Title of Invention

"NOVEL PYRROLO[2,1-C] [1,4] BENZODIAZEPINES USEFUL AS ANTITUMOUR AGENTS"

Abstract A novel pyrrolo[2,l-c][l,4]benzodiazepine useful as antitumour agent, of formula IV as shown below wherein R is H, OH, R1 is OCH3, R2 is H, NH2 and n is 2 to 15.
Full Text The present invention relates to novel pyrrolo[2,l-c][l,4]beuzodiazpines useful as antitumour agents. This invention also relates to a process for preparation of novel pyirolo[2,l-c][l,4] benzodiazepines useful as antirumour agents. More particularly, it provides a process for the preparation of 2-(3-[7-methoxy-5~oxo-(llaS)-2,3,5,lla-teti ahydro-5H-pyrrolo[2,1 -c][ 1,4]bem:odiazepme-8-yloxy]alkyl]-2,3 -dihydro- lH-benzo (de]isoquinolin-l,3-dione, with aliphatic chain length variations for the compounds, The structural formula of novel pyrrolo[2,l-c][l,4]benzodiazepine is as follows wherein R is H, OH, R1 is OCH3, R2 is H, NH2 and n is 2 to 15.
Pyrrolo[2,l-c][l,4]benzodiazepine antitumour antibiotics are commonly known as anthramycin class of compounds. A large number of these compounds have been studied for their in vitro anticancer activity. These compounds exhibit their anticancer or antitumour activity by covalently binding with the DNA at N2 of guanine. These Pyrrolobenzodiazepine antibiotics exhibit their antitumour activity by forming an adduct in the minor groove of DNA, thus interfering with DNA processing. After insertion in the minor groove an aminal bond is formed through nucleophilic attack of the N2 of a guanine base at the electrophilic Cll position. (Ref. Kunimoto, S. Masuda, T. Kanbayashi, N. Hamada, M. Naganawa, H. Miyamoto, M. Takeuchi, T. and Unezawa, H. J. Antibiot., 1980,33, 665.; Kohn, K.W. and Speous, C.L. J. Mol. Biol, 1970, 57, 551.; Hurley, L.H. Gairpla, C. and Zmijewski, M. Biochem. Biophys. Acta., 1977, 475, 521.; Kaplan, D.J. and Hurley, L.H. Biochmestry, 1981,20,7572.)
However, the clinical efficacy for these antibiotics is hindered by several limitations, such as poor water solubility and cardiotoxicity and development of dmg resistance and me abolic inactivation.
The main object of the present invention is to provide novel pyrrolo[2,l-c][l,4]benzodiazepine useful as antitumour agenti. Another objective of the present invention is to provide a process for the preparation of novel pyrrolo[2,l-c][l,4]benzodiazepines useful as antitumour agents.
The compounds of present invention has a difference with C-8 linked napthalimides congeners by the replacement of phenolic hydrogen of (1 laS)-8-Hydroxy-7-methoxy-1,2,3,11 a-tetrahydro-5H-pyrrolo[2,1 -c][ 1,4]benzodiazepine-5-one(DC-81). Naphthalimides are a class of compounds that bind to DNA by intercalation of the chromophore group and show high antitumour activity against a variety of murine and human tumour ceils. These are a new class of DNA interacting agents, which exerf their antitumour activity through the inhibition of topoisomerase-II.
Accordingly the present invention relates to a novel pyrrolo[2,l-c][l,4]benzodiazepine of formula IV wherein R is H, OH, RI is OCH3, R2 is H, NH2 and n is 2 to 15.
(Formula Removed)
Accordingly the present invention provides a process for preparation of pyrrolo[2,l-c][l,4]benzodiazepines of formula IV of the drawing accompanying this specification wherein R is H, OH, RI is OCH3, R2 is H, NH2 and n is 2 to 15 which comprises: reacting nitrodiethyl thioacetal of formula I, wherein R is H, OH RI is OCH3 with haloalkyl
napthalimide in water soluble aprotic organic solvent in presence of a organic base upto refluxing temperature for a period in the range of 24 to 48 hours, affording the C-8 linked napthalimide compound of formula II wherein R, R1 and R2 as stated above by conventional methods, reacting above said C-8 linked naphthalimide compounds with reducing agent at a temperature up to reflux, affording the C-8 linked naphthalimide amino compounds of formula III wherein R is H, OH RI is OH, OCH}, R2 is NH2 by conventional methods, reacting the above said amino compound of formula III with known deprotecting agents in a conventional manner, recovering the pyrrolo[2,l-c][l,4]benzodiazepines of formula IV wherein R, RI and R2 and n are as stated above. In an embodiment of the present invention the haloalkyl naphthalimide used may be such as 2-bromo-N-ethyl-l,8-naphthalimide, 3-bromo-N-propyl-l,8-naphthalimide, 4-bromo-N-butyl-1,8-naphthalimide, 5-bromo-N-pentyl-1,8-naphthalimide, 6-bromo-N-hexyl-1,8-naphthalimide and 8-bromo-N-octyl-l,8-naphthalimide. In another embodiment of the invention the organic solvents used may be aprotic water miscible such as acetone, THF and DMF. In yet another embodiment of the invention the inorganic base used may be such as K2C03, NaH and Na2CO3 up to refluxing temperature for a period in the range of 24 to 48 hours, affording the C-8 linked naphthalimide compound of formula II wherein R, RI and R2 as stated above by conventional methods, reacting above said C-8 linked naphthalimide compounds with the reducing agent used may be such as SnCl2-2H2O Zn-acetic acid in presence of organic solvents such as MeOH, DMF and dioxane and their mixture at a temperature up to reflux, affording the C-8 linked naphthalimide amino compound of formula III, wherein R is H, OH, R1 is OCH3, R2 is NH2, by conventional methods, reacting the above said amino compound of formula III with known deprotecting agents used may be such as HgCL2/HgO, HgCl2/CaCO3 in presence of organic solvents such as acetonitrile, MeOH in a conventional manner recovering the pyrrolo[2,l- c][l,4]bem:odiadiazepines of formula IV, from the above reaction mixture by know methods wherein R, R|, Ri and n are as stated above.
In an embodiment of the present invention the haloalkyl naphthalimide used may be such as 2-bromo-N-ethyl-l,8-naphthalimide, 3-bromo-N-propyl-l,8-naphth.).limide, 4-bromo-N-butyl-1,8-naphthalimide, 5-bromo-N-pentyl-1,8-naphthalimide, 6-bromo-N-hexyl-l,8-naphthalimide and 8-bromo-N-octyl-l,8-naphthalimide.
In another embodiment of the invention the organic solvents used may be aprotic water miscible such as acetone, THF and DMF.
