AU2006200519A1 - New compound - Google Patents

Description

Regulation 3.2

AUSTRALIA

Patents Act 1990 COMPLETE SPECIFICATION FOR A STANDARD PATENT

(ORIGINAL)

Name of Applicant: Actual Inventors: Address for Service: Invention Title: Astellas Pharma Inc. of 3-11, Nihonbashi-Honcho 2-chome, Chuo-ku, Tokyo 103-8411, Japan Eisaku TUJII, Katsuyuki MAKI, Yukiko YAMAGISHI, Seiji YOSHIMURA, Kazuhiko OSODA, Kazuo NAKAI and Hiroshi

SASAKI

DAVIES COLLISON CAVE, Patent Attorneys, 1 Little Collins Street, Melbourne 3000, Victoria, Australia "New compound" Details of Associated Provisional Application Nos: 2005901287 filed 16 March 2005 2005901288 filed 16 March 2005 The following statement is a full description of this invention, including the best method of performing it known to us:

DESCRIPTION

NEW COMPOUND TECHNICAL FIELD The present invention relates to a new compound. More particularly, the present invention relates to a new cyclic peptide compound or a salt thereof having inhibitory activity against the RNA replication of hepatitis C virus (hereafter referred to as HCV) replicon. In particular, the present invention relates to a new peptide compound or a salt thereof, to a process for preparation thereof, to a pharmaceutical composition comprising the new cyclic peptide compound or a salt thereof, and to a method for the prophylactic and/or therapeutic treatment of hepatitis C in a human being or animal.

BACKGROUND ART The estimated number of HCV carriers is about 170 million worldwide (about and about 1.5 million in Japan.

Even in the combination therapy of using interferon (hereafter referred to as IFN) and ribavirin (Virazole), available as a first option for treatment, its effectiveness is 40% for all types of HCV. Furthermore, its effectiveness is only 15 to 20% for IFN-resistant virus (genotype Ib), particularly abundantly found in Japan. On the other hand, the combination therapy has side effects frequently. It is thus difficult to get rid of the virus completely by using currently available treatment methods. In the case when chronic hepatitis cannot be cured completely, the hepatitis will securely develop into cirrhosis hepatitis or hepatocellular carcinoma In Europe and the United States, hepatitis C has been a major indication for liver transplant. However, the redevelopment of HCV occurs frequently even in transplanted livers. For these reasons, the needs for new agents being improved in both effectiveness and safety, having higher antiviral effects and capable of inhibiting hepatitis C are very strong in society.

HCV is a virus having plus-strand RNA as a gene and is classified into Flaviviridae in accordance with the analysis of the base sequence of the gene. According to Fields Virology fourth edition, D. Knipe et al ed., Philadelphia, Lippincott Williams Wilkins 2001, 1127-1161, although the existence of HCV was anticipated in 1970s, the discovery of HCV was very difficult. HCV was called non-A non-B hepatitis virus for many years. In 1989, according to Choo Q-L et al., Science 244, 359-362 (1989), part of the gene of this virus was cloned from the serum of an infected laboratory animal, and its cDNA sequence was identified and confirmed, whereby the virus was named "HCV".

DISCLOSURE OF THE INVENTION Cyclosporin A is used as an immunosuppressant for organ transplant. M. Thali et al., Nature 372, 363-365 (1994) reported that Cyclosporin A had anti-HIV activity by inhibiting the interaction between Cyclosporin A and the virus particle forming protein of Human Immunodeficiency Virus Type 1 (HIV-1). Furthermore, K. Inoue et al., 6th International Symposium on Hepatitis C and Related Virus, 3-6 June (2000) Bethesda, MD, USA reported that Cyclosporin A had an anti-HCV activity. However, reports for supporting this finding are not presented by other groups up until now.

M. Berenguer et al., J. Hepatol 32, 673-684 (2000) reported that the clinical use of Cyclosporin A serving as an immunosuppressant caused HCV to multiply in transplant patients.

Hence, an anti-hepatitis C agent improved in the activity, transition in blood, selectivity and the side effects, for example, in comparison with Cyclosporin A, has been demanded because of the above-mentioned reasons.

The object cyclic peptide compound in the present invention is a new compound, and can be represented by the following general formula R1 0R HN N 0 0 0 N o N

(I)

o H 0 H o N N N N N 0 wherein

R

1 is lower alkyl which may be substituted with one or more suitable substituent(s) or lower alkenyl which may be substituted with one or more suitable substituent(s), higher alkyl which may be substituted with one or more suitable substituent(s) or heterocyclic group which may be substituted with one or more suitable substituent(s), and

R

2 is hydrogen, hydroxy, oxo, lower alkanoyloxy, aryloxy, lower alkoxycarbonyloxy, aryloxythiocarbonyloxy, or carbamoyloxy which may be substituted with one or more suitable substituent(s), or R1 and R2 are linked together to form heterocyclic ring which may be substituted with one or more suitable substituent(s);

R

X is C-R 4 or C

R

3 is hydrogen, and

R

4 is hydrogen or lower alkyl substituted with one or more suitable substituent(s) selected from the group consisting of hydroxy, oxo, acetyloxy, aryloxythiocarbonyloxy, aryloxycarbonyloxy which may be substituted with one or more suitable substituent(s), lower alkoxycarbonyloxy, carbamoyloxy which may be substituted with one or more suitable substituent(s) and heterocycliccarbonyloxy which may be substituted with one or more suitable substituent(s), or

R

3 and R 4 are linked together to form heterocyclic ring which may be substituted with one or more suitable substituent(s);

R

5 is hydrogen, hydroxy, oxo, lower alkoxy, lower alkanoyloxy which may be substituted with one or more suitable substituent(s), aroyloxy, amino, lower alkoxyimino or aryloxyimino; and is single bond or double bond, Providing that When R 1 is 1-propenyl and R 2 is hydroxy or acetyloxy, then R 5 is not hydroxy or acetyloxy, or a salt thereof.

The compound or a salt thereof in the present invention can be prepared by the processes as illustrated in the following reaction schemes.

to be continued on the next page Process 1 Nucleophilic substitution reaction 0

\N

^o (Ia) or a salt thereof (Ib) or a salt thereof Process 2 Elimination reaction (Ic) or a salt thereof (Id) or a salt thereof Process 3 Nucleophilic substitution reaction (Ie) or a salt thereof (If) or a salt thereof Process 4 Nucleophilic substitution reaction (Ig) or a salt thereof (Ih) or a salt thereof Process Nucleophilic substitution reaction (Ii) or a salt thereof (Ii) or a salt thereof Process 6

R

6 R 7

H

3 CO OCH 3 Cyclization reaction (Ik) or a salt thereof (Ii) or a salt thereof' Process 7 Cyclization reaction (Im) or a salt thereof (In) or a salt thereof Process 8 Elimination reaction or a salt thereof (1p) or a salt thereof Process 9 Reduction (Iq) or a salt thereof (Ir) or a salt thereof Process Oxidation Reduction

(II)

or a salt thereof (Is) or a salt thereof Process 11 Oxidation (It) or a salt thereof (Iu) or a salt thereof Process 12 Metathesis

(II)

or a salt thereof (Iv) or a salt thereof Process 13 Acylation (1w) or a salt thereof (lx) or a salt thereof Process 14 (1y) or a salt thereof (Iz) or a salt thereof Process Oxidation (Iq) or a salt thereof (Iaa) or a salt thereof Process 16 (laa) or a salt thereof (lab) or a salt thereof Process 17 Cyclization

CN

'N

(Iac) or a salt thereof (lad) or a salt thereof Process 18 Halogenat ion (Iaa) or a salt thereof (Iae) or a salt thereof Process 19 Esterification '(Iq) or a salt thereof (laf) or a salt thereof Process Acylation (lag) or a salt thereof (Iah) or a salt thereof Process 21 Dehydrogenation (It) or a salt thereof (Iai) or a salt thereof Process 22 Metathesis (Iq) or a salt thereof (laj) or a salt thereof Process 23 Rlh

R

2 3 O I Acylation 0 0 a-- 0 NO H 0 H 0 2 Ri N' y N N OR 3

R

0 ON 0 NN 0 0 H N N N Nf NI YR 3

R

00 °O N (Iaj) 0 0 or a salt thereof f H H -N N N N N H 0 01 (lak) or a salt thereof wherein

R

1

R

2

R

3

R

4 and R 5 are as defined above, Rla is lower alkyl substituted with hydroxy or lower alkenyl substituted with hydroxy; RIb is lower alkyl substituted with one or more suitable substituent(s) except hydroxy or lower alkenyl substituted with one or more suitable substituent(s) except hydroxy; RId is lower alkenyl which may be one or more suitable substituent(s),

R

le is lower alkyl which may be one or more suitable substituent(s), RIf is lower alkyl substituted with oxo or lower alkenyl substituted with oxo,

R

1 g is lower alkyl substituted with lower alkoxyimino or aryloxyimino, or lower alkenyl substituted with lower alkoxyimino or aryloxyimino,

R

lh is lower alkyl substituted with one or more hydroxy or lower alkenyl substituted with one or more hydroxy, Rii is lower alkyl substituted with heterocyclic group which may be substituted or lower alkenyl substituted with heterocyclic group which may be substituted,

R

lj is lower alkyl substituted with one or more halogen or lower alkenyl which may be one or more halogen, RIk is lower alkenyl substituted with lower alkoxycarbonyl or lower alkyl substituted with lower alkoxycarbonyl, R11 is lower alkenyl substituted with lower alkanoyloxy or lower alkyl substituted with lower alkanoyloxy, R2a is carbamoyloxy which may be substituted with one or more suitable substituent(s), lower alkoxycarbonyloxy or aryloxythiocarbonyloxy;

R

2 b is hydroxy;

R

1c and R 2 c are linked together to form heterocyclic ring which may be substituted with one or more suitable substituent(s);

R

4 a is lower alkyl substituted with hydroxy,

R

4 b is lower alkyl substituted with a substituent selected from the group consisting of aryloxythiocarbonyloxy, aryloxycarbonyloxy which may be substituted with one or more suitable substituent(s) and lower alkoxycarbonyloxy, R4c is lower alkyl substituted with aryloxycarbonyloxy which may be substituted with one or more suitable substituent(s), R4d is lower alkyl substituted with carbamoyloxy which may be .substituted with one or more suitable substituent(s);

R

4 e is lower alkyl; 4f is lower alkyl substituted with oxo;

R

4 g is hydrogen;

R

5a is lower alkanoyloxy which may be substituted;

R

5 b is amino, lower alkoxyimino or aryloxyimino;

R

5c is hydroxy;

R

5d is lower alkanoyloxy or aryloxy; and

R

6 and R7 are independently hydrogen, lower alkyl or aryl.

The compound (II) can be produced by fermentation of fungus (Stachybotrys chartaum No. 19392: deposit number FERM BP-3364) according to the method described in Japanese Laidopen Patent Application Hei 5-271267, for example.

