WO2004087712A1 - NEW PYRIMIDINE LINKED PYRROLO[2,1-c][1,4]BENZODIAZEPINES AS POTENTIAL ANTITUMOUR AGENTS - Google Patents

NEW PYRIMIDINE LINKED PYRROLO[2,1-c][1,4]BENZODIAZEPINES AS POTENTIAL ANTITUMOUR AGENTS Download PDF

Info

Publication number
WO2004087712A1
WO2004087712A1 PCT/IN2003/000122 IN0300122W WO2004087712A1 WO 2004087712 A1 WO2004087712 A1 WO 2004087712A1 IN 0300122 W IN0300122 W IN 0300122W WO 2004087712 A1 WO2004087712 A1 WO 2004087712A1
Authority
WO
WIPO (PCT)
Prior art keywords
formula
mmol
methoxy
fluorophenyl
methylpyrimidine
Prior art date
Application number
PCT/IN2003/000122
Other languages
French (fr)
Inventor
Ahmed Kamal
Karnati Laxma Reddy
Original Assignee
Council Of Scientific And Industrial Research
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Council Of Scientific And Industrial Research filed Critical Council Of Scientific And Industrial Research
Priority to PCT/IN2003/000122 priority Critical patent/WO2004087712A1/en
Priority to AU2003222444A priority patent/AU2003222444A1/en
Publication of WO2004087712A1 publication Critical patent/WO2004087712A1/en

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D487/00—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00
    • C07D487/02—Heterocyclic compounds containing nitrogen atoms as the only ring hetero atoms in the condensed system, not provided for by groups C07D451/00 - C07D477/00 in which the condensed system contains two hetero rings
    • C07D487/04—Ortho-condensed systems
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

Definitions

  • the present invention relates to a process for the preparation of novel pyrimidine linked pyrrolo [2,l-c][l,4]benzodiazepines useful as potential antitumour agents.
  • This invention also relates to a process for the preparation of new pyrimidine linked pyrrolo[2,l- c][l,4]benzodiazepines as potential antitumour agents.
  • Pyrrolo[2,l-c][l,4]benzodiazepines anti-tumour antibiotics are commonly known as anthramycin class of compounds.
  • PBDs pyrrolo [2, 1-c] [l,4]benzodiazepines
  • PBD dimers have been developed that comprises two C2-exo-methylene- substituted DC-81 subunits tethered through their C-8 position via an inert propanedioxy linker (Gregson, S. I; Howard, P. W.; Hartely, J. A.; Brooks, N. A.; Adams, L. J.; Jenkins, T. C; Kelland, L. R. and Thurston, D.
  • PBD dimers have been synthesized which have significant DNA binding ability and potent anti tumour activitiy.
  • Naturally occurring pyrrolo[2,l-c][l,4]benzodiazepines belong to a group of antitumour antibiotics derived from Streptomyces species. Recently, there is much impetus for the PBD systems as they can recognize and bind to specific sequence of DNA. Examples of naturally occurring PBD's include anthramycin, DC-81, tomaymycin, sibiromycin and neothramycin. However, the clinical efficacy for these antibiotics is hindered by several limitations, such as poor water solubility and cardiotoxicity and development of drug resistance and metabolic inactivation. Objects of the invention
  • the main object of the present invention is to provide new pyrrolo[2,l-c][l,4] benzodiazepines useful as antitumour agents.
  • Another object 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. Summary of the invention
  • acetylchloride (100 mg, 1.20 mmol) was added drop wise at same temperature and reaction mixture was allowed to stir at room temperature for overnight and then poured into water, extracted with dichloromethane and dried over Na 2 SO 4 and evaporated in vacuum to afforded the corresponding compound (4R)-acetyloxy-(2S)-N-[ ⁇ 4- [6 ' -(4 " -fluorophenyl)-2 ' -methylpyrimidine-4 ' -yloxy]butoxy ⁇ -5 -methoxy-2-nitrobenzoyl] pyrrohdine-2-carboxaldehyde diethyl thioacetal.
  • Cytotoxicity Compounds Via and VIb were evaluated in vitro against sixty human tumour cells derived from nine cancer types (leukemia, non-small-cell lung, colon, CNS, melanoma, ovarian, prostate, and breast cancer). For each compound, dose response curves for each cell line were measured at a minimum of five concentrations at 10 fold dilutions. A protocol of 48 h continuous drug exposure was used and a sulforhodamine B (SRB) protein assay was used to estimate cell viability or growth. The concentration causing 50% cell growth inhibition (GI50), total cell growth inhibition (TGI 0% growth) and 50% cell death (LC50, -50% growth) compared with the control was calculated.
  • GI50 50% cell growth inhibition
  • TGI 0% growth total cell growth inhibition
  • LC50 50% cell death
  • the mean graph midpoint values of log 10 TGI and log 10 LC50 as well as log 10 GI50 for Via and VIb are listed in Table 1. As demonstrated by mean graph pattern, compound VIb exhibits an interesting profile of activity and selectivity for various cell lines. The mean graph mid point of log 10 TGI and log 10 LC50 showed similar pattern to the log 10 GI50 mean graph mid points.
  • log 10 GI50 log 10 TGI and log 10 LC50 mean graphs midpoints(MG_MLD) of in vitro cytotoxicity data for the compounds Via and VI against human tumour cell lines.
  • Each cancer type represents the average of six to eight different cancer cell lines.