In yet another embodiment of the invention the inorganic base used may be such as K2CO3, NaH and Na2CO3 up to refluxing temperature for a period in the range of 24 to 48 hours, affording the C-8 linked naphthalimide compound of formula II wherin R, RI and Ri as stated above by conventional methods, reacting above said C-8 linked naphthalimide compounds with the reducing agent used may be such as SnCl2-2H2O Zn-acetic acid in presence of organic solvents such as MeOH, DMF and dioxane and their mixture at a temperature up to reflux, affording the C-8 linked naphthalimide amino compound of formula III, wherein R is H, OH R1 is OCH3, R2 is NH2, by conventional methods, reacting the above said amino compound of formula III with known deprotecting agents used may be such as HgCk/HgO, HgCl2/CaCCl3 in presence of organic solvents such as acetonitrile, MeOH in a conventional manner recovering the pyrrolo[2,l-c][l,4]benzodiadiazepines of formula IV, from the above reaction mixture by know methods wherein R, RI, R2 and n are as stated above.
These precursors, (2S)-N-(4-Hydroxy-5-methoxy-2-nitrobenzoyl)pyrrolidine-2-carboxaldehyde diethyl thioacetal (intermediates of DC-81) (formula I) and bromo-N-alkyl-l,8-napthalimide have been prepared by literature methods. (Ref. Langley, D.R. and Thurston, D.E. J. Org. Chem., 1987, 52, 91.)
Some of the compounds of present invention are given below:
1) 2-(2-[7-Methoxy-5-oxo-(l laS)-2,3,5,l la tetrahydro-5H-pyrrolo[2,l-c][l,4]
benzodiazepine-8-yloxy]ethyl)-2,3-dihydro-lH-benzo[de]isoquinoline-1,3-dione.
2) 2-(3-[7-Methoxy-5-oxo-(l laS)-2,3,5,l la-tetrahydro-5H'-pyiTolo[2,l-c][l,4]
benzodiazepine-8-yloxy]propyl)-2,3-dihydro-lH-benzo[(de]isoquinoline-l,3-dione.
3) 2-(4-[7-Methoxy-5-oxo-(l laS)-2,3,5,l la-tetrahydro-5H-pyrrolo[2,l-c][l,4]
benzodiazepine-8-yloxy]butyl)-2,3-dihydro-lH-benzo[Je]isoquinoline-l,3"dione.
4) 2-(5-[7-Methoxy-5-oxo-(l laS)-2,3,5,l la-tetrahydro-5H-pyrrolo[2,l-c][l,4]
benzodiazepine-8-yloxy]pentyl)-2,3-dihydro-lH-benzo[de]isoquinoline-l,3-dione.
5) 2-(6-[7-Methoxy-5-oxo-(l laS)-2,3,5,l la-tetrahydro-5H-pyrrolo[2,l-c][l,4]
benzodiazepine-8-yloxy]hexyl)-2,3-dihydro-1H-benzo[de]isoquinoline-f,3-dione.
6) 2-(8-[7-Methoxy-5-oxo-(l laS)-2,3,5,l la,tetrahydro-5H-pyrrolo[2,l-c][l,4]
benzodiazepine-8-yloxy]octyl)-2,3-dihydro-lJ7-benzo[ 7) 2-{4-[2-Hydroxy-7-methoxy-5-oxo(2/2,l laS)-2,3,5,l la-tetrahydro-5H-pyrrolo
[2,1 -c] [ 1,4]benzodiazepine-8-yloxl]butyl} -2,3-dihydro- lH-benzo[de]isoquinolin-
1,3-dione
The process of preparation of novel pyrrolo[2,l-e][l,4]benzodiazepines is disclosed & claimed in our cope;nding patent application no.
These new analogues of pyrrolo[2,l-c][l,4]benzodiazepines linked at C-8 position have shown promising antitumour activity in various cell lines particularly in small celt lung, colon, CNS, ovarian, renal, breast cancers and leukeamia. It is anticipated that these compounds may have improved drug resistance profile. This resulted in design and synthesis of new congeners as illustrated in scheme-1 which comprise. 1 The ether linkage at C-8 position of DC-81 intermediate with bromo-N-alkyl-1,8-napthalimides.
2) Refluxing the reaction mixture for 24-48 h.
3) Synthesis of C-8 linked PBD antitumour antibiotic imines.
4) Purification by column chromatography.
The following examples are given by way of illustration and should not be construed the limit and the scope of the invention. Preparation of C-8 naphthalimides linked PBD's
Example 1
To a stirred solution of 2-bromo-N-pentyl-l,8-napthalimide (0.303g,1.0 mmol) in dry acetone (25 mL) was added , anhydrous potassium carbonate (1.0 gm, 7.22 mmol) and the (formula I) (0.4 g, 1.0 mmol), the reaction mixture was refluxed in an oil bath for 48 h. The reaction was monitored by TLC using ethyl acetate-hexane (1:1) as a solvent system. The potassium carbonate was removed by suction filtration and the solvent was removed under vacuum. A solution of ethyl acetate-hexane (1:1) was added to the resulting solid and the nonsoluble component removed by suction filtration. 2-{2-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro- 1H-1 -pyrrolylcarbonyl)-2-methoxy-5-nitro-
phenoxy]ethyl}-2,3-dihydro-lH-benzo [d]- isoquinclin-l,.3-dione(formula II) was purified
by column chromatography using ethyl acetate-hexane (4:6) as eluent.
'H NMR (CDCl3): δ 1.20-1.40 (m, 6H), 2.21-2.45 (m, 4H), 2.60-2.85 (m, 4H), 3.10-3.35
(m, 2H), 3.40-3.50 (s, 3H), 4.20-4.32 (m, 2H), 4.40-4.50 (m, 2H), 4.60-4.75 (m, 1H), 4.80-
4.90 (d, 1H, J=4.5Hz), 6.60 (s, 1H), 7.60-7.65 (s, 1H), 7.70-7.85 (t, 2H, J=9.8Kz), 8.15-
8.25 (d, 2H, J=10.0Hz), 8.51-8.64 (d, 2H, J=10.0Hz).
FABMS: m/z 624 (M+l).
The 2-{2-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro-lH-l-pyrrolylcarbonyl)-2-methoxy-5-nitrophenoxy]ethyl} -2,3-dihydro-1H-benzo[de]-isoquinolin-1,3-dione(formula II) (0.623 g, 1.0 mmol) was dissolved in dichloromethane (5 mL), methanol (10 mL) and added SnCI2.2H2O (1.124 g, 5.0 mmol) was refluxed for 1.5 h. The reaction mixture was then carefully adjusted to pH 8 with saturated NaHCO3 solution and then extracted with ethyl acetate (3x20 mL). The combined organic phase was dried over Na2S04 and evaporated under vacuum to afford the crude 2-{2-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro- 1H-1 -pyrrolylcarbonyl)-2-methoxy-5-aminophenoxy]ethyl}-2,3-dihydro-lH-benzo [de] isoquinolin-l,3-dione(IH) yield is 70%. 'H NMR (CDC13): δ 1.25-1.40 (m, 6H), 2.20-2.40 (m, 4H), 2.65-2.85 (m, 4H), 3.12-3.33 (m, 2H), 3.40-3.50 (s, 3H), 4.20-4.32 (m, 2H), 4.40-4.50 (m, 2H), 4.60-4.75 (m, 1H), 4.85-4.90 (m, 1H), 6.60 (s, 1H), 7.60-7.65 (s, 1H), 7.7-7.85 (t, 2H, J=9.8Hz), 8.15-8.25 (d, 2H, J=10.0Hz), 8.50-8.56 (d, 2H, J=10.0Hz). FABMS: m/z 594 (M+l).
A stirred solution of 2-{2-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro-lH-l-pyrrolylcarbonyl)-2-methoxy-5-aminophenoxy]ethyl}-2,3-dihydro-lH-benzo[de]-isoquinolin-l,3-dione(formula III) (0.593 g, 1.0 mmol), HgCl2 (0.8 g, 2.94 mmol, 2.26 eq.) and HgO (0.32 g, 3.2 mmol, 2.46 eq.) in CH3CN/H20 (4:1) (15 mL) was stirred slowly at
room temperature for about 24 h. The reaction mixture was diluted with ethyl acetate (20 mL), filtered through a sintered funnel and then centrifuged at 10,000 rpm for 10 minutes. The clear yellow organic supernatant was extracted with saturated aq.NaHCO3 (2x20 mL), brine (2x20 mL) the combined aqueous phase was back extracted with ethyl acetate (2x20 mL). The combined organic phase was dried over Na2S04 and then filtered through a short bed of celite. The filtrate was evaporated under vacuum to afford 2-(2-[7-Methoxy-5-oxo-(1 laS)-2,3,5,l la tetrahydro-5H--pyrrolo[2,l-c][l,4]benzodiazepine-8-yloxy]ethyl)-2,3-dihydro-lH-benzo- [de]isoquinoline-l,3-dione(formula IV) as yellow oil which was purified by flash chromatography using ethyl acetate as eluent.
