| Title of Invention | "TRICYCLIC DERIVATIVES OR PHARMACEUTICALLY ACCEPTABLE SALTS THEREOF, THEIR PREPARATIONS AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM" |
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| Abstract | The present invention relates to tricyclic derivatives or pharmaceutically acceptable salts thereof, their preparations and pharmaceutical compositions containing them. More precisely, the present invention relates to tricyclic derivatives as colchicine derivatives, pharmaceutically acceptable salts thereof, their preparations and pharmaceutical compositions containing them. Tricyclic derivatives of the present invention show very powerful cytotoxicity to cancer cell lines but were much less toxic than colchicine or taxol, confirmed through animal toxicity test. Tricyclic derivatives of the invention also decrease the volume and weight of a tumor and have a strong angiogenesis inhibiting activity in HUVEC cells. Thus, tricyclic derivatives of the present invention can effectively be used as an anticancer agent, anti-proliferation agent and an angiogenesis inhibitor. |
| Full Text | TRICYCLIC DERIVATIVES OR PHARMACEUTICALLY ACCEPTABLE SALTS THEREOF, THEIR PREPARATIONS AND PHARMACEUTICAL COMPOSITIONS CONTAINING THEM FIELD OF THE INVENTION The present invention relates to tricyclic derivatives represented by following pharmaceutically acceptable salts thereof, their preparations and pharmaceutical compositions containing them. (Wherein; RI, R2/ the description. ) -i and X are as defined in BACKGROUND One of pseudo-alkaloid compounds, colchicine has an anti-inflammation action, making it a therapeutic agent for rheumatoid arthritis [Internal Medicine, 86, 1 Mo.2, 342-345, 2000] . Colchicine and thiocolchicine derivatives have functions of muscle relaxation and anti-inflammation (USP 5 973 204, EP 0870761 Al) . Thiocolchicoside has been used for the treatment of contracture and inflammation in skeletal muscles. Also, » colchicine inhibits infiltration of monocytes and Tcells in a transplanted organ in animal experiment and at the same time restrains the production of TNF-a, IL- 1 and IL-6, inflammatory cytokines, suggesting an inhibiting effect on immune response [ Proceedings, 32, 2091-2092, 2002] . Thus, colchicine is very attractive candidate for the development of an immune response inhibitor (WO 02/100824) . Colchicine inhibits a inicrotubule assembly by the interaction with tubulin, resulting in the suppression of cell division [The Alkaloids, 1991, 41, 125-176; USP 4 533 675] . Such colchicine has been used for the treatment of gout and other inflammatory diseases related to gout. However, the • use of colchicine is limited to an acute inflammatory .disease because of the limitation in therapeutic index and toxicity to gastrointestinal tract [Pharmacotherapy, 11, 3, 196-211, 1991]. All the endeavors to develop colchiciue derivatives as an anbicancer drug have not been successful so far [USP 3 222 253; USP 00/6080739; WO 97/01570] , and only demecolcine has been used for the treatment of leukemia. However, toxicity to gastrointestinal tract and limitation in therapeutic index are still problems of demecolcine. The present inventors have completed this invention by developing colchicine derivatives having excellent activities of anticancer, anti-proliferation and angiogenesis inhibition that have now stable therapeutic index resulted from decreased toxicity. SUMMARY OF THE INVENTION It is an object of the present invention to provide tricyclic derivatives or pharmaceutically acceptable salts thereof having excellent activities of anti-cancer, anti-proliferation and angiogenesis inhibition with stable therapeutic index by reduced toxicity. It is also an object of this invention to provide a preparation method for- tricyclic derivatives or pharmaceutically acceptable salts thereof. It is a further object of this invention to provide a pharmaceutical composition containing tricyclic derivatives or pharmaceutically acceptable salts thereof as an effective ingredient. BRIEF DESCRIPTION OF THE DRAWINGS PIG. 1 is a graph showing the changes of the volume of a tumor in a BALB/c nude mouse transplanted with human lung cancer cell line NCI-H460 after the administration of tricyclic derivatives of the present invention (Example 8), FIG. 2 is a graph showing the changes of the body weight of a BALB/c nude mouse transplanted with human lung cancer cell line NCI-H4GO after the administration of tricyclic derivatives of the present invention (Example 8), FIG. 3 is a graph showing the changes of the volume of a tumor in a BALB/c nude mouse transplanted with human lung cancer cell line NCI-H460 after the administration of tricyclic derivatives of the present invention (Example 12) by different concentrations (1, 3, 10 ing/kg) , v. FIG. 4 is a graph showing the changes of the body # weight of a BALB/c nude mouse transplanted with human lung cancer cell line NCI-H460 after the administration of tricyclic derivatives of the present invention (Example 12) by different concentrations (1, 3, 10 rag/kg) , FIG. 5 is a set of photographs showing the volume of a tumor growing in a BALB/c nude mouse transplanted with human lung cancer cell line NCI-H460, which was separated on the 14th day after the administration of tricyclic derivatives of the present invention, PIG. 6 is a set. of photographs showing the activity of tricyclic derivatives of the present invention to inhibit angiogenesis in HUVEC cells. DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS The present invention relates to tricyclic derivatives represented by following pharmaceutically acceptable salts '.thereof. (Wherein, (1) R1 is -T1-B1; in which T1 is -X1-, -X1-C(X2)-, -N(R5)-/ -N(R5)C (X3) -, - N(R5)S(0)n1-, -N(R5)C(0) -X1- or -N(RS) C (X1)NH-, in that X1 and X2 are each 0 or S, R5 is each H or C1-C5 alkyl group, nx is an integer of 1-2; and Bx is selected from a group consisting of following (a) ~ (j) , (a) & (CH2)n3-R7 — (CH2)n2 R» ~( CHj)n2 -ZfR7 (b) (c) (CH2)n3-T2-B2 -CCH-CH—o)n4-R7 (d) Z2 Z3 -(CH2)n5-CH-(CH2)n6-R7 ^( CH2)n2 ZfT2-B2 —(CH-CH-0)n4-T2-B2 (g) (h) z, z3 ~-(CII2)n5-CII-(CH2)n6-T2-B2 Wherein, Rs and Ra are each H, halogen, hydroxy, Ci~C3 alkoxy, amino, nitro, cyano or Ci^Ca lower alkyl group; R7 and R9 are each independently halogen, hydroxy, mercapto, -ONO, -ON02 or SNO, in which R7 and R9 are same or different; is C5-C6 membered saturated or unsaturated heterocyclic ring containing 1~2 of hetero atom, in which the hetero atom is selected from a group S and N, preferably, v vj -v~ ~v Y Y Cl C! C3 01 cs C6 /r-i N H C7 _jr-N ^ ~y~ H Cti / — ^ fi C9 /— NH CIO Cll C12 more preferably, Cl (pyridyl group) substituted at position 2 and 6 or position 2 and 5, C7 (pyrrolyl group) substituted at position 2 and 5 or position 2 and 4, Cll (thiophenyl group) or C12 (furanyl group) ; Z1 is C1 - C10 straight -chain or branched-chain alkyl group, preferably C2-Cs straight-chain, or branchedchain alkyl group or cycloalkyl group having substituent; Z2 and Z3 are each independently H or methyl group, in which Z3 is H when Z2 is methyl group, Z2 is H when Z3 is methyl group; T2 is -X1- or -X1-C(X2)-, in that X1 and X2 are each independently O or S; B2 is selected from a group consisting of said (a) , (b) , (c) , (d) or (e) ; n2 is an integer of 0-3, n3 is an integer of 0~5, n4 is an integer of 1-5, ns and n6 are each independently an integer of 1-6; (2) R2 and R3 are each independently H, -P03H2, phosphonate, sulfate, C3~C7 cycloalkyl, C2 ~ C7 alkenyl, C2~C7 alkynyl,, Cx ~ C7 alkanoyl , GI ~ C7 straight-chain or branched-chain alkyl or sugar, in which sugar is a 8 monosaccharide such as glucuronyl, glucosyl or galactosyl; (3) R4 is OCH3, SCH3 or NR10R11, in which R10 and R1X are each independently H or C1-5 alkyl; (4) X is 0 or S.) Preferably in the compound of (1) R1 is -T1-B1; in which TO. is -N (R5) C (X2) - , -N(RS) C (0) -Xx- or - N(R5)C(X1)NH-, in that X1 and X2 are each O, R5 is each H or C1~C5 alkyl group; and B1 is selected from a group consisting of following (a) ~ (j) , (a) (Cl I2)n3-T2-B2 — (CH2)n2 (CH2)n3-R7 CH2)n2 -ZfR7 (b) (c) -(CH-CH-0)n4-R7 (d) 22 Z3 (CH2)n3-T2-B2 (8) CH2)n2 ZfT2-B2 -(CH-CH—O)n4-T2-B2 Z2 Z3 00 -(CH2)n5— CH-(CH2)n6-R7 (CH2)n5—CH— (CH2)n6-T2-B2 Wherein, R6 and RB are each H, halogen, hydroxy, C1~C3 alkoxy, amino, nitro, cyano or C1~C3 lower alkyl group; R7 and R9 are each independently halogen, hydroxy, mercapto(thiol) , -ONO, -ON02 or SNO, in which R7 and R9 are same or different} is C5-C6 rnembeied saturated or unsaturated heterocyclic ring containing 1-2 of hetero atom, in which the hetero atom is selected from a group consisting of 0, S and N, preferably, • N N ^N' Cl C2 C3 /r-N ,—v V* C4 /— NH V ~V V H H C5 C6 / ^ "V" "V C7 C8 C9 CIO Cll C12 more preferably, Cl (pyridyl group) substituted at position 2 and 6 or position 2 and 5, C7 (pyrrolyl group) substituted at position 2 and 5 or position 2 and 4, Cll (thiophenyl group) or C12 (furanyl group),a bond of substituents may be at symmetrical or asymmetrical position; Za is C1~C10 straight-chain or branched-chain alkyl group, preferably C2-CS straightchain or branched-chain alkyl group or cycloalkyl group 10 having substituent; Z2 and Z3 are each independently H or methyl group, in which Z3 is H when Z2 is methyl group, Z2 is H when Z3 is methyl group; T2 is -X1- or - V X1-C(X2)-, in that X1 and X2 are each O or S; B2 is selected from a group consisting of said (a), (b), (c), (d) or (e) ; n2 is an integer of 0-3,. n3 is an integer of 0-5, n4 is an integer of l~3, ns and n6 are each independently an integer of 1-3; (2) R2 and R3 are each independently C3 ~ C7 cycloalkyl or C1~C7 alkyl; (3) R4 is SCH3 or OCH3; (4) X is 0 or S. Preferably, the compounds of comprise: 1) 6-nitrooxymethyl-N-[(7S)-1,2,3-trimethoxy-10- methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen -7-yl]-nicotineamide; " 2) 5-nitrooxymethyl-furan-2-carboxylic acid- [ (7S) -l^^-trimethoxy-lO-methylsulfanyl-g-oxo-SfS,?, 9- tetrahydro-benzo[a]heptalen-7-yl]-amide; 3) N-[(7S)-3-isopropoxy-l,2-dimethoxy-10-methylsulfanyl- 9-oxo-5, 6,7, 9-tetrahydro-benzo [a] heptalen-7- yl] -3-nitrooxymethyl-benzamide; 11 4) N-[(7S)-3-ethoxy-l,2-dimethoxy-10-methylaulfanyl- 9-oxo-5,6,7,9-tetrahydro-benzo[a]heptalen-7- yl]-3-nitrooxymethyl-benzamide; 5) 6-nitrooxymethyl-pyridine-2-carboxylic acid- [ (7S) -1, 2 , 3-trimethoxy-10~rnethylsulf anyl-9-oxo-5, 6,7,9- * tetrahydro-benzo[a]heptalen-7-yl]-amide; 6) 5-nitrooxymethyl-thiophene-2-carboxylic acid- [(7S)-l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptalen-7-yl] -amide; 7) N- [ (7S) -3-cycloperityloxy-l, 2-dimethoxy-10- methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo [a] heptalen -7-yl]-3-nitrooxymethyl-benzamide; 8) N-[(73)-3-ethoxy-l,2-dimethoxy-10- methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo [a]heptalen-7-yl]-2-fluoro-3-nitrooxymethylbenzamide; 9) 2-fluoro-N-[(7S)-3-isopropoxy-l,2-dimethoxy- 10-methylsulfanyl-9-oxo-5,6,7, 9-tetrahydrobenzo [a]heptalen-7-yl]-3-nitrooxymethyl-benzamide; 10) 2-fluoro-3-nitrooxymethyl-N-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo [a] heptalen-7-yl ] -benzatnide ; 11) N-[(7S)-3-cyclopentyloxy-l,2-dimethoxy-10- methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- 12 benzo [a]heptalen-7-yl] -2-fluoro-3-nitrooxytnethylbensamide ,- 12) 3-fluoro-5-nitrooxymethyl-N-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl]-benzamide ;• 13) N- [ ( I S ) -3-ethoxy-l,2-ditnethoxy-10- methylsulfanyl-9-oxo-5, 6,7,9-tetrahydrobenzo[ a]heptalen-7-yl]-3-fluoro-5-nitrooxymethylbenzamide; 14) 3-fluoro-N-[(7S) -3-isopropoxy-l,2-dimethoxy- 10-methylsulfanyl-9-oxo-5, 6,7, 9-tetrahydrobenzo [a] heptalen-7-yl] -5-nitrooxymethyl-benzamide; 15) N- [ (7S) -3-cyclopentyloxy-l, 2-dimethoxy-10- methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl] -3-fluoro-5-nitrooxymethylbenzamide ; 16) 4-fluoro-3-nitrooxymethyl-N-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl]-benzamide; 17) 2-fluoro-5-nitrooxymethyl-N-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl]-benzamide; 18) 3-hydroxy-5-nitrooxymethyl-N-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl]-benzamide; 13 19) 3,5-Jbis-nitrooxymethyl-N- [ (IS) -1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl]-benzamide; 20) 2-hydroxy-4-nitrooxymethyl-N- [ (IS)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,*6,7, 9-tetrahydro- * benzo[a]heptalen-7-yl]-benzamide; 21) 4-nitrooxymethyl-thiophene-2-carboxylic acid [(IS)-l,2,3-trimethpxy-10-methylsulfanyl-9-oxo-5,6,7,2- tetrahydro-benzo[a]heptalen-7-yl]-amide; 22) 3-nitrooxymethyl-thiophene-2-carboxylic acid [(IS)-1,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptalen-7-yl]-amide; 23) 2-(3-nitrooxymethyl-phenyl)~N- [ (7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl]-acetamide; 24) 3-(2-nitrooxy-ethyl)-N-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl]-benzamide; 25) 3-nitrooxybenzoic acid-5-[(7S)-1,2,3- tritnethoxy-10-methylsulfanyl-9-oxo-5, 6,7, 9-tetrahydrobenzo[ a]heptalen-7-yl-carbamoyl]-pyridine-2-ylmethylester; ; 26) 4-nitrooxybutyric acid-5-[ (7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- 14 benzo [a]heptalen-7-yl-carbamoyl]-pyridine-2-ylmethylester; 27) 3-nitrooxymethyl-benzole acid-6-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl-carbamoyl]-pyridine-2-yl- » methylester; 28) 4-nitrooxybutyric acid-6-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl-carbamoyl]-pyridine-2-yltnethylester; 29) 3-nitrooxymethyl-benzole acid-2-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl-carbamoyl]-phenylester; 30) 4-nitrooxybutyric acid-2-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl-carbamoyl]-phenylester; 31) 3-nitrooxymethyl-benzole acid-3-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo [a]heptalen-7-yl-carbamoyl]-phenylester; 32) 4-nitrooxybutyric acid-3-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl-carbamoyl]-phenyleater; 33) 3-nitrooxymethyl-benzoic acid-3-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl-carbamoyl]-benzylester; 15 34) 4-nitrooxybutyric acid-3-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7, 9-tetrahydrobenzo [a]heptalen-7-yl-carbamoyl] -benzylester; 35) 2-nitrosothio-N-[(7S)-1,2,3-trimethoxy-10- Tnethylsulfanyl-9-oxo-5, 6 , 7 , 9-tetrahydro- " ' • benzo[a]heptalen-7-yl]-benzamide; 36) 3-nitrosooxymethyl-N-[(7S)-1,2,3-trimethoxy- 10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl]-benzamide; 37) 3-fluoro-5-nitrosooxymethyl-N-[(73)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl]-benzamide; 38) 3-nitroaothiomethyl-N-[(7S)-1,2,3-trimethoxy- 10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo [a] heptalen-7 -yl] -beirzam.ide ; 39) 3-fluoro-5-nitrosothiomethyl-N- [ (7S) -1,2,3;- triraethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl]-benzamide; . 