The processes for the preparation of the object compounds are described below.

Process 1 The object compound (Ib) or a salt thereof can be prepared by subjecting the compound (Ia) or a salt thereof to nucleophilic substitution reaction in the presence of base.

The reaction is usually carried out in a conventional solvent which does not adversely influence the reaction such as an alcohol methanol, ethanol, propanol, isopropanol), tetrahydrofuran, dioxane, diethylether, amide N,N-dimethylformamide, N,N-dimethylacetamide), nitrile acetonitrile), or any other organic solvent. The reaction may be usually carried out under cooling to heating since the reaction temperature is not critical.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 2 The object compound (Id) or a salt thereof can be prepared by subjecting the compound (Ic) or a salt thereof to elimination reaction.

The reaction is usually carried out in a conventional solvent which does not adversely influence the reaction such as an alcohol methanol, ethanol, propanol, isopropanol), tetrahydrofuran, dioxane, diethylether, amide N,N-dimethylformamide, N,N-dimethylacetamide), nitrile acetonitrile), or any other organic solvent. The reaction may be usually carried out under cooling to heating since the reaction temperature is not critical.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 3 The object compound (If) or a salt thereof can be prepared by subjecting the compound (Ie) or a salt thereof to nucleophilic substitution reaction in the presence of base.

This reaction can be carried out in a similar manner to that of the aforementioned Process i.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 4 The object compound (Ih) or a salt thereof can be prepared by subjecting the compound (Ig) or a salt thereof to nucleophilic substitution reaction in the presence of base.

This reaction can be carried out in a similar manner to that of the aforementioned Process i.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process The object compound (Ij) or a salt thereof can be prepared by subjecting the compound (Ii) or a salt thereof to nucleophilic substitution reaction in the presence of base.

This reaction can be carried out in a similar manner to that of the aforementioned Process 1.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 6 The object compound (Ii) or a salt thereof can be prepared by reacting the compound (Ik) or a salt thereof with the compound (II) or a salt thereof to the cyclization reaction.

The reaction is usually carried out in a conventional solvent which does not adversely influence the reaction such as water, methanol, ethanol, propanol, isopropanol, N,Ndimethylformamide, N,N-dimethylacetamide, acetonitrile, diethyl ether, dioxane, tetrahydrofuran, etc., or a mixture thereof. The reaction temperature of this reduction is not critical and the reaction is usually carried out under cooling to warming.

Process 7 The object compound (In) or a salt thereof can be prepared by subjecting the compound (Im) or a salt thereof to cyclization reaction in the presence of base. This reaction can be carried out in a similar manner to that of the aforementioned Process 6, and therefore the reagents to be used and the reaction conditions solvent, reaction temperature, etc.) can be referred to those of the Process 6.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 8 The object compound (Ip) or a salt thereof can be prepared by subjecting the compound (Io) or a salt thereof to elimination reaction. This reaction can be carried out in a similar manner to that of the aforementioned Process 2, and therefore the reagents to be used and the reaction conditions solvent, reaction temperature, etc.) can be referred to those of the Process 2.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 9 The object compound (Ir) or a salt thereof can be prepared by reducing the compound (Iq) or a salt thereof.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process The object compound (Is) or a salt thereof can be prepared by oxidizing the compound (II) with Os04, and then NaIO 4 and then by reducing the compound (II) with BaBH 4 More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 11 The object compound (Iu) or a salt thereof can be prepared by oxidizing the compound (It) or a salt thereof.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 12 The object compound (Iv) or a salt thereof can be prepared by subjecting the compound (II) to a metathesis reaction. More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 13 The object compound (Ix) or a salt thereof can be Sprepared by subjecting the compound (Iw) or a salt thereof'to acylation reaction. More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 14 The object compound (Iz) or a salt thereof can be prepared by reacting the compound (Iy) or a salt thereof with hydroxylamine derivative. More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process The object compound (Iaa) or a salt thereof can be prepared by oxidizing the compound (Iq) or a salt thereof.

This reaction can be carried out in a similar manner to that of Process 11 in the above.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 16 The object compound (Iab) or a salt thereof can be prepared by reacting the compound (Iaa) or a salt thereof with hydroxylamine derivative.

This reaction can be carried out in a similar manner to that of Process 14 in the above.

More specifically, the object compound can be prepared by the'processes described in Examples in the present application or similar processes.

Process 17 The object compound (Iad) or a salt thereof can be prepared by subjecting the compound (Iac) or a salt thereof to cyclization reaction.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 18 The object compound (Iae) or a salt thereof can be prepared by halogenating the compound (Iaa) or a salt thereof.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 19 The object compound (Iaf) or a salt thereof can be prepared by subjecting the compound (Iq) or a salt thereof to the esterification reaction.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process The object compound (Iah) or a salt thereof can be prepared by subjecting the compound (Iag) or a salt thereof to the acylation reaction.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 21 The object compound (Iai) or a salt thereof can be prepared by subjecting the compound (It) or a salt thereof to the dehydrogenation reaction.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 22 The object compound (Iaj) or a salt thereof can be prepared by subjecting the compound (Iq) or a salt thereof to metathesis reaction.

This reaction can be carried out in a similar manner to that of Process 12 in the above.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

Process 23 The object compound (lak) or a salt thereof can be prepared by subjecting the compound (Iaj) or a salt thereof to acylation reaction.

This reaction can be carried out in a similar manner to that of Process 13 in the above.

More specifically, the object compound can be prepared by the processes described in Examples in the present application or similar processes.

The compounds obtained by the above-mentioned processes 1 to 23 can be isolated and purified by a conventional method, such as pulverization, recrystallization, column chromatography, high performance liquid chromatography, reprecipitation and demineralized resin column chromatography.

Suitable salts of the object compound are conventional pharmaceutically acceptable and non-toxic salts, and may be a salt with a base or an acid addition salt, for example, a salt with an inorganic base (such as an alkali metal salt, e.g. sodium salt, potassium salt, etc., an alkaline earth metal salt, e.g. calcium salt, magnesium salt, etc., an ammonium salt), a salt with an organic base (such as an organic amine salt, e.g. triethylamine salt, diisopropyl ethylamine salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N'N'dibenzylethylenediamine salt, etc.), an inorganic acid addition salt (such as hydrochloride, hydrobromide, sulfate, phosphate, etc.), an organic carboxylic acid or sulfonic acid addition salt (such as formate, acetate, trifluoroacetate, maleate, tartrate, gluconate, fumarate, methanesulfonate, benzenesulfonate, toluenesulfonate, etc.), a salt with a basic or acidic amino acid (such as arginine, aspartic acid, glutamic acid, etc.) and the like.

In the above and subsequent descriptions of the present specification, suitable examples and illustrations of the various definitions to be included within the scope of the invention are explained in detail as follows.

The term "lower" is intended to mean 1 to 6 carbon atom(s), unless otherwise indicated.

The term "higher" is intended to mean 7 to 20 carbon atoms, unless otherwise indicated.

Suitable examples of "halogen" may include fluoro, chloro, bromo, iodo, etc.

Suitable examples of "lower alkoxy" may include a straight or branched one, such as methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, tert-butoxy, pentyloxy, tertpentyloxy, neopentyloxy, hexyloxy, isohexyloxy, and the like.

Suitable examples of "higher alkoxy" may include a straight or branched one, such as heptyloxy, octyloxy, dimethyloctyloxy, 3,7-dimethyloctyloxy, nonyloxy, decyloxy, undecyloxy, dodecyloxy, tridecyloxy, tetradecyloxy, hexadecyloxy, heptadecyloxy, octadecyloxy, nonadecyloxy, icosyloxy, and the like, in which a preferable one may be,

(C

7

-C

1 4)alkoxy.

Suitable examples of "lower alkyl" may include a straight or branched one having 1 to 6 carbon atom(s), such as methyl, ethyl, propyl, isopropyl, butyl, isobutyl, secbutyl, tert-butyl, pentyl, tert-pentyl, neo-pentyl, hexyl, isohexyl, and the like.

Suitable examples of "lower alkenyl" may include a straight or branched one having 2 to 6 carbon atoms, such as vinyl, 1- or 2-propenyl, isopropenyl, 1- or 2- or 3-butenyl, isobutenyl, sec-butenyl, tert-butenyl, pentenyl, tertpentenyl, neopentenyl, hexenyl, isohexenyl, and the like.

Suitable examples of "lower alkynyl" may include a straight or branched one having 2 to 6 carbon atoms, such as ethynyl, 1- or 2-propynyl, 1- or 2- or 3-butynyl, pentynyl, hexynyl, and the like.

Suitable examples of "higher alkyl" may include a straight or branched one, such as heptyl, octyl, dimethyloctyl, 3,7-dimethyloctyl, nonyl, decyl, undecyl, dodecyl, tridecyl, tetradecyl, hexadecyl, heptadecyl, octadecyl, nonadecyl, icosyl, and the like, in which a preferable one may be (C 7

-C

14 )alkyl.

Suitable examples of "cyclo(lower)alkyl" may include cyclic alkyl having 3 to 6 carbon atom, such as cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl, and this cyclo(lower) alkyl may be substituted with one or more suitable substituent(s) such as lower alkyl, hydroxy, and the like.

Suitable examples of "aryl" and "ar" moiety may include phenyl which may be substituted with lower alkyl phenyl, mesityl, tolyl, etc.), naphthyl, anthryl, tetrahydronaphthyl, indenyl, tetrahydroindenyl, and the like, and this "aryl" and "ar" moiety may be substituted with one or more suitable substituent(s) such as nitro, optionally substituted heterocyclic group, halogen, lower alkyl, and the like.

Suitable example of "aroyl" moiety in the "aroyloxy" may include benzoyl, toluoyl, naphthoyl, anthrylcarbonyl, and the like.

Suitable examples of "lower alkoxycarbonyloxy" may include a straight or branched one, such as methoxycarbonyloxy, ethoxycarbonyloxy, propoxycarbonyloxy, isopropoxycarbonyloxy, butoxycarbonyloxy, isobutoxycarbonyloxy, sec-butoxycarbonyloxy, tertbutoxycarbonyloxy, pentyloxycarbonyloxy, tertpentyloxycarbonyloxy, neopentyloxycarbonyloxy, hexyloxycarbonyloxy, isohexyloxycarbonyloxy, and the like.

Suitable example of "lower alkanoyl" or "lower alkanoyl" moiety may include C 1

-C

6 alkanoyl, in which the preferred one may be formyl, acetyl, propanoyl, butanoyl, tert-butanoyl, 2-methylpropanoyl, pentanoyl, 2,2dimethylpropanoyl, hexanoyl, and the like.

Suitable example of "lower alkanoyloxy" may include C 1

C

6 alkanoyloxy, in which the preferred one may be formyloxy, acetyloxy, propanoyloxy, butanoyloxy, tert-butanoyloxy, 2methylpropanoyloxy, pentanoyloxy, 2,2-dimethylpropanoyloxy.

hexanoyloxy, and the like, and this lower alkanoyloxy may be substituted with lower alkoxy.