'H NMR (CDC13): δ 1.20-1.45 (m, 4H), 2.22-2.40 (m, 2H), 3.10-3.22 (m, 1H), 3.30-3.40 (s, 2H), 4.20-4.30 (m, 3H), 4.35-4.40 (m, 2H), 6.95 (s, 1H), 7.25 (s, 1H), 7.65-7.80 (m, 3H), 8.12-8.22 (d, 2H, J=10.0Hz), 8.45-8.60 (d, 2H, J=10.0Hz). FABMS: m/z 470 (M+l).
Example 2
To a stirred solution of 3-bromo-N-pentyl-l,8-napthalimide (0.317g,1.0 mmol) in dry acetone (25 mL) was added , anhydrous potassium carbonate (1.0 gm, 7.22 mmol) and the (2,S)-N-(4-Hydroxy-5-methoxy-2-nitrobenzoyl)pyrrolidine-2-carboxaldehyde diethyl thioacetal(formula I) (0.4 g, 1.0 mmol), the reaction mixture was refluxed in an oil bath for 48 h. The reaction was monitored by TLC using ethyl acetate-hexane (1:1) as a solvent system. The potassium carbonate was removed by suction filtration and the solvent was removed under vacuum. A solution of ethyl acetate-hexane (1:1) was added to the resulting solid and the nonsoluble component removed by suction filtration. 2-{3-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro- 1H-1 -pyrrolylcarbonyl)-2-methoxy-5-nitro-phenoxy]propyl}-2,3-dihydro-lH-benzo[de]-isoquinolin-l,3-dione(formula II) was purified by column chromatography using ethyl acetate-hexane (4:6) as eluent.
'H NMR (CDCl3): 5 1.20-1.40 (m, 8H), 1.50-1.80 (m, 4H), 1.90-2.05 (m. 4H). 2.70-2.90 (m, 4H), 120-3.35 (m, 2H), 3.95 (s, 3H), 4.10-4.30 (m, 2H), 4.85-4.90 (d, IH, J=4.7Hz), 6.80 (s, IH), 7.60-7.65 (s, IH), 7.70-7.85 (t, 2H, J=9.8Hz), 8.15-8.25 (d, 2H, J=10.0Hz), 8.60-8.65 (d, 2H, J=10.0Hz). FABMS:m/z 638(M+l).
The 2- {3-[4-(2-di(ethylsulphanyl)methyl-(2H)-tetrahydro-1H-1-pyrrolyl-carbonyl)-2-methoxy-5-nitrophenoxy]propyl} -2,3-dihydro- lH-benzo[de]-isoquinolin-1,3-dione-(formula II) (0.637g, 1.0 mmol) was dissolved in dichloromethane (5 mL), methanol (10 mL) and added SnCl2.2H2O (1.124 g, 5.0 mmol) was refluxed for 1.5 h. The reaction mixture was then carefully adjusted to pH 8 with saturated NaHCO3 solution and then extracted with ethyl acetate (3x20 mL). The combined organic phase was dried over Na2SO4 and evaporated under vacuum to afford the crude 2-{3-[4-(2-di(ethylsulphanyl) methyl-(2R)-tetrahydro-lH-l-pyrrolyl carbonyl)-2-methoxy-5-aminophenoxy]propyl}-2,3-dihydro-1H-benzo[de] isoquinolin-1,3-dione(IH).
'H NMR (CDC13): δ 1.15-1.0 (m, 8H), 1.80-2.15 (m, 2H), 2.15-2.35 (m, 3H), 2.55-2.80 (m, 4H), 3.55-3.60 (m, 3H), 3.70-3.75 (s, 2H), 4.0-4.05 (t, 2H, J=4.6Hz), 4.15-4.27 (t, 2H, J=4.6Hz), 4.50-4.70 (m, 2H), 6.21 (s, IH), 6.80 (s, IH), 7.65-7.80 (t, 2H, J=9.8Hz), 8.10-8.20 (d, 2H, J=10.0Hz), 8.50-8.60 (d, 2H, J=10.0Hz). FABMS: m/z 608 (M+l).
A stirred section of 2-{3-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro-] iY-1-pyrrolyl-carbonyl)-2-methoxy-5-aminophenoxy]propyl}-2,3-dihydro-lH-benzo[de]-isoquinolin-l,3-dione(formula III) (0.607 g, 1.0 mmol), HgCl2 (0.8 g, 2.94 mmol, 2.26 eq.) and HgO (0.32 g, 3.2 mmol, 2.46 eq.) in CH3CN/H2O (4:1) (15 mL) was stirred slowly at room temperature for about 24 h or until TLC indicates the completion of the reaction. The reaction mixture was diluted with ethyl acetate (20 mL), filtered through a sintered funnel
and then centrifuged at 10,000 rpm for 10 minutes. The clear yellow organic supernatant was extracted with saturated aq.NaHCO3 (2x20 mL), brine (2x20 mL) the combined aqueous phase was back extracted with ethyl acetate (2x20 mL). The combined organic phase was dried over Na2SO4 and then filtered through a short bed of celite. The filtrate was evaporated under vacuum to afford 2-(3-[7-Methoxy-5-oxo-(llaS)-2;3,5,lla-tetrahydro-5H-pyrrolo[2,1 -c][ 1,4]benzodiazepine-8-yloxy]propyl)-2,3-dihydro- lH-benzo [de]isoqui-noline-l,3-dione(formula IV) as yellow oil yield is 65% which was purified by flash chromatography using ethyl acetate as eluent.
H NMR (CDC13): δ 1.40-1.60 (s, 6H), 2.20-2.41 (m, 2H), 3.05-3.20 (t, 1H, J=4.5Hz), 3.30-3.40 (s, 2H), 4.20-4.30 (m, 3H), 4.34-4.50 (m, 2H), 6.95 (s, 1H), 7.22 (s, 1H), 7.65-7.80 (m, 3H), 8.13-8.22 (d, 2H, J=10.0Hz), 8.46-8.60 (d, 2H, J=10.0Hz). FABMS: m/z 484 (M+l).
Example 3
To a stirred solution of 4-bromo-N-pentyl-l,8-napthalimide (0.33Ig, 1.0 mmol) in dry acetone (25 mL) was added , anhydrous potassium carbonate (1.0 gm, 7.22 mmol) and the (2S)-N-(4-Hydroxy-5-methoxy-2-nitroberizoyl)pyrrolidine-2-carboxaldehyde diethyl thioacetal(formula I) (0.4 g, 1.0 mmol), the reaction mixture was refluxed in an oil bath for 48 h. The reaction was monitored by TLC using ethyl acetate-hexane (1:1) as a solvent system. The potassium carbonate was removed by suction filtration and the solvent was removed under vacuum. A solution of ethyl acetate-hexane (1:1) was added to the resulting solid and the nonsoluble component removed by suction filtration. 2-{4-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro- 1H-1 -pyrrolylcarbonyl)-2-methoxy-5-nitro-phenoxy]butyl}-2,3-dihydro-lH-benzo[de]-isoquinolin-l,3-dione(formula II) was purified by column chromatography using ethyl acetate-hexane (4:6) as eluent.
'H NMR (CDCl3): 5 1.22-1.45 (m, 6H), 1.50-1.72 (m, 4H), 1.81-2.05 (m, 4H), 2.70-2.92 (m, 4H), 3.15-3.30 (m, 2H), 3.90 (s, 3H), 4.0-4.30 (m, 4H), 4.6.5-4.78 (m, IH), 4.80-4.90 (d, IH), 6.75-6.82 (s, IH), 7.50-7.55 (s, IH), 7.70-7.80 (t, 2H, J=9.8Hz), 8.25-8.37 (d, 2H, J=10.0Hz), 8.55-8.65 (d, 2H, J=10.0Hz). FABNS: m/z 652 (M+l).
The 2- {4-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro-IH-1 -pyrrolylcarbonyl)-2-methoxy-5-nitrophenoxy]butyl} -2,3-dihydro-lH-benzo[de]-isoquinolin-1,3-dione (formula II) (0.651g, 1.0 mmol) was dissolved in dichloromethane (5 mL), methanol (10 mL) and added SnCl2.2H20 (1.124 g, 5.0 mmol) was refluxed for 1.5 h or until the TLC indicated that the reaction is complete. The reaction mixture was then carefully adjusted to pH S with saturated NaHCO3 solution and then extracted with ethyl acetate (3x20 mL). The combined organic phase was dried over Na2SO and evaporated under vacuum to afford the crude 2-{4-[4-(2-di(ethylsulphanyl)methyl-(2R-tetrahydro-1H-l-pyrrolyl-carbonyl)-2-methoxy-5-aminophenoxy]butyl) -2,3-dihydro-lH-benzo[de] isoquinolin-1,3-dione(III).