40) 3-fluoro-5-nitrooxymethyl-N-[(7S)-1,2,3,10- tetramethoxy-9-oxo-5,6,7,9-tetrahydro-benzo[a]heptalen- 7-yl] -benzamide; 41) 3-nitrooxymethyl-N-methyl-N- [(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo [a] heptalen-7-yl] -benzamide; 16 42) 3-fluoro-N-methyl-5-nitrooxymethyl-N-[(7S)- 1,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptalen-7-yl]-benzamide; 43) 2- (3-fluoro-5-nitrooxymethyl-phenyl)-N-[(7S)- 1,2, 3 -trimethoxy-10-methylsulf anyl -9 -oxo-5,6,7,9- * tetrahydro-benzo[a]heptalen-7-yl]-acetamide; or 44) 2- (2-fluoro-5-nitrooxymethyl-phenyl)-N-[(7S)- l,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6, 7,9- tetrahydro-benzo[a]heptalen-7-yl]-acetamide. The present invention also provides pharmaceutically acceptable salts of the compound represented by salts of the present invention can include acid addition salt of a compound according to the invention when the compound is fully basic. Such acid addition salt includes salts holding inorganic acid providing pharmaceutically acceptable anion such as hydrogen halide, or organic acid, or salts holding sulfuric acid or phosphoric acid, or salts holding trifluoroacetic acid, citric acid or maleic acid. And, it include for example hydrochlorides, hydrobromides, phosphonates, sulfates, alkylsulfonates, arylsulfonates, acetates, benzoates, citrates, maleates, fumarates, succinates, lactates and tartarates by suitable salts. When a 17 compound of the present invention . is fully acidic, pharmaceutically acceptable salts can include inorganic salts or organic salts providing pharmaceutically acceptable cation. Said inorganic salts include sodium salts, potassium salts, calcium salts or magnesium salts, etc., said organic salts include methylamine salts, dimethylamine salts, trimethylamine salts, piperidine salts or morpholine salts, etc. The present invention also provides a preparation method for tricyciic derivatives represented by the derivatives of the present invention is described in the below Scheme 1 ~ Scheme 8. Precisely, in the (b) , (c) , (d) and (e), the derivatives are prepared according to the method of Scheme 1 ~ Scheme 6. In the meantime, in the is one of said (f) , (g) , (h) , (i) and (j), the derivatives are prepared by the method of Scheme 7 and Scheme 8. And a concrete compound of the is represented by general formulas (IIa), (IIb), (IIe), (IId), (IIe), (IIf), (IIg), (IIh), (IIi), (II j), (IIk), (II1) , (IIm) , (IIn) , (IIo) and (IIp) in Scheme 1 ~ Scheme 8. 18 ,ON(0)n7 lla Hal—Y-OH XI XII RS ,^SNOn d 19 D-X2H Hal XIV VI1 ,OP XVII rv 1 JL XVI > D-A xvm ,JCON(0)n: lie JL. XIX Xv:; ^vY'SNO nr D-NHR5 m XX (0)n, II XXI (0)ns •^'S"Y ON(0)n7 (OK xxn IIli 20 D-NHR, 0 nrV OP ra ii) deprotecting o D-NAxr XXIV XXV o Hi Uj m ii) deprotecting xxvn R5 xxvm 1 Ilk ni 21 XXXIII •'?« j- Y'^° Ilk In the above Scheme, E is following El ~ E6 respectively; 22 x El E2 E3 (O)n7 O ?M -N Y ~N X, ^ N E4 E5 E6 Wherein, Xi( X2 and X3 are each 0 or S. In the above Scheme, D i s , a n d R2,R3, R4 and X are same as defined in the R5 is H or lower alkyl; X1, X2 and X3 are each independently 0 or S; Hal1 and Hal2 are halogens; Hal1 and Hal2 of general formula (IV) and (IX) are each same or different halogens, for example F, Cl, Br or I; P is conventional protecting group of hydroxy such as methoxymethyl, t-butyldimethylsilyl or benzyl; Y and Y1 are same or different, and indicate following general formula (a1), (b1); (c1), (d1) and (e1) respectively, 23 -(CH2)n2 (a1) -(CH-CH—0)n4- Z, Z3 (d') zf~ (c1) -(CH2)n5-CH~(CH2)n6- (e') Wherein, R6, R8, R9, Z1, Z2, Z3, n2, n3, n4, n5 and n6 are same as defined in the and na are integers of 1-2. The preparation method for tricyclic derivatives of the present invention is illustrated more precisely hereinafter. According to method 1 of the present invention for the preparation of compounds of formula (lla) and (l1b), the compound represented by formula (V) is prepared by amidation reaction making amine compound of formula (III) be reacted with halogen compound of formula (IV), which is step 1. In step 1, a base might be excluded, but the reaction is generally performed with a solvent such as dichloromethane, chloroform, 24 tetrahydrofuran, diethylether, toluene or dimethylformamide etc., which have no influence on amidation reaction, in the presence of pyridine, triethylamine, diethylisopropylamine or Nmethylmorpholine etc., a base that can be acceptable » for amidation reaction in general. Reaction temperature is not limited in particular, but generally reaction can performed under cold temperature or elevated temperature, is performed preferably at room temperature. In step 2, conversion of the compound of formula (V) prepared in the above step 1 to nitrooxy compound (n7 = 2) of formula (IIa) and to nitrosooxy compound (n7 = 1 ) of formula (IIa) was accomplished by nitration reaction and nitrosation reaction, respectively. Nitration reaction needs a compound that is able to convert halogen into nitrate, arid is performed using silver nitrate (AgN03) , t-butylammonium nitrate (Bu4NN03) , etc., in the presence of chloroform, acetonitrile, a mixture of acetonit.rile and aqueous solution, or dichloromethane, which are all solvents not affecting the reaction. Nitrosation reaction might use a compound that is able to convert halogen into nitrosate, too, and is performed preferably using silver nitrite (AgN02) or sodium nitrite (NaN02) in the 25 presence of chloroform, acetonitrile, a mixture of acetonitrile and aqueous solution, aqueous solution, or dichloromethane, which are also solvents not affecting the reaction. Reaction temperature is not limited in particular, but generally reaction can performed under » cold temperature or elevated temperature, is performed preferably at room temperature. Another way to give the compound of formula (IIa) is as follows; reaction of the compound of formula (III) with the compound of formula (VI) is performed to give the compound of formula (VII) , and then, conversion of the compound of formula (VII) to the compound of formula (Ila) is accomplished. The reaction of the compound of formula (III) and the compound of formula (VI) is performed in the presence of a coupling agent such as 1-(3-ditnethylaminopropyl) - 3-ethylcarbodiimide(EDGI), 1-hydroxybenzotriazole hydrate(HOBT) or 1,3-dicyclohexyl carbodiimide(DCC). This reaction might be performed without a base, but generally with a base such as 4-dimethylaminopyridine, pyridine, triethylamine, diethylisopropylamine, Nmethylmorpholine or dimethylphenylamine etc., which can be used in amidation reaction, in a solvent having no negative effect on the reaction, for example acetonitrile, dimethylformamide, dichloromethane, etc. 26 Reaction temperature is not limited in particular, but generally reaction can performed under cold temperature or elevated temperature, is performed preferably at room temperature. Direct conversion of the compound of formula (VII) to the compound of formula (Ila) is » accomplished by the reaction of alcohol with triphenylphosphin (PPh3) , N-bromosuccineimide (NBS) and silver nitrate, or silver nitrite. The reaction is performed in a solvent having no effect on the reaction such as chloroform, acetonitrile, dichloromethane, a mixture of acetonitrile and dichloromethane, etc. Reaction temperature is not limited in particular, but generally reaction is performed under cold temperature or at room temperature. Another way for conversion of the compound of formula (VII) to the compound of formula (Ila) is as follows; conversion of the compound of formula (VII) to halogen compound of formula (V) is accomplished first, and then conversion thereof to the compound of formula (Ila) is accomplished again. At this time, the conversion into halogen compound is performed by using a reagent that generally converts hydroxy group to halogen, for example tribromophosphin, tetrabromomethane etc., in the presence of chloroform* acetonitrile, dichloromethane etc., which are solvents having no negative effect on the reaction. Reaction 27 temperature is not limited in particular, but generally reaction is performed under cold temperature or at room temperature. Processes for preparing the compound of formula (IIb) of the method 1 of the present invention are as follows; conversion of hydrogen included in alcohol of formula (VII) to a leaving group such as mesylate, tosylate or triplate is accomplished, followed by reaction with potassium thioacetate, to give thioacetate ester compound. Hydrolysis of the compound in the presence of a base is accomplished to give the compound of formula (VIII) . At this time, a base is selected among general bases that are able to hydrolyze an ester compound, for example sodium hydroxide, potassium hydroxide or sodium thiomethoxide. And an alcohol solution such as methanol or ethanol is preferred as a solvent for the reaction. Reaction temperature is not limited in particular, but generally reaction can performed under cold temperature or elevated temperature, is performed preferably at room temperature. Reaction of the compound of formula (VIII) with sodium nitrite under an acidic condition, leads to the conversion of the compound to nitrosothio compound of formula (IIb) . A solvent for the reaction is selected from a group consisting of methanol, ethanol, 28 acetonitrile, a mixture of acetonitrile and aqueous solution, or dichloromethane etc., which is not to affect the reaction. Reaction temperature is not limited in particular, but generally reaction can performed under cold temperature "or elevated » temperature, is performed preferably at room temperature. According to method 2 of the present invention, compounds of formula (IIe) and (IId) are prepared. Particularly, in step 1, reaction of the compound of formula (III) with the compound of formula (IX) is accomplished to give the compound of formula (X) . This reaction is performed in analogy to the procedure described in method 1 in which conversion of the compound of formula (III) to the compound of formula (V) was accomplished by amidation reaction. In step 2, conversion of the compound of formula (X) prepared in step 1 to the compound of formula (IIe) is accomplished by nitration reaction along with nitrosation reaction. This reaction is performed in analogy to the procedure described in method 1 in which conversion of the compound of formula (V) to the compound of formula (IIa) was accomplished. 29 Another way to give the compound of formula (IIe) is as follows; reaction of the compound of formula (III) with the compound of formula (XI.) is performed to give the compound of formula (XII) , and then, conversion of . the compound of formula (XII) to the compound of formula (IIe) is accomplished. Reaction of the compound of formula (XI) with the compound of formula (III) is performed in analogy to the procedure described in method 1 in which conversion of the compound of formula (III) to the compound of formula (V) was accomplished by amidation reaction. Conversion of the compound of formula (XII) to the compound of formula (IIe) is accomplished under the same condition as provided for the conversion of the compound of formula (VII) to the compound of formula (IIa) in method 1. Conversion of the compound of formula (XII) to the compound of formula (IId) is accomplished under the same condition as provided for the conversion of the compound of formula (VII) to the compound of formula (IIb) in method 1. According to method 3 of the present invention, compounds of formula (IIe) and (IIf) are prepared. 30 Particularly, in step 1, reaction of the compound of formula (XIV) with the compound of formula (IV) is accomplished to give the compound of formula (XV). The reaction in this method is esterification reaction of .alcohol (X2=0) or thioalcohol (X2=S) with acyl or thioacyl halide, which is performed in the presence of a base that is generally acceptable for esterification reaction. Preferable bases are pyridine, 4- dimethylaminopyridine, triethylamine, diethylisopropylamine, 2,6-lutidine, sodium hydride (NaH) , cesium carbonate, or sodium hydroxide and can be used along with a phase transfer catalyst such as benzyltriethylammoniumchloride. Also, above reaction is preferably performed in a solvent having no negative effect on the reaction, for example dichloromethane, chloroform, tetrahydrofuran, diethylether, toluene, dimethylformamide, acetonitrile or aqueous solution. Reaction temperature is not limited in particular, but generally reaction can performed under cold temperature. or elevated temperature, is performed preferably at room temperature. In step 2, conversion of the compound of formula (XV) prepared in step 1 to the compound of formula (IIe) is accomplished by nitration reaction along with nitrosation reaction. This reaction is performed in 31 analogy to the procedure described in method 1 in which conversion of the compound of formula (V) to the compound of formula (IIa) was accomplished. Another way to give the compound of formula (IIe) is as follows; reaction of the compound of formula (XIV) with the compound of formula (VI} having a protective group in alcohol group is performed to give the compound of formula (XVII), followed by deprotection reaction to give the compound of formula (XVIII). Conversion of the resultant compound to the compound of formula (lie) is accomplished. The reaction of the compound of formula (XIV) with the compound of formula (VI') is processed by esterification reaction of alcohol (X2=O) or thioalcohol (X2=S) and carboxylic acid or thiocarboxylic acid. The reaction is performed either in an aqueous solution supplemented with an acid such as hydrochloric acid, sulfuric acid, dodecylbenzene sulfonic acid or p-toluenesulfonic acid, at room temperature or under elevated temperature, or under the same condition provided for conversion of the compound of formula (III) to the compound of formula (VII) in method 1. Another esterification reaction is performed by Misunobu reaction using triphenylphosphine and diethyl azodicarboxylate in a solvent not affecting the 32 reaction. And the solvent is preferably selected from a group consisting of dichloromethane, chloroform, tetrahydrofuran, diethylether, toluene or acetonitrile. Reaction temperature is not limited in particular, but general-ly reaction is performed under cold temperature or at room temperature. Protecting and deprotecting reaction of alcohol group is performed by known method in general organic synthesis. Reaction of the compound of formula (XIV) with the compound of formula (XVI) is performed to give the compound of formula (XVIII) in analogy to the procedure described in method 3 in which conversion of the compound of formula (XIV) to the compound of formula (XV) was accomplished. Conversion of the compound of formula (XVIII) to the compound of formula (Ie) is performed under the same condition provided for conversion of the compound of formula (VII) to the compound ' of formula (IIa) in method 1. Conversion of the compound of formula (XVIII) to the compound of formula (IIf) is performed under the same condition provided for conversion, of the compound of formula (VII) to the compound of formula (IIb) in method 1. 