Suitable examples of "heterocyclic group" and "heterocyclic" moiety may include unsaturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing 1 to 4 nitrogen atom(s), for example, pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridyl, dihydropyridyl, pyrimidyl, pyrazinyl, pyridazinyl, triazolyl 4H-1,2,4-triazolyl, 1H-1,2,3triazolyl, 2H-1,2,3-triazolyl, etc.), tetrazolyl 1Htetrazolyl, 2H-tetrazolyl, etc.), azepinyl, etc.; saturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing 1 to 4 nitrogen atom(s), for example, aziridinyl, azetinyl, pyrrolidinyl, imidazolidinyl, piperidyl, piperazinyl, methanopiperazinyl, hexahydroazepinyl, etc.; unsaturated condensed heterocyclic group containing 1 to 4 nitrogen atom(s), for example, indolyl, isoindolyl, indolinyl, indolizinyl, benzimidazolyl, quinolyl, isoquinolyl, indazolyl, benzotriazolyl, tetrahydroquinolyl, tetrahydroindolyl, dihydroindazolyl, etc.; unsaturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing 1 or 2 oxygen atom(s) and 1 to 3 nitrogen atom(s), for example, oxazolyl, isoxazolyl, oxadiazolyl 1,2,4-oxadiazolyl, 1,3,4oxadiazolyl, 1,2,5-oxadiazolyl, etc.), etc.; saturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing 1 or 2 oxygen atom(s) and 1 to 3 nitrogen atom(s), for example, morpholinyl, sydnonyl, etc.; unsaturated condensed heterocyclic group containing 1 or 2 oxygen atom(s) and 1 to 3 nitrogen atom(s), for example, benzoxazolyl, benzoxadiazolyl, etc.; unsaturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing 1 or 2 sulfur atom(s) and 1 to 3 nitrogen atom(s), for example, thiazolyl, isothiazolyl, thiadiazolyl 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl, 1,2,5-thiadiazolyl, etc.), dihydrothiazinyl, etc.; saturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing 1 or 2 sulfur atom(s) and 1 to 3 nitrogen atom(s), for example thiazolidinyl, etc.; unsaturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing 1 or 2 sulfur atom(s), for example, thienyl, dihydrodithiinyl, dihydrodithionyl, etc.; unsaturated condensed heterocyclic group containing 1 or 2 sulfur atom(s) and 1 to 3 nitrogen atom(s), for example, benzothiazolyl, benzothiadiazolyl, imidazothiadiazolyl, etc.; unsaturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing an oxygen atom, for example, furyl etc.; saturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing 1 or 2 oxygen atom(s), for example, oxiranyl, 1,3-dioxolanyl, tetrahydrofuranyl, tetrahydropyranyl, etc unsaturated 3 to 8-membered (more preferably 5 or 6membered) heteromonocyclic group containing an oxygen atom and 1 or 2 sulfur atom(s), for example, dihydrooxathiinyl, etc.; unsaturated condensed heterocyclic group containing 1 or 2 sulfur atom(s), for example benzothienyl, benzodithiinyl, etc.; and unsaturated condensed heterocyclic group containing an oxygen atom and 1 or 2 sulfur atom(s), for example, benzoxathiinyl, etc.; saturated condensed heteromonocyclic group containing 1 to 3 nitrogen atom(s), for example, tetrahydropridopyrrolidinyl, etc.; and the like, this "heterocyclic group" and "heterocyclic" moiety may be substituted with one or more suitable substituent(s) such as lower alkyl, hydroxy, lower alkoxy, aryl, aryl(lower)alkyl, oxo, halogen, lower alkylsulfonyl, optionally substituted cyclo(lower)alkyl, mesyl, and the like.

Suitable example of "suitable substituent(s)" may include optionally substituted aryloxycarbonyloxy, lower alkylcarbamoyloxy, optionally substituted aryl, lower alkyl, aryl(lower)alkyl, nitro, lower alkoxycarbonyl(lower)alkyl, cyano(lower)alkyl, lower alkanoylamino(lower)alkyl, lower alkoxy, heterocyclic(lower)alkyl, optionally substituted cyclo(lower)alkyl, mono- or di-lower alkylamino(lower)alkyl, lower alkynyl(lower)alkyl, lower alkenyl(lower)alkyl, cyclo(lower)alkyl(lower)alkyl, aryl(lower)alkoxy, optionally substituted heterocyclic group, lower alkanoyloxy, lower alkoxyimino, aryloxyimino, lower alkanoylamino, oxo, hydroxy, lower alkylsulfonylamino, heterocycliccarbonyl, N- (lower)alkylamino-N-(lower)alkoxy(lower)alkylamino, heterocyclicthio, lower alkoxy(lower)alkyl, carbamoyl, lower alkoxy(lower)alkyl, halogen, optionally substituted spiroheterocyclic group, lower alkoxycarbonyl, aryl(lower)alkoxycarbonyl, optionally substituted arylthio, in which the preferred one may be hydroxy, oxo, cyclo(C 3 Cs)alkyl, (C 1

-C

4 )alkyl, (C 1

-C

4 )alkanoyloxy, (C 1

-C

4 )alkoxyimino, phenyloxyimino, halogen, (C 1

-C

4 )alkoxy or phenyl.

V. Lohmann et al., Science 285, 110-113 (1999) reported that they prepared human hepatoma cell lines (Huh-7) in which subgenomic HCV RNA molecules were introduced, and found that subgenomic HCV RNA was replicated in the cells at a high rate.

It is thought that the replication mechanism of the subgenomic HCV RNA in these cell lines is extremely similar to the replication of full length HCV RNA genome in hepatic cells infected with HCV. Hence, the method for evaluating the activity of the compound for inhibiting RNA replication in accordance with the present invention is based on the cellular assay method that uses Huh-7 cells in which subgenomic HCV RNA is introduced.

In order to show the usefulness of the compound or a salt thereof in the present invention, a pharmacological test example of a representative compound in the present application is shown as follows.

Test example HCV replicon assay The inhibitory action against the replication of HCV replicon was evaluated by directly determining the amount of replicon RNA, purified from cell lysate with a conventional column method, by using real-time RT-PCR based on Taq-Man chemistry. The assay was carried out with the modified method reported by Lohmann et al., Science 285: 100 (1999) and Takeuchi et al., Gastroenterology 116: 636-642 (1999).

The details thereof are described in the following.

1) Addition of agent to cells x 106 HCV replicon cells in 2 ml of D-MEM medium containing 5% fetal bovine serum and 300 [g/ml of G418 (Kishine et al., B.B.R.C. 293: 993-999 (2002)) were seeded in each well of a 6-well microtiter plate (Corning Inc.). After the plate was incubated at 37 0 C for 16 hours in 5% CO2, the medium was replaced with the above-mentioned medium in which the test compound was dissolved.

2) Extraction of RNA from cells After cultivation for two more days, total RNA was extracted from the cells according to the protocol of the RNA extraction column RNeasy Mini (Qiagen Inc.). The total RNA amount was determined by measuring the absorbance ratio (260 nm/280 nm) using the DU800 ultraviolet-visible spectroscopic analysis system (Beckman Inc.).

3) Determination of the amount of replicon RNA according to the real-time RT-PCR method The real-time RT-PCR was carried out by adding the appropriate primer set for amplifying the part of the HCV gene sequence and the complementary probe (all of these were produced by Takara Shuzo Co., Ltd.).

The RNA extracted at 2) was diluted with NF water containing RNase inhibitor to 25 ng/Rl, and dispensed to a 384-well PCR plate, 2 tl in each well. As the reaction solution for RT-PCR, TaqMan Ez RT-PCR Core Reagent (Applied Biosystems Inc.) was mixed according to the protocol and added, 8 il in each well.

RT-PCR was carried out by using the ABI PRISM 7900HT sequence detection system (Applied Biosystems Inc.), and the gene copy number of the HCV replicon RNA in each cell was determined. Various concentrations of HCV RNA standard solutions, 10 times serially diluted were used for the calibration curve creation. The reaction for negative control was carried out without RNA.

4) Measurement of intrinsic control RNA The amount of Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA as intrinsic control was determined according to the TagMan GAPDH control reagent protocol purchased from Applied Biosystems Inc.

Estimation of cellular toxicity HCV replicon cells were cultured in the same conditions as those of the evaluation system of the replicon inhibitory activity. The cells were dispersed by using 0.05% trypsin mMEDTA, and stained with 0.1% ERYTHROCIN-PBS(-). The number of living cells was counted under a microscope.

Test result The copy numbers of the replicon RNA in replicon cell treated at each concentrations of the compound, corrected by the intrinsic control GAPDH values at each point, were employed for the calculation of EC50 value of the each compound, which gave the compound concentration indicating RNA level to the control (no drug group, containing only DMSO). The cellular toxicity was calculated by plotting the drug concentration indicating 50% level to the control (no drug group, containing only DMSO) and obtained as a value.

Test compound HCV replicon replication inhibitory activity:

(M)

Object compound of Example 4 Object compound of Example 26 Object compound of Example 29 Object compound of Example 37 Object compound of Example 39 Object compound of Example 40 From the result of the above-mentioned test example, it is realized that the compound or a salt thereof of the present invention possesses an anti-hepatitis C virus activity.

The anti-HCV agent in the present invention, containing the compound or a salt thereof as an active ingredient, can be used in the form of a pharmaceutical preparation, for example, in a solid, semisolid or liquid form, in admixture with an organic or inorganic carrier or excipient suitable for oral; sublingual; buccal; nasal; respiratory; parenteral (intracutaneous, intraorgan, subcutaneous, intradermal, intramuscular, intra-articular, central-venous, hepaticvenous, peripheral-venous, lymphatic, cardiovascular, arterial, ocular including injection around eye or intravenous drip around eye);.intravenous drip into eyeball, augen structure or augen layer; aural including auditory canal, papillary chamber, external and internal auditory canals, drum membrane, tympanum, internal-auditory including spiralis cochleae ganglion, labyrinth, etc.; intestinal; rectal; vaginal; ureteral; and vesical administration. With respect to intrauterine and perinatal adaptation diseases, parenteral administration is preferable since administration is carried out in maternal blood vessels, or in vacancies, such as maternal organs including uterus, uterine cervix and vagina; fetal embryo, fetus, neonate, and combination tissue; and amnion, umbilical cord, umbilical artery and vein; placenta, and the like. Use of these passages is changed depending on the condition of each patient.