1H NMR (CDC13): δ 1.20-1.45 (m, IH), 1.50-1.75 (m, 4H), 1.80-1.95 (m, 4H), 2.15-2.30 (m, IH), 2.60-2.80 (m, 4H), 3.54-3.75 (m, 3H), 3.95-4.0 (m, 2H), 4.10-4.40 (m, 2H), 4.60-4.72 (brs, 2H), 6.20 (s, IH), 6.80 (s, IH), 7.65-7.80 (t, 2H, J=9.8Hz), 8.15-8.20 (d, 2H, J=10.0Hz), 8.50-8.60 (d, 2H, J=10.0Hz). FABMS:m./z 632(M+l).
A stirred solution of 2-{4-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro-lH-l-pyrrolylcarbonyl)-2-methoxy-5-aminophenoxy]butyl}-2,3-dihydro-lH-benzo[de]-isoquinolin-l,3-dione(formula III) (0.631 g, 1.0 mmol), HgCl2 (0.8 g, 2.94 mmol, 2.26 eq.) and HgO (0.32 g, 3.2 mmol, 2.46 eq.) in CH3CN/H20 (4:1) (15 mL) was stirred slowly at room temperature for about 24 h. The reaction mixture was diluted with ethyl acetate (20
mL), filtered through a sinter d funnel and then centrifuged at 10,000 rpm for 10 minutes. The clear yellow organic supernatant was extracted with saturated aq.NaHCO3 (2x20 mL), brine (2x20 mL) the combined aqueous phase was back extracted with ethyl acetate (2x20 mL). The combined organic phase was dried over Na2SO4 and then filtered through a short bed of celite. The filtrate was evaporated under vacuum to afford 2-(4-[7-Methoxy-5-oxo-(1 laS)-2,3,5,11 a-tetrahydro-5H-pyrrolo[2,l-c][l,4]benzodiaze-pine-8-yloxy]buryl)-2,3-dihydro-lH-benzo- [de]isoquinoline-l,3-dione(formula IV) as yellow oil which was purified by flash chromatography using ethyl acetate as eluent.
'H NMR (CDCl3): δ 1.15-1.42 (m, 4H), 1.56-1.70 (m, 4H), 2.22-2.40 (m, 2H), 3.32-3.40 (s, 2H), 3.80 (m, 1H), 4.20-4.30 (m, 3H), 4.35-4.40 (m, 2H), 6.95 (s, 1H), 7.22 (s, 1H), 7.60-7.75 (m, 3H), 8.10-8.20 (d, 2H, J=10.0Hz), 8.45-8.55 (d, 2H, J=10.0Hz). FABMS:m/z 498(M+l).
Example 4
To a stirred solution of 5-bromo-N-pentyl-l,8-napthalimide (0.684 g, 2 mmol) in dry acetone (25 mL) was added , anhydrous potassium carbonate (1.0 gm, 7.22 mmol) and the (25)-N-(4-Hydroxy-5-methoxy-2-nitrobenzoyl)pyrrolidine-2-carboxaldehyde diethyl thioacetal(formula I) (0.8 g, 2.0 mmol), the reaction mixture was refluxed in an oil bath for 48 h. The reaction was monitored by TLC using ethyl acetate-hexane (1:1) as a solvent system. The potassium carbonate was removed by suction filtration and the solvent was removed under vacuum. A solution of ethyl acetate-hexane (1:1) was added to the resulting solid and the nonsoluble component removed by suction filtration. 2-{5-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro-lH-l-pyrrolylcarbonyl)-2-methoxy-5-nitro-phenoxy]pentyl}-2,3-dihydro-lH-benzo[de]-isoquinolin-l,3-dione(formula II) was purified by column chromatography using ethyl acetate-hexane (4:6) as eluent.
'H NMR (CDCl3): 1.20-1.40(m, 6H), 1.50-1.70(m, 6H), l.SO-2.0(m, 4H), 2.70-2.90(m, 4H), 3.20-3.30(m, 2H), 3.90(s, 3H), 4.05-4.30(m, 4H), 4.65-4.78(m, 1H), 4.80-4.90(d, 1H), 6.20(s, 1H) 6.75-6.80(s, 1H), 7.70-7.85(t, 2H, J=9.8Hz), 8.28-8.37(d, 2H, J=10Hz), 8.55-8.65(d,2H,J=10Hz). FABMS : 666 (M+H).
The 2-{5-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro-lH-l-pyrrolyl-carbonyl)-2-methoxy-5-nitrophenoxy]pentyl}-2,3-dihydro-lH-benzo[de]-isoquinolin-l,3-dione (formula II) (0.665 g, 1.0 mmol) was dissolved in dichloromethane (5 mL), methanol (10 mL) and added SnCl2.2H20 (1.124 g, 5.0 mmol) was refluxed for 1.5 h. The reaction mixture was then carefully adjusted to pH 8 with saturated NaHCO3 solution and then extracted with ethyl acetate (3x20 mL). The combined organic phase was dried over Na2SO4 and evaporated under vacuum to afford the crude 2-{5-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro- 1H-1 -pyrrolyl-carbonyl)-2-methoxy-5-amino-phenoxy]pentyl}-2,3-dihydro-lH-benzo[de] isoquinolin-l,3-dione(III) yield is 70%. 'H NMR(CDC13): 1.20-1.40(m, 12H), 1.50-1.70(m, 4H), 1.80-2.0(m, 4H), 2.15-2.32(m, 1H), 2.60-2.80(m, 4H), 3.55-3.75(m, 3H), 3.95-4.0(t, 2H), 4.10-4.22(1, 2H), 4.6-4.70(brs, 2H), 6.25(s, 1H), 6.75(s, 1H), 7.65-7.80(t, 2H, J=9.8Hz), 8.10-8.20(d, 2H, J=10Hz), 8.50-8.60(d,2H,J=10Hz). FABMS : 636 (M+H).
A stirred solution of 2-{5-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro-lH-l-pyrrolyl-carbonyl)-2-methoxy-5-aminophenoxy]pentyl} -2,3-dihydro-1 H-benzo[de]-isoquinolin-l,3-dione(formula III) (0.635 g, 1.0 mmol), HgCl2 (0.8 g, 2.94 mmol, 2.26 eq.) and HgO (0.32 g, 3.2 mmol, 2.46 eq.) in CH3CN/H20 (4:1) (15 mL) was stirred slowly at room temperature for about 24 h. The reaction mixture was diluted with ethyl acetate (20 mL), filtered through a sintered funnel and then centrifiiged at 10,000 rpm for 10 minutes.
The clear yellow organic supernatant was extracted with saturated aq.NaHCO3 (2x20 mL), brine (2x20 mL) the combined aqueous phase was back extracted with ethyl acetate (2x20 mL). The combined organic phase was dried over Na2SO4 and then filtered through a short bed of celite. The filtrate was evaporated under vacuum to afford 2-(5-[7-Methoxy-5-oxo-(11 as) 2,3,5,1 la-tetrahydro-5H-pyrroio[2,l-c][l,4]benzodiazepine-8-ylcxy]pentyl)-2,3-dihydro-lH-benzo [de]isoquinoline-l,3-dione(formula IV) as yellow oil which was purified by flash chromatography using ethyl acetate as eluent.
'H NMR (CDCl2): 0.80-0.95(m, 2H), 1.20-11.35(s, 1H), 1.50-2.0(m, 4H), 3.40-3.50(s, 3H), 3.70-3.90(m, 2H), 3.95-4.0(m, 1H), 4.03-4.20(m, 2H), 7.20-7.30(m, 3H), 7.30-7.40(m, 2H), 7.65-7.80(m, 3H), 8.10-8.25(m, 2H), 8.40-8.60(m, 2H). FABMS : 512 (M+H).
Example 5
To a stirred solution of 6-bromo-N-pentyl-l,8-napthalimide (0.684 g, 2 mmol) in dry acetone (25 mL) was added , anhydrous potassium carbonate (1.0 gm, 7.22 mmol) and the (2S)-N-(4-Hydroxy-5-methoxy-2-nitrobenzoyl)pyrrolidine-2-carboxaldehyde diethyl thioacetal(formula I) (0.8 g, 2.0 mmol), the reaction mixture was refluxed in an oil bath for 48 h. The reaction was monitored by TLC using ethyl acetate-hexane (1:1) as a solvent system. The potassium carbonate was removed by suction filtration and the solvent was removed under vacuum. A solution of ethyl acetate-hexane (1:1) was added to the resulting solid and the nonsoluble component removed by suction filtration. 2-{6-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro- 1H1 -pyrrolylcarbonyl)-2-methoxy-5-nitro-phenoxy]hexyl}-2,3-dihydro-lH-benzo[de]- isoquinolin-l,3-dione(fomiula H) was purified by column chromatography using ethyl acetate-hexane (4:6) as eluent.
The 2- {6-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro-1 H-1 -pyrrolylcarbonyl)-2-methoxy-5-nitrophenoxy]hexyl}-2,3-dihydro-lH-benzo[de]-isoquinolin-l,3-dione