33 According to method 4 of the present invention, compounds of formula (IIg) and (IIh) are prepared. Particularly, in step 1, reaction of the compound of formula (III) with the compound of formula (XX) is accomplished to give the compound of formula (XXI). When the compound of formula (IIg) is sulfinylamide (na=l) , the reaction of the compound of formula (III) with sulfinylhalide of formula (XX) is performed without a base or with a base that is applicable to amidation reaction, for example pyridine, triethylamine, diethylisopropylamine, Nmethylmorpholine or dimethylphenylamine, in a solvent having no negative effect on the reaction such ns dichloromethane, chloroform, tetrahydrofuran, diethylether, toluene or dimethylformamide. Reaction temperature is not limited in particular, but generally reaction can performed under cold temperature or elevated temperature, is performed preferably at room temperature. When the compound of formula (Ilg) is sulfonylamide (nB=2) , the reaction of the compound of formula (III) with sulfonylhalide of formula (XX) is performed either without a base or with a base that is applicable to amidation reaction in general, for 34 example pyridine, triethylamine, diethylisopropylamine, N-methylmorpholine, sodium hydroxide, sodium carbonate or potassium carbonate, in a solvent having no negative effect on the reaction such as dichloromethane, chloroform, tstrahydrofuran, diethylether, toluene or dimethylformaraide. Reaction temperature is not limited in particular, but generally reaction can performed under cold temperature or elevated temperature, is performed preferably at room temperature. In step 2, conversion of the compound of formula (XXI) prepared in step 1 to the compound of formula (IIg) is performed under the same condition provided for conversion of the compound of formula (VII) to the • compound of formula (IIa) in method 1. Conversion of the compound of formula (XXI) to the compound of formula (IIh) is performed under the same condition provided for conversion of the compound of formula (VII) to the compound of formula (IIb) in method l. According to method 5 of the present invention, compounds of formula (Hi) and formula (IIj) are prepared as follows. In step 1, reaction of the compound of formula (III) with the compound of formula 35 (XXIII) having a protecting group to alcohol group is performed, followed by deprotection reaction. Conversion of the compound of formula (XXIV) resulted from the above reaction to the compound of formula (Hi) is accomplished. At this time, the reaction of the compound of formula (III) with the compound of formula (XXIII) is performed by using a coupling reagent such as carbonyl dichloride, triphosgen, di-tbutyl dicarbonate or 1,1'-carbonyl diimidazole etc. The reaction can be performed either without a base or with a base that is generally acceptable for amidation reaction, for example pyridine, triethylamine, diethylisopropylamine, N-methylmorpholine or dimethylphenylamine, in a solvent having no negative effect on the reaction such as dichloromethane, chloroform, tetrahydrofuran, diethylether, ethanol or dimethylformamide. Reaction temperature is not limited in particular, but generally reaction is performed under cold temperature or at room temperature. And deprotection reaction is performed by known method in general organic synthesis. In step 2, conversion of the compound of formula (XXIV) prepared in step 1 to the compound of formula (Hi) is performed under the same condition provided 36 for conversion of the compound of formula (VII) to the compound o£ formula (IIa) in method 1. Conversion of the compound of formula (XXIV) to the compound of formula (IIj) is performed under the same condition provided for conversion of the compound of formula (VII) to the compound of formula (IIb) in method 1. According to method 6 of the present invention, compounds of formula (IIk) and formula (II1) are prepared as follows. In step 1, reaction of the compound of formula (III) with the compound of formula (XXVI) having a protecting group to alcohol group is performed, followed by deprotection reaction. Then, conversion of the resultant compound of formula (XXVII) to the compound of formula (IIk) is accomplished. At this time, the reaction of the compound of formula (III) with the compound of formula (XXVI) is performed either without a base or with a base that is acceptable for amidation reaction, for example pyridine, triethylamine, diethylisopropylamine or Nmethylmorpholine etc, in a solvent having no negative effect on the reaction such as dichloromethane, chloroform, tetrahydrofuran, diethylether, benzene, 37 acetonitrile, etc. Reaction temperature is not limited, in particular, but generally reaction is performed under cold temperature or at room temperature. Protecting and deprotecting reaction of alcohol group is performed by known method in general organic synthesis. In step 2, conversion of the compound of formula (XXVII) prepared in step l to the compound of formula (IIk) is performed under the same condition provided for conversion of the compound of formula (VII) to the compound of formula (IIa) in method 1. Conversion of the compound of formula (XXVII) to the compound of formula (II1) is performed under the same condition provided for conversion of the compound of formula (VII) to the compound of formula (IIb) in method 1. According to method 7 of the present invention, compounds of formula (IIm) and formula (IIn) are prepared from the compound of formula (XXIX) in analogy to the procedure described in method 3. 38 According to method 8 of the present invention, compounds of formula (IIo) and formula (IIp) are prepared as follows. In step 1, reaction of the compound of formula (XXIX) with the compound of formula (IX) is performed to give a compound of formula (XXXIV). This reaction is performed by etherification reaction of alcohol (X2=0) or thioalcohol (X2=S) with alkylhalide in the presence of a base acceptable for etherification reaction. As a base suitable for that purpose, sodium hydride (NaH), t-potassium butoxide (t- BuOK), n-BuLi, sodium hydroxide, potassium hydroxide and a phase transfer catalyst such as benzyltriethylammoniumchloride etc, or crown ether is preferred. The reaction is performed preferably in a solvent having no negative effect on the reaction, for example dichloromethane, chloroform, tetrahydrofuran, diethylether, toluene, dimethylformamide, aqueous solution, dimethylsufoxide or benzene, etc. Reaction temperature is not limited in particular, but generally reaction can performed under cold temperature or elevated temperature, is performed under cold temperature or at room temperature. In step 2, conversion of the compound of formula (XXXIV), prepared in the above step 1, to the compound of formula (IIo) is accomplished by nitration reaction 39 or nitrosation reaction. This reaction is performed in analogy to the procedure described in method 1 in which, conversion of the compound of formula (V) to the compound of formula (IIa) was accomplished. Another way to prepare the compound of formula (IIo) is as follows. Reaction of the compound of formula (XXIX) with the compound of formula (XI') having a protecting group to alcohol group is performed, followed by deprotection reaction to give the compound of. formula (XXXV) . Conversion of the compound of formula (XXXV) to the compound of formula (IIo) is accomplished. Reaction of the compound of formula (XXIX) with the compound of formula (XI') is performed under the same condition given for conversion of the compound of formula (XXIX) to the compound of formula (XXXIV) accomplished by etherification reaction in method 8. Conversion of the compound of formula (XXXV) to the compound of formula (IIo) is performed under the same condition provided for conversion of the compound i of formula (VII) to the compound of formula (Ila) in method 1. Conversion of the compound of formula (XXXV) to the compound of formula (IIp) is performed under the same condition provided for conversion of the compound 40 of formula (VII) to the compound of formula (IIb) in method 1. The target compounds given by the above reactions can be separated and purified by general methods such as column chromatography, recrystallisation, etc. The present invention provides also a pharmaceutical composition containing tricyclic derivatives represented by the pharmaceutically acceptable salts thereof as an effective ingredient. Tricyclic derivatives according to the present invention or pharmaceutically acceptable salts thereof show very strong cytotoxicity to cancer cell lines but have much less toxicity 'to test animals than colchicine or taxol injection has. When tricyclic derivatives of the present invention were administered to a BALB/c nude mouse transplanted with human lung cancer cell line NCI-H460, \the size and the weight of a tumor were remarkably decreased in proportion to the dosage. Tricyclic derivatives of the present invention also have a strong activity of antiangiogenesis in HUVEC cells. 41 Therefore, tricyclic derivatives of the present invention or pharmaceutically acceptable salts thereof can be effectively used as an anticancer agent, an anti-proliferation agent and an angiogenesis inhibitor. The composition of the present invention might additionally include, in addition to tricyclic derivatives or pharmaceutically acceptable salts thereof, at least one of active ingredients having the same or similar function to the mentioned tricyclic derivatives or pharmaceutically acceptable salts thereof. The said tricyclic derivatives or pharmaceutically acceptable salts thereof can be administered orally or parenterally and be prepared in general forms of pharmaceutical formulation. The tricyclic derivatives of the present invention or pharmaceutically acceptable salts thereof can be prepared for oral or parenteral administration by mixing with generally used fillers, extenders, binders, wetting agents, disintegrant, diluents such as surfactants, or excipients. Solid formulations for oral administration are tablets, pills, powders, granules and capsules. These solid formulations are prepared by mixing one or more suitable excipients such 42 as starch, calcium carbonate, sucrose, lactose and galatin, etc. Except for the simple excipients, lubricants, for example magnesium stearate, talc, etc, can be used. Liquid formulations for oral administrations are suspensions, solutions, emulsions and syrups, and the above mentioned formulations can contain various excipients such as wetting agents, sweeteners, aromatics and preservatives in addition to generally used simple diluents such as water and liquid paraffin. Formulations for parenteral administration are sterilized aqueous solutions, water-insoluble excipients, suspensions, emulsions, lyophilized agent and suppositories. Water insoluble excipients and suspensions can contain, in addition to the active compound or compounds, propylene glycol, polyethylene glycol, vegetable oil like olive oil, injectable ester like ethylolate, etc. Suppositories can contain witepsol, macrogol, tween 61, cacao butter, laurin butter, glycerol and gelatin. The composition of the present invention can be prepared for either oral or parenteral administration (for example, intravenous, subcutaneous, intraperitoneal or local injection), and dosage is determined by weight, age, gender, condition of health and diet of a patient and administration method, 43 excretion rate and severity of a disease. The preferable effective dosage of the tricyclic derivatives of the present invention is 3-300 nig/kg (body weight), and administration times are once or several times per day. EXAMPLES Practical and presently preferred embodiments of the present invention are illustrative as shown in the following Examples. However, the present invention is not limited by following examples. 7-Deacetylcolchicine used in the below examples was prepared by the method described in [EP 0493064; Synthetic Communications 1997, 27(2), 293-296]. 7-Amino-l,2,3-trimethoxy-10-methylsulfanyl-6,7- dihydro-5H-benzo[a]-heptalen-9-one was prepared by the method described in (WO 9421598; Bioorganic & Medicinal Chemistry, Vol 5, No. 12, pp 2277-2282, 1997). Thiodemecolcine was prepared by the method described in (J. Med. Chem, 1985, 28, 1204-1208). (7S)-7-Amino-3-cyclopentyloxy-l,2-dimethoxy-10- methylsulfanyl-6,7-dihydro-5H-benzo[a]heptalen-9-one, 44 (7S)-7-amino-3-isopropoxy-l,2-dimethoxy-lOmethylsulfanyl- 6,7-dihydro-5H-benzo[a]heptalen-9-one, (7S)_7_amino-3-ethoxy-l,2-dimethoxy-10-methylsulfanyl- 6,7-dihydro-5H-benzo[a]heptalen-9-one were prepared by the method described in (WO 9611184). Example 1: Preparation of 6-n:.trooxymethyl-,N- [ (7S) - l,2,3-trimethoxy-10-methyleulfanyl-9-oxo--5, 6,7,9- tetrahydro-benzo[a]heptalen-7-yl]-nicotineamide tritnethoxy-10-tnethylsulfanyl-9-oxo-5, 6,7, 9-tetrahydrobenzo [a]heptalen-7-yl3-nicotineamide CO2H EDCI, DMAP MeCN MeS ° MeS 6-Hydroxymethyl-nicotinic acid was synthesized by the method described in (Bioorg. Med. Chem. Lett, 1996, 6, 3025-3028) . 45 To a solution of 7-amino-l,2,3-trimethoxy-10- methylsulfanyl-6, 7-dihydro-5H-benzo [a] -heptalen-9-one (300 mg, 0.80 tnmol) , 6-hydroxymethylnicotinic acid (135 mg, 0.88 tnmol) and DMAP (60 rag, 0.48 tnmol) in 10 mfi of acetonitrile was added EDCI (308 mg, 1.60 mmol) at 0°C. * The reaction mixture was stirred at room temperature for 2 hours. Water was added to quench the reaction, and aqueous layer was extracted with ethyl acetate. Combined organic layer was dried over anhydrous, sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chroma tography (ethyl acetate: methanol = 8:1), to give 244 mg (yield: 60%, solid having yellow color) of the target compound. 