The compound or a salt thereof can be administered independently as a therapeutic agent or may be desired to be used as part of prescribed drugs. The "anti-HCV. agent" in accordance with the present invention can be used in the form of a pharmaceutical preparation, for example, in a solid, semisolid or liquid form, in admixture with at least one or some suitable organic or inorganic carriers or excipients, or other pharmacological therapeutic agents. The active ingredient can be compounded with, for example, usual pharmacologically acceptable and non-toxic carriers in a solid form, such as granules, tablets, pellets, troches, capsules or suppositories; creams; ointments: aerosols; powders for insufflation; in a liquid form, such as solutions, emulsions or suspensions for injection; oral ingestion; eye drops; and any other forms suitable for use. And, if necessary, there may be included in the above preparations auxiliary substances, such as stabilizing, thickening, wetting, hardening and coloring agents; perfumes or buffers; or any other additives used commonly.

The compound or a pharmaceutically acceptable salt thereof is/are included in the pharmaceutical composition in an amount sufficient to produce the desired anti-hepatitis C effect upon the process or condition of diseases.

The combination use of IFN and/or ribavirin with the compound or a salt thereof is effective against hepatitis

C.

For applying the composition to humans, it is preferable to apply it by intravenous, intramuscular, pulmonary, oral administration, eye drop administration or insufflation.

While the dosage of therapeutically effective amount of the compound varies from and also depends upon the age and condition of each individual patient to be treated, in the case of intravenous administration, a daily dose of 0.001-400 mg of the compound per kg weight of human being in the case of intramuscular administration, a daily dose of 0.1-20 mg of the compound per kg weight of human being, in case of oral administration, a daily dose of 0.5-50 mg of the compound per kg weight of human being is generally given for treating or preventing hepatitis C. However, these doses may be required to exceed the limit thereof to obtain therapeutic results.

The amount of the lipopeptide compound or its pharmaceutically acceptable salt contained in the composition for a single unit dosage of the present invention is 0.1 to 400 mg, more preferably 1 to 200 mg, still more preferably to 100 mg, specifically 5, 10, 15, 20, 25, 30, 35, 40, 45, 60, 65, 70, 75, 80, 85, 90, 95 and 100 mg.

An article of manufacture, comprising packaging material and the compound identified in the above contained within said packaging material, wherein said the compound is therapeutically effective for preventing or treating hepatitis C, and wherein said packaging material comprises a label or a written material which indicates that said compound can or should be used for preventing or treating hepatitis C.

A commercial package comprising the pharmaceutical composition containing the compound identified in the above and a written matter associated therewith, wherein the written matter states that the compound can or should be used for preventing or treating hepatitis C.

It is to be noted that the compound or a salt thereof may include one or more stereoisomer(s), such as optical isomer(s) and geometrical isomer(s), due to asymmetric carbon atom(s) and double bond(s), and that all such isomers and the mixture thereof are included within the scope of the present invention.

The compound or a salt thereof may include solvated compound hydrate, ethanolate, etc.).

The compound or a salt thereof may include both the crystal form and non-crystal form.

The compound or a salt thereof may include the prodrug form.

The patent specifications and publications cited herein are incorporated in this specification by reference.

The following Preparations and Examples are given for the purpose of illustrating the present invention. However, the present invention is not limited to these Preparations and Examples.

The Starting Compounds used and the Object Compounds obtained in the following Preparations 1 and 2, and Examples 1 to 80 are given in the table as below, in which the formulas of the starting compounds are in the upper column, and the formulas of the object compounds are in the lower column, respectively.

Abbreviations and Acronyms Abbreviations and Full Name Acronyms AcOEt or EtOAc ethyl acetate AcOH acetic acid BuOH, t-BuOH, etc. butanol, tert-butyl alcohol, etc.

DME 1,2-dimethoxyethane DMF N,N-dimethylformamide DMSO Dimethyl sulfoxide Et3N triethylamine EtOH Ethanol IPE diisopropyl ether MeOH Methanol PrOH, i-PrOH, etc. Propanol, isopropyl alcohol, etc.

TFA trifluoroacetic acid THF tetrahydrofuran EDCI l-ethyl-3-[3'- (dimethylamino)propyl]carbodiimide HOBt or HOBT l-hydroxybenztriazole BSU bis(trimethylsilyl)urea MSA mono(trimethylsilyl)acetamide Pd/C palladium on carbon deg C=degree centigrade min minute(s) hr or h hour(s) conc. concentrated aq aqueous (ex. aq NaHCO3 solution) OTBS tert-butylsilyloxy OBn benzyloxy OAc acetyloxy to be continued on the next page Preparation Formula No.

Preparation F ormnul1a No.

TB SO,,.

0

H

N N HN N OTBS 0 0 0 N 0 H H0 N N N N N H 0 0 2 Example No. Formula

I

F ormul1a minor major Example No. Formula

HO,,

0

H

N N OAc 0 0 0 0 N" N 0 N

N=

0 0 3 0I HN NN N OH major n 0 0 n M minor F ormul1a 43 Example No. Formula

HO,.

0

H

NN OAc 0 0 0 0 N 0 N"

A

0 H 0 N -N NM N N Hl H 0

H

NN N OH

HNN

O0 NN H 0 N

N,

N NT N H00 44 Example No. Formula

I

Example No. F o rmu 1 a Example No. Formula 0 -0

I

Example No. Formula S

HO,,

0 I-IN N OH 00 0 N 0 N 0 H -0

NN

H0 0 8

)CO

2 Me Example No. Formula S

HO,,

0 IHNN N OH 0 0 0 N0 H 0 0 9 Example No. Formula S

HO,,

0 HN N OH 0 0 0 N 0 N 0 H H 0 NH ",kyN NN N N Example No.1 Formula -0

N

-0 i Example No. Formula Cq D* Example No.J Formula Example No. Formula

HO

HO,,

S 0 HNY N OH 0 0 0 0 N 0 0 H 1

N

N"N N H 0 0 14 0 Example No. Formula 0 2

N

fc2 Example No. Formula S

HO,,

0 HN NOH O 0 0O 0 O N

H

N

NN

00 16 Ph

HO,,,

HT"J N N Me 0 0 N 0 H N N

N

H0 Example No.1 Formula

'N

Example No. Formula Example No.j Formula Example No. Formula

I

Example No. Formula

OH

HW-

HO,,.

0 J

H

H;N N" O 0 0 0 0 N

N~

0 H H =0 N N -N 0 7 0 21 j Example No. Formula

HO

O HH N N SHNN OH 0 0 0 N 0 H =0

N

H

22 MeO Example No. Formula minor major Example No. Formula

O/Q

0 "N Example No. J Formula

NI-

-0

N

64 Example No. Formula S

NO

2

N

HNN 0A"I N 0 -0 0 0 N 0 N 0 H II 0

NN

N N 26

H

Example No. Formula HO,, 02 00 0 N~ 0 H 0 N

NN

H 0 .0 27 Example No. Formula S 0 'NO 2 NN N 0K 0 0 0 0 N 0 N 0 H Ii-0

NN

N CN 0N H00 28 Example No. Formula S

HO,,

0 N 1 HN N OH 00 0 N0 0 N0 0 H H N N

H

j 68 Example No. Formula Example No. Formula Example No.

Formula Example No. Formula 4- 'aN0 2 0

N

Example No. I Formula I Example No.] Formula

J

73 Example No. 1 Formula .1 Example No. Formula Example No. Formula 1 0', 1 =0 0 4kN-

H

Example No. Formula

HO,,

0H HN N OH 0 -0 0 0 N 0-1 Example No. Formula

HO,.

0H N N HMJ N OH 00 0H N 0 N~ 0 H H 0_ N N NH N N

I

Example No.- oml

HO,,

0

H

HN "KN OH O 0 N11 O N .0 H H 0 N N N N N H 0 0 39 0 S 0 HN N OH 00 0 O 0 0 H N N N

HN

J

Example No. Formula 79 Exampe No.Formula

HO,.

K

HN N OH S0 0 0 0 N 0 N~ 0 =0

N

N N H N 01 41 0 HO,, MN N OH 0 N~ 0 H 0

N

N N' 01 01 S HO., 0 HN IJ OH 0 0 N minor 0 N~ H 0=0N N N N H 0 i 0 Example No. Formula

HO,

HN N OH 0 O0 N 0- N-\ Example No. Formula Example No. Formula S

HO,,.

0

HNN

0* 0~ 0

H

N

N Ii' 0 44

I

83 E~amn1~ No. Formula Example No.

I

~I

Example No. jFormula

I

Example No. Formula

J

Example No.1 Formula

I

Example No.

Formula 4 88 Example No. T Formula

I

Example No. Formula AcO,,

NO

HIN N 0 00 0 .0 N 0 N~ 0 H N

NN

N0 0 51 -0

N

Example No. Formula .1 Example No. Formula S

HO,,

0 HN N OH 0 0 0 N 0 N 0 Ht H 0

J

Example No.1 Formula Example No. j Formula 4- Example No. Formula Example No. Formula N Me AcO,, 0 ~Jq H HN N OAc 0 0 0 0 N 0 N N =0 NA NN H 0

I

0 I 0 57 HO.

Example No. F ormul1a Example No. Formula

I

Example No. Formula Example No. Formula AcO,,, 0H HN N GAc 0 0 0 0 0 N- 0 0 H H -0 NN N N

H

0 0 61 100 Example No.

Formula

-I

-0

N

101 Example No. 1 Formula 102 Example No. Formula

I

103 Example No. Formula OMe

.I

104 Example No. Formula Ii 105 Example No. Formula

HO

O

H

Y0 Ho'' N N HN N OH 0 N0 0 0 N 0, I

I

106 Example No.J Formula 107 Example No.I Formula

I

108 Example No.1 Formula 109 Example No. Formula

I

110 Example No. Formula ill Example No. Formula

HO,,

H

N N HN N OH 0 0) 0 NN 0 H H a0 H 00 73 112 Example No. Formula 0HO,, HN N 0 0 0 N '1 0 .0 H N N

NI

H

0 74

I

113 Exmle No. Formula 114 Example No.] Formula 115 Example No. Formula

HOI--

0HO,,.

H N N

OH

0 H H 0 N0 0 77 AcO

HO,,,

0o N H N N HN N OH 0 0mao n n I-11majo minor 116 Example No. j Formula

I

117 Example No. Formula 0

N

.1 118 Example No.1 Formula 0 0.1-

I

119 Preparation 1 To a solution of starting compound (P1)(24.2 mg) and TBSOTf(19.9 mg) in CH 2 C12 (0.5 ml) was added 2,6lutidine(8.51 mg) with cooling on an ice bath. The reaction mixture was stirred at room temperature for 0.5 hour. The mixture was concentrated in vacuo, and the residue was partitioned between ethyl acetate and water. The mixture was extracted with ethyl acetate. The organic layer was washed with aqueous IN hydrogen chloride solution, saturated aqueous sodium bicarbonate solution and saturated aqueous sodium chloride solution, dried over magnesium sulfate, and concentrated in vacuo. The residue was purified by preparative thin layer chromatography eluted with a mixture of hexane and acetone to give the object compound (PI) (28.8 mg) as an amorphous.

MS m/e 1447.0.

