro Ser Ala Asp Val65 70 75 80Gln Ile Phe Gly Ser Phe Lys Thr Gly Leu Tyr Leu Pro Thr Ser Asp 85 90 95Ile Asp Leu Val Val Phe Gly Lys Trp Glu Asn Leu Pro Leu Trp Thr 100 105 110Leu Glu Glu Ala Leu Arg Lys His Lys Val Ala Asp Glu Asp Ser Val 115 120 125Lys Val Leu Asp Lys Ala Thr Val Pro Ile Ile Lys Leu Thr Asp Ser 130 135 140Phe Thr Glu Val Lys Val Asp Ile Ser Phe Asn Val Gln Asn Gly Val145 150 155 160Arg Ala Ala Asp Leu Ile Lys Asp Phe Thr Lys Lys Tyr Pro Val Leu 165 170 175Pro Tyr Leu Val Leu Val Leu Lys Gln Phe Leu Leu Gln Arg Asp Leu 180 185 190Asn Glu Val Phe Thr Gly Gly Ile Gly Ser Tyr Ser Leu Phe Leu Met 195 200 205Ala Val Ser Phe Leu Gln Leu His Pro Arg Glu Asp Ala Cys Ile Pro 210 215 220Asn Thr Asn Tyr Gly Val Leu Leu Ile Glu Phe Phe Glu Leu Tyr Gly225 230 235 240Arg His Phe Asn Tyr Leu Lys Thr Gly Ile Arg Ile Lys Asp Gly Gly 245 250 255Ser Tyr Val Ala Lys Asp Glu Val Gln Lys Asn Met Leu Asp Gly Tyr 260 265 270Arg Pro Ser Met Leu Tyr Ile Glu Asp Pro Leu Gln Pro Gly Asn Asp 275 280 285Val Gly Arg Ser Ser Tyr Gly Ala Met Gln Val Lys Gln Ala Phe Asp 290 295 300Tyr Ala Tyr Val Val Leu Ser His Ala Val Ser Pro Ile Ala Lys Tyr305 310 315 320Tyr Pro Asn Asn Glu Thr Glu Ser Ile Leu Gly Arg Ile Ile Arg Val 325 330 335Thr Asp Glu Val Ala Thr Tyr Arg Asp Trp Ile Ser Lys Gln Trp Gly 340 345 350Leu Lys Asn Arg Pro Glu Pro Ser Cys Asn Gly Pro Val Ser Ser Ser 355 360 365Ser Ala Thr Gln Ser Ser Ser Ser Asp Val Asp Ser Asp Ala Thr Pro 370 375 380Cys Lys Thr Pro Lys Gln Leu Leu Cys Arg Pro Ser Thr Gly Asn Arg385 390 395 400Val Gly Ser Gln Asp Val Ser Leu Glu Ser Ser Gln Ala Val Gly Lys 405 410 415Met Gln Ser9320PRTHomo sapiens 9Tyr Ser Pro Gly Ile Gln Gly Leu His Glu Glu Ile Ile Asp Phe Tyr1 5 10 15Asn Phe Met Ser Pro Cys Pro Glu Glu Ala Ala Met Arg Arg Glu Val 20 25 30Val Lys Arg Ile Glu Thr Val Val Lys Asp Leu Trp Pro Thr Ala Asp 35 40 45Val Gln Ile Phe Gly Ser Phe Ser Thr Gly Leu Tyr Leu Pro Thr Ser 50 55 60Asp Ile Asp Leu Val Val Phe Gly Lys Trp Glu Arg Pro Pro Leu Gln65 70 75 80Leu Leu Glu Gln Ala Leu Arg Lys His Asn Val Ala Glu Pro Cys Ser 85 90 95Ile Lys Val Leu Asp Lys Ala Thr Val Pro Ile Ile Lys Leu Thr Asp 100 105 110Gln Glu Thr Glu Val Lys Val Asp Ile Ser Phe Asn Met Glu Thr Gly 115 120 125Val Arg Ala Ala Glu Phe Ile Lys Asn Tyr Met Lys Lys Tyr Ser Leu 130 135 140Leu Pro Tyr Leu Ile Leu Val Leu Lys Gln Phe Leu Leu Gln Arg Asp145 150 155 160Leu Asn Glu Val Phe Thr Gly Gly Ile Ser Ser Tyr Ser Leu Ile Leu 165 170 175Met Ala Ile Ser Phe Leu Gln Leu His Pro Arg Ile Asp Ala Arg Arg 180 185 190Ala Asp Glu Asn Leu Gly Met Leu Leu Val Glu Phe Phe Glu Leu Tyr 195 200 205Gly Arg Asn Phe Asn Tyr Leu Lys Thr Gly Ile Arg Ile Lys Glu Gly 210 215 220Gly Ala Tyr Ile Ala Lys Glu Glu Ile Met Lys Ala Met Thr Ser Gly225 230 235 240Tyr Arg Pro Ser Met Leu Cys Ile Glu Asp Pro Leu Leu Pro Gly Asn 245 250 255Asp Val Gly Arg Ser Ser Tyr Gly Ala Met Gln Val Lys Gln Val Phe 260 265 270Asp Tyr Ala Tyr Ile Val Leu Ser His Ala Val Ser Pro Leu Ala Arg 275 280 285Ser Tyr Pro Asn Arg Asp Ala Glu Ser Thr Leu Gly Arg Ile Ile Lys 290 295 300Val Thr Gln Glu Val Ile Asp Tyr Arg Arg Trp Ile Lys Glu Lys Trp305 310 315 3201019RNAartificialHuman PAPD5 target RNA 10caucaaugcu uuauaucga 191119RNAartificialHuman PAPD5 target RNA 11ggacgacacu ucaauuauu 191219RNAartificialHuman PAPD5 RNA target sequence 12gauaaaggau ggugguuca 191319RNAartificialhuman PAPD5 Target mRNA sequence 13gaauagaccu gagccuuca 191419RNAartificialhuman PAPD7 target mRNA sequence 14ggagugacgu ugauucaga 191519RNAartificialhuman PAPD7 target mRNA sequence 15cggaguucau caagaauua 191619RNAartificialhuman PAPD7 target RNA sequence 16cggaguucau caagaauua 191719RNAartificialhuman PAPD7 target mRNA sequence 17gcgaauagcc acaugcaau 191819DNAartificialPrimer 18ctgtgccttg ggtggcttt 191924DNAArtificialprimer 19aaggaaagaa gtcagaaggc aaaa 242025DNAartificialprobe 20ttctttataa gggtcgatgt ccatg 25