Abstract

The present invention relates to a process for the preparation of novel pyrrolo [2,1­c][1,4]benzodiazepines useful as potential antitumour agents. This invention also relates to a process for the preparation of new pyrrolo[2,1-c][1,4]benzodiazepines as potential antitumour agents. More particularly, it provides a process for the preparation of 7-methoxy-8-[6'-(4'-fluorophenyl)-2'-methylpyrimidine-4'-yloxy] alkoxy-(l 1aS)-1,2,3,11 a-tetrahydro-5H­-pyrrolo[2,1,c][1,4]benzodiazepin-5-one with aliphatic chain length variations for the compounds and it also describes the anticancer (antitumour) activity. The structural formula (I) of novel pyrrolo[2,1-c] [ 1,4]benzodiazepine is as follows.

Description

NEW PYRIMIDINE LINKED PYRROLO [2,l-c][l,4]BENZODIAZEPINES AS POTENTIAL ANTITUMOUR AGENTS
Field of the invention
The present invention relates to a process for the preparation of novel pyrimidine linked pyrrolo [2,l-c][l,4]benzodiazepines useful as potential antitumour agents. This invention also relates to a process for the preparation of new pyrimidine linked pyrrolo[2,l- c][l,4]benzodiazepines as potential antitumour agents. More particularly, it provides a process for the preparation of 7-methoxy-8-[6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'- yloxy] alkoxy-(l laS)-l,2,3, 1 la-tetrahydro-5H-pyrrolo[2, l,c][l ,4]benzαdiazepin-5-one with aliphatic chain length variations for the compounds and it also describes the anticancer (antitumour) activity. The structural formula of novel pyrrolo [2, 1-c] [l,4]benzodiazepine is as follows.
Figure imgf000002_0001
Background of the invention
Pyrrolo[2,l-c][l,4]benzodiazepines anti-tumour antibiotics are commonly known as anthramycin class of compounds. In the last few years, a growing interest has been shown in the development of new pyrrolo [2, 1-c] [l,4]benzodiazepines (PBDs). These antibiotics react covalently with DNA to form an N2-guanine adduct that Ues within the minor groove of duplex DNA via an acid-labile aminal bond to the electrophilic imine at the N10-C11 position (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, 51, 551.; Hurley, L. H.; Gairpla, C. and Zmijewski, M. Biochem. Biophys. Ada., 1977, 475, 521.; Kaplan, D. J. and Hurley, L. H. Biochmestry, 1981, 20, 7572). The molecules have a right-handed twist, which allows them to follow the curvature of the minor groove of B-form double-stranded DNA spanning three base pairs. Recently, PBD dimers have been developed that comprises two C2-exo-methylene- substituted DC-81 subunits tethered through their C-8 position via an inert propanedioxy linker (Gregson, S. I; Howard, P. W.; Hartely, J. A.; Brooks, N. A.; Adams, L. J.; Jenkins, T. C; Kelland, L. R. and Thurston, D. E. JMedChem. 2001, 44, 131). A recent development has been the linking of two PBD units through their C-8 positions to give bisfunctional alkylating agents capable of cross-linking DNA (Thurston, D. E.; Bose, D. S.; Thomson, A.
S.; Howard, P. W.; Leoni, A.; Croker, S. J.; Jenkins, T. C; Neidle, S. and Hurley, L. H. J.
Org. Chem., 1996, 61, 81 1-8147). Recently, a non -crass - linking - mixed imine -amide