Preparation 2 While a mixed solution of starting compound (P2)(100 mg) and N-methylmorpholine N-oxide (32.4 mg) in a solvent (0.6 ml) of acetonitrile and water was stirred, 4% Os0 4 aqueous solution (176 il) was added thereto. After the solution was stirred for 18 hours at room temperature, ethyl acetate was added to the solution. The organic layer was washed with water, 0.1N hydrochloric acid, saturated NaHCO 3 aqueous solution and saturated brine, dried with magnesium sulfate, and removed under reduced pressure to give the object compound (P2)(100 mg) as white amorphous.

ESI-MS: 1480.81(M+H) Example 1 A solution of starting compound (10 mg) in pyridine (0.1 ml) was added isocyanic acid phenyl ester (4.31 at room temperature. After being stirred at 60 0 C for 18 hours, the mixture was poured into ice-water and extracted with ethyl acetate. The extract was washed with brine and dried over sodium sulfate. The solution was evaporated under reduced pressure to give a crude oil, which was purified by 120 preparative thin layer chromatography using ethyl acetate to give 6 mg of the object compound as white amorphous.

ESI-MS: 1379.17 (M+H) HRESI-MS: 1379.8918 (M+H) Example 2 A solution of starting compound (4 mg) in methanol (0.1 ml) was added potassium carbonate (2 mg) at room temperature. After being stirred at room temperature for 18 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 1.6 mg of the object major compound (2) and 1.1 mg of the object minor compound as white amorphous.

Major compound ESI-MS: 1200.31 (M+H) HRESI-MS: 1200.8335 (M+H) Minor compound ESI-MS: 1337.91 (M+H) HRESI-MS: 1337.8812 (M+H) Example 3 A solution of the object compound (20 mg) in dichloromethane (0.15 ml) were added pyridine (6.42 p1) and trichloroacetylisocyanate (9.41 pil) at 0°C. After being stirred at room temperature for 1 hour, the mixture was evaporated under reduced pressure to give a crude oil, which was dissolved into methanol (1 ml). To the mixture was added silica gel (150 mg) at room temperature and stirred at room temperature for 24 hours. After removal of the silica gel by filtration, the filtrate was evaporated under reduced pressure. The residue was dissolved into methanol (0.3 ml) and to the reaction mixture was added potassium carbonate (5.49 mg) at 0°C. After being stirred at room temperature for 18 hours, the mixture was evaporated under reduced 121 pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 4.1 mg of the major object compound as white amorphous and 2.6 mg of the minor object compound as white amorphous.

major compound ESI-MS 1261.15 (M+H) HRESI-MS 1261.8499 (M+H) minor compound ESI-MS 1200.31 (M+H) HRESI-MS 1200.8335 (M+H) Example 4 A solution of the starting compound (10 mg) in toluene (0.1 ml) and dichloromethane (0.1 ml) were added 2tert-butylimino-2-diethylamino-1,3-dimethyl-perhydro-1,3,2diazaphosphorine (4.14 il) and isocyanic acid ethyl ester (9.42 il) at room temperature. After being stirred at room temperature for 18 hours, the mixture was poured into icewater and extracted with ethyl acetate. The extract was washed with brine and dried over sodium sulfate. The solution was evaporated under reduced pressure to give a crude oil, which was dissolved into methanol (0.2 ml). To the mixture was added potassium carbonate (2.74 mg) at 0°C.

After being stirred at 0°C for 2 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 2.7 mg of the object compound as white amorphous.

ESI-MS: 1289.33 (M+H) HRESI-MS: 1289.8812 (M+H) Example A solution of the starting compound (15 mg) in pyridine (0.15 ml) and dichloromethane (0.15 ml) were added 122 diisopropylethylamine (5.18 il), 4-dimethylaminopyridine (3.64 mg) and diethyl pyrocarbonate (52.6 over 10 minutes at 0°C. The reaction mixture was stirred at room temperature for 4 hours and then further stirred at 70 0 C for 18 hours.

The mixture was poured into ice-water and extracted with ethyl acetate. The extract was washed with brine and dried over sodium sulfate. The solution was evaporated under reduced pressure to give a crude oil, which was dissolved into methanol (0.2 ml). To the mixture was added potassium carbonate (3.31 mg) at OC. After being stirred at 0°C for 3 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 1.4 mg of the object compound as white amorphous.

ESI-MS: 1290.13 (M+H) HRESI-MS: 1290.8652 (M+H) Example 6 Chloroformic acid 4-nitrophenyl ester (166 mg) was added portionwise to a stirred solution of the starting compound (100 mg) in pyridine (1 ml) at o0C. After being stirred at room temperature for 18 hours, the mixture was poured into ice-water and extracted with ethyl acetate. The extract was washed with IN hydrochloric acid and brine and dried over sodium sulfate. The solution was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of hexane and acetone (25:20) to give 85 mg of the object compound as white amorphous.

ESI-MS: 1383.14 (M+H) Example 7 The object compound was obtained according to a similar manner to that of Example 6.

ESI-MS: 1385.29 (M+H) 123 Example 8 A solution of the starting compound (10 mg) in dichloromethane (0.2 ml) were added pyridine (13.26,1), 4dimethylaminopyridine (5.0 mg) and chloroformic acid methyl ester (9.5 pl) at room temperature. After being stirred at room temperature for 18 hours, the mixture was poured into ice-water and extracted with ethyl acetate, which was washed with brine and dried over sodium sulfate. The solution was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 3.1 mg of the object compound as white amorphous.

HRESI-MS: 1278.8652 (M+H) Example 9 To a solution of the starting compound (5.95 mg) in pyridine (0.2 ml) was added a solution of phenyl chlorothioformylate (1.69 mg) at room temperature. The reaction mixture was stirred at room temperature for overnight. The solvent was removed under nitrogen gas flow and the residue was purified by preparative thin layer chromatography eluted with a mixture of hexane and ethyl acetate to give the object compound as an amorphous (5.77 mg).

MS(ES+)z/e: 1354.9 Example The compound (10) was obtained according to a similar manner to that of Example 9.

MS(ES+)z/e: 1356.9 Example 11 To a solution of the starting compound (11) (123 mg) and diisopropylamine (57 mg) in THF (5 ml) was added a 1.58M hexane solution of n-butyl lithium (581 mg) dropwise at -78 0

C.

124 The reaction mixture was stirred at -78 0 C for 1 hour and then phenyl chlorothioformate (112 mg) was added to the reaction mixture. The reaction mixture was stirred at -78 0 C for 1 hour and then warmed to room temperature. The mixture was poured into the mixture of ethyl acetate and saturated aqueous ammonium chloride solution. The mixture was extracted with ethyl acetate. The organic layer was washed with aqueous IN hydrogen chloride solution, saturated aqueous sodium bicarbonate solution and saturated aqueous sodium chloride solution, dried over magnesium sulfate, and concentrated in vacuo. The crude product was used to the next reaction without purification.

Example 12 To a solution of starting compound (12) and 2,2'- Azobisisobutyronitrile(15.1 mg) in toluene (0.5 ml) was added a solution of tributyltin hydride (267 mg) at room temperature. The reaction mixture was stirred at 100°C for hour. The solvent was removed and the residue was purified by preparative thin layer chromatography eluted with a mixture of hexane and ethyl acetone to give the object compound (12) as an oil.

MS z/e: 1317.0 Example 13 To a solution of the starting compound (13) (125 mg) in acetonitrile (1 ml) was added a solution of 5% HF in acetonitrile dropwise at room temperature. The reaction mixture was stirred at room temperature for 40 minutes. The mixture was concentrated in vacuo, and the residue was partitioned between ethyl acetate and 10% calcium chloride solution. The mixture was extracted with ethyl acetate. The organic layer was washed with aqueous IN hydrogen chloride solution, saturated aqueous sodium bicarbonate solution and saturated aqueous sodium chloride solution, dried over magnesium sulfate, and concentrated in vacuo. The residue 125 was purified by preparative thin layer chromatography eluted with a mixture of hexane and acetone to give the object compound (13) as an amorphous (3.35 mg).

MS(ES+)z/e: 1202.9 Example 14 To an ice-cooled solution of the starting compound (14) (34 mg) and N-methylmorpholine (14 mg) was added pnitrophenyl chloroformate (7.5 mg). After stirring at that temperature for one hour, N-methylmorpholine (14 mg) was added and the resulting mixture was stirred at room temperature for 14 hours. The reaction mixture was diluted with ethyl acetate, and the organic phase was washed with 0.1N hydrochloric acid, saturated aqueous sodium bicarbonate, water and brine. The resulting mixture was purified by preparative thin layer chromatography on SiO 2 to give 19 mg of the object compound (14).

ESI-MS: 1400 (M+H) Example To a solution of the starting compound (15) (3.3 mg) in THF (0.5 ml) was added a 2M solution of ethylamine in THF (59 Al). The mixture was stood at room temperature for 14 hours, and then evaporated to dryness. The residue was purified by preparative thin layer chromatography on SiO 2 to give the object compound (15) (2.8 mg).

ESI-MS: 1306 (M+H) Example 16 A solution of the starting compound (16) (15 mg) in toluene (0.2 ml) were added benzaldehyde dimethyl acetal (92 ti) and pyridinium p-toluenesulfonate (93.7 mg) at room temperature. After being stirred at 100 0 C for 20 minutes, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using ethyl acetate to give 6.5 mg of the 126 object compound (16) as white amorphous.

ESI-MS: 1306.05 (M+H) HRESI-MS: 1306.8754 (M+H) Example 17 The compound (17) was obtained according to a similar manner to that of Example 16.

ESI-MS: 1258.17 (M+H) HRESI-MS: 1258.8754 (M+H) Example 18 A solution of the starting compound (18) (5 mg) in methanol (0.1 ml) was added potassium carbonate (1.57 mg) at room temperature. After being stirred at room temperature for 18 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 2.3 mg of the object compound (18) as white amorphous.

ESI-MS: 1231.96 (M-H) HRESI-MS: 1234.8390 1256.8210 (M+Na) Example 19 A solution of the starting compound (19) (1.4 mg) in methanol (0.2 ml) was added 0.1N hydrochloric acid (0.02 ml) at room temperature. After being stirred at room temperature for 18 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 1.1 mg of the object compound (19) as white amorphous.

HRESI-MS: 1248.8546 1270.8366 (M+Na) Example A mixture of the starting compound (20) (4.6 mg), DMAP mg), pyridine (3 mg) and p-toluenesulfonyl chloride 127 mg) in dichloromethane (0.5 ml) was stirred at room temperature for 22 hours. The mixture was partitioned between ethyl acetate and 0.1N hydrochloric acid, and the organic phase was washed with water, saturated aqueous sodium bicarbonate and brine, dried over MgSO 4 and evaporated to dryness. The residue was purified by preparative thin layer chromatography methanol in chloroform) to give 3.0 mg of the object compound HRESI-MS: 1190.8140 (found), 1190.8128 (calcd for M+H) Example 21 The object compound (21) was obtained according to a similar manner to that of Example ESI-MS: 1204.62 (M-H) HRESI-MS: 1206.8077 1228.7897 (M+Na) Example 22 A solution of the starting compound (22) (2.1 mg) in methanol (0.2 ml) was added 0.1N hydrochloric acid (0.02 ml) at room temperature. After being stirred at room temperature for 18 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 1.8 mg of the object compound (22) as white amorphous.