1H'NMR (400MHz, CDC13) : 52.07-2.15(m, 1H) , 2.31- 2.44(m, 2H) , 2.45(s, 3H) , 2.56-2.59(m, 1H) , 3.75(s, 3H) 3.91(s, 3H) , 3.97(s, 3H) , 4.66(q, J=10.2Hz, 2H) , 4.90- 4.93(m, 1H) , 6.56(s, 1H) , 7.13(t, J=9.1Hz, 2H) , 7.40(d, J=10.2Hz, 1H) , 7.52(8, 1H) , B.15(dd, J=2.2, 5.8Hz, 8.80(d, J=6.9Hz, 1H) , 8.96(s, 46 trimethoxy-10-methylsulf anyl-9-oxo-S ,6,7 , 9~tetrahydrobenzo[ a]heptalen-7-yl]-nicotineamide MeQ ON02 The compound (100 mg, 0.19 mmol) prepared in the step 1 of Example 1 and triphenylphosphine (57 rag, 0.21 mmol) were dissolved in acetonitrile/dichloromethane (1.25 ml/0.5 ml), therein NBS (42 mg, 0.23 mmol) was added at -35°C. The reaction mixture was stirred for 20 minutes. Thereafter, therein silver nitrate (40 mg, 0.23 mmol) was slowly added dropwise at room temperature , the reaction mixture was stirred at room temperature for 18 hours. Water was added to quench the reaction, and aqueous layer was extracted with chloroform. Combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chroma tography ( chloroform rmethanol = 10:1), to 47 give 18 mg (yield: 35%, solid having yellow color) of the target compound. NMR (400MHz, CDC13): 52. 09-2 .13.(m, 1H) , 2.31- 2.43(m, 2H) , 2.46(s, 3H) , 2 .55-2 . 64 (m, 1H) , 3.75(s, 3H) , 3.91(s, 3H) , 3.97(s, 311), 4.93-4.98 (m, *1H) , 5.50(s, 2H) , S.56(s, 1H) , 7.16(t, J=10.9Hz, 1H) , 7.26(d, J=8.8Hz, 1H), 7.40(d, J=10.6Hz, 1H), 7.59(s, 8.27(dd, J=2.2, 5.8Hz, 1H), 8.77(d, J=7.3Hz, 9.08(s, Example 2-4 Compounds of Example 2 - Example 4 were synthesized in analogy to the procedure as described in Example 1, and intermediates were prepared by the method described as follows. 2-carboxylic acid 5 r-Hydroxymethyl-f uran-2 -carboxylic acid was synthesized by the method described in (Helv. Chim. Acta, 1926, 9, 10S8) . 48 acid nw 2NNaOH OH EtOH Isophthalic acid diethylester (9.100 g, 40.95 mmol) was dissolved in tetrahydrofuran (20ml). Thereafter, therein lithiumborohydride. (11.26 ml, 22.52 mmol, 2M tetrahydrofuran solution) was slowly added dropwise, and the reaction mixture was refluxed for 3 hours. Water was added to quench the reaction, and aqueous layer was extracted with ethyl acetate. Combined organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (hexane:ethyl acetate = 2:1), to give 5.76 g (yield: 77.1%, colorless liquid) of 3- hydroxymethylbenzoic acid ethylester. Ester compound (1.317 g, 7.311 mmol) prepared above was dissolved in ethanol (6 ml) . Thereafter, therein 2N NaOH aqueous solution (11.0 ml, 21.93 mmol) was slowly added dropwise, and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was neutralized with 1% HCl aqueous solution, 49 extracted with ethyl acetate, and washed with saturated NaCl solution. Combined organic layer was dried over anhydrous magnesium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (ethyl acetate:methyl alcohol = 5:1), to give 1.03 g (yield: 99.2%, white solid) of the target compound. H NMR (400MHz, CD3OD) : 54.66(s, 2H) , 7 . 4 4 ( t , J=7.7HZ, 1H) , 7.5G(d, J=7.7Hz, 1H) , 7.92(d, J-=7.7Hz, 1H) , 8.04(3, 1H) * Example 2; 5-nitrooxymethyl-furan-2-carboxylic acid- [ (7S) -1,2, 3-trimethoxy-10-methylsulfanyl-9-oxo-5, 6, 7,_9- tetrahydro-benzo [a] heptalen-7-yl] -amide Me.O;- -QM02 aH NMR (400MHz, CDG13) : 52. 37-2 . 51 (m, 3H) , 2.46(s, 3H) , 2.61-2.92(m, 1H) , 3.74(s, 3H) , 3.93(s, 3H), 3.98(s, 3H) , 4. 82-4. 85 (m, 1H) , 4.85(d, J=13.2Hz, 1H) , 4.90(d, 50 J=13.2Hz, 1H), 6.39(3, 1H) , 6.58(s, 1H) , 6.64(8, 1H) , 7.16(d, J=10.6Hz, 1H) , 7.42(d, J=10.2Hz, 1H) , 7.72 (s, 1H) , 8.99(s, 1H) Example _ 3j _ N- [ (7S) -3-isopropoxy-l, 2-^dimethoxy-10- methylsulf anyl-9-oxo-5 , 6 ,1 , 9-tetrahydrobenzo [a] heptalen-7-yl] -3 -nitrooxymethyl-benzamlde fylpO? Q,N'Q lll NMR (400MHz, CDC13) : 61,42 (t, ; J=6.6Hz, 6H), 2 . 2 2 - 2 . 2 7 ( m , 1H) , 2 . 3 4 - 2 . 4 9 ( s , 2H) , 2.54-2.57(m, 1H) , 3.75(s, 3H) , 3.96(3, 3H) , 4 . 59-4 . 63 (m, 1H) , 4.93-4.97(01, 1H) , 5.23(q, J=13.0H2, 2H) , 6.56(s, 1H) , 7.15(d, J=10.6Hz, Hi), 7. 18-7.25 (m, 1H) , 7.33(cl, J=O.OHz, 2H) , 7.46(d, J=10.2Hz, 1H) , 7.64(s, lH),7.71.(d, J=8.0Hz, 7.88 (s,. 1H) , 8.40 (s, 1H) 51 Example 4 : N- [ (_7_s) -3 -ethgxy_-1, 2 -dimethoxy-1 Qjmethylsulf anyl- Q-oxo- 5,6,7, 9 -tetrarahydrobenzo [a] heptalen-7-yl]-3-nitr-oxymethyl-benzamide •0" XH NMR (400MH2, CDC13) : 61.49(t, J=6.9Hz, 3H) , 2.17-2. 24. (m, 1H) , 2. 34-2. 47 (in, 211) , 2.45(s, 3H) , 2.49- 2.58(m, 1H) , 3.75(3, 3H) , U.iT/U, 311), 4 .12-4 .15 (m, 2H) , 4.89-4.96 (tn, 1H) , 5.24(q, J=12.0Hz, 2H) , 6.56(s, III), 7.15(d/ J=10.2Hz, 1H) , 7.21-7.25(m/ III), 7.31-7.35(01, 1H) , 7.41(d, J=10.6Hz, 1H) , 7.60(s, 1H) , 7.71(d, J=7.3Hz, 1H) , 7.78(s, 111), 0.23(3, 1H) 52 Example 5; Preparation of __6-nitrooxymethyl_-pyridine-2- carboxylic acid- [(75) -1,2,3-trimethoxy-lQmethylsulfanyl- 9-oxo-5, 6,7,9-tetrahydrpbenzolfa] heptal en - 7 -yl ] - amide carboxylic acid- [ (75) -l,2,3-trimethoxy-10- methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen -7-yl]-amide EDCI.DMAP MeCN MeS According to the similar procedure in the step l of Example 1l, by using 6-hydroxymethyl pyridine-2- carboxylic acid (23 mg, 0.17 mmol), 35 mg (yield: 53%, yellow solid) of the target compound was obtained. XH NMR (400MHz, CDC13) : 52.45(s, 3H), 2.33-2.50(m, 3H) 2.52-2.73 (m, 1H) , 3.75(s, 3H) , 3.92(s, 3H) , 3.97(s", 53 3H) , 4.28(d, J-14Hz., Ill), 4 , 4 4 ( d , J-l-lHz, 1H) 4 . f - ; G - - l . D 2 ( m , HI), C . b V ; , , , : : . . , . •, . 1 ' ( a , J- : 1 0 . 4 z . , 1.H) , V . . 7 - 7.48(m, 4H) , 7.73(3, 111), 9 . 80 (d, J^CHK, 111) carboxylic acid-1 l-iLLzi-JL-r': trimethoxy - I 0 - methylsul£anyl-9-oxo-5, 6 , 7 , 9 - tetrahydroObenzo [a] heptalen -7 - vlj - MeS PBl'3 CH-.CI, MeO—V/ --r t'Tf MeS Br A compound (50,45,mmol prepared in the step 1 was dissolved in dichlorometharie (6 ml) . Thereafter, therein tribromophosphine (PBr3, 0.005 ml, 0.05 mmol) was slowly added dropwise at 0°C, and the reaction mixture was stirred at room temperature for 3 hours. Methanol was added to quenen the reaction. Combined organic layer was dried over anhydrous magnesium sulfate, filtered arid and concentrated under reduced pressure. The residence was purified by column chromatography (chloroform:methanol = 99:1), to give 40 mg (yield: 71%, yellow solid) of the target compound. XH NMR (400MHz, cnc!3): 52 . 06- 2 . 14 (in, 1H) , 2.33- 2.41(m, 1H). , 2.4l(s, 3H) , 2. 46-2. 54 (m, 1H) 2 . 58-2 . 63 (m, 1H) , 3.72(s, 3H), 3.90(3, 3H) , 3.95(S,*3H), 4.55(d, J=2.8Hz, 2H) , 4. 77-4, 83 (m, 1H) , 6.57(s, 1H) , 7.04(d, •7-10. 4Hz, 1H) , 7.29(s, 1H) , 7.31(d, J-10.4HZ, III), 7.58(d, J=7.6Hs, 1H) , 7.78(t, J = 7 . GHz , • HI), 7.90(d, i7=»8.0Hz, 1H) , 8.53(d, J"=7.$Hz, 1H) carboxylic acid- [ (7S) -1, 2 , 3 -r.rimethoxy-lQmethylsulfanyl- 9-oxo-5,6,7,9-tetrahydrobenzo [a]heptalen -7-yl]-amide AgNOa MeCN MeS 'ON02 MeS A compound (40 mg, 0.070 utmol) prepared in the step 2 was dissolved in acetonitrile (3rnl) . Thereafter, therein silver nitrate (23 mg, 0.14 mmol) was slowly 55 added dropwise, and the reaction mixture was stirred at room temperature for 12 hours. The reaction mixture was washed with water and concentrated under reduced pressure. The residue was purified by column chromatography ( chloroform: met: hano 1 = 99:1), to give 17 mg (yield: 44.7%, yellow solid) of the target compound. XH NMR (400MHz, CDC13) : 52. 03-2.10 (m, 1H) , 2.33- 2.41(m, 1H) , 2.43(s, 3H) , 2.46-2.54(m, 1H) 2.58-2.65(m, 1H) , 3.73(s, 3H) , 3.92(3, 3H) , 3.95(S, 3H) , 4.77-4.83(m, 1H) , 5.63(d, i7=3.2Hz:, 2H) , 6.58(s, 1H) , 7.06(d, J=10.4Hz, 1H) , 1.21(3, 1H) , 7.31(d, J=»10.4Hz, 1H) , 7.53(d, J=7.6Hz, 1H) , 7.88(t, J=B.OHz, 1H) , 8.02(d, J=7.2Hz, 1H) , 8.39(d, J-7.2Hz, Example 6-24 Compounds of Example 6 - Example 24 were synthesized in analogy to the procedure as described in Example 5, and intermediates were prepared by the method described as follows. 56 2-carboxylic acid NaOH MeOH 6- Hydroxyme thyl- pyr idine-2-carboxy lie acid ethylester (200 mg, 1.1 rtimol) ( Thereafter, therein 2N NaOH aqueous solution (1 ml) was slowly added dropwise, and the reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was acidified (pH=3) with 2N HC1 . Solvent was concentrated under reduced pressure, then, dissolved inmethanol and filtered. The filtrate was concentrated under reduced pressure to give 150 mg (yield: 89%, white solid) of the target compound. XH NMR (400MHz, CD3OD) : 55.05(s, 2H) , 8.34(d, J=8.0Hz, 1H) , 8.47(d, J=8.0Hz, 1H) 8.73(d, J=8.0Hz, 57 2-carboxylic acid MeOH Me° )Me THF o 2N-NaOH, HO' MeOH Thiophene-2 , 5-dicarboxylic acid (4 g, 23.3 mmol) was dissolved in methanol (300ml) . Catcilytic amount of sulfuric acid was slowly added therein. The reaction mixture was re fluxed to give 3.8 g (yield: 81.7%, white solid) of thiophene-2 , 5-dicarboxylic acid dimethylester . The thiophene-2, 5-dicarboxylic acid dimethylester (3.7 g, 18.84 mmol) was dissolved in anhydrous tetrahydrofuran (50 ml) at room temperature under a nitrogen atmosphere. 2 . OM Lithiutnborohydrn.de tetrahydrofuran solution (5.5 ml, 11 mmol) was slowly added therein at 0°C. The reaction mixture was refluxed for 3 hours to give 2.1 g (yield: 64. 7 r , white solid) of 5-hydroxymethyl-thiophene-2-carboxy ic acid methylester . 5 -Hydroxymethyl -thiophene-2 -corboxylic acid methyl ester (2.1 g, 12.2 mmol) was dissolved in methanol (20 ml) . 2N NaOH aqueous solution (15ml) was slowly added therein. The reaction mixture was stirred 58 at room temperature for l hour to give 1.75 g (yield: 89%, white solid) of the target compound. 1H NMR (400MHz, CDC13): 53.90(Br, 1H) , 4.79(d, J-O.BHz,, 2H) , 6.97(d, J=4Hz, 1H) , 7 .66 (d, J=4Hz, 1H)J hydroxymethyl-benzoic acid XT F O: OF' 1 nU ^. urn'-^1. T.r 2-Fluoroisophthalic acid (3 g, 16.3 inmol) (J". Amer. Chem. Soc. , 1943, 65, 2308) was dissolved in methanol (150 ml) . Catalytic amount of sulfuric acid was slowly added therein. The reaction mixture was refluxed. The resultant was concentrated under reduced pressure to remove solvent, and dissolved in ethyl acetate. Combined organic layer was washed with saturated sodium carbonate, dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by column chrolmatography (ethyl acetaterhexane = 1:2), to give 3.1 g (yield: 08%, white solid), of 2-f luoro-isophthalic acid dimethylester. 59 Thiophene-2,5-dicarboxylic acid dimethylester (3.1 g, 14.6 mmol) was dissolved in anhydrous tetrahydrofuran (50 ml) at room temperature under a nitrogen atmosphere. 2.0M Lithiumborohydride tetrahydrofuran solution (4.4 ml, 8.7 mmol) • was slowly added therein at 0°C. The reaction mixture was refluxed for 3 hours. The reaction mixture was acidified with IN HCl aqueous solution, concentrated under reduced pressure to remove solvent, and extracted with chloroform. Combined organic layer was dried over anhydrous sodium sulfate, filtered, and solvent was concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane: methanol = 99:1}, to give 1.5 g (yield: 58%, white solid) of 2-f luor o -3- hydr oxyme thy 1- benzoic acid methylester. The 2-fluoro-3~ hydroxymethyl-benzpic acid methylester (1.3 g, 7i6 mmol) was dissolved in methanol (20 ml) . 2N NaOH aqueous solution (14 ml) was slowly added therein. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixture was acidified (pH=3) with 2N HCl. Solvent was concentrated under reduced pressure, then, dissolved in methanol and filtered. The filtrate was concentrated under reduced pressure to give 1.15 g (yield: 88%, white solid) of the target compound. 60 MMR (400MHz, DMSO-d6-) : 54.58(d, J=5.2Hz, 2H),5.37(t, J-=5.2HZ, 1H) 7.28(t, J=SHz, 1H) , 7.68(t, , 7.74(t, J=8Hz, Preparation __ of 3-£luoro-5- hydroxymethyl -benzole acid acid methyleater 5-Fluoroisophthalic acid dimetliylester (1.6 g, 7.54 mmol) (J. Org. Chem; 1969, 34, 1960-1961) was dissolved in tetrahydrofuran (15 ml) . Thereafter, therein 2.0 M lithiutnborohydr ide tetrahydrofuran solution (2.6 ml, 5.27 mmol) was slowly added dropwise at 0°C, and the reaction mixture was refluxed for 3 hours. The reaction mixture was acidified with IN HC1 aqueous solution, concentrated under reduced pressure to remove solvent, and extracted with chloroform. 61 Combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane : methanol = 99:1), to give 800 mg (yield: 57%, white solid) of the target compound . XH NMR (400MHz, CDC13): 63.92(d, J"=1.6Hz, 3H) , 4.75(d, J=4Hz, 2H) , 7.29-7.32(m, 1H) , 7.61(dd, J=9.2, I 1.6Hz, 1H) , 7.8(d, iJ=0.8ITz, benzoic acid MeO. NaOH MeOH According to the similar procedure in the method of intermediate 3, by using a compound prepared in the step 1, 1.6 g (yield: 94%, white solid) of the target compound was obtained. XH NMR(400MHz, CD3OD) : 5-1 . 66 (d, J=O.QHs, 211), 7.33-7.36(m/ 1H), 7.56-7.59(m, 1H), 7.03-7.84(ra/ 62 hydroxymethyl-benzole acid KMn04 HO H20, reflux 4-Fluoro-3-methyl-benzole acid (2.52 g, 16.346 tnmol) and potassium permanganate (10.33 g, 65.382 mmol) were dissolved in aqueous solution (300 ml) , and the mixture was refluxed for 1 day. The reaction mixture was filtered and the resultant solution was cooled down at room temperature, then cone. HC1 solution was added thereto. The produced solid was heated until it was completely melted. The temperature was lowered again into room temperature and then the solid was filtered, to give 2.08 g (yield: 69.1%, white solid) of the target compound. H NMR (400MHz, CD3OD) : 57.32(dd, 17=10.4, 8. GHz, 1H), 8.21-8.25(m, 1H) , 8.59(dd, J=7 . 0, .2 . 4Hz, 1H) 63 dimethylester O cat. H2SO4 OH MeO. MeOH, reflux ,OMe O A compound prepared in the step 1 (2.08 g, 11.29 mmol) was dissolved in methanol (30 ml) . Then, 10 drops of cone, sulfuric acid were added therein. The reaction mixture was refluxed for 1 day, neutralized with saturated sodium hydrogen carbonate aqueous solution, and extracted with chloroform. Combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure, to give 2.21 g (yield: 92.2%, white solid) of the target compound . XH NMR (400MHz, CDC13) : 53.96(s, 3H) , 3.97(s, 3H) , 7.22(dd, J-10.3, 8.8Hz, 1H) , 8. 20-8. 23 (m, 1H) , 8.64(dd, J=7.0, 2.2Hz, 64 benzoic acid methylester and 2-fluoro-5-hydroxymethylbenzoic acid methylester LiBH4 MeO THF. reflux A compound prepared in the step 2 (104.5 mg, 0.493 mmol) was dissolved in tetrahydrofuran solution (4 ml) . Then, 2M lithiumborohydride tetrahydrofuran solution (0.123 ml, 0.246 mmol) was slowly added therein. The reaction mixture was refluxed for 1 day. The reaction was quenched by water. Then, pH was adjusted to 5 with 1M HGl solution at 0°C. Extraction with ethyl acetate was performed. Combined organic layer was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by column chromatography (hexane:ethylacetate = 2:1), to give 4-fluoro-3- hydroxymethyl-benzoic acid methylester (45.4 mg, yield: 50.1%, colorless liquid) and 2-fluoro-5-hydroxymethylbenzoic acid methylester (15.4 mg, yield: 17.0%, colorless liquid). 65 4-fluoro-3-hydroxymethyl-benzoic acid methylester: XH NMR (400MHz, CDC13) : 52.73(t, J-S.lHz, 1H) , 3.90(0, 3H) , 4.78(d, J-5.1HZ, 2H) , 7.07(dd, JW9.2, 9.2Hz, 1H) , 7.93-7-.97(m, 1H) , 8.14(dd, J=7.1, 2.2Hz, 1H) 2-fluoro-5-hydroxymethyl-benzoic acid methylester: 1H NMR (4 0 0MHz , CDCl3) : 51.96(1;,. J*=4 . 