PBD dimers have been synthesized which have significant DNA binding ability and potent anti tumour activitiy. (Kamal, A.; Ramesh, G.; Laxman, N.; Ramulu, p.; Srinivas, O.;
Neelima, K.; Kumar, K.K.; Srinu, V.B.; Nagarajaram, H.M. J. Med. Chem. 2002, 45, 4679).
Figure imgf000003_0001
anthramycin C2-exo-metlιylene-substituted DC-81
Figure imgf000003_0002
DC-81 dimers (n= 3-5); DSB-120 (n= 3)
Figure imgf000003_0003
imine-amide PBD dimers; n = 3 - 5
Naturally occurring pyrrolo[2,l-c][l,4]benzodiazepines belong to a group of antitumour antibiotics derived from Streptomyces species. Recently, there is much impetus for the PBD systems as they can recognize and bind to specific sequence of DNA. Examples of naturally occurring PBD's include anthramycin, DC-81, tomaymycin, sibiromycin and neothramycin. However, the clinical efficacy for these antibiotics is hindered by several limitations, such as poor water solubility and cardiotoxicity and development of drug resistance and metabolic inactivation. Objects of the invention
The main object of the present invention is to provide new pyrrolo[2,l-c][l,4] benzodiazepines useful as antitumour agents. Another object 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. Summary of the invention
Accordingly the present invention provides a process for the preparation of a novel pyrrolo[2,l-c][l,4]benzodiazepine of formula VI wherein R = H, OH, OAc and n is 3 - 5.
Figure imgf000004_0001
FORMULA VI
The present invention also provides a process for preparation of pyrrolo[2,l- c][l,4]benzodiazepines of formula VI above wherein R=H, OH, OAc and n is 3 to 5 which comprises reacting 6-4-(4-fluorophenyl)-2-methyl-4-pyrimidinol of formula I with dibromoalkanes in an aprotic water miscible organic solvents like acetone, THF and DMF in presence of mild inorganic bases like K2C03, CsCO3 and BaCO3 upto refluxing temperature for a period up to 48 h, isolating n-bromoalkyl-6-(4-flurophenyl)-2-methyl-4-pyrimidinyl ether of formula II with (2S)-N-(4-hydroxy-5-methoxy-2-nitrobenzoyl)pyrrolidine-2- carboxaldehyde diethyl thioacetal of formula III in presence of mild inorganic bases like K2CO3, CsCO3) and BaC03 in presence of aprotic water miscible organic solvents up to refluxing for a period of 48 h isolating (2S) - N - [{4 - [6" - (4' -fluorophenyl) - 2' - methylpyrimidine - 4' - yloxy] alkoxy} - 5 - methoxy - 2 - nitrobenzoyl] pyrrolidine - 2 - carboxaldehyde diethyl thioacetal IV where n is 3 to 5 by conventional methods, reducing the above nitro compounds of formula IV with SnCl2.2H2O in presence of organic solvent up to a reflux temperature, isolating the (2S) - N - [{4 - [6' - (4" - fluorophenyl) - 2' - methylpyrimidine - 4' - yloxy] alkoxy} - 5 - methoxy - 2 - amonobenzoyl] pyrrolidine - 2 - carboxaldehyde diethyl thioacetal of formula V where n is 3 to 5 by known methods, reacting the above said amino compound of formula V with known deprotecting agents in a conventional manner to give novel pyrrolo[2,l-c][l,4]benzodiazepines of formula VI wherein n is as stated above.
The precursor, (2S)-N-(4-hydroxy-5-methoxy-2-nitrobenzoyl)pyrrolidine-2- carboxaldehyde diethyl thioacetal of formula I (intermediates of DC-81) is prepared by literature methods (Thurston, D. E.; Murthy, V. S.; Langley, D. R.; Jones, G. B. Synthesis,