ESI-MS: 1218.47 (M-H) HRESI-MS: 1220.8233 1242.8053 (M+Na) Example 23 A solution of the starting compound (23) (10 mg) in methanol (0.1 ml) was added potassium carbonate (4.39 mg) at room temperature. After being stirred at room temperature for 18 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 7.17 mg of the object 128 compound (23) as white amorphous.

ESI-MS: 1176.57 (M+H) HRESI-MS: 1176.8335 (M+H) Example 24 To a solution of the starting compound (24) (5.77 mg) and 2,2'-azobisisobutyronitrile (0.14 mg) in toluene (0.5 ml) was added a solution of tributyltin hydride (6.2 mg) at room temperature. The reaction mixture was stirred at 100 0 C for 1.5 hour. The solvent was removed and the residue was purified by preparative thin layer chromatography eluted with a mixture of hexane and ethyl acetate to give the object compound (24) as an oil (3.0 mg).

MS(ES+)z/e: 1202.9 Example The object compound (25) was obtained according to a similar manner to that of Example 24.

MS(ES+)z/e: 1204.9 Example 26 A solution of the starting compound (26) (6 mg) in tetrahydrofuran (0.1 ml) were added N,N-diisopropylamine (2.27 pl) and n-propylamine (11.13 il) at room temperature.

After being stirred at room temperature for 18 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 4.1 mg of the object compound (26) as white amorphous.

ESI-MS: 1302.96 (M+H) HRE.SI-MS: 1303.8968 (M+H) Example 27 The object compound (27) was obtained according to a similar manner to that of Example 26.

129 ESI-MS: 1389.18 (M+H) HRESI-MS: 1389.9336 (M+H) Example 28 The object compound (28) was obtained according to a similar manner to that of Example 26.

ESI-MS: 1317.75 (M+H) HRESI-MS: 1317.9125 (M+H) Example 29 A solution of the starting compound (29) (10 mg) in pyridine (0.1 ml) was added isocyanic acid phenyl ester (4.46 at room temperature. After being stirred at 60 0 C for 18 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of chloroform and methanol (20:1) to give 9.1 mg of the object compound (29) as white amorphous.

ESI-MS: 1337.37 (M+H) HRESI-MS: 1337.8812 (M+H) Example The object compound (30) was obtained according to a similar manner to that of Example 26.

ESI-MS: 1305.9 (M+H) HRESI-MS: 1305.9125 (M+H) Example 31 The object compound (31) was obtained according to a similar manner to that of Example 26.

ESI-MS: 1319.08 (M+H) HRESI-MS: 1319.9281 (M+H) Example 32 The object compound (32) was obtained according to a similar manner to that of Example 26.

130 ESI-MS: 1263.78 (M+H) HRESI-MS: 1263.8655 (M+H) Example 33 A solution of the starting compound (33) (500 mg) in dichloromethane (3.5 ml) was added 1,1'-carbonylbis-1Himidazole (133 mg) at 0°C. After being stirred at 0 C for hours, n-propylamine (0.338 ml) was added to the mixture at 0°C. The mixture was poured into ice-water and extracted with ethyl acetate. The extract was washed with IN hydrochloric acid and brine and dried over sodium sulfate.

The solution was evaporated under reduced pressure to give a crude oil, which was purified by column chromatography on silica gel using a mixed solvent of n-hexane and acetone to give 506 mg of the object compound (33) as white amorphous.

ESI-MS: 1302.96 (M+H) HRESI-MS: 1303.8968 (M+H) Example 34 A solution of the starting compound (34) (10 mg) in 1,2-dichloroethane (0.1 ml) were added 2-tert-butylimino-2diethylamino-1,3-dimethyl-perhydro-1,3,2-diazaphosphorine (1.45 il) and isocyanic acid ethyl ester (9.11 il) at room temperature. After being stirred at room temperature for 18 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using ethyl acetate to give 4.6 mg of the object compound (34) as white amorphous.

ESI-MS: 1374 (M+H) HRESI-MS: 1374.9340 (M+H) Example The object compound (35) was obtained according to a similar manner to that of Example 34.

ESI-MS: 1436.7 (M+H) HRESI-MS: 1436.9496 (M+H) Example 36 The object compound (36) was obtained according to a similar manner to that of Example 34.

ESI-MS: 1422.8 (M+H) HRESI-MS: 1422.9304 (M+H) Example 37 A mixed solution of methanol (1 ml), the starting compound (37) (100 mg) and platinum dioxide (7.5 mg) was stirred for 1 hour under hydrogen atmosphere (1 atom). The platinum dioxide was filtered and the filtrate was removed under reduced pressure. After dissolution in ethyl acetate (0.3 ml), the solution was diluted with n-hexane (1.2 ml).

The resultant precipitate was filtered, washed with n-hexane and dried under reduced pressure to obtain the object compound (37) (86 mg) as white powder.

HRESI-MS: 1220.8604 (found), 1220.8598 (calcd for M+H) Example 38 While a mixed solution of the starting compound (38) (10.7 mg) and N-methylmorpholine N-oxide (3 mg) in a solvent (1 ml) of acetonitrile and water was stirred, 4% Os04 aqueous solution (11 [il) was added thereto. After the solution was stirred for 16 hours at room temperature, ethyl acetate was added to the solution. The organic layer was washed with water, 0.1N hydrochloric acid, saturated NaHCO 3 aqueous solution and saturated brine, dried with barium sulfate, and removed under reduced pressure. The resultant residue was dissolved in ethyl acetate (1 ml) and water ml), and NaI0 4 (5.7 mg) was added thereto. After the solution was stirred for 24 hours at room temperature, the organic layer was extracted with ethyl acetate and washed with water and saturated brine. This solution was concentrated and dissolved in a mixed solvent (0.8 ml) of methanol and water NaBH 4 (1 mg) was added thereto, and the mixture was stirred for 15 hours. After extraction with ethyl acetate, washing with saturated brine, drying with 132 barium sulfate and removal under reduced pressure, the resultant residue was purified by PTLC methanolchloroform solution) to obtain the object compound (38) (6.7 mg).

HRESI-MS: 1208.8248 (found), 1208.8234 (calcd for M+H) Example 39 Under ice cooling, Dess-Martin periodinane (2.3 mg) was added to a solution of the starting compound (39) (6 mg) in dichloromethane (0.5 ml). After stirred for 2 hours under ice cooling, the mixed solution was purified directly with PTLC methanol-chloroform solution) to the obtain object compound (39) (4.8 mg) as a tautomer.

HRESI-MS: 1216.8284 (found), 1216.8285 (calcd for M+H) Example Under ice cooling, Dess-Martin periodinane (2.7 mg) was added to a solution of the starting compound (40) (3.7 mg) in dichloromethane (0.5 ml). After the mixed solution was stirred for 6 hours at room temperature, Dess-Martin periodinane (2.0 mg) was added further. After 14 hours, the mixed solution was purified directly by PTLC methanolchloroform solution) to obtain the object compound (40) mg) as a tautomer.

HRESI-MS: 1214.8121 (found), 1214.8128 (calcd for M+H) Example 41 Isobutane gas was bubbled into a solution of the starting compound (41) (11 mg) and tricyclohexylphosphine[1,3-bis(2,4,6-trimethylphenyl)-4,5dihydroimidazole-2-ylidene][benzylidene] ruthenium (IV) dichloride (2.3 mg) in toluene (1 ml) for 1 minute. The mixed solution was heated in a sealed tube at 0 C for 1 hour, cooled to room temperature and roughly purified by silica gel (1 The resultant residue was purified by HPLC to obtain the major object compound (4.1 mg) and the minor object compound (41) (2.3 mg).

Major object compound: HRESI-MS: 1232.8601 (found), 1232.8598 133 (calcd for M+H) Minor object compound: HRESI-MS: 1204.8287 (found), 1204.8285 (calcd for M+H) Example 42 The object compound (42) was obtained according to a similar manner to that of Example 39.

ESI-MS: 1218.82 (M+H) HRESI-MS: 1218.8441 (M+H) Example 43 A solution of the starting compound (43) (10 mg) in methanol (0.1 ml) was added potassium carbonate (2.19 mg) at room temperature. After being stirred at room temperature for 18 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 6.2 mg of the object compound (43) as white amorphous.

ESI-MS: 1218.82 (M+H) HRESI-MS: 1218.8441 (M+H) Example 44 The object compound (44) was obtained according to a similar manner to that of Example 39.

HRESI-MS: 1260.8546 (M+H) Example The object compound (45) was obtained according to a similar manner to that of Example ESI-MS: 1216.48 (M+H) HRESI-MS: 1216.8284 (M+H) Example 46 A solution of the starting compound (46) (20 mg) in tetrahydrofuran (0.2 ml) was added 1,1'-carbonylbis-1H- 134 imidazole (6.57 mg) at 0°C. After being stirred at room temperature for 18 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was dissolved into tetrahydrofuran (0.4 ml). To the mixture was added tetrabutyl ammonium fluoride (35.3 mg) at room temperature and stirred at room temperature for 18 hours. The solution was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of chloroform and methanol (20:1) to give 7 mg of the object compound (46) as white amorphous.

ESI-MS: .1278.03 (M+H) HRESI-MS: 1278.8288 (M+H) Example 47 The object compound (47) was obtained according to a similar manner to that of Example 38.

ESI-MS: 1252.36 (M+H) HRESI-MS: 1252.8496 (M+H) Example 48 A solution of the starting compound (48) (20 mg) in chloroform (0.2 ml) was added 3-chloroperoxybenzoic acid (11.3 mg) at 0°C. After being stirred at room temperature for 18 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of chloroform and methanol (15:1) to give 11 mg of the object compound (48) as white amorphous.

ESI-MS: 1234.54 (M+H) HRESI-MS: 1234.8390 (M+H) Example 49 The object compound (49) was obtained according to a similar manner to that of Example 39.

ESI-MS: 1260.22 (M+H) HRESI-MS: 1260.8546 (M+H) 135 Example A solution of the starting compound (50) (5 mg) in methanol (0.2 ml) were added pyridine (0.02 ml) and omethylhydroxylamine hydrochloride (3.3 mg) at room temperature. After being stirred at 50 0 C for 8 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 2.9 mg of the object compound (50) as white amorphous.