4Hz , 1H} , 3.94(S, 3H) , 4.70(d, J-4.4Hz, 2H) , 7.13(dd, J=10.6, 8.4Hz, 1H) , 7.52-7.56(m, 1H) , 7.92(dd, J=7.0, 2.2Hz, 1H) acid 2N NaOl-l / MeOl-l - i-io According to the similar procedure in the method of intermediate 3, by using a compound prepared in the step 3 (1.074 g, 5.380 mmol), 0.906 g (yield: 91.3%, white solid) of the target compound was obtained. 66 XH NMR (400MHz, CD3OD) : 54.69(s, 2H) , 7.13 J=9.9, 8.8Hz, 1H) , 7. 90-7. 97 (m, 1H) , 8.16(dd, J=7.3, 2.2Hz, _ Pi-eparation _ of _ 2-fluoro-5- hydroxymethyl -benzole acid F 2N NaOH / MeOH 'Ma HO According to the similar procedure in the method of intermediate 3, by using 2-fluoro-5-hydroxymethylbenzole acid methylester prepared in the step 3 of intermediate 7, a target compound was obtained. XH NMR (400 MHz, CD3OD) : 54.61(s, 2H) , 7.17(dd, J-11.0, 8.4Hz, 1H) , 7.55-7.59(m, 1H) , 7.93(dd, J=7.l; 2.4Hz, 67 hydr oxyme t hy 1 - ben z ojic_ o NaOH / MeOH H| OH 5-Hydroxy-isophthalic acid raethylester (300 mg, 1.42 mmol) was dissolved in tetrahydrofuran (20 ml) at O°C under a nitrogen atmosphere. Lithium aluminium hydride (30 mg, 0.7 mmol) was added therein. The reaction mixture was stirred at room temperature for 3 hours. Water was added to quench the reaction, and aqueous layer was extracted with ethyl acetate. Combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane:methanol = 20:1). ABbove-obtained 3-hydroxy-5-hydroxymethyl-benzoic acid methyleater (170 mg, 0.93 mmol) was dissolved in methanol (1 ml) . IN NaOH aqueous solution (1 ml) was added therein. The reaction mixture was stirred at room temperature for 1 hour. The reaction mixtux'e was acidified (pH=3) with 2N HC1. Solvent was concentrated under reduced pressure, and then, dissolved in methanol 68 and filtered. The filtrate was concentrated under reduced pressure to give 150 mg (yield: 96%, white solid) of the target compound. XH NMR (400MHz, CD3OD) : 53.87(3, 3H),, 4.57(s, 2H) , 7.02(s, 1H) , 7.31(s, 1H) , 7.48(s, ^Intermediate 10> Preparation of 3,5-Ms-hydroxymethylbenzoic acid acid methylester OMe 1 eq. LAH HO' THF OMe Benzene-1 , 3 , 5-tricarboxylic acid trimethylester (1.010 g, 4.003 mmol) was dissolved in tetrahydrofuran (15ml) . Thereafter, the temperature was lowered into 0°C, and therein lithium aluminium hydride (0.160 g, 4.003 mmol) was slowly added, and the reaction mixture was stirred at room temperature for 3 hours. Water 69 (0.15 ml) and 15% NaOH aqueous solution (0.15 ml) were elowly added to quench the reaction. Aqueous solution (0.45 ml) was added again. Solvent was concentrated under reduced pressure. The residue was purified by column chromatography (hexane:ethyl acetate = 1:2), to • give 0.34 g (yield: 43.1%, colorless liquid) of the target compound. XH NMR (400MHz, CD3OD) : 53.91(s, 3H) , 4.66(s, 4H), 7.59(3, 1H) , 7.93(3, 2H) r acid According to the similar procedure in the method of intermediate 3, by using a compound prepared in the Step 1 (1.50 g, 7.65 mmol) , 0.617 g (yijfld: 44.3%, white solid) of the target compound was obtained. H NMR (400MHz, CD3OD) : 55.21(s, 4H) , 7.55(s, 7.92(3, 2H) 70 hydroxymethyl-benzole acid benzoic acid methylester HO. i) NBS, Benzoyl peroxide, CGI 2) H2O, Dioxane MeO. 2-Hydroxy-4-methyl-benzole acid methylester (166 mg, 1 mmol) was dissolved in CC14 (1.5ml). NBS (177 mg, 1 mmol) and benzoyl peroxide (5 mg, 0.02 mmol) were added therein. The reaction mixture was stirred at 70°C for 12 hours. Then, the reaction mixture was washed with water, dried over anhydrous sodium sulfate, and filtered. Solvent was concentrated under reduced pressure. The residue was purified by column chromatography (dichloromethane:hexane =1:4), to give 4-bromomethyl-2-hydroxy-benzoic acid methylester (130 mg, yield: 53%, white solid). The resultant compound i1 (130 mg, 0.53 mmol) was dissolved in aqueous solution (1.5. ml) and 1,4-dioxane (1.5 ml), and the mixture was stirred at 90°C for 12 hours. The reaction mixture was 71 extracted with chloroform. Combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (ethyl acetate:hexane - 1:4), to give 55 nig (yield: 57%, white Solid) of the barget compound. H NMR (400MHz, CDC13): 53.95(s, 311) , 4.71(d, J=6Hz, 2H) , 6.88(d, J=8Hz, III), 6.99 (a, IE), 7.82(d, J=8Hz, 1H) , 10.79(3, 1H) benzoic acid x--^ ^"^ NaOH MeOH/HoO According to the similar procedure in the method of intermediate 3, by using a compound prepared in the Step 1, 45 mg (yield: 90%, white solid) of the target compound was obtained.. 72 2-carboxcylic acid B; Preparation of 3-methyl-thiophene-_2- carboxylic acid i)n-Bu Li/Ether 2.5M n-Butyllithium (12.2 ml/ 30.55 nunol) was V, dissolved in diethyl ether (1 ml) at room temperature under a nitrogen atmosphere. 3-Methyl-thiophene (3 g, 30.55 mmol) dissolved in cliethyl ether was slowly added therein. The mixture was refluxed for 2 hours. Reaction chamber was cooled to 0°C and dry-ice was slowly added therein. Reaction was quenched with 45 ml of water, Diethyl ether layer was extracted and removed. Water layer was acidified with IN HCl solution and extracted with ethylacetate. Combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chroma tography 73 (hexane:ethyl acetate = 9:1), to give the target compound A (900 mg, white solid) and B (650 mg, white solid). carboxylic acid methyl ester 4-Methyl-thiophene-2-carboxylic acid (900 mg, 6.33 mmol) prepared in the step 1 was dissolved in methanol (15 ml) . Catalytic amount of sulfuric acid was slowly added therein. The reaction mixture was refluxed to .give 4 -methyl -thiophene -2 -carboxylic acid methylester (890 mg, yield: 90%, white solid) . 4- methyl -thiophene -2 -carboxylic acid methyl ester (200 mg, 1.28 mmol) and NBS (215 mg, 1.216 mmol) , and benzoyl peroxide of catalytic amount were dissolved in tetrachloromethaiie solution (5 ml) . The reaction mixture was refluxed (70°C) for 3 hours to give 165 mg (yield: 55%, white solid) of the target compound. T3. NMR(400MHz, CDC13) ; 53.89(s, 3H) , 4.46(s, 2H) , 7.49(S, 1H) , 7.80(s, 1H) 74 carboxylic acid fit • /—.Oil ' " H2Q/M-pio-xariB" Jr.' $ MeO.H/HizO A compound (150 mg, 0.638 mmol) prepared in the step 2 was dissolved in 1,4-dioxane (1.5 ml) and water (1.5 ml). Silver nitrate (130 mg, 0.765 mmol) was slowly added therein. The reaction mixture was stirred at room temperature for 12 hours to give 4- hydroxymethyl-thiophene-2-carboxylic acid methyl ester (60 mg, yield: 55%, white solid). This compound (60 mg, 0.348 mmol) was dissolved in methanol (1 ml) at room temperature. IN NaOH aqueous solution (1 ml) was slowly added therein, and the reaction mixture was stirred at room temperature for 1 hour, to give 50 mg (yield: 95%, white solid) of the target compound. 75 intermediate 13 > Preparation of 3-hydroxytnethylthiophene- 2-carboxylic acid carboxylic acid methyl ester E>f NBS,-Den?pyl peroxide;- , r - . - " H3CO . S Me.OH If S' CCL,; 1 Q .U Q According to the similar procedure in the method of step 2 of intermediate 12, by using 3-methylthiophene -2 -carboxyl ic acid (650 mg, 4.57 mmol) prepared in the step 1 of intermediate 12, 750 mg (yield: 79%, white solid) of the target compound was obtained. H NMR (400MHz, CDC13); 53.90(s, 3H) , 4.91(s, 2H) , 7.18(d, J=5.2Hz, 1H) , 7 . 4 6 ( d , J=5.2Hz, carboxylic acid 76 Br HUO/'M-Dfoxane: IT ':SX 'MeOH/.H2-0 ' • According to the similar procedure in the method of step 3 of intermediate 12, by using a compound prepared in the step 1 (750 mg, 3.19 mmol) , 210 mg (yield: 92%, white solid) of the target compound was obtained. acetic acid CaC.03; H20/Me.QH.- m-Tolyl acetic acid ethyl ester (1 g, 5.6 mmol), NBS (948 mg, 5.33 mmol) and benzoyl peroxide of catalytic amount were dissolved in tetrachloromethane (15 ml) . The reaction mixture was refluxed (70°C) for 3 hours to give (3-bromomethyl-phenyl)-acetic acid ethyl ester (GOO mg, yield: 42%, white solid). And (3- bromomethyl-phenyl)-acetic acid ethyl ester (130 mg, 0.50 mmol) and calcium carbonate (300 mg, 3 mmol) were 77 dissolved in water (2 ml) and 1,4-dioxane (2 ml). The reaction mixture was refluxed to give (3-hydroxymethylphenyl)- acetic acid ethyl ester (85 mg, yield: 87%, white solid). (3-Hydroxymethyl-phenyl)-acetic acid ethyl ester (85 mg, 0.43 mmol) was dissolved in methanol (1 ml) at room temperature. IN NaOH aqueous solution (1 ml) was added therein. The reaction mixture was stirred at room temperature for 1 hour, to give 65 mg (yield: 91%, white solid) of the target compound. H NMR(400MHz, CDC13) ; S3.66(s, 2H) , 4.57{s, 2H) , 7.21-7. 32 (m, 4H) •^Intermediate 15> Preparation of 3- (2-hydroxy-ethyl) - benzoic acid pec THF. NaOK M^a-LAlr * ^NX^k^W^^h |"|pf_J •f'NaHMDS. «)• (Metivoxyrnelhyl)triphe hyl rphbsplidniurh Chlafide" TftF BOH OMe M00 78 Isophthalic acid (5 g, 30 mmol) was dissolved in methanol (50 ml) . Catalytic amount of sulfuric acid was added therein. The reaction mixture was stirred at reflux for 12 hours to give isophthalic acid dimethyl ester (5.2 g, yield: 90%, white solid). Isophthalic acid dimethyl ester (5.2 g, 26.7rnmol) was dissolved in tetrahydrofuran (30 ml) . 2M Li thiumborohydr ide tetrahydrofuran (13 ml, 26.7 mmol) was added therein. The reaction mixture was refluxed to give 3- hydroxymethyl'-benzoic acid methyl ester (2.7 g, yield: 63%, colorless liquid), and 3-hydroxymethyl-benzoic acid methyl ester (200 mg, 1.20 mmol) was dissolved in dichloromethane (5 ml) . PCC (388 mg, 1.8 mmol) was added therein. The reaction mixture was stirred at room temperature for 3 hours to give 3-formyl-bensbic acid methyl ester (140 mg, yield: 72%, white solid). (Methoxymethyl)triphenylphosphonium chloride (770 mg, 2.24 mmol) was dissolved in tetrahydrofuran (5ml) under a nitrogen atmosphere. 1M NaHMDS tetrahydrofuran (2 ml, 2.04 mmol) was slowly added therein at -78"C, and the reaction mixture was stirred for 1 hour. 3-Formylbenzoic acid methyl ester (160 mg, 0.975 mmol) dissolved in tetrahydrofuran (2 ml) was slowly added therein. The reaction mixture was stirred at room 79 temperature for 24 hours to give 3- (2-methoxyvinyl) - benzole acid methyl ester (121 mg, yield: 65%, white solid). 3- (2-Methoxyvinyl) -benzole acid methyl ester (120 mg, 0.63 mmol) was dissolved in tetrahydrofuran (3 ml) . 4M HCl (2 ml) was added therein, and the mixture was stirred at room temperature for 24 hours, to give 3- (2-oxo-ethyl) -benzole acid methyl eater (60 mg, yield: 54%, white solid). 3- (2-Oxo-ethyl) -benzole acid methyl ester (60 .mg, 0.036 mmol) was dissolved in ethanol (2 ml). MaBIli (25 ing, 0 , G7 mmol) was added therein at 0°C, and the mixture was stirred at1 hour, to give 3- (2-hydroxyethyl) -benzole acid methyl ester (50 mg, yield: 84%, white solid). 3 - (2-Hydroxyethyl) - benzole acid methyl ester (50 mg, 0.28 mmol) was dissolved in methanol (1 ml) . IN NaOH aqueous solution (1 ml) was added therein. The reaction mixture was stirred at room temperature for 1 hour to give 43 mg (yield: 95%, white solid) of the target compound. 1II NMR(400MH2, CDC13) ; 62 . 32 (br, 1H) , 2.89(t, J=6.4HZ, 2H) , 3.85(t, J=6.4Hz, 2H) , 7.36(t, i7=7.6Hz, 1H) , 7.42(d, ,J=5.6Hz, 1H) , 7.88(d, .7=8. 8Hz, 1H) , 7. 89 (a, 1H) 80 Example 6: 5-nitrooxymethyl-thiophene-2-carboxylic acid- [ (7S) -1, 2 , 3-trimethoxy-lO-methylsxilf anyl-9-oxo- 5,6,7,9-tetrahydro~benzo[a]heptalen-7-yl]-amide MeS ZH NMR(400MHz, CDC13) ; 62 . 07-2 .11 (m, 1H) , 2.32- 2.45(01, 2H) , 2.45(s, 3H) , 2.53-2.56(m, 1H) , 3.71(s, 3H) , 3.91(3, 3H) , 3.96(S, 3H) , 4. 84-4. 91 (m, 1H) , 5.46(s, 2H) , 6.56(s, 1H) , 6.95(d, J=3.6Hs, 1H) , 7.13(d, J-10.8Hzf 1H), 7.37(d, J=10.8Hz, 1H)/ 7.58(d, J«4.0Hz, 7.61(S, 1H), 8.50(d, J=6.SHz, 81 Example 7 : N- [ (7S) -3-cyclopentyjoxy-1-2-dlmethoxy-10 methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo [a]heptalen-7-yl] -S-nitrooxymethyl-benzamlde fvleS ^•H NMR(400MHz, CDC13) ; 61. 61-1.73 (m, 2H) , 1.85- 2.00(m, 6H) , 2.21-2.65(m, 4H) , 2.44(s, 3H) , 3.75(s, 3H), 3.93(S, 3H) , 4 . 82-4.88 (m, 1H) , 4.?2-5.00(m, 1H) , 5.16(dd, J=12, 31.2H2, 2H) , 6.55(s, 1H) , 7.13-7.17 (m, 2H) , 7.28(d, J=6.4Hz, 1H), 7.41(d, J=10.8Hz, 1H) , 7.68- 7.70{m, 1H) , 7.72(s, 1H) , 7.78(s, 1H), 8.78(d, J»7.2Hz, 1H) Example 8: N- [ (75)-3-ethoxy-l,2-dimethoxy-lQmethylsulfanyl- 9-oxo-5,6,7,9-tetrahydro- 82 benzo[a]heptalen-7-yl3 -2-fluoro-3-nitrooxyTnethylbenzamide XH NMR(400MHz, CDC13) ; 51.49(t, J=6.9Hz, 3H) , 1.93-1.98 (m, 1H) , 2.31-2.39(m, 1H) , 2.43(e, 3H) , 2.46- 2.49(m, 1H) , 2.51-2.59 (m, l.H) , 3.73(s, 3H) , 3.96(s, 3H) , 4.11-4.14(111, 2H) , 4.83-4.86(01, 1H) , 5.53(d, J=12.8Hz, 1H) , 5.59(d, ,7=12.4Hz, 1H) , G.56(s, 1H) , 7.07(d, j=10.2Hz, 1H) , 7.10-7.15(m, III), 7.20-7.28(m, 1H) , 7.31(d, J-=10.2Hz, 1H) , 7.57(t, J"=6.4Hz7 1H) , 7.97(t, J=6.9Ez, IE) 83 Example 9; 2-fluoro-N- [ (7S) -3-isopropoxy-1,2-dimethoxy- 10-methylBulf anyl-9-oxo-5 , 6 , 7 , 9-tetrahydro— benzo[a]heptalen-7-yl]-3-nitrooxymethyl-benzamide Me:a *H NMR (400MHz, CDC13) ; 51.41(q, J=6.0Hz, 6H) , 2-1.99(m, 1H), 2.31-2.40(m, 1H) , 2.43(s, 3H) , 2.46- 2,52(m, 1H) , 2.55-2.60(m, 1H) , 3 . 7 2 ( s ; 3H) , 3.94(s, 3H) 4.57-4.63(m1, IE), 4.83-4.89 (m, Hi), 5.53(d, J=12.8Hz, 1H), ' 5.59(d, J=12.4Hz, 1H), 6.57(s, 1H) , 7.07(d, JV.10.6Hz, 1H) , • 7.15-7.19(m, 1H) , 7.23-7.27(m, 1H) , 7.34(d,. J=10.2Hz, 1H) , 7.56(t, J=7.1Hz, 1H) , 7 . 7 ( t , 84 Example 10: 2 -f luoro-g-i-i t-CKjxymgLh 1-- [(75) -l:,j, tritnethoxy- 10 -me thylsulf anyl-9-oxo_--_5, _6 , 7 , 9-tetrahydro benzo[a]heptalen-7-yl]-benzamide - NMR (400MHz, GDC13) ; 61. 95-2 . 02 (m, 1H) , 2.31- 2.54(m, 2H) , 2.43(s, 3H) , 2.59-2.63(m, 1H) , 3.73(s, 3H), 3.87(s, 3H) , 3.92(s, 3H) , 4.82-4.88(^1, 1H) , 5.51(d, J-=12.8Hz, 1H) , 5.60(d, .J=12 . QIIz, 1H) , 6.50(s, 1H) , 7.06(d, J=10.6Hz, 1H) , 7.22-7.27'(m, 2H) , 7.28(s, 1H) , 7.32(d, J=10.6Hz, 1H) , 7.54-7.58(m, 1H) , 7.93- 7.98(m, Example 11; N- [ (7S) -3-cycloperityloxy-1, 2 -dlmethoxy-10- methylsulfanyl-9-oxo-5,6,7,9-betrahydro- 85 benzo [a]heptalen-7'yl]-2-fluoro-3-nitrooxymethylbenzamide TH NMR(400MHz, CDC13) ; 51. 61-1. 73 (m, 2H) , 1.85- 2.00(10, 6H) , 2.35-2.52 (m, . 4H) , 2.42(s, 3H) , 3.72(s, 3H) , 3.93(3, 3H) , 4. 82-4. 88 (m, 2H) , 5.57(dd, J"=12 . 8 , 37.6HZ, 2H), 6.55(s, 1H), 7.05(d, J=10.4Hz, 1H), 7.09- 7.13(m, 1H) , 7.26(d, i7=7.2Hz, 2I-I) , 7.32(d, .7=10.4PIz, 1H), 7.55-7.59(m, 1H), 7.96-8.00(m, 1H) Example 12; S-fluoro-S-nitrooxymethyl-Jf-[ (7S)-1,2 , 3- trimethoxy-10-methylsul£anyl-9-oxo-5,6,7,9-tetrahydrobenzo [a]heptalen-7-yl] -benzamlclf 86 •fvleS F XH NMR(400MHz, CDC13); 62.31-2.45(m, 3H) , 2.46(3, 3H) , 2.59-2.63(m, 1H) , 3.75(g, 3H) , 3.93(s, 3H) , 3.98(S, 3H) , 4.90-4.95(m, 1H) , 5.11(dd, J=12.8, 45.6Hz, 2H) , 6.57(s, 1H) , 7.03(d, iJ=7 . 