(formula II) (0.665 g, 1.0 mmol) was dissolved in dichloromethane (5 mL), methanol (10 mL) and added SnCl2.2H20 (1.124 g, 5.0 mmol) was refluxed for 1.5 h or until the TLC indicated that the reaction is complete. The reaction mixture was then carefully adjusted to pH 8 with saturated NaHCO3 solution and then extracted with ethyl acetate (3x20 mL). The combined organic phase was dried over Na2SO4 and evaporated under vacuum to afford the crude 2-{6-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro-1H-l-pyrrolyl-carbonyl)-2-methoxy-5-aminophenoxy]hexyl}-2,3-dihydro-lH-benzo[de] isoquinolin-1,3-dione(III).
A stirred solution of 2-{6-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro-lH-l-pyrrolylcarbonyl)-2-methoxy-5-aminophenoxy]hexyl}-2,3-dihydro-lH-benzo[de]-isoquinolin-l,3-dione(formula III) (0.635 g, 1.0 mmol), HgCl2 (0 8 g, 2.94 mmol, 2.26 eq.) and HgO (0.32 g, 3.2 mmol, 2.46 eq.) in CH3CN/H20 (4:1) (15 mL) was stirred slowly at room temperature for about 24 h. The reaction mixture was diluted with ethyl acetate (20 mL), filtered through a sintered funnel and then centriruged at 10,000 rpm for 10 minutes. The clear yellow organic supernatant was extracted with saturated aq.NaHCO3 (2x20 mL), brine (2x20 mL) the combined aqueous phase was back extracted with ethyl acetate (2x20 mL). The combined organic phase was dried over Na2SO4 and then filtered through a short bed of celite. The filtrate was evaporated under vacuum to afford 2-(6-[7-Methoxy-5-oxo-(11 aS)-2,3,5,11 a-tetrahydro-5H-pyrrolo[2,1 -c] [ 1,4]benzodiazepine-8-yloxy]hexyl)-2,3-dihydro-lH-benzo [de]isoquinoline-l,3-dione(formula IV) as yellow oil which was purified by flash chromatography using ethyl acetate as eluent.
Example 6
To a stirred solution of 8-bromo-N-pentyl-l,8-napthalimide (0.387g,1.0 mmol) in dry acetone (25 mL) was added , anhydrous potassium carbonate (1.0 gm, 7.22 mmol) and the (2,S)-N-(4-Hydroxy-5-methoxy-2-nitrobenzoyl)pyrrolidine-2-carboxaldehyde diethyl
thioacetal(formula I) (0.4 g, 1.0 mmol), the reaction mixture was refluxed in an oil bath for 48 h. The reaction was moniiored by TLC using ethyl acetate-hexane (1:1) as a solvent system. The potassium carbonate was removed by suction filtration and the solvent was removed under vacuum. A solution of ethyl acetate-hexane (1:1) was added to the resulting solid and the nonsoluble component removed by suction filtration. 2-{8-[4-(2-di(ethylsulphanyl)methyl-(ZK)-tetrahydro- 1H-1 -pyrrolylcarbonyl)-2-methoxy-5-nitro-phenoxy]octyl}-2,3-dihydro-lH-benzo [de]- isoquinolin-l,3-dione(formula II) was purified by column chromatography using ethyl acetate-hexane (4:6) as eluent. 1H NMR (CDC13): δ 1.20-1.60 (m, 6H), 1.65-1.90 (m, 12H), 1.80-2.0 (m, 4H), 2.70-2.85 (m, 4H), 3.20-3.30 (m, 2H), 3.60 (s, 3H), 4.02-4.20 (m, 4H), 4.40-4.60 (m, 2H), 4.62-4.70 (m, IH), 4.85 (s, IH), 6.75 (s, IH), 7.60 (r,, IH), 7.75 (t, 2H, J=9.7Hz), 8.20 (d, 2H, J=10.0Hz), 8.60 (d, 2H, J=10.0Hz). FABMS: m/z 708 (M+l).
The 2- {8-[4-(2-di(ethylsulphanyl)memyl-(2R)-tetrahydro-1H-1 -pyrrolylcarbonyl)-
2-methoxy-5-nitrophenoxy]octyl}-2,3-dihydro-lH-benzo[de]-isoquinolin-l,3-dione
(formula II) (0.707g, 1.0 mmol) was dissolved in dichloromethane (5 mL), methanol (10
mL) and added SnCl2.2H2O (1.124 g, 5.0 mmol) was refluxed for 1.5 h. The reaction
mixture was then carefully adjusted to pH 8 with saturated NaHCO3 solution and then
extracted with ethyl acetate (3x20 mL). The combined organic phase was dried over
Na2SO4 and evaporated under vacuum to afford the crude 2-{8-[4-(2-
di(ethylsulphanyl)methyl-(2^)-tetrahydro-lH-l-pyrrolyl- carbonyl)-2-methoxy-5-
aminophenoxy]octyl}-2,3-dihydro-lH-benzo[de] isoquinolin-l,3-dione(III). 1H NMR (CDC13): δ 1.20-1.40 (m, 6H), 1.50-1.70 (m, 12H), 1.80-2.10 (m, 4H), 2.60-2.75 (m, IH), 3.40-3.50 (s, 3H), 3.95-4.0 (m, 2H), 4.10-4.22 (m, 2H), 4.60-4.70 (brs, 2H), 6.25
(s, 1H), 6.70 (s, 1H), 7.60-7.80 (t, 2H, J=10.0Hz), 8.15-8.20 (d, 2H, J=10.0Hz), 8.50-8.60 (d, 2H, J=10.0Hz). FABMS: m/z 678 (M+l).
A stirred solution of 2-{8-[4-(2-di(ethylsulphanyl)methyl-(2R)-tetrahydro-lH-l-pyrrolylcarbonyl)-2-methoxy-5-aminophenoxy]octyl}-2,3-dihydro-lH-benzo[de]isoqui-nolin-l,3-dione(formula III) (0.677 g, 1.0 mmol), HgCl2 (0.8 g, 2.94 mmol, 2.26 eq.) and HgO (0.32 g, 3.2 mmol, 2.46 eq.) in CH3CN/H2O (4:1) (15 mL) was stirred slowly at room temperature for about 24 h. The reaction mixture was diluted with ethyl acetate (20 mL), filtered through a sintered runnel and then centrifuged at 10,000 rpm for 10 minutes. The clear yellow organic supernatant was extracted with saturated aq.NaHCO3 (2x20 mL), brine (2x20 mL) the combined aqueous phase was back extracted with ethyl acetate (2x20 mL). The combined organic phase was dried over N32SO4 and then filtered through a short bed of celite. The filtrate was evaporated under vacuum to afford 2-(8-[7-Methoxy-5-oxo-(11 aS)-2,3,5,11 a-tetrahydro-5H-pyrrolo[2,1 -c][ 1,4]benzodiazepine-8-yloxy]octyl)-2,3-dihydro-l//-benzo (de] isoquinoline-l,3-dione as yellow oil which was purified by flash chromatography.
1H NMR (CDC13): δ 0.80-0.90 (m, 2H), 1.20-1.35 (m, 2H), 1.40-1.65 (m, 12H), 3.50 (s, 3H), 3.70-3.90 (m, 2H), 3.91-3.95 (m, 1H), 4.0-4.20 (m, 2H), 7.20-7.30 (m, 2H), 7.32-7.40 (m, 2H), 7.65-7.80 (m, 3H), 8.10-8.25 (m, 2H), 8.40-8.60 (m, 2H). FABMS : m/z 554 (M+l).
Example 7-
To a stirred solution of 4-bromo-N-pentyl-l,8-napthalimide (0.631 g, 2 mmol) in dry acetone (25 mL) was added , anhydrous potassium carbonate (1.0 gm, 7.22 mmol) and the(25)-N-(4-Hydroxy-5-methoxy-2-nitrobenzoyl)-4-hydroxypyrrolidine-2-carboxalde-hyde diethyl thioacetal(formula I) (0.416g, 1.0 mmol), the reaction mixture was refluxed in