ESI-MS: 1289.08 (M+H) HRESI-MS: 1289.8812 (M+H) Example 51 The object compound (51) was obtained according to a similar manner to that of Example ESI-MS: 1365.37 (M+H) HRESI-MS: 1365.9125 (M+H) Example 52 A solution of the starting compound (52) (6 mg) in methanol (0.2 ml) were added 2.0 M ammonia in ethanol (0.0238 ml) and sodium cyanoborohydride (1.49 mg) at 5 0 C. After being stirred at room temperature for 18 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of chloroform and methanol (10:1) to give 4.2 mg of the object compound (52) as white amorphous.

ESI-MS: 1261.17 (M+H) HRESI-MS: 12261.8863 (M+H) Example 53 A solution of the starting compound (53) (13 mg) in acetic anhydride (0.15 ml) was added 4-dimethylaminopyridine (13 mg) at room temperature. After being stirred at room 136 temperature for 18 hours, the mixture was poured into aqueous saturated sodium hydrogen carbonate solution and extracted with ethyl acetate. The extract was washed with brine and dried over sodium sulfate. The solution was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 10 mg of the object compound (53) as white amorphous.

ESI-MS: 1304.14 (M+H) HRESI-MS: 1304.8809 (M+H) Example 54 The object compound (54) was obtained according to a similar manner to that of Example 79.

ESI-MS: 1263(M+H) Example A solution of the starting compound (55) (40 mg) in a mixed solvent of 1-methyl-2-pyrrolidone (0.3 ml) and water (0.05 ml) were added palladium(II) chloride (16.3 mg) and cuprous chloride (27.4 mg) at room temperature. After being stirred at room temperature for 18 hours, the mixture was poured into ice-water and extracted with ethyl acetate. The extract was washed with brine and dried over sodium sulfate.

The solution was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 32.5 mg of the object compound (55) as white amorphous.

ESI-MS: 1318.04 (M+H) HRESI-MS: 1318.8601 (M+H) Example 56 A solution of the starting compound (56) (7 mg) in methanol (0.2 ml) were added pyridine (0.02 ml) and omethylhydroxylamine hydrochloride (5.32 mg) at room 137 temperature. After being stirred at 50 0 C for 2 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using ethyl acetate to give 5.1 mg of the object compound (56) as white amorphous.

ESI-MS: 1347.32 (M+H) HRESI-MS: 1347.8867 (M+H) Example 57 A solution of the starting compound (57) (6 mg) in methanol (0.1 ml) was added potassium carbonate (2.46 mg) at room temperature. After being stirred at room temperature for 18 hours, the mixture was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using a mixed solvent of n-hexane and acetone to give 3.9 mg of the object compound (57) white amorphous.

ESI-MS: 1263.27 (M+H) HRESI-MS: 1263.8655 (M+H) Example 58 The object compound (58) was obtained according to a similar manner to that of Example 56.

ESI-MS: 1423.81 (M+H) HRESI-MS: 1423.9180 (M+H) Example 59 The object compound (59) was obtained according to a similar manner to that of Example 57.

ESI-MS: 1339.71 (M+H) HRESI-MS: 1339.8968 (M+H) Example A solution of the starting compound (60) (10 mg) in 1,2-dichloroethane (0.15 ml) was added (diethylamino)sulfur trifluoride (27.8 Rl) at room temperature. After being 138 stirred at 60 0 C for 18 hours, the mixture was poured into aqueous saturated sodium hydrogen carbonate solution and extracted with ethyl acetate. The extract was washed with brine and dried over sodium sulfate. The solution was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using ethyl acetate to give 3.6 mg of the object compound (60) as white amorphous.

ESI-MS: 1340.72 (M+H) HRESI-MS: 1340.8620 (M+H) Example 61 Through a cooled mixture of the starting compound (61)(5mg) and sodium bicarbonate (0.15mg) in a mixture of dichloromethane and methanol (2ml, 1:1) was passed ozone gas at -78 0 C until the mixture slightly colored blue. The mixture was treated with dimethylsulfide (6ml), and then gradually warmed to room temperature. The resulting mixture was diluted with ethyl acetate, and the organic phase was washed with water and brine, dried over magnesium sulfate, and evaporated to dryness. The residue was purified by PTLC to give the object compound (61)(3.6mg).

ESI-MS: ESI-MS 1291 (M+H) Example 62 The object compound (62) was obtained according to a similar manner to that of Example ESI-MS: 1312.56 (M+H) HRESI-MS: 1312.8307 (M+H) Example 63 A solution of the starting compound (63) (20 mg) in pyridine (0.3 ml) were added disopropylethylamine (6.91 tl), 4-dimethylaminopyridine (4.85 mg) and benzoic anhydride (18 mg) at room temperature. After being stirred at 80 0 C for 18 hours, the mixture was poured into ice-water and extracted 139 with ethyl acetate. The extract was washed with brine and dried over sodium sulfate. The solution was evaporated under reduced pressure to give a crude oil, which was purified by preparative thin layer chromatography using ethyl acetate to give 6.3 mg of the object compound (63) as white amorphous.

ESI-MS: 1364.41 (M+H) HRESI-MS: 1364.8809 (M+H) Example 64 To a solution of the starting compound (64) (8 mg), methyl acrylate (1.98 mg) in methylene chloride (0.6 ml) was added a Grubb's catalyst (1.11 mg) at room temperature. The reaction mixture was stirred at room temperature for hours. The solvent was removed under reduced pressure. The residue was purified by silica gel column chromatography eluted with a mixture of chloroform and methanol (95:5) to give the object compound (64) (4.7 mg) as an amorphous.

MS(ES+)z/e: 1263.4 Example To a solution of the starting compound (65) (3.3 mg) in MeOH (0.5 ml) was added a suspension of PtO 2 (1.31 mg) in MeOH (0.5 ml) at room temperature. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was filtered through celite and the solvent of the filtrate was removed and the residue was purified by preparative thin layer chromatography eluted with a mixture of chloroform and methanol (95:5) to give the object compound (3.3 mg) as a powder.

MS(ES+)z/e: 1264.8 Example 66 The object compound (66) was obtained according to a similar manner to that of Example 64.

MS(ES+)z/e: 1280.9 140 Example 67 The object compound (57) was obtained according to a similar manner to that of Example MS(ES+)z/e: 1282.9 Example 68 To a solution of the starting compound (68) (458 mg) in pyridine (5 ml) was added a pivaloyl chloride (453 mg) at room temperature. The reaction mixture was stirred at 60 0

C

for 4 hours. The mixture was concentrated in vacuo, and the residue was partitioned between ethyl acetate and water. The mixture was extracted with ethyl acetate. The organic layer was washed with aqueous IN hydrogen chloride solution, saturated aqueous sodium bicarbonate solution and saturated aqueous sodium chloride solution, dried over magnesium sulfate, and concentrated in vacuo. The residue was purified by silica gel column chromatography eluted with a mixture of hexane and acetone to give the object compound (68) (413 mg) as a powder.

MS(ES+)z/e: 1032.9 Example 69 A solution of the starting compound (69) (2.1 mg) and Pd/C (0.322 mg) in MeOH (0.8 ml) was stirred at room temperature under hydrogen (1 atm) for 2 hours. The reaction mixture was stirred at room temperature for 2 hours. The reaction mixture was filtered through celite and the solvent of the filtrate was removed and the residue was purified by preparative thin layer chromatography eluted with a mixture of chloroform and methanol (95:5) to give the object compound (69) (2.0 mg) as a powder.

MS(ES+)z/e: 1304.9 Example To a solution of the starting compound (70) (20 mg) in

CH

2 C12 (0.2 ml) was added diethylaminosulfur trifluoride 141 (25.5 mg) dropwise with cooling on an ice bath. The reaction mixture was stirred at room temperature for 4 hours. The mixture was concentrated in vacuo, and the residue was partitioned between ethyl acetate and water. The mixture was extracted with ethyl acetate. The organic layer was washed with aqueous IN hydrogen chloride solution, saturated aqueous sodium bicarbonate solution and saturated aqueous sodium chloride solution, dried over magnesium sulfate, and concentrated in vacuo. The residue was purified by preparative thin layer chromatography eluted with a mixture of hexane and acetone to give the object compound mg) as a powder.

MS(ES+)z/e: 1244.7 Example 71 The object compound (71) was obtained according to a similar manner to that of Example MS(ES+)z/e: 1184.9 Example 72 To a solution of the starting compound (72) (8 mg) in MeOH (0.1 ml) was added a solution of K 2 C0 3 (1.78 mg) at room temperature. The reaction mixture was stirred at room temperature for 30 minutes. The mixture was concentrated in vacuo, and the residue was partitioned between ethyl acetate and water. The mixture was extracted with ethyl acetate.

The organic layer was washed with aqueous IN hydrogen chloride solution, saturated aqueous sodium bicarbonate solution and saturated aqueous sodium chloride solution, dried over magnesium sulfate, and concentrated in vacuo. The residue was purified by preparative thin layer chromatography eluted with a mixture of hexane and ethyl acetone to give the object compound (72) (6.0 mg) as a powder.

MS(ES+)z/e: 1202.9 Example 73 To a solution of the starting compound (73) (11 mg) and 1,8-diaminonaphthalene (2.9 mg) in CH 2

CI

2 (0.5 ml) was added 142 a trimethyloxonium tetrafluoroborate (1.6 mg) with cooling on an ice bath. The reaction mixture was stirred at room temperature for 4 hours. The mixture was concentrated in vacuo, and the residue was partitioned between ethyl acetate and water. The mixture was extracted with ethyl acetate.

The organic layer was washed with aqueous IN hydrogen chloride solution, saturated aqueous sodium bicarbonate solution and saturated aqueous sodium chloride solution, dried over magnesium sulfate, and concentrated in vacuo. The residue was purified by preparative thin layer chromatography eluted with a mixture of chloroform and methanol (95:5) to give the object compound (73) (6.1 mg) as a powder.

MS(ES+)z/e: 1234.9 Example 74 To a solution of the starting compound (74) (113 mg) in pyridine (0.5 ml) was added a benzoic acid anhydride (83.8 mg) at room temperature. The reaction mixture was stirred at 0 C for 2 days. The mixture was concentrated in vacuo, and the residue was partitioned between ethyl acetate and water.

The mixture was extracted with ethyl acetate. The organic layer was washed with aqueous IN hydrogen chloride solution, saturated aqueous sodium bicarbonate solution and saturated aqueous sodium chloride solution, dried over magnesium sulfate, and concentrated in vacuo. The residue was purified by preparative thin layer chromatography eluted with a mixture of hexane and acetone to give the object compound (74) (100 mg) as a powder.

MS(ES+)z/e: 1324.88 Example The object compound (75) was obtained according to a similar manner to that of Example 73.

MS(ES+)z/e: 1248.9 Example 76 143 To a solution of the starting compound (76) (50 mg) and ally alcohol (48 mg) in toluene (1 ml) was added tricyclohexylphosphine [1,3-bis(2,4,6-trimethylphenyl)-4,5dihydroimidazol-2-ylidene][benzylidene]ruthenium

(IV)

dichloride (14 mg). The mixture was stirred under nitrogen for 19 hours. The resulting mixture was directly purified by SiO 2 to yield 41 mg of the object compound (76).