Claims

1-16. (canceled)

17. An inhibitor of PAPD5 and/or PAPD7 for use in treating a HBV infection, wherein said inhibitor is a. a small molecule that binds to PAPD5 and/or PAPD7; or b. an antibody that specifically binds to PAPD5 and/or PAPD7.

18. The inhibitor according to claim 17, which a. binds to PAPD5 and/or PAPD7 polypeptide; and/or b. inhibits expression and/or activity of PAPD5 and/or PAPD7.

19. The inhibitor according to claim 17, wherein the inhibitor reduces secretion of HBsAg and HBeAg.

20. The inhibitor according to claim 17, wherein the inhibitor inhibits development of chronic HBV infection and/or reduces the infectiousness of a HBV infected person.

21. The inhibitor according to claim 17, wherein the inhibitor is the compound of formula (II): ##STR00020## wherein R.sup.1 is C.sub.1-6alkyl, C.sub.3-7cycloalkyl, haloC.sub.1-6alkyl, hydroxyC.sub.1-6alkyl, nitroC.sub.1-6alkyl, C.sub.1-6alkoxycarbonylC.sub.1-6alkyl, carboxyC.sub.1-6alkyl, di(C.sub.1-6alkoxycarbonyl)methylenyl, cyanoC.sub.1-6alkyl, C.sub.3-7cycloalkylC.sub.1-6alkyl, phenylC.sub.1-6alkyl, C.sub.1-6alkylsufanylC.sub.1-6alkyl, C.sub.1-6alkylsufonylC.sub.1-6alkyl, aminoC.sub.1-6alkyl, C.sub.1-6alkylcarbonylaminoC.sub.1-6alkyl, C.sub.1-6alkylsufonylaminoC.sub.1-6alkyl, C.sub.1-6alkoxycarbonyl aminoC.sub.1-6alkyl, aminocarbonylC.sub.1-6alkyl, diC.sub.1-6alkylaminocarbonylC.sub.1-6alkyl, monocyclic heterocycloalkylC.sub.1-6alkyl or imidazolylC.sub.1-6alkyl; R.sup.2 is aryl or heteroaryl, said aryl or heteroaryl being unsubstituted, or substituted by one, two, three or four substituents independently selected from C.sub.1-6alkyl, C.sub.3-7cycloalkyl, halogen, haloC.sub.1-6alkyl, cyano, nitro, hydroxy, haloC.sub.1-6alkoxy, --O--C.sub.xH.sub.2x--R.sup.3, --O--C.sub.yH.sub.2--NHR.sup.6, --NR.sup.9R.sup.10, --SO.sub.2--R.sup.11, --SO.sub.2--NR.sup.12R.sup.13, carboxy, C.sub.1-6alkoxycarbonyl, --C(.dbd.O)--NR.sup.12R.sup.13, aryl, heteroaryl, monocyclic heterocycloalkyl and --O-monocyclic heterocycloalkyl; wherein monocyclic heterocycloalkyl is unsubstituted or substituted by C.sub.1-6alkyl, C.sub.3-7cycloalkyl, C.sub.1-6alkylcarbonyl, C.sub.1-6alkylsufonyl or C.sub.1-6alkoxycarbonyl; R.sup.3 is hydrogen; C.sub.3-7cycloalkyl; haloC.sub.3-7cycloalkyl; hydroxy; hydroxyC.sub.1-6alkylC.sub.3-7cycloalkyl; C.sub.1-6alkoxy; monocyclic heterocycloalkyl; monocyclic heterocycloalkyl substituted by C.sub.1-6alkyl, C.sub.1-6alkylcarbonyl, C.sub.1-6alkylsufonyl, C.sub.3-7cycloalkyl or C.sub.1-6alkoxycarbonyl; --C(.dbd.O)--R.sup.4; C.sub.1-6alkylsulfinyl; --SO.sub.2--R.sup.5; --C(NHR.sup.7)--C(.dbd.O)--R.sup.8; carboxyC.sub.1-6alkoxy or aminocarbonylC.sub.1-6alkoxy; wherein R.sup.4 is hydroxy, C.sub.1-6alkoxy, amino, C.sub.1-6alkylamino, diC.sub.1-6alkylamino, tetrahydrofuranylamino, pyrrolidinyl or morpholinyl; R.sup.5 is C.sub.1-6alkyl, C.sub.3-7cycloalkyl, hydroxy, amino, C.sub.1-6alkylamino or diC.sub.1-6alkylamino; R.sup.7 