an oil bath for 48 h. The reaction was monitored by TLC using ethyl acetate-hexane (1:1) as a solvent system. The pofassium carbonate was removed by suction filtration and the solvent was removed under vacuum. A solution of ethyl acetate-hexane (1:1) was added to the resulting solid and the nonsoluble component removed by suction filtration. 2-{4-[4-(2-di(ethylsulphanyl)methyl-4-hydroxy-(4S)-tetrahydro-1H-1 -pyrrolylcarbonyl)-2-methoxy-5-nitrophenoxy]butyl}-2,3-dihydro-lH-benzo[de]-isoquinolin-l,3-dione(formula II) was purified by column chromatography using ethyl acetate-hexane (4:6) as eluent. 1H NMR (CDC13): δ 1.20-1.42 (m, 6H), 1.52-1.68 (m, 4H), 2.20-2.40 (m, 2H), 2.60-2.85 (m, 4H), 3.10-3.30 (m, 2H), 3.42-3.51 (s, 3H), 4.20-4.32 (t, 3H, J=4.6Hz), 4.40-4.50 (t, 2H, J=4.6Hz), 4.53 (brs, 1H), 4.60-4.75 (m, 1H), 4.80-4.90 (d, 1H), 6.60 (s, 1H), 7.60-7.65 (s, 1H), 7.70-7.85 (t, 2H, J=9.8Hz), 8.15-S.25 (d, 2H, J=10.0Hz), 8.50-8.65 (d, 2H, J=10.0Hz). The 2- {4-[4-(2-di(ethylsulphanyl)methyl-4-hydroxy-(45)-tetrahydro-1H-1 -pyrrolyl carbonyl)-2-methoxy-5-nitrophenoxy]butyl}-2,3-dihydro-lH-benzo[de]-isoquinolin-l,3-dione(formula II) (0.662 g, 1.0 mmol) was dissolved in dichloromethane (5 mL), methanol (10 mL) and added SnCl2.2K2O (1.124 g, 5.0 mmol) was refluxed for 1.5 h. The reaction mixture was then carefully adjusted to pH 8 with saturated NaHCO3 solution and then extracted with ethyl acetate (3x20 mL). The combined organic phase was dried over Na2SO4 and evaporated under vacuum to afford the crude 2-{4-[4-(2-di(ethylsulphanyl)methyl-4-hydroxy-(4S)-tetrahydro-1H-1 -pyrrolyl carbonyl)-2-methoxy-5-aminophenoxy]butyl}-2,3-dihydro-1H-benzo[de] isoquinolin-l,3-dione(III). 1H NMR (CDC13): δ 1.20-1.40 (m, 6H), 1.80-2.15 (m, 2H), 2.65-2.85 (m, 4H), 3.32-3.89 (m, 5H), 3.92 (s, 3H), 4.20-4.32 (t, 2H, J=4.6Hz), 4.40-4.50 (t, 2H, J=4.6Hz), 4.60-4.75 (m, 1H), 4.80-4.90 (d, 1H), 6.60 (s, 1H), 7.60-7.65 (s, 1H), 7.70-7.85 (t, 2H, J=9.8Hz), 8.15-8.20 (d, 2H, J=10.0Hz), 8.52-8.62 (d, 2H, J=10.0Hz). FABMS: m/z 638 (M+l).
A stirred solution of 2-{4-[4-(2-di(ethylsulphanyl)methyl-4-hydroxy-(1S)-tetrahydro-1H-1 -pyrroly lcarbonyl)-2-methoxy-5 - aminophenoxyjbutyl} -2,3 -dihydro-1H-benzo[de]-isoquinolin-l,3-dione(formula III) (0.632 g, 1.0 mmol), HgCl2 (0.8 g, 2.94 mmol, 2.26 eq.) and HgO (0.32 g, 3.2 mmol, 2.46 eq.) in CH3CN/H2O (4:1) (15 mL) was stirred slowly at room temperature for about 24 h. The reaction mixture was diluted with ethyl acetate (20 mL), filtered through a sintered funnel and then centrifuged at 10,000 rpm for 10 minutes. The clear yellow organic supernatant was extracted with saturated aq.NaHCO3 (2x20 mL), brine (2x20 mL) the combined aqueous phase was back extracted with ethyl acetate (2x20 mL). The combined organic phase was dried over Na2S04 and then filtered through a short bed of celite. The filtrate was evaporated under vacuum to afford2- {4-[2-Hydroxy-7-methoxy-5-oxo(2R, 11 a-tetrahydro-5H-pyyyolo {2,1 -c][ 1,4]benzodiazepine-8-yloxy]butyl} -2,3-dihydro- lH-benzo[de]isoquinolin-1,3-dione (formula IV) as yellow oil which was purified by flash chromatography using ethyl acetate as eluent.
1H NMR (CDC13): δ 1.15-1.42 (m, 4H), 1.56-1.70 (m, 4H), 2.2-2.4 (m, 2H), 3.32-3.40 (s, 2H), 3.80 (m, 3H), 4.20-4.30 (m, 3H), 4.35-4.40 (m, 2H), 6.90 (s, 1H), 7.22 (s, 1H), 7.60-7.75 (m, 3H), 8.10-8.20 (d, 2H, J=10.0Hz), 8.45-8.55 (d, 2H, J=10.0Hz). FABMS:m/z 514 (M+l).
Biological Activity: In rirro biological activity studies were carried out at National Cancer Institute (USA).
In vitro human cell line assay: For each compound dose response curves for each cell line were measured with five different concentrations and the concentration causing 50% cell growth inhibitions observed. The dose-response curves of some tumour carcinoma sensitive to the compounds (IVa, IVc) are shown and discussed below. These dose response curves were plotted, the percentage growth of each cell line against different concentrations of the compounds in logarithmic values.
Dose response curves: Compound IVa is effective towards the (e.g. NCI-H23 & NCI-H522) from small cell lung cancer, (e.g. HT 29) from colon cancer, (e.g. SF-539) from CNS cancer, (e.g. RPMI-8226 & SR) from leukaemia, (e.g. MALME-3M & SK-MEL-2) from melanoma cancer, (e.g. UO-31, RXF-393, SN 12C, ACHN & CAKI-1) from renal cancer and (e.g. MIDA-N) from breast cancer.
Compound IVc is effective towards the (e.g. NCI, HOP-92, H322M, HOP-62 & NCI-H522) from small cell lung cancer, (e.g. KM12 & COLO205) from colon cancer, (e.g. SNB-75 & SF-539) from CNS cancer, (e.g. RXF393, UO-31, & A498) from renal cancer, (e.g. MALME-3M, LOXIMVI, SK-MEL-5 & SK-MEL-2) from melanoma cancer, (e.g. BT-549 & NCyADR-RES) from breast cancer and (e.g. OVCEAR-3, OVCEAR-4 & IGROVI) from ovarian cancer. These compounds showed total growth inhibition of all the cell lines at 10-6 molar concentration.
Mean Graph: The mean graph of the compound IVa shown and discussed below. In these mean graphs, the log10 GI 50 of compound expressed in the form of mean graphs. In these graphs, the mean logarithmic value of GI 50 in all cell lines for each compound is used as mid point of the bar graph. Bars extending to the right represent sensitivity of the cell lines to the test agent in excess of the average sensitivity of all tested cell lines. The bar scale is
logarithmic, therefore, a bar two units to the right shows the compound achieved the GI 50 for the cell line at a concentration one-hundredth the mean concentration required overall the cell lines thus, the cell line is unusually sensitive to the compound. Bars to the left correspondingly imply sensitivity less than the mean. The log 10 TGI and log10 LC 50 values for these compounds were also measured and expressed with similar bar mean graphs.
Mean Graph for the Compound: Figure logio GI 50 mean graph of compound IVa the in vitro antitumour activity against human tumour cell lines [logio GI 50: The logarithmic concentration required to produce 50% of the growth inhibition]. Responses to the right of the mean logio GI 50 line (the mid point of the bar graph) are more sensitive than the mean, while those on the left are more resistant, (data from NCI)
As demonstrated by the mean graph pattern these compounds have similar selectivity over leukaemia (e.g. PML-8226, SR), non small cell lung cancer (e.g. NCI-H23 & NCI-H522), renal cancer (e.g. UO-31, RXF393, SN12C, ACHN & CAKI-1), breast cancer (e.g. MIDA-N), melanoma cancer (e.g. MALME-3M & SK-MEL-2), and colon cancer (e.g. HT29) cell lines. However some compounds shown increased selectivity towards renal cancer cell lines (e.g. VO-31, RXF393, SN12C, ACHN & CAKI-1). In general all the compounds shown greater activity towards all cell lines. Mid points of logio TGI and logio LC 50 showed similar patterns to the logio GI 50 mean graph mid points.