6Hz , Hi) , 7.19(d, iJ=8.8Hz, Hi) , 7.30-7.33(111, 1H) , 7.42(d, J=10.0Hz, 1H) , 7.55(s, 1H) , 7.68(s, 1H), 8.7S(d, J=7.2Hz, Example 13: N- [ (75) -3 - e L hqxy--1, 2-dimethoxy-10- methylsulfanyl-9-oxo-5, 6, 7, 9-tetrahydrobenzo[ a]heptalen-7-yl) -3-£luoro-5-nitrooxymethylbenzamide 'MeS 87 XH NMR(400MHz, CDC13) ; 51.50(t, J=6.9, 3H) , 2.33-2.42 (m, 3H) , 2.46(s, 311), 2 . 51-2 . 59 (m, 1H) , 3.75(s, 3H) , 3.98(s, 3H) , 4.11-4.16(m, 2H) , 4.90-4.93(m, 1H) , 5.05{d,. J=12.4, 1H) , 5.17 (d, J=12.4, 1H) , 6.56(s, 1H) / 7.03(d, J=7.3, 1H) , 7.20(d, J=10.6, 1H), 7.31(d, J=9.1, » 1H) , 7.45(d, J=10.6, 1H) , 7.55(s, 1H) , 7.69(s, 8.88(d, J=7.3, 1PI) Example 14: 3-fluoro-N-[(75)-3-isopropoxy-l,2- dimethoxy-lO-methylsulfanyl-g-oxo-S,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl]-5-nitrooxymethyl-benzamide LH NMR(400MHz, CDC13) ; 61. 37-1. 44 (m, 6H) , 2.34-2.41(m, 3H), 2.46(s/3H), 2.51-2.58(m, 1H), 3.75(s, 3H) , 3.96(3, 3H) , 4. 57-4. 62 (m, III), 4. 91-4. 95 (m, 1H) , 5.05(d, J=12.4, 1H), 5.17(d, J=12.4, 1H), 6.56(s, 1H), 7.03(d, J=8.4, 1H), 7.20(d, J=10.6, 1H) , 7.31(d, J=7.3, 1H) , 7.45(d, J=10.6, III) , 7 . 5 : 5 ( 3 , 1H) , 7 . 7 0 ( s , 1I-I) , 8.91(d, J-7.3, III) Example 15; N- [ (75) -3-cyclopentyloxy-l,2-dimethoxy-10- methyleulfanyl-9-oxo-5,6,7, 9-tetrahycIrobenzo[ a]heptalen-7-yl3 -3-fluoro-5-nitrooxyjnethylbenzamide WeS 0. •QNO>> XH NMR(400MHz, CDC13) ; 61.64-1.69(m, 2H) , 1.83-1.98(m, 6H) , 2.35-2.43(m7 3H) , 2.46(s, 3H) , 2.51-2.58(m, 1H) , 3.74(s, 3H) , 3 . 9 4 ( s , 3H) , 4 . 8 2 ~ 4 . 8 4 ( m , 1H), 4.91-4. 94 (m, III) ,. 5.04(d, ,7=12.4, III), 5.16(d, J-12.4, 1H) , 6.55(S j III), 7 . 0 2 ( d , J=8.4, 1H) , 7.20(d, J=10.6, 1H) , 7.31(d, J=7.3f 1H) , 7.45(d, J=10.2, 7.55(S, 1H) , 7.70(s, III), 8.93(d, J=6.9, 1H) 89 Example 16: 4-fluoro-3-nitrooxymethyl-N- [ (75) -1, 2, 3- trimethoxy-lO-methylsulfanyl-9-oxo-5,6,7,9-tetrahydro- *beh2oIa]heptalen.-7-yl] -benzamide H NMR (400MHz, CDC13) ; 62 . 22-2 .51 (m, 3H) , 2.46(s, 3H)2.55-2.63(m, 1H) , 3.76(s, 3H) , 3 . 9 2 ( s , 3H) , 3 . 9 8 ( s , 3H)4.90-4.97 (m, 1H) , 5.10(d, J=12.1Hz, 1H) , 5.35(d, J=12.1Hz, 1H) , 6 . 5 7 ( s , 1H), 6 . 8 6 ( d d , J=9.2, 8.8Hz, 1H), 7.18(d, J=10.3Hz, 1H) , 7.43(d, J=10.3B2, 1H) , 7.72(s, 1H) , 7.76-7.80 (m, 1H) , 7.91(dd, J=6.80, 2.2Hz, HI), a.93(d, J=7.2Hz, Example 17: 2-fluoro-5-nitrooxymethyl-JJ'- [ (7S) -1,2,3- tritnethoxy-lO-methylsulfanyl-g-oxo-S, 6,7,9-tetrahydrobenzo [a]heptalen-7-yl]-benzamide 90 •f XH NMR(400MHz, CDC13) ; 51.94-2.01(m, 1H) , 2.31- 2.54{m, 2H) , 2.43(s, 3H) , 2 . 50-2 . 63 (m, 1H) , 3.73(s, 3H) , 3.92(S, 3H) , 3.97(s, 3H) , 4.81-4.87 (m, 1H) , 5.36(s, 2H) , 6.5.7(8, 1H) , 7.06(d, J=10.3Hz, HI), 7.16-7.28 (m, 2H) , 7.26(s, 1H) , 7.33(d, J=10.3H-z, I I I ) , 7 . 51-7.55 (m, I I I ) , 7.99(dd, j=7.3, 2.6Hz, II!) Example 18: 3-hydroxy-5-nitrooxymethyl-J7- [ (75) -1, 2, 3- triTnetho3cy-10-methylsulfanyl-9-oxo-5, 6 , 7 , 9-tetrahydrobenzo [a]heptalen-7-yl] -benzamide MeS ON.09' 91 4H NMR(400MHz, CDC13) ; 52 . 30-2 . 43 (m, 6H) , 2.56- 2.57(m, 1H) , 3.51(s, 3H) , 3 . 9 0 ( s , 311) , 3.92(s, 3H) , 4.82-4.85(m, 1H), 5.00(dd, J=30.8Hz, 12.8Hz, 2H) , 6.56(s, 1H) , 6.62(s, 1H) , 7.11(s, 2H) , 7.16-7.19(m, 2H) , 7.41(d, J=10.8Hz, 1H), 7 . 6 6 ( s , 1H), 8.57(brs, 1H, NH), 8.78(brs, 1H, OH) Example 19: 3,5-bia-nitrooxymebhyl-Jf- [ (7S) -1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5, 6,7, 9-tetrahydrobenzo[ a]heptalen-7-yl]-benzamide ON.O-> XH NMR (400MHz, CDC13) ; 52 . 29-2 . 51 {m, 3H) , 2.46(3, 3H), 2.60-2.62(m, 1H), 3.77(s, 3H), 3.93{s, 3H), 3.99(s, 3H) , 4.94-4.98 (m, 1H) , 5.18(s, 411), 6.58(s, 1H) , 7 . 2 0 ( d , J=10.6Hz, 1H) , 7.26(s, III), 7.45(d, J=10.6Hz, 1H) , 7-.77(s, 2H) , 7.81(8, 1H) , 9.12(d, J=7.0Hz, 1H) Example 20 : 2-hydroxy-4-riitrooxymethyl-N- [ (7S)-1, 2, 3 - trimethoxy-10-methyl3ulfanYl-9-oxo-5 , 6 , 7 , g-tetrahydrobenzo[ a]heptalen-7-yl]-benzamide MeS a'H NMR (400MHz, CDC13); 52 . 25-2 . 63 (m, 4H) , 2.47(s, 3H), 3.74(s, 3H) , 3.91(s, 3H) , 3.97{S, 3H), 4.85-4.92(111, , 5.27{d, J=4.8Hz, 2H), 6.53(d, , 7.63(s, IE) , 7.72(d, J=8Hz, Hi), 9.13(d, ^7=6.8112, 1H) , 12.29(3, 1H) Example 21: 4-nitrooxyrnethyl-t hiophene - 2 - carboxyl i c acid [ (7S) -1,2, 3-trimethoxy-l0-methylsulf ariyl-9-oxo- 5,6,7,9-tetrahydro-benzo[a]heptalen-7-yl]-amide 93 Hi NMR(400MHz, CDC13); 52.21-2.29(m, 1H) , 2.34- 2.46(m, 5H), 2.51-2.58(m, 1H), 3.71(s, 3H) , 3.91(s, 3H), 3.96(5, 3H), 4.88-4.95(m, 1H) , 5.18(dd, J-12.8, 20Hz, 2H), 6.56(s, 1H) , 7.17(d, J=10.4Hz, 1H) , 7.30(s, 1H) , j7.41(d, J=10.4Hz, 1H) , 7.61 (a, III), 7.74(s, 1H) , 8.81(d, j=6.4Hz, 1H) Example 22; 3-nitrooxymethyl-thiophene-2-carboxylic acid [(7S)-1,2,3-trimethoxy-10-methylsulfanyl-9-oxo- 5,6,7,9-tetrahydro-benzq[a]heptalen-7-yl]-amide 94 XH NMR (400MHz, CDC13) ; 52 . 04-2 .17 (m, 1H) , 2.30- 2.45(m, 5H) , 2.57-2.64(m, 1H) , 3.73(s, 3H) , 3.91(s, 1H) , 3.96(S, 3H) , 4.67-4.88(m, 1H) , 5.66(d, J-=»13.6Hz, 1H) , i 5.80(d, J=13.6Hz, 1H) , 6.56(s, 1H) , 7 . 02 (d, .7=4.4112, 1H) , 7.08(d, J=l0.4Hs, Hi), 7.26(d, iJ=4.4Hz, 1H) , * 7.29(d, i7=10.4Hz, 1H) , 7.43(s, 1H) , 7.45(d, J=7.2H2, 1H) Example 23 ; 2- (3-nitrooxymethyl-phenyl)-N-[(7S) -1,2,3- trinlethoxy-lQ-Tnethylsulfanyl-9-oxo-5, 6, 7, 9~tetrahydrobenzo [a] heptalen-7-yl-] -acetamide NMR(400MHz, CDC13) ; 51.82-1.95(m, 1H) , 2.21- 2.28(m, 1H) , 2.31-2.42(01, 1H) , 2.45(s/3H), 2,47-2.53(m, 1H), 3.51(d, J=14.0Hz, 1H) , 3.64(s, 3H) , 3.66(d, J=14.0Hz, 1H) , 3.88(s, 3H) , 3.93(s, 3H) , 4.66-4.72(m, 5.40(s, 2H) , 6.51(s, 1H) , 7.11(d, J=10.4Hz, 7.24-7.38(tn, 5H) , 7.48(3, 1H) , 7.90(3, J=7.2Hz, 1H) 95 Example 24: 3-(2-nitrooxy-ethyl)-N-[(75)-1,2,3- trimethoxy-lO-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl] -benzamide MeS NMR (400MHz, CDC13) ; 52 . 05-2 .11 (m, 1H) , 2.17- , 2H) , 2.46(s, 3H), 2.56-2.60(m, 1H), 2.81-2.91(m, 2H), 3.69(s, 3H) , 3.91(s, '3H), 3.97(s, 3H) , 4.45(t, J=6.a,Hz, 2H) , 4.89-4.95(m, 1H) , 6.56(s, 1H) , 7.12- 7.24(m, 3H), 7.39(d, J=10.4Hz, 1H), 7.61-7.64(m, 2H), 7.69(8, 1H) , 8.22(d, J-=6.8Hz, 1H) 96 Example 25: Preparation of 3 -nihrooxy- benzole acid-S- [(7S) -1,2, 3-trimethoxy-10-Tnethylsulfanyl-9-oxo-5, 6,7,9- tetrahydro-benzo [a] heptalen-7-yl-carbamoyl] -pyridine-2 - l -Methyles t er _[_{7S) - 1 ,2 , 3 - trime thoxy- 1 0 -me t hyJLsu- If a n.yl- 9 -oxo- 5 ,6,7, 9 ^ tetrahydro-benzo [a] heptalen-7 -yl-carbamonyl] -pyridine- 2~yl } -methylester MeO WeO MeO A compotond prepared in step 1 of the Example, i (100 mg, 0.19 mmol) was dissolved in 3 uifi of dlchlororaethane. 3-(Chloromethyl) benzoylchloride (0.030 ml, 0.21 mmol) and triethylamine (0.082 ml, 0.59 mmol) were slowly added therein, and the mixture was reacted at room temperature for 10 minutes. Water was. added to quench the reaction, arid aqueous layer was 97 extracted with dichlorornethane. Combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatoqraphy (ethyl acetate) , to give 99 mg (yield: 79%, yellow solid) of the target compound . aH NMR (400MHz, CDC13) : 62 . 14-2 .16 (m, 1H) , 2.30- 2.39(m, 1H) , 2. 43-2. 45 (m, 1H) , 2. 48 (a, 3H) , 2. 68-2. 96 (m, 1H) , 3.67(s, 3H) , 3.90(sf 3H) , 3.91(s, 3H) , 4.83-4.93(m, 1H), 4.88(8, 2H) , 5.51(8, 2H) , 6.77(s, 1H) , 7.27(s, 1H) , 7.52(t, J=7.7Hz, 1H) , 7. 65-7. 70 (m, 2H) , 8.05(d, J=7.7Hz, 8.14(8, 1H) , 8.28(d, J=10.6Hz, l H ) , - 9 . 0 2 ( s , [(7S) -1 ,2,3 -trimethoxy- 10 -methylsulfanyl-g-oxo-S, 6,7,9 - fcetrahydro-benzo [a] heptalen-7-yl-car-bamoyl] -pyridine-2- yl } -methylester // 1)Nal, acetone 2)AgNO3, MeCN 98 A compound prepared in the step 1 (90 mg, 0.13 mmol) and sodium iodide (31 mg, 0.20 mmol) were dissolved in 3 ml of acetone, and the mixture was reacted at room temperature for 1 day. Water was added to quench the reaction, and aqueous layer was extracted with ethyl acetate. Solvent was concentrated under reduced pressure. The reaction concentrate and silver nitrate (3 Omg, 0.045 mmol) were dissolved in 5 ml of • acetonitrile, and the mixture was reacted at room temperature for l hour. Water was added to quench the reaction, and aqueous layer war; extracted with ethyl acetate. Solvent was concentrated under reduced pressure. The residue was purified by short column nhromatography (ethyl acetate) and PLC, to give 26 mg [yield: 29%, yellow solid) of the target compound. XH NMR (400MH2, CDC13) : 52 . 09-2 .17 (m, 1H) , 2.35- 2.41(m, 2H) , 2.44(s, 3H) , 2.55-2.58(m, 1H) , 3.74(s, 3H) , 43.91(8, 3H) , 3.96(3, 3H) , 4.92-4.95 (m, 1H) , 5.45(8, 2H) , 5.47(s, 2H) , 6.56(s, 1H) , 7.13(d, J=10.2Hz, 1H) , 7.35(t, J-9.1HZ, 2H), 7.50(t, J=Q.5Hz, 1H), 7.54(s, 1H), 7.62(d, J=7.3Hz, 1H) , 8.14(d, J=B.3Hz, 2H) , 8.24(dd, J«2.2, 6.2Hz, 1H, ArH), 8.36(d, J-6.9Hz, 1H) , 9.09(s, 1H) 99 Example 26~34 Compounds of Example 26 - Example 34 were synthesized in analogy to the procedure as described in Example 25, and intermediates were prepared by the method described as follows. intermediate 16> Preparation of 2-hyebco3cy-N- [ (7S) - 1,2/3-trimsthoxy-10-methylsulfanyl-9-oxo-5,6,7,9- tetrahydro-benzo [a]I heptalen-7 - yl] - be nz amide MeC .. ,, x EDCI, HOBt, Et3N J—4 ] n I OH DMF MeS ° MeS ° 2-Hydroxy-benzoic acid (203 mg, 1.47 mmol) , EDCI (385 mg/ 2.01 mmol) and HOBt (271 mg, 2.01 mmol) were dissolved in dimethylforniamide (10 ml) . Triethylamine (0.37 ml, 2.67 mmol) was added therein, and the mixture was stirred at room temperature for 1 day. 7-Aminol, 2,3-trimethoxy-10-methylsulfonyl-6,7-dihydro-5Hbenzo[ al-heptalen-9-one (500 mg, 1.34 mmol) was added 100 therein, and the mixture was stirred at room temperature for l day. Water was added to quench the reaction, and aqueous layer was extracted with diethyl ether. Combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (hexane: ethyl acetate = 1:5) and recrystallized in methanol, to give 516 mg (yield: 78%, yellow solid) of the target compound. XH NMR (400MHz, CDCl3) : 52 . 14-2 . 17 (m, 1H) , 2.31- 2.37(m, 1H) , 2.40-2.44(3, 1H) , 2.45(s, 3H) , 2.56-2.60(m, ev 3.74(S, 3H) , 3.91(8, 3H) , 3.97(s, 3H) , 4.85- , 6.56(8, 1H) , 6.64(d, J=7.7Hz, 1H) , 6.77(d, , 7.15(d, J=10.6Hz, 1H) , 7.22(d, J-7.3H3, 1H) , 7.38(d, J=10.6Hz, III), 7.57(s, 1H) , 7.71(d, J=8.0Hz, 1, 2,3-trimethoxy-lO-methylsulf anyl-9-oxo-5, 6,7,9- tetrahydro-benzo [a] heptalen-7-yl 3 -benzamide 101 MeQ EDCI, HOST •OH / DMF MeS Me! According to the similar procedure in the preparation method of intermediate 16, by using 3- hydroxybenzoicacid (497 mg, 1.33 mmol), 558 mg (yield: 85%, yellow solid) of the target compound was obtained. XH NMR (400MHz, DMSO-d6): 62.07-2.15 (m, 1H) , 2.10- 2.35(m, 6H) , 3.55(s, 3H) , 3.80(s, 3H) , 3.84(s, 3H) , 4.53-4.56(m, 1H) , 6.81(s, 1H) , 6.92 (d, J-6.8HZ, 1H) , 7.10(8, 1H), 7.15-7.31(m, 5H) , 8.98(d, J-7.6H2, 1H, - NH), 9.70(s, 1H, -OH) Example 26; 4-nitrooxybutyric acid-5- [ (_7S) -1,2,3- trimethoxy-lO-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo [a] heptalen-7-Yl-cairbainoyl ] -- p y r i d .1 tie - 2 - y 1 - methylester 102 aH NMR (400MHz, CDC13) : 52 . 06-2 . 16 (m, 1H) , 2.08- 2.11(m, 2H) , 2.31-2.49{m, 1H) , 2.45(3, 3H) , 2.57(t, J-7.1HZ, 2H), 3.75(s, 3H), 3.91(s, 3H) , 3.96(s, 3H) , 4.52(t, J=6.4Hz, 2H) , 4.91-4.94(m, 1H) , 5.21(s, 2H) , -6.56(8, 1H) , 7.13(d, J=i0.y][-., 1H) , 7.21-7.25 ( m , 1H) , 7.38(01, J=10.6Hz, 111} , 7.52(s, 1H) , 8.22 (a., J-8.0HZ, 1H), 9.05(s, 1H) 27 : 3 -nitrooxymethyl -benzoicacid- 6- [ (7S) -1,2,3- triraethoxy-10-methylsulfanyl-9-oxo-5, 6,1, 9-tetrahydrobenzo[ a]heptalen-7-yl-carbamoyl]-pyridine-2-ylmethylester XH NMR (400MH2, CDCla) : SI. 84-1.92 (m, 1H) , 2.33- 2.60(m, 1H), 2.43(s, 3H), 2.46-2.54(m, 1H) 2.55-2.60(m, 1H), 3.73(S, 3H) , 3.92(3, 3H) , 3.96(3, 3H) , 4.74-4 .81 (m, 1H), 5.51(s, 2H) 5.57(d, J=4.0Hz, 1H) , 6.56(s, HI), » 7.04(d, J=10.4Hz, 1H) , 7.25(d, J=10.4Hz, 1H) , 7.54- 7.60(m, 2H), 7.65(d, J=7.GHz, 1H), 7.87(t, J-8.0HZ, 1H), 7.99(d, J-=7.6Hz, 1H) , 8.21(s, 2H) , 8.22(8, 1H) , 8.41(d, J«7.2Hz, Example 28; 4-nitrooxybutyric acid-6- [ (7S)-1,2,3- trimethoxy-10-methylsul£anyl-9-oxo-5,6,7,9~tetrahydrobenzo[ a]heptalen-7-yl-carbamoyl]-pyridine-2-ylme, thyl ester MeS XH NMR (400MHz, CDC13) : 52 . 02-2 . 09 (tn, 1H) , 2.11- 2.18(m, 2H), 2.21-2.40(m, 2H), 2 . 4 3 ( s , 3H) , 2.47-2.56(m, 1H) . 2.63(t, J=7.2Hz, 2H) , 3.73(s, 311), 3 . 9 2 ( s , 311), 104 3.96(S, 3H) , 4.57(t, J=6.4Hz, 2H) , 4.77-4.83(m, 5.30(d, J-A.8llz, 2H) , 6.58(s, 1H) , 7.05(d, J=10.4Hz, 1H), 7.25(S, 1H) , 7.32(d, J=10.4Hz, 1H) , 7.52(d, J-8.0HZ, 1H) , 7.84(t, J=a.OHz, 1H) , 7.98(d, J=8.0Hz, 1H) , 8.45(d, i7=6.8Hz, IB) Example 29; 3-nitrooxymethyl-benzole acid-2-[(7S)- 1,2,3-trimethoxy-10-methylsu1fanyl-9-oxo-5,6,7,9- tetrahydro-benzo[a]heptalen-7-yl-carbamoyl]-phenylester •Me XH NMR (400MHz, CDC13): 51. 48-1.55 (m, 1H) , 1.83- 1.89(m, 1H) , 2.22 -2.3 9 (m, 211), 2.42(s, 3H) f 3.62(s, 3H) , 3.87(8, 3H) , 3.92(g, 3H) , 4 .63-4 . 70 (m, 1H) , 5.54(s, 2H) , 6.47(s, 1H) , 6.89(d, J=7.4Hz, 1H) , 7.03(d/ J=10.2Hz, 1H), 7.17(s, 1H) , 7.23(d, J=10.1Hz, 1H) , 7.35(t, J-7.3HZ, 1H) , 7.54(t, J=7.4Hz, III), 7.58(t, J=7.6I!::, 1H) , 7.72(d, J=8.0I-I2, III) ,7.03 (cl, J=7.7Hz, 1H) , 8.26(m, 2H) 105 Example 30: 4-nitrooxybutyric acid-2-[(7S)-1,2,3- trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl-carbamoyl] -phenylesfrer XH NMR (400MHz, CDG13) : 51.60-1.90 (m, 1H) , 2.10- 2.14(m, 2H) , 2.26-2.39(m, 1H) , 2.44(s, 3H) , 2.46-2.51(m, 1H) .2.56-2.61(01, 1H) , 2.80(t, J=7.1Hz, 2H) , 3.70(s, 3H), 3.92(s, 3H) , 3.96(s, 3H) , 4.55(t, J=6.2Hz, 2H) , 4.74-4.81 (m, 1H) , G.57(s, 1I-I) , 6 . 82 (d, J=6.9Hz, IK), 7.08(d, J=10.6Hz, 1H) , 7.13(d, J=9.1Hz, 1H) , 7.23(s, 1H), 7.28-7.36 (m, III), 7.47 ( t , J=9.9Hz, 1H) , 7.68(d, J=9.5Hz, 106 Example 31: 3-nitrogoxyrnebhyl-benzole acid-3- [ (7S) - 1,2,3-trimethoxy-10-methylsulfanyl-9-oxo-5,6,7,9- tetrahydro-benzo [a]heptalen-7-yl-carbamoyl]-phenylesber XH NMR (400MHz, CDC13) : 62 . 05-2 .15 (m, 1H) , 2.23- 2.59(m, 6H) , 3.75(s, 3H) , 3.91(3', 3H) , 3.97(s, 3H) , 4,90-5.00(01, 1H), 5.49(8, 2H) , 6.56(3, 1H) , 7.03(d, J-10.4HZ, 1H) , 7.27(s, 2H) , 7.34(d, J-8.8HZ, 2H) 7.53- 7.65(m, 2H), 7.76(3, 1H) , 7 . 7 7 ( d , J-6.4HZ, 1H), 8.13(m, 3H) Example 32; 4-nitrooxybutyric acid-3-[(7S)-1,2,3- trimethoxy-lO-methylsulfanyl-S-oxo-S,6,7,9-tetrahydrobenzo[ a]heptalen-7-yl-carbamoyl]-phenylester 107 XH NMR (400MHz, CDC13) : 62.04-2.18 (m, 3H) , 2.25- 2.60(m, 6H) , 2.65(t, iT=6.8Hs, 211), 3.74 (a, 3H) , 3.91(5, 3 H ) , - 3 . 9 6 ( s , 3H) , 4 . 5 6 ( t , J-6.4HZ, 2H) , 4 . 8 7 - 4 . 9 4 ( m , 1H), .6.56(3, 1H) , 7.08-7.12 (m, 2H) , 7.26-7.35(m, 2H) , 7.48(S7 1H) , 7.59(8, 1H) , 7.7l(d/ J=8.0Hz, 1H), 7.75 (d, J=7.6Hz, 1H, -NH) Example 33; 3-nitrooxymethyl-benzole acid-3-[ (7S)- 1/2,3-triTnethoxy-lO-methyl-sulfanyl-9-oxo-5y 6,7, 9- tietrahydro-benzo [a] heptalen-7-yl-carbamoyl] -benzylester "0- 108 XH NMR (400MHz, CDC13) : 52 . 05-2 .18 (m, 1H) , 2.31- 2H) , 2.40(s, 3H) , 2.55-2.59 (m, 1H) , 3.75(8, 3H) , 3.91(s, 3H) , 3.97(s, 3H) , 4 . 90--1 . 