is hydrogen or C.sub.1-6alkoxycarbonyl; R.sup.8 is hydroxy or C.sub.1-6alkoxy; R.sup.6 is hydrogen, C.sub.1-6alkylcarbonyl, haloC.sub.1-6alkylcarbonyl, C.sub.1-6alkoxycarbonyl, C.sub.1-6alkylsulfonyl, C.sub.3-7cycloalkylsulfonyl or C.sub.1-6alkoxyC.sub.1-6alkylsulfonyl; R.sup.9 and R.sup.10 are independently selected from hydrogen, C.sub.1-6alkyl, C.sub.3-7cycloalkyl, C.sub.1-6alkylcarbonyl, C.sub.1-6alkylsulfonyl, C.sub.3-7cycloalkylcarbonyl and C.sub.3-7cycloalkylsulfonyl; or R.sup.9 and R.sup.10 together with the nitrogen to which they are attached form monocyclic heterocycloalkyl; R.sup.11 is C.sub.1-6alkyl, haloC.sub.1-6alkyl, C.sub.3-7cycloalkyl, haloC.sub.3-7cycloalkyl, hydroxyC.sub.1-6alkyl, C.sub.1-6alkoxyC.sub.1-6alkyl, haloC.sub.1-6alkoxyC.sub.1-6alkyl, C.sub.3-7cycloalkylC.sub.1-6alkyl, aminoC.sub.1-6alkyl, C.sub.1-6alkylaminoC.sub.1-6alkyl, diC.sub.1-6alkylaminoC.sub.1-6alkyl, C.sub.1-6alkylcarbonylaminoC.sub.1-6alkyl, C.sub.1-6alkylsulfonylaminoC.sub.1-6alkyl, C.sub.1-6alkoxycarbonylaminoC.sub.1-6alkyl, C.sub.1-6alkylsulfenylC.sub.1-6alkyl, C.sub.1-6alkylsulfanylC.sub.1-6alkyl or C.sub.1-6alkylsulfonylC.sub.1-6alkyl; R.sup.12 and R.sup.13 are independently selected from hydrogen, C.sub.1-6alkyl, C.sub.1-6alkoxyC.sub.1-6alkyl, haloC.sub.1-6alkyl, C.sub.3-7cycloalkyl and haloC.sub.3-7cycloalkyl; or R.sup.12 and R.sup.13 together with the nitrogen to which they are attached form monocyclic heterocycloalkyl; x is 1, 2, 3, 4, 5, 6, 7 or 8; y is 1, 2, 3, 4, 5, 6, 7 or 8; U, W and Z are independently selected from CH and N; one of X and Y is N, and the other one is CH or N.

22. The inhibitor for the use according to claim 17, wherein the inhibitor is an antibody that specifically binds to the amino acid stretch of any one of SEQ ID NOs: 7, 8 or 9.

23. A pharmaceutical composition for use in the treatment of a HBV infection, wherein the pharmaceutical composition comprises a. the inhibitor for the use according to claim 17; and b. a pharmaceutically acceptable carrier.

24. A method for monitoring the therapeutic success during the treatment of a HBV infection, wherein the method comprises: a. analyzing in a sample obtained from a test subject the amount and/or activity of PAPD5 and/or PAPD7; b. comparing said amount and/or activity with reference data corresponding to the amount and/or activity of PAPD5 and/or PAPD7 of at least one reference subject; and c. predicting therapeutic success based on the comparison step (b).

25. A pharmaceutical composition for use in the treatment of a HBV infection, wherein the pharmaceutical composition comprises: a. the inhibitor according to claim 21; and b. a pharmaceutically acceptable carrier.

26. A pharmaceutical composition for use in the treatment of a HBV infection, wherein the pharmaceutical composition comprises: a. the inhibitor according to claim 21; and b. a pharmaceutically acceptable carrier.

Images