We Claim:
1) A novel pyrrolo[2,l-c][l,4]benzodiazepine useful as antitumour agent, of formula IV as shown below wherein R is H, OH, R1I is OCH3, R2 is H, NH2 and n is 2 to 15.

(Formula Removed)
2) A novel pyrrolobenzodiazepine as disclosed in claim 1 has structural formula shown below.
(Formula Removed)
3) A novel pyrrolobenzodiazepine as disclosed in claim 1 has structural formula shown below.
(Formula Removed)
4) A novel pyrrolobenzodiazepine as disclosed in claim 1 has structural formula shown below.
(Formula Removed)
5) A novel pyrrolobenzodiazepine as disclosed in claim 1 has structural formula shown below.
(Formula Removed)
6) A novel pyrrolobenzodiazepine as disclosed in claim 1 has structural formula shown below.
(Formula Removed)
7) A novel pyrrolobenzodiazepine as disclosed in claim 1 has structural formula shown below.
(Formula Removed)
8) A novel pyrrolobenzodiazepine as disclosed in claim 1 has structural formula shown below.
(Formula Removed)
10) A novel pyrrolo[2,l-c][l,4]benzodiazepine useful as antitumour agent, of formula IV wherein R is H, OH, R1 is OCH3, R2 is H, NH2 and n is 2 to 15 substantially as herein described with reference to the examples.