96 (m, 1H) , 5.24(s, 211} , 5.44(s, 2H) , 6.56(s, 1H) , 7.09(d, J=10.6Hz, 7.27(dd, J=7.S, 7.6Hz, 1H), 7.36(d, J=10.6Hz, » 7.44(d, J"=7-.6Hz, 1H) , 7.45(dd, J=8..0, 7.6HZ, 7.58(d, J=8.0Hz, 1H) , 7.59(s, 1H) , 7.79(d, J=7.6Hz, 1H) 7.91(3, 1H) , 8.05(cl, J=7.6IIa, I I I ) , 0 . 0 6 ( 3 , 1H) , 8.10(d, J=7.2Hz, Example 34; 4-nitrooxybutyric acid-3- [(7S)-1,2,3- trimethO3cy-10-methylsulfa.nyl-9-oxp-5, 6 , 7, 9-tetrahydrobenzo[ a]heptalen-7-yl-carbamoyl]-benzylester .MeO 'B; JH NMR (400MHz, CDC13) : 52 . 00-2 .14 (m, 3H) , 2.31- 2.51(01, 4H) , 2 . 4 4 ( s , 3H) , 2 . 5 6 - 2 . 6 0 ( m , 1H) , 3.75(s, 3PI) , 3.92(8, 3H) , 3.97(s, 3H) , 4.49(t, J=6.2HZ, 2H) , 4.89- 4.95(m, 1H) , 4.99(d, J=12.4Hz, 1H) , 5.04(d, J-12.4HZ, 109 6.56(s,'lH), 7.11(d, J=10.6Hz, 1H) , 7.26(dd, J=7.6, 7.6H2, 1H) , 7. 35-7. 37 (m, 211), 7.54(s, 1H) ; 7.76(d, J-B.OHz, 1H) , 7.81(S, 1H) , 8.00(d. J=7.6Hz, 1H) Example 35 ; Preparation of 2 -nitrosothio-N- [ (7S) -1 , 2 , 3 - trimethoxy- 10 -me thyl sul f anyl-9-oxo-5, 6,7, 9-tetrahydrobengo [a] heptalen- 7 -yl] -benzamide trimethoxy-10 -me thyl sul fanyl - 9 - oxo -5,6,7,9 -1 e trahydro - benzo[a]heptalen-7-yl]-benzamide M OH SH , 2) Et3N, CH2CI2 Excess thionylchloride was added in thiosalycilic acid (115 mg, 0.75 mmol) , and the mixture was stirred with heating for 1 day. The reaction mixture was concentrated under reduced pressure to remove thionylchloride, and to give a chloride compound. 7- 110 amino-l,2,3-trimethoxy-10-methylGulfonyl-6,7-dihydro- Eff-benzo [a]-hepcalen-9-one (243 mg, 0.62 BBlol) was dissolved in purified dichloromethane. Triethylamine (0.26 ml, 1.86 mmol) was slowly added therein. Thiosalycilic chloride dissolved in dichloromethane was also added therein at 0°C, and the mixture was stirred for 30 minutes. Water was added to quench t&ft reaction, and aqueous layer was extracted with chloroform. Combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (chloroform: me thanol = 12:1), to give 210 mg (yield: 66%, white solid) of the target compound. XH NMR (400MH2, CDCl3) : 52. 06-2 .17 (m, 1H) , 2.33- 2.49(m, 2H) , 2.42(s, 3H) , 2.54-2.59(m, 1H) , 3.72(s, 3H) , 3.92(S, 3H) , 3.96(8, 3H) , 4.86-4.93 (in, 1H) , 6.56(s, 1H), 7.06(d, J=10.6Hz, 1H) , 7.14(t, J=7.5Hz, !7.24(d/ J«=7.7Hz, 1H) , 7.31(d, J=10.6Hz, 7.51(s, 1H) , 7.58(d, J=7.3Hz, Hi), 7.63(t, J=8 2H) 111 trimetho3cy-10-Tnethylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo [a] heptalen-7-yl] -benzamide r~\ I MeO ^^' 1A/-HCI. NaN02 Me°H MeS MeS Compound prepared in step 1 (40 mg/ 0.078 mmol) was dissolved in methanol. 1 N HC1 aqueous solution (3 ml) was added therein. Sodium nitrite (NalNO2, 6.5 mg, 0.094 mmol) dissolved in water (1.5 ml) was also added therein; and the mixture was stirred at room temperature for 1 hour. Sodium hydrogen carbonate was added to quench the reaction, and aqueous layer was extracted with chloroform. Combined, organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (chlorbform:methanol 12:1) and recrystallized in methanol, to give 34.1 mg (yield: 81%, yellow solid) of the target compound. 112 XH NMR (400MHz, CDC13) : S2 . 07-2 .14 (m, 1H) , 2.31- 2.37(m, 1H), 2 . 4 2 ( s , 3H), 2 . 4 5 - 2 . 4 8 ( m , 1H), 2.54-2.59(ra, 1H) , 3.72(s, 311), 3.91(s, 311), 3.96(s, 3H) . 4.88- 4.94(m, 1H) , 6.56(s, 1H) , 7.06(d, J=10.6Hz, 1H) , 7.10(t, J=7.5Hz, 1H) , 7.21(t, J=6.9Hz, 1H) , 7.31(d, J»10.6Hz, » 1H), 7.54(8, 1H) , 7.64(t, J=8.6Hz, 2H) , 7.72(d, J-7.3HZ, 1H) Example 36; Preparation of 3-nitroBooxytnatfcarly-[ (7S)- l, 2,3-trimethoxy-10-methylsulf anyl-9-oxo-5, 6,7,9- tetrahydro-benzo [a] heptalen -7- yl ] - ben garni de Me -M-x 1) Nal, Acetone 2) AgNO2, MeCN MeS 3-Chloromethyl-W- [(7S)-1,2,3-trimethoxy-10- methylsulfanyl-S-oxo-S, 6, 7, 9-tet.rahydrobenzo [a]heptalen -7-yl] -benzamide (119.8 mg, 0.22.8 mmol) and sodium iodide (136,5 mg, 0.911 mmol) were dissolved in acetone (15 ml) , and the mixture was stirred at 55°C for 1 day. The reaction mixture was extracted with chloroform and 113 washed with saturated sodium chloride aqueous solution. Combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue and silver nitrite (125.5 mg, 0.812 mmol) were dissolved in acetonitrile (5 ml) , and the mixture was stirred at room temperature for 1 day. The reaction mixture was extracted with chloroform. Combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (ethyl acetate:chloroform = 4:1), to give 35.4 mg (yield: 32.4%, yellow solid) of the target compound. XH NMR (400MHz, CDC13) : 52 . 35-2 . 55 (m, 3H) , 2.48{s, 3H) , 2.62-2.66 (m, 1H) , 3.76(s, 311) , 3.84(d, •J=12.ailz, 1H), 3.92(s, 3H) , 3.98(3, 3H) , 4.26{d, J=»12.8Hz, 1H) , 4.91-4.97(m, 1H) , 6.58(a, 1H) , 6.96(dd, J^7.7, 7.7Hz, 1H), 7.22(d, J=10.6Hz, 1H) , 7.26-7.30 (m, 2H) , 7.44- 7.47(m, 2H) , 7.86(s, 1H) , 9.30(rl, J=6 114 Example 37 ; 3 -f luoro-S-iiitrosooxymethyl-jy- [ (75) -1, 2 ,.3 - t r ime thoxy-10 - me thy 1 sul f anyl - 9 - oxo -5,6,7,9- tetrahydr o - benzo[a]heptalen-7-yl] -benzamide A target compound was synthesized in analogy to the procedure as described in the Example 36. •.Meo; Me a H NMR (400 MHz, CDC13) : 52 . 04-2 . 20 (m, 1H) , 2.47(s, 3H), 2. 52-2. 62 (m, 2H) , 2. 67-2. 72 (m, 1H) , 3.81(s, 3H) , 3.91(S, 3H), 3.97(s, 3H), 4.71(s, 2H) , 4.93-4.96(m, 1H), 6.42(d, J=6.2, 1H), 6.58(s, III), 7.14-7.21 (m, 2H) , 7.30(d, J=8.4, 1H), 7.42(s, 1H), 7.53(d, J=10.6, 115 Example 38; Preparation of 3-nitrosothiomethyl-jy- [ (75) - 1,2,3-trimethoxy-10-methylsulfanyl- 9-oxo-5,6,7,9- tetrahydro-benzo[a]heptalen-7-yl]-benzamide 1,2,3-trimethoxy-10-methylsulfanyl- 9 -oxo-5,6,7,9- tetrahydro-benzo[a]heptalen-7-yl-carbamoyl]-benzyl ester M MeS MeQ MfiO O MeO MeS MsO 3-Hydroxymethyl-N-[(7S)-1,2,3 -trimethoxy-10- methylsul£anyl-9-oxo-5,6,7,9-tetrahydrobenzo [a]heptalen-7-yl]-benzamide (252.5 rag, 0.497 mmol) was dissolved in dichloromethane (10 ml), and the temperature was lowered into o°C. Methanesulfonylchloride (42.4 ìl 0.547 mmol) and triethylamine (0.104 ml, 0.745 mmol) were added therein, and the mixture was stirred at room temperature for 2 116 hours. The reaction mixture was extracted with chloroform. combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (ethyl acetate: chloroform a 3:2), to give 182.3 mg (yield: 62.6%, yellow solid) of the target compound. XH NMR (400 MHz, CDC13) : 62 . 14-2 .21 (m, 1H) , 2.31- 2.50(m, 2H) , 2.46(3, 3H) , 2. 57-2. 62 (m, 1H) , 2.85(s, 3H) , 3.76(S, 3H) , 3.92(s, 3H) , 3. 97 (a, 3H) , 4.87-4.94(m, 1H) , 5.06(d, i7=12.7Hz, III), 5.10(d, J-.i2.7Hz, 1H) , 6.57(s, 1H) , 7.14(d, J-10.6HZ, HI), 7.28(dd, J=7 .7 , 7.7HZ, 1H) , 1.38-7. 42 (m, 2H) , 7.56(s, 1H) , 7.'74(d, J=8HZ1H) , df J=7.7Hz, 1, 2, 3-trimethoxy- 10 -methylsulfanyl- 9-oxo-5, 6, 7,9- tetrahydro-benzp [a] heptalen-7-yl-carbamoyl] -benzyl ester 117 MeQ A compound prepared in the step 1 (162.3 mg, 0.311 mol) was dissolved in acetone (6 ml) , and the temperature was lowered into 0°C. Potassium thioacetate (53.2 mg, 0.467 mmol) was slowly added therein at 0°C, and the mixture was stirred for 1 hour. The reaction mixture was extracted with chloroform. Combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure, to give 173.6 mg (yield: 90.6%, yellow solid) of the target compound. *H NMR (400MHz, CDC13) : 52 . 05-2 . 14 (m, 1H) , 2.30(s, 2H) , 2.32-2.49(m, 211), 2.44(0, 311), 2 . 52-2 . 61 (m, III), 3.74(s, 3H), 3.92(s, 3H) , 3.97(s, 3H) , 4.0l(d, 47=13. 9Hz, 1H) , 4.05(d, JXL3.9HZ, 1H) , 4. 87-4. 93 (m, |£) , 6.56(s, 7.10(d, J=10.3Hz, 1H) , 7.21(dd, J=7.7, 7.7H2, .36(m, 2H) , 7.49(s, 1H) , 7. 64 -7. 71 (m, 3H) 118 trimethoxy-10-methylsulfanyl-9-oxo-5,6,7, 9-tetrahydrobenzo [a] heptalen-7-yl] -benzaml.de MeS HS A compound prepared in the step 2 (165.2 mg, 0.292 mol) was dissolved in methanol (6 ml) , and the temperature was lowered into o"0. Sodium thiomethoxide (21.5 mg, 0.307 mmol) was slowly added therein at 0°C, and the mixture was stirred at room temperature for 30 minutes. Reaction was quenched by adding 0.1N HCl aqueous solution. The reaction mixture was extracted with chloroform and washed with saturated sodium chloride aqueous solution. Combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (ethyl acetate:chloroform = 1:1), to give 182.3 mg (yield: 62.6%, yellow solid) of the target: compound. 119 H NMR (400MHz, CDC13): 51.70(t, J-7.3HZ, HI), 2.12-2.19(m, 1H) , 2.31-2.50(m, 2H) , 2.18(s, 3H) , 2.57- 2.61(m, 1H) , 3.58(d, J-=7.3Hz, 1H) , 3.'74(s, 3H) , 3.92(s, 3H), 3. 97 (a, 3H) , 4 .89-4 .95 (m, 1H) , 6.57(s/ 1H|, 7.12(d, |7-10.3HZ, 1H) , 7.19(dd, J"=7.7, 7.7Hz, 1H) , 7.34(d, r»7.7H2, 1H), 7.37(d, J-10.3II7., Ill), 7.55(s, I I I ) , 7.59(d, J=7.7Hz, 1H) , 7 . 7 0 ( s , 1H) , 7,93(d, J=7.3Hz, HI) 1,2,3-trimethoxy-lO-methylsulf any 1 - 9 - oxo -5,6,7,9- tetrahydro-benzo [a] heptalen-7-yl] -benzamide MeQ 1N-HCI/NaN02 MeOH O MeS MeS A compound prepared in the step 3 (101.2 mg, 0.193 tnol) was dissolved in rnethanol (3 ml) and dimethylformamide (3ml) , and the temperature was lowered into 0°C. 0.1N HCl aqueous solution (3 ml) and 120 sodium nitrite (16.0 mg) were slowly added therein at 0°C, and the mixture was stirred at room temperature for 2 hours. The reaction mixture was extracted with Chloroform and washed with saturated sodium carbonate solution. Combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (ethyl acetate:chloroform:hexane 3:2:1.5), to give 19.5 mg (yield: 18.3%, yellow solid) of the target compound. aH NMR (400MHz, CDC13) : 52.08-2 .16 (mf 1H) , 2.31- 2.51(m, 2H) , 2.45(8, 3H), 2.57-2.61(m, 1H) , 3.57(s, 3H), 3.80(s, 2H), 3.92(s, 3H) , 3.97(s, 3H) , 4.89-4.96(m, 1H) , 6.57(S, 1H), 7.10(d, J=10.3Hz, 1H) , 7.25(dd, J=7.7, 7.7HZ, 1H) , 7.36(d, J=10.3I-Iz, 1.11), 7.38(d, J=7.7Hz, III), 7.53(3, 1H) , 7.68(d, J=7.7Il2'./ 111), 7.77(8, 1H) , 7.79(d; i7«7.0Hz, 121 Example 39; 3-fluoro-5-nitrosothiomethyl-N- f (78) -1,2,3 - trimethoxy- 1 0 -me thy 1 sul f anyl - 9 - oxo -5,6,7,9- te trahydro - benzo [a] heptalen-7-yl] -benzamide A target compound of Example 39 was , synthesized in analogy to the procedure as described iti^trHBi Example 38. McD- •.Me H NMR (400MHz, CDC13): 62.30-2.48(m, 3H) , 2.44(s, 3H), 2.56-2. 59 (m, 1H) , 3.57(q, J=13.9, 16.5, 2H) , 3.73(s, 3H), 3.91(3, 3H) , 3.97(s, 3H) , 4.97-5.03(m, 1H) 6.57(8, 1H), 7.02(d, J=8.4, 1H) , 7.15(d, J«10.6, 1H) , 7.39(d, J=10.2, 1H) , 7.43(s, 1H) , 7.64 (d, J=9.1, HI), 7.73(s, 1H) , B.96(d, J=7.3, III) 122 Example 40; Preparation of 3-fluorg-j-nitrooxymethyl-M- [(7S) -l,2,3,10-tetrametho:Ky-9-o:x:o-5, 6,7, 9-tetrahydrobenzo[ a]heptalen-7-yl]-benzamide a]heptalen-7-yl]-benzamide DH :DMF Jpeacetylcolchicine (150 mg, 0.42 mmol), 3-fluoroithyl- benzoic acid (85 mg, 0.50 nvmol) and HOBt (67 mg, 0.50 mmol) were dissolved in dimethylformamide solution (2 ml) , and the temperature was lowered into 0"C. EDCI (95 mg, 0.50 mmol) was slowly added therein at 0°C, and the mixture was Stirred at room temperature. Water was added to quench *^ ' • the reaction, and aqueous layer was extracted with ethyl acetate. Combined organic layer was dried over anhydrous sodium sulfate, filtered and 123 under reduced pressure. The residue was purified by column chromatography (chloroform: ethyl acetate = 2:1), to give 110 mg (yield: 52%, yellow solid) of the target compound. 1H NMR (400MHz, CDC13) : 52 . 40-2 . 58 (m, 3H) , 2.62- 2.69(tn, 1H) , 3.64-3.70 (in, 1H) , 3.74(s, 3H) , 3.92(s, 311), 3.98(8, 3H) , 4.07(S, 311), 4 . 23-4 . 20 (m, 1H) , 4 .81-4 . 88 (m, 1H), 6.58(a, 1H) , 6.87(d, J=9.2Hz, 1H), 6.97(d, J=9.2Hz, 1H), 7.03(d, i7=10.4Hz, 1H) , 7.38(8, 1H) , 7.50(d, •7-10.4Hz, 1H) , 7.96(s, 1H) , 9.64 (A, J=6.0Hz, 1H) [ (78) -1,2,3,10-tetramethoxy-9_-oxo-5_/ 6,_7 / 9-tetrahydrobenzo[ a]heptalen-7-yl]-benzamidc MeO •^QFf MeO According to the similar procedure in the step 2 and 3 of Example 5, by using a compound prepare in the 124 step 1 (90 mg, 0.10 mmol), 30 mg (yield: 30%, yellow solid) of the target compound was obtained. H NMR (400MHz, CDC13) : 52.30-2.49(m, 3H) , 2.58- 2.59(m, 1H) , 3.75(8, 3H) , 3.92(s, 311)3.98(8, 311), 4.05(S, 3H) , 4.89-4.93(m, 1H) , 4.94(d, J=12.8Hz, 1H) , 5.10(d, ,J=12.eHz:, 1H) , 6.58(s, 1H) , 6.96 (d, J-8.4H2, 1H) , 7.01(d, J=10.4Hz, 1H) , 7.17(d, J=8.4Hz, 1H) , ,7.45(8, 1H) , 7.48(d, J=10.4Hz, 1H) , 7.89{s, 1H) , 9.25(d, Example 41; Preparation of 3-nitrooxymethyl-i^i^thyl-N- [ (75) -l^^-trimethoxy-lO-methylsulfanyl-g-oxo-S^,?, 9- tetrahydro-benzo [a] heptal «3n~_7_-yl_] - be viz amide Preparation of 3-chloromethy|^i^|ethyl-N- *-l,2,3-brimethpxy-10-methylsulfanyl-9-oxo-5<.6> tetrahydro-benzo[a] heptalen-7-yl]-benzamide PJridih.il- MC Ma Mei 125 Thiodemecolcine (50 mg, 0.129 mmol) and pyridine (0.012 ml, 0.154 mmol) were dissolved in dichloromethane, and the temperature was lowered into O°C. 3-(Chloromethyl)benzoyl chloride (0.022 ml, 0.154 mmol) was slowly added therein at 0°C, and the mixture was stirred at room temperature. Water was added to guench the reaction, and aqueous layer was extracted with ethyl acetate. Combined organic layer was dried over anhydrous sodium sulfate, filtered and concentrated under reduced pressure. The residue was purified by column chromatography (chloroform:ethyl -acetate solid) of the target compound. aH NMR (400MHz, CDC13) : 52.24-2.38 (m, 2H), 2.44(s, 3H),2.45-2.58(m, 1H) , 2.61-2.74(m, 1H) , 3.25{s, 3H) , 3.72(8, 3H) , 3.91(s, 3H) , 3.92(s, 3H) , 4.54(s, 2H) , 5.05(br, 1H) , 6.57(3, 1H) , 7.06(3, J=10.0Hz, 1PI) , 7.09(S, 1H) , 7.22-7.41(m, 511) 126 [(7S)-1,2,3-trimethoxy-lQ-methylsulfanyl-9-oxo-5y6,7,9- tetrahydro-benzo[a3heptalen-7-yl3-benzamide iJNal/Acetone •ii)AaN03/;c:H3CN According to the similar procedure in the step 2 of Example 25, by using a compound prepared in the step 1 (45 mg, 0.085 mmol), 40 mg (yield: 83%, yellow solid) pf the target compound was obtained. XH NMR {400MHz, CDC13): 52.24-2.38(m, 2H), 2.44(a; 3H) ,2.45-2.58 (m, 1H) , 2 . 61-2 . 74 (in, Hi), 3.24(s, 311), 3.71(8, 3H) , 3.91(s, 311), 3.92(s, 3H) , 4.77(br, 1H) , 5.58(s, 2H), 6.85(s, 2H), 7.21(d, J=10.0Hz, 1H), 7.29{d, J-10.4HZ, 1H), 7.40-7.53(m, 4H) Example 42: Preparation of 3 -fluoro-N-methyl-5- taltrooxymethyl -N- [ (75) -1,2, 3-l:rimethoxy-10- 127 mebhylsulfanyl-9-oxo-5,6,7,9-tetrahydrobenzo [a]heptalen-7-yl]-benzamide N- [(75)-1,2,3-trimethoxy-10-methylsulfanyl-9- oxo-5,6,1, 9-tetrahydro-ben2O[a]heptalen-7-yl}-benzamide MoO HO >=0 i)Bhyl diloroforma(e,.TEA ' lOPjfriiljha .MP HO' 3-Pluoro-5-hydroxymel:hyl-benzoic acid (52 mg, 0.309 mmol) was dissolved in dichloromethane (3 ml) under a nitrogen atmosphere. Ethylchloroformate (0.022 ml, 0.231 mmol) and TEA (0.042 ml, 0.309 mmol) were Slowly added therein at 0°C, and the mixture was Stirred at 0°C for 30 minutes. Pyridine (0.012 ml, 0.154 mmol) and thiodemecolcine (60 mg, 0.154 mmol) were also added therein, and the mixture was stirred at room temperature f or 3 hours. Water was added to guench the reaction, and aqueous layer was extracted with ethyl acetate. Combined organic layer was dried over anhydrous sodium sulfate, filtered and 128 concentrated under reduced pressure. The residue was purified by column chromatography (chloroform: ethyl acetate = 2:1), to give 30 rag (yield: 21%, yellow solid) of the target compound. aH NMR (400MHz, CDC13) : 51.76(br, 1H) , 2.24-2.38(m, 2H) , 2.44(s, 3H) ,2.45-2.58 (m, III), 2 . 