ESI-MS: 1235 (M+H) Example 77 To s solution of the starting compound (77) (6.8 mg) and pyridine (2.2 mg) in dichloromethane (0.5 ml) was added acetic anhydride (0.7 mg). After stirring for 20 minutes, DMAP (0.2 mg) was added, and then the mixture was stirred for 2 hours. The resulting mixture was diluted with ethyl acetate, and the organic solution was washed with 0.1N hydrochloric acid, saturated aqueous sodium bicarbonate and brine. The organic phase was dried over MgSO 4 and evaporated to dryness. The residue was purified by preparative thin layer chromatography to give 2.5 mg of the major product (77) and 1.2 mg of the minor produce (77).

ESI-MS of the major produce 1277 (M+H) ESI-MS of the minor product 1319 (M+H) Example 78 To a solution of the starting compound (78) (22 mg) and 1-decene (25 mg) in toluene (2 ml) was added tricyclohexylphosphine [1,3-bis(2,4,6-trimethylphenyl)-4,5dihydroimidazol-2-ylidene][benzylidene]ruthenium

(IV)

dichloride (4.6 mg). The mixture was stirred at room temperature for 24 hours. The resulting mixture was passed through a pad of silica gel, and purified by PTLC. The purified olefin was then hydrogenated under atmospheric hydrogen with PtO 2 (1 mg) for 2 hours. The catalyst was removed off by filtration, and the filtrate was evaporated.

The residue was purified by preparative thin layer 144 chromatography to give the object compound (78) (10.2 mg).

ESI-MS: 1319 Example 79 To a solution of the starting compound (79) (10 mg) and pyridine (1.3 mg) was added caproyl chloride (1.7 mg). After 2 hours, DMAP (2 mg) and caproyl chloride (7.5 mg) was added.

The mixture was stirred for additional 22 hours. The resulting mixture was diluted with ethyl acetate, and the organic phase was washed with 0.1N hydrochloric acid, saturated aqueous sodium bicarbonate and brine, dried over MgSO 4 and evaporated. The residue was purified by preparative thin layer chromatography to yield 3.8 mg of the object compound (79).

'ESI-MS: 1317 Example The object compound (80) was obtained according to a similar manner to that of Example 79.

ESI-MS: 1293 The Starting Compounds in the following Examples 81 to 177 and The Object Compounds (81) to (167) in the following Examples 81 to 167 are illustrated by chemical formulae as below.

145 The Starting Compound (the same in Examples 81 to 167) The Object Compounds (81) to (167) In the following Examples, The Object Compound [e.g.

The Object Compound means the object compound of Example Example 146 The following object compounds (81) to (167) were obtained according to a similar manner to that of Example 26.

Example No. R 81 H N N

CH

3 82

H

H

83 N-CH 3 84 O H84 N CH H 3 0 -N N

CH

3

N

-N

86 H N/v -N N H 87

-N

H 88

N

H

89 -N -N N CH 3 H3 147 Example No. R 91 -N 0 H C H 92 /CH 3 93

H

94

H

Me

H/

N_

Me Ho 96

-N

97

-N

Me

H

Me 98

M

99

-C

H__

100 101 H 148 Example No.-

R

H

1020 103 -N -No 104N

HC

105NN

H..

106

_N

107

HN_

H

108 109

N

110-NM NH

CH

3 149 Example No.

R

N

112 0- N

H

N

113N 114 -NO -OH

OH

115

N&H

116 0HN N 117 3/ -No N 0\

H

118 -N /-\N-CH 3 119

OHK

OH

150 Example No._ R 0 120

H

3C

N)

121

NH

122 H

N

123N 124

N;

0 125

H

3 C0 126

N

0\H 127N_

N

151 Example No. R /CH 3 128 No/

H

N

129H3/N

H

3 C3 1301

HNN

13 0 132 NI

N

133- N-\0

NN

15.2 Example No. R 137-N I

H

138 -N N 139 -No NC 0

WO-

140 141

-N

H

F

142

CK

143

H

3 Cl 0 144 j 153 Example No.- R 145 H 3 C N CH 3 146 H N 0 147 NJ 0' c

NJ

N

148

N

U

CH

3 149 -N

NO

150

N,

H3C\ 151N 152 -N

H

153 154 Example No. R 154 -NO CH 3 1550 156 1

CH

3 1570

CH

3

NJ

158 159 -No 160 155 Example No. R

N

161

CH

3 162

N-

H

163C

O

NH

164 165 166 156 Example No.

167 N- The Mass spectrum data of the object compounds (81) to (1.67) are as follows.

Example 81 1301.58 Example 82

MS(M+):

Example 83

MS(M+):

1347.63 1276.57 Example 84 1292.60 Example

MS(M+):

Example 86

MS(M+):

Example 87

MS(M+):

1373.70 1353.66 1353.65 Example 88 MS(M+): 1353.66 157 Example 89

MS(M+):

Example 1 MS(M+): Example 91

MS(M+):

Example 92

MS(M+):

Example 93

MS(M+):

1302.64 1333.69 1331.88 1300.62 1290.63 Example 94 MS(M+): 1302.64 Example

MS(M+):

Example 96

MS(M+):

Example 97

MS(M+):

Example 98

MS(M+):

1304.65 1316.65 1316.65 1318.68 Example 99 MS(M+): 1316.66 Example 100 1330.67 158 Example 101

MS(M+):

Example 102

MS(M+):

Example 103

MS(M+):

Example 104

MS(M+):

Example 105

MS(M+):

1352.67 1368.72 1413.83 1330.71 1435.83 Example 106

MS(M+):

Example 107

MS(M+):

Example 108

MS(M+):

1344.74 1366.73 1380.75 Example 109

MS(M+):

Example 110

MS(M+):

Example 111

MS(M+):

1421.80 1424.87 1372.17 Example 112 MS(M+): 1344.16 159 Example 113

MS(M+):

Example 114

MS(M+):

1431.50 Example 115

MS(M+):

Example 116

MS(M+):

Example 117

MS(M+):

1345.19 1359.19 1475.14 1422.14 Example 118 MS(M+): 1344.18 Example 119

MS(M+):

Example 120

MS(M+):

Example 121

MS(M+):

1362.15 1379.14 1367.18 Example 122

MS(M+):

Example 123

MS(M+):

Example 124

MS(M+):

1374.20 1386.20 1424.19 160 Example 125

MS(M+):

Example 126

MS(M+):

Example 127

MS(M+):

Example 128

MS(M+):

Example 129

MS(M+):

1388.23 1460.20 1461.16 1416.26 1380.19 Example 130

MS(M+):

Example 131

MS(M+):

1438.18 1375.18 Example 132

MS(M+):

Example 133

MS(M+):

Example 134

MS(M+):

Example 135

MS(M+):

Example 136

MS(M+):

1455.23 1442.27 1370.25 1333.24 1408.24 161 Example 137

MS(M+):

Example 138

MS(M+):

1341.24 Example 139

MS(M+):

Example 140

MS(M+):

Example 141

MS(M+):

1407.26 1398.30 1499.23 1404.21 Example 142

MS(M+):

Example 143

MS(M+):

Example 144

MS(M+):

1365.25 1359.27 1510.21 Example 145

MS(M+):

Example 146

MS(M+):

Example 147

MS(M+):

1317.30 1372.27 1464.24 Example 148 MS(M+): 1523.17 162 Example 149 1377.25 Example 150 1432.29 Example 151 1379.28 Example 152 1406.30 Example 153 1377.34 Example 154 1343.43 Example 155 1343.39 Example 156 1377.33 Example 157 1519.39 Example 158 1403.34 Example 159.

1405.40 Example 160 1455.34 163 Example 161

MS(M+):

Example 162

MS(M+):

Example 163

MS(M+):

Example 164

MS(M+):

Example 165

MS(M+):

1458.48 1391.42 1427.43 1420.52 1426.50 Example 166

MS(M+):

Example 167

MS(M+):

1442.54 1497.35

Claims (7)

1. A compound of the following general formula R1 R2 N N 1 is lower alkyl which may be substituted with one or 0 HN ON O 0 1 0 (I) 0 H 0 H o N N N H 0 wherein R1 is lower alkyl which may be substituted with one or. more suitable substituent(s) or lower alkenyl which may be substituted with one or more suitable substituent(s), higher alkyl which may be substituted with one or more suitable substituent(s) or heterocyclic group which may be substituted with one or more suitable substituent(s), and R 2 is hydrogen, hydroxy, oxo, lower alkanoyloxy, aryloxy, lower alkoxycarbonyloxy, aryloxythiocarbonyloxy, or carbamoyloxy which may be substituted with one or more suitable substituent(s), or R 1 and R 2 are linked together to form heterocyclic ring which may be substituted with one or more suitable substituent(s); R X is C-R 4 or C R3 is hydrogen, and 165 R 4 is hydrogen or lower alkyl substituted with one or more suitable substituent(s) selected from the group consisting of hydroxy, oxo, acetyloxy, aryloxythiocarbonyloxy, aryloxycarbonyloxy which may be substituted with one or more suitable substituent(s), lower alkoxycarbonyloxy, carbamoyloxy which may be substituted with one or more suitable substituent(s) and heterocycliccarbonyloxy which may be substituted with one or more suitable substituent(s), or R 3 and R 4 are linked together to form heterocyclic ring which may be substituted with one or more suitable substituent(s); R 5 is hydrogen, hydroxy, oxo, lower alkoxy, lower alkanoyloxy which may be substituted with one or more suitable substituent(s), aroyloxy, amino, lower alkoxyimino or aryloxyimino; and is single bond or double bond, Providing that When R 1 is 1-propenyl and R 2 is hydroxy or acetyloxy, then R 5 is not hydroxy or acetyloxy, or a salt thereof.

2. A pharmaceutical composition which comprises, as an active ingredient, a compound of claim 1 or a pharmaceutically acceptable salt thereof in admixture with pharmaceutically acceptable carriers or excipients.

3. Use of a compound of claim 1 or a pharmaceutically acceptable salt thereof for the manufacture of a medicament.

4. A compound of claim 1 or a pharmaceutically acceptable salt thereof for use as a medicament.

A method for the prophylactic and/or therapeutic treatment 166 of hepatitis C, which comprises administering a compound of claim 1 or a pharmaceutically acceptable salt thereof to a human being or an animal.

6. A commercial package comprising the pharmaceutical composition of claim 2 and a written matter associated therewith, wherein the written matter states that the pharmaceutical composition can or should be used for preventing or treating hepatitis C.

7. An article of manufacture, comprising packaging material and the compound identified in claim 1 contained within said packaging material, wherein said the compound is therapeutically effective for preventing or treating hepatitis C, and wherein said packaging material comprises a label or a written material which indicates that said compound can or should be used for preventing or treating hepatitis C.