Documents:

210-del-2000-abstract.pdf

210-del-2000-claims.pdf

210-del-2000-correspondence-others.pdf

210-del-2000-correspondence-po.pdf

210-del-2000-description (complete).pdf

210-del-2000-form-1.pdf

210-del-2000-form-19.pdf

210-del-2000-form-2.pdf

210-DEL-2000-Form-3.pdf

abstract.jpg


Patent Number 212061
Indian Patent Application Number 210/DEL/2000
PG Journal Number 47/2007
Publication Date 23-Nov-2007
Grant Date 14-Nov-2007
Date of Filing 09-Mar-2000
Name of Patentee COUNCIL OF SCIENTIFIC AND INDUSTRIAL RESEARCH
Applicant Address RAFI MARG, NEW DLEHI-110 001, INDIA.
Inventors:
# Inventor's Name Inventor's Address
1 AHMED KAMAL INDIAN INSTITUTE OF CHEMICAL TECHNOLOGY, HYDERABAD - 500 007, ANDHRA PRADESH, INDIA.
2 SATYANARAYANA REDDY INDINA INSTITUTE OF CHEMICAL TECHNOLOGY, HYDERABAD - 500 007, ANDHRA PRADESH, INDIA.
PCT International Classification Number C07D 519/00
PCT International Application Number N/A
PCT International Filing date
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 NA