61-2 . 74 (m, Hi), 3.23(8, 3H) , 3.72(s, 311), 3.93(s, 3H) , 3.97(s, 3H) , 4.65(s, 2H) , 5.02(br, 1H) , 6.57(8, 1H) , 6.92(s, 1H) , 6.99-7.24 (m, 4H) , 7.34(s, III) nitrooxymethyl-N- [ (7S) -1,2,3-trimethoxy-lOmethylsuIf anyl-9-oxo-5,6,7, 9-1etrahydrobenzo [a] heptalen-7 -yl] -benzamicle ^CH- 0'. According to the similar procedure in the step 2 and 3 of Example 5, by using a compound prepared in the 129 step 1 (55 mg, 0.101 mmol) , 20 mg (yield: 37%, yellow solid) of the target compound was obtained. XH NMR (400MHz, CDC13) : 52. 22-2 .41 (m, 2H) , 2.45(s, 3H) , 2.45-2. 58 (m, 1H) , 2 . 61-2 . 74 (in, 1H) , 3.24(s, 3H) , » 3.75(s, 3H) , 3.91(8, 311), 3.95 ( a , 3H) , 5.02(br, 1H) , 5.39(s, 2H) , 6.57(s, 1H) , 7 . 0 5 ~ 7 . 2 4 ( m , 5H) , 7.34(s, Example 43 : _Prepcirati_o_n. of 2- (3-£luoro:1 bnitrooxymethyl- phenyl) -N- [ (7S) -l, 2, S-trimethQanr-lOmethylsulfanyl- 9-oxo-5,6, 7, 9-tetrahydrobenzo[ a]heptalen-7 -y1]- acetamide acetic acid Matt THF KCM, iQ-'Crovvn-B.- Br — —- - MeO: BOH/:-H2o; H0_, OH A compound prepared in the step 1 of intermediate 6 (2.5 g, 13.57 mmol) was dissolved in dichloromethane 130 (30 ml). PBr3 (1.15 ml, 12.21 mmol) was slowly added therein, and the m.ixture wan stirred at room temperature for 3 hours to give 3-bromomethyl-5-fluorobenzoic acid methyl ester (1.6 g, yield: 50%, white solid). 3-Bromomethyl-5-fluoro-benzoic acid methyl » ester (1.5 g, 6.1 mmol), KCM (l.G g, 24.4 mmol) and 18- crown-6 (820 mg, 3.1 rmnol) were dissolved In acetonitrile (10 nil) . The re si CM. ion mixture was stirred at room temperature ior 10 houra to give 3-cyanomethyl- 5-fluoro-benzoic acid methyl ester (900 mg, yield: 77%, white solid). 3-Cyanomethyl-5-fluoro-benzoic acid methyl ester (900 mg, 4.65 mmol) was dissolved in tetrahydrofuran (10ml) . 2M Lithiutnborohydride tetrahydrofuran (2.3 ml, 4.6 mmol) was slowly added therein, and the mixture was re fluxed, to give (3- fluoro-5-hydroxymethyl-phenyl)-acetonitrile (430 mg, yield: 56%, white Kc.licl) . (3 1'luoro-5-hydroxymethylphenyl) -acetonitrile (400 ny, 2.42 mmol) and potassiumhydroxide (1.34 g, 23.8 mmol) were dissolved in ethanol (10 ml) and water (5 ml) . The reaction mixture was re fluxed for 24 hour;:: to give 356 mq (yield: 80%, white uolid) of the target compound. 1H NMR(400MHz, CD3OD); 53.62(s, 2H), 4.59(3, 2H), 6.94(d, J=9.6Hz, 1H), 7.04(d, J=9.6Hz, 1H), 7.08(s, 131 N-[(7S)-1,2,3-trimethoxy-lO-methylsulfanyl-9- oxo-5,6,7,g-tetrahydro-benzo[a]heptalen-7-yl3-acetamide Me OH >+ V fr HO^ :0 EDCI,.H0Bt DMF MeS According to the similar procedure in the step 1 of Example 40, by using a compound prepared in the step 1 (30 mg, 0.16 mmol) , 58 mg (yield: 70%, yellow solid) of the target compound was obtained. :H NMR (400MHz, CDC13) : 51 . 92-2 . 01 (m, 1H) , 2.19- 2.40(m, 2H) , 2.4l(s, 3H) , 2 . 46-2 . 52 (m, 1H) , 3.4S(d, J=14.0,Hz, 1H) , 3.52(d, J=14.0,Hz, 1H) , 3.63(s, 3H) , 3.89(s, 3H) , 3.91(0, 3H) , 4 . 64-4 . 74 (m, 1H) , 4.75(s, 2H) , 6.53(s, 1H) , 6.86-6.94(m, 3H) , 7.07(d, J-=10.8Hz, III), 7.13(s, 1H) , 7.30(d, J=10.8Hz, 1H) , 7.39(s, 132 5-[(7S)-(l,2,3-trimethoxy-lO-methylsul£anyl-9-oxo- 5,6,7, 9 - tet r ahydro - ben20 [a ]lie;p t: ci1 on ~ 7-ylcarbamoyl) - methyl]-benzyl ester MSCI;TEA MC. 'Mi According to the similar procedure in the step 1 of Example 38, by using a compound prepared in the step 2 (58 mg, 0.107 mmol) , GO mg (yield: 90%, yellow solid) of the target compound was obtained. phenyl) -N- [ (7S) -.1 ,_2 , 3 - triiuethoxy-10 - me thy 1 sul f anyl - 9 - oxo-5,6,7,9-tetrahydro-benzo[a] heptalen-7-yl]-acetamide 133 Me'p 'i)|\lal/Acetone- MeQ-if — According to the similar procedure in the step 2 of Example 25, by using a compound prepared in the step 3 (60 mg, 0.097 mmol) , 30 nig (y:u-:ld: 83%, yellow solid) of the target compound was obtained. H NMR (400MHz, CDC13) : 51. 92-1.98 {m, 1H) , 2.22- 2.42(m, 2H) , 2.46(s, 311), 2.49-2.52(m, 1H) , 3.50(d, J=14.0,Hz, 1H) , 3.651::;, 3ii:, 3 . b6 (d, iJ=14 . 0 , Hz 1H) , 3.89(s, 311), 3.93(s, 311), 4. 64-4. 74 (m, 111), 5.37(s, 2H) , 6. 52 (a, 1H) , 6.97(d, Jr=8.8Hz/ 1H) , 7.11- 7.17(m, 3H) , 7.35(d, J=10.8Hz, 1H) , 7. 50 (a, 1H) , 8.04(d, J=7 Example 44 ; 2- (2-f luoro-5-niti-ooxymethyl-phenyl) -N- [{7S) -l/Z/B-trimethoxy-lO-methylsulfanyl-g-oxp-S, 6, 7, 9- tetrahydro-benzo [a ] hep I; a 1 e 1 1 - 7 - y 1 ]_-acetaamide A compound of Example 44 was synthesized in t analogy to the procedure as described in Example 43, 134 and an intermediate was prepared by the described as follows. Me XH NMR (400MHz, CDC13) : 61. 09-1. 93 (m, 1H) , 2.26- 2.29(m, 1H), 2.34-2.40(m, Hi), 2.43(8, 3H), 2.49-2.54(m, 1H) , 3.61(s, 311), 3.G2(d, .T. l-;, . lil, h i 1 H ) , 3.72(0, J=15.2Hz, 1H) , 3.89(s, 311), 3.92(0, 311), 4.70-4.75(m, 1H) , 5.49(s, 2H) , 6.53(s, 1H) , 7.06-7.11(01, 2H) , 7.25- 7.31(01, 2H) , 7.35(t, J=7.6Hz/ 1H) , 7.44(s, 1H) , 7.57(d, J«7.2Hz, hydroxymethyl-phenyl)-acetic acid MeO XOH 135 According to the similar procedure in the step 1 of Example 43, by using a compound prepared in the intermediate 5 (2.2 g, 11.94 mmol), 400 mg (yield: 90%, white solid) of the target compound was obtained. Experimental Example 1: Cytotoxicity test to cancer cell lines Cytotoxicity to A549 (Korea Research Institute of Chemical Technology) , SK-OV-3 (Korea Research Institute of Chemical Technology), SK-MEL-2 (Korea Research Institute of Chemical Technology), HCT-15 (Korea Research Institute of Chemical Technology) and MCF7 (Korean Cell Line Bank, Seoul National University School of medicine} cells was measured by Sulforhodamin-B (SRB) method (1909, National Cancer Institute (NCI)) which was developed for the measurement of in vitro anticancer activity of a drug. Cells were separated using 0.25% trypsin-EDTA solution, leading to the preparation of a cell suspension by 5xl03 - 2xl04 cells/well. Then, the suspension was distributed into a 96 well plate by 100 ìl/well, which Was cultured in a 37°C, 5% CO-, incubator for 24 hours. Compounds prepared in examples of the present invention were used as a sample. Precisely, the compound way 136 dissolved in dimethylsulfoxide and diluted with RPMI 1640 medium before being uso.d a.s a sample. The final concentration of the sample used varied ranging 1 ìM ~ 0.00001 ìM. The medium was removed from the 96 well plate and then diluted sample solution was added by 100 ìl/well, followed by further culture in a 37 C, 5% C02 incubator for 48 hours. Tz (time zero) plate was collected from the point of adding the sample. Upon completing the culture, the medium was removed from each well along with Tz plate, then 10% trichloroacetic acid (TCA) was added by 100 ìl/well. The plate was left at 4 C for 1 hour to let cells be fixed on the floor of the plate. After cells were fixed completely, the plate was washed with water 5-6 times to remove the remaining trichloroacetic acid solution completely and moisture was completely dried at room temperature. 0.4% Sulforhodamine-B was dissolved in 1% acetic acid solution to prepare a staining solution. Cell.s were stained for 30 minutes with a dye added by 100 ^ to each well of the completely dried plate. Then, the plate was washed with 1% acetic acid solution 5-6 times to remove sulforhodamine-B remained uncombined with cells. Then, the plate was dried at room temperuture. 10 mM Tris solution was added by 100 ìl/well thereto to dissolve 137 the dye, and optical density (OD) was measured ' at 520 nm with a micro plate reader. ED50 (concentration that inhibits cancer cell growth 50%, 50% effective dose., nM/ml) of the sample to cancer cell was calculated aa follows. OD value at the point of beginning the culture with the sample was determined as Tz (time zero) value. OD value of a well to be cultured without the cample was determined as a control value (C) . OD vaJue of a well pretreated with the sample was determined as experimental value (T) . After calculating T?. , C and T, cytotoxicity of the sample was measured by the: below ^Mathematical 'Formula T£ Tz, (T-Tz) / (C-Tz) .x 100 T > Tz., (T-T2.) /Tz -A 100 ED50, the concentration that can inhibit cancer cell growth by 50%, wan calculated by using, a regression analysis of .lotus program based on the degree of cytotoxicity obtained by the 138 Each EDSO of paclitaxed, doxorubicin and colchicine, which were used as controls, was also calculated by the same way as described in the above. Results were presented in Table 1. Example Paclitaxel Doxorub i c i n Colchicine Example 1 Example 2 'Example 3 Example 4 Example 5 Example 6 Example 7 Example 8 Example 9 Example 10 Example 11 Example 12 Example 13 Example 14 Example 15 Cytotoxicity [ED50 : nM] A549 0.3 13 .0 21.0 1.8 15.8 2.4 0. 01 .1. . 0 0.70 >50 0.02 0.49 0 .20 >50 0.05 0.13 28 .2 30.2 SK-OV-3 1.2 47.0 18.0 1.0 9.3 4.0 0 . 01 1 . U 0.37 >50 0.04 0.34 0.23 >50 0.27 0.10 30.6 >50 SK-MEL-2 0.1 16.0 6.0 0.4 ' 5.4 1.0 0 . 0 1 .1 . 1 0 .08 >50 0.01 0 .22 0 .12 >50 0.11 0.04 32. 0 22.5 HCT15 0.1 25.0 9.0 0.6 2.3 4.8 0.03 4.4 0.27 >50 0.03 0.64 0.39 >50 0.05 0.11 9.9 23.4 MCF-7 0.9 50.0 NT NT 2.9 2.3 0.01 NT 0.16 >50 0.01 0.13 0.09 39.0 0.03 0.02 11.2 18.8 139 OH os e-Z.fr S'OI O'S .IN e foi •IN L'TrZ T O ' O ,IN •LN ,IN UN iN IN JIN £N £N e-e T Z 'O O T ' O S T ' T 2'8 J.JST 80'0 IM OS >'9e E ' f r S 6'6 0' E S'Z, / L ' O 6*02 T O ' O O ' T 80 '0 8*0 Z'S 8*0 o- e 0' Z 0'OS OVSI fr-e S6**0 ei'o E O ' T T'OI £ 'S S I 'O O ' T os os L' 09 OG E' E T > ' 6 T ' O 6 'T17 TO' 0 n i , 5 0 ' 0 6 • 0 . 9 ' £ 6 ' 0 0' t' 0 ' /. Q'LT? O ' T r T S'e /L8'0 62 - 0 L I- • Z 0 ' T E 0 ' 1 LI' 0 0 ' T os os 9'S!> Z ' K Z I' :l7 8'8E T ' T OS TO' 0 9 ' D O ' T 0' E Z ' B O ' E 0 ' S'l ( ) ' 9 't: 0'OS fr'S 9 'S T^'T G S ' O E O ' T O ' ^ E 8' e G T ' O O ' T os os 6 ' £ ^ ^,'01> 17' 2 os 6'Z 05 9 0 ' 0 » ' E Z'S O T ' T S ' E T ' T 0 ' 9 o - g O ' T C 8'8 fr'9 T T ' T L G ' O E 6 ' E S'ES T ' 8 T S ' O 9 ' T Zf. stduiBxg Tfr siduiBxg 0^ s"[duiBxg 6E sjdujBxg 8£ aiduiBXg ^e STduiBxa 9E ajduiBXH 5E siduiHxa ^e 3T.durexg EE ajduiHxg SE aiduiexg IE s-[durexa oe siduiexg 62 9-[duiBX3 82 sfduiexg Z^S 9-[duiexg 92 afdurexg 92 atduiexg E2 aiduiHxg ZZ ai;duiexg is ajduiexg OZ aidurexg 6T sidui^xa 8T STduiBxg LI 9-[duiBxg 9T aiduiexg As shown in Table 1, tricyclic derivatives according to the present invention showed very strong cytotoxicity to cancer cell lines. Experimental Example 2: Inhibiting effect of tricyclic derivatives of the present invention on tumor growth In order to investigate inhibiting effect of tricyclic derivatives of the invention on tumor growth, following experiments were performed. Samples for the experiment were stored in a refrigerator all the time. Different dosages of compounds were administered to a test animal; dosage of a compound prepared in the Example 8 was 10 mg/kg, dosage of a compound prepared in the Example 12 was 1, 3, 10 mg/kg respectively and dosage of a positive control was 2.5 mg/kg (Taxol) and 2 mg/kg (Adriamycin). The compound of the Example 12 was dissolved in 4% tween 80, and the positive control Taxol was dissolved in a mixed solvent of 5% athanol + 25% cremophor + 75% PBS. The prepared sample is subject to be precipitated, so that tip sonication was performed right before the administration to disperse it well. 7 week-old female S.P.F. BALB/c nude mice, provided by Charles Rivet: Co., Japan, were used as teat 141 animals. The test animals were adapted in a Hepafilter room over a week before the test. The temperature was 21±2°C, humidity was 55±5% and 12-hour light and dark cycle was automatically repeated in that lab. Solid feed (CheilJedang) was sterilized by radioactive rays and drinking water was also sterilized by autoclave. Feed and water were taken by the animal freely. Cancer cell line used .in this experiment was NCI-H460 (human lung tumor cell line) provided by Korea Research Institute of Bioscience and Biotechnology. The tumor cell line, stored in liquid nitrogen, was thawed and cultured in a 37 °C, 5% C02 incubator for a required time. Upon completing the culture, all the cells were recovered and cell concentration of the culture fluid was adjusted using PBS to 3xl07 cells/ml. The adjusted cell culture solution was injected hypodermically into armpit between right shoulder girdle and chest wall by 0.3 ml per mouse. From the next day of grafting, NCI-H460 xenografted nude mice were administered intraperitoneal everyday with the sample solution by 0.2 ml/20 g of weight, once a day. After the grafting of tumor cells, the volume of a tumor in each individual was measured in three dimensions by using a vernier caliper, which was represented in the below Mathematical Formula 2>. 142 Mathematical Formula 2> Tumor volume = (lengUixwidthxhoight) /2 Body weight changes of animal were measured three times a week. Each xenografted nude mouse was sacrificed to separate a tumor, which was then weighed. All the test results of experimental groups were compared with those of control groups by t-Test to see if there is any significant difference between the two groups. Changes of the volume and the weight of a tumor were shown in Table 2 and in FIG. 1, 3, and 5, and changes of the body weight of a mouse were shown in Table 3, in FIG. 2 and in FIG. 4. Best View in Resolution of 1024x768 or later. Enable Javascript for Better Performance. |
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| Patent Number | 233339 | |||||||||||||||||||||||||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Indian Patent Application Number | 384/DELNP/2006 | |||||||||||||||||||||||||||
| PG Journal Number | 13/2009 | |||||||||||||||||||||||||||
| Publication Date | 27-Mar-2009 | |||||||||||||||||||||||||||
| Grant Date | 28-Mar-2009 | |||||||||||||||||||||||||||
| Date of Filing | 20-Jan-2006 | |||||||||||||||||||||||||||
| Name of Patentee | JE IL PHARMACEUTICAL CO., LTD | |||||||||||||||||||||||||||
| Applicant Address | 745-4 BANPO-I DONG, SEOCHO-GU, SEOUL 137-810, REPUBLIC OF KOREA. | |||||||||||||||||||||||||||
Inventors:
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| PCT International Classification Number | C07C 317/30 | |||||||||||||||||||||||||||
| PCT International Application Number | PCT/KR2004/001518 | |||||||||||||||||||||||||||
| PCT International Filing date | 2004-06-23 | |||||||||||||||||||||||||||
PCT Conventions:
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