1990, 81)
Detailed description of the invention
Some representative compounds of formula VI present invention are given below:
1. 7-methoxy-8-[6'-(4"-fluorophenyl)-2'-methylpyrinιidine-4'-yloxy]propoxy-(l 1 aS)- 1,2,3,1 la-tetrahydro-5H-pyrrolo[2,l,c][l,4]benzodiazepin-5-one.
2. 7-methoxy-8-[6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'-yloxy]butoxy-(l 1 aS)- 1,2,3,1 la-tetrahydro5H-pyrrolo[2,l,c][l,4]benzodiazepin-5-one.
3. 7-methoxy-8-[6'-(4''-fluorophenyl)-2'-methylpyrimidine-4'-yloxy]pentoxy-(l 1 aS)- 1,2,3, 1 la-tetrahydro-5H-pyrrolo[2, l,c][l,4]benzodiazepin-5-one.
4. 7-methoxy-8-[6'-(4"-fluorophenyl)-2'-methylpyrirnidine-4'-yloxy]propoxy-(4R)- hydroxy-( 11 aS)- 1 ,2,3 , 11 a-tetrahydro-5H-pyrrolo[2, 1 ,c] [ 1 ,4]benzodiazepin-5-one
5. 7-methoxy-8-[6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'-yloxy]butoxy-(4R)- hydroxy-(l laS)-l,2,3,l la-tetrahydro-5H-pyrrolo[2,l,c][l,4]benzodiazepin-5-one
6. 7-methoxy-8-[6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'-yloxy]pentoxy-(4R)- hydroxy-(l laS)-l,2,3,lla-tetrahydro-5H-pyrrolo[2,l,c][l,4]benzodiazepin-5-one
7. 7-methoxy-8-[6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'-yloxy]propoxy-(4R)- acetyloxy-(l laS)-l,2,3, 1 la-tetrahydro-5H-pyrrolo[2, l,c][l,4]benzodiazepin-5-one
8. 7-methoxy-8-[6' -(4"-fluorophenyl)-2' -methylpyrimidine-4 ' -yloxy]butoxy-(4R)- acetyloxy-(l 1 aS)- 1 ,2,3 , 11 a-tetrahydro-5H-pyrrolo[2, 1 ,c] [ 1 ,4]benzodiazepin-5-one
9. 7-methoxy-8-[6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'-yloxy]pentoxy- (4R) - acetyloxy-(l laS)-l,2,3,l la tetrahydro5H-pyrrolo[2,l,c][l,4]benzodiazepin-5-one. These new analogues of pyrrolo[2,l-c][l,4]benzodiazepine hybrids linked at C-8 position have shown promising anticancer activity in various cell lines. The molecules synthesized are of immense biological significance with potential sequence selective DNA- binding property. This resulted in design and synthesis of new congeners as illustrated in Scheme- 1, which comprise:
1. ether linkage at C-8 position of DC-81 intermediates with pyrimidine ring moiety.
2. Refluxing the reaction mixture for 24-48 h.
3. Synthesis of C-8 linked PBD antitumour antibiotic hybrid imines.
4. Purification by column chromatography using different solvents like ethyl acetate, hexane, dichloromethane and methanol.
The following examples are given by way of illustration and therefore should not be construed as limiting the scope of invention.
Figure imgf000006_0001
R = H, OH, OAc n = 3, 4, 5
Scheme I
Example 1
Solution of 6-(4-fluorophenyl)-2-methyl-4-pyridinol of formula I (1000 mg 4.90 mmol), 1, 3-dibromopropane (2475 mg, 12.25 mmol) and K2CO3 (2030 mg, 14.7 mmol) in dry acetone (40 mL) was refluxed for 48 h. After the completion of reaction as indicated by
TLC, EtOAc-hexane (2:8), the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crude product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (5:95) gave the pure 3-bromopropyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II. 1HNMR: (CDC13) 2.1-2.2 (q, 2H), 2.6 (s, 3H), 4.1-4.15 (t, 2H), 4.45-4.55 (t, 2H), 6.6-6.8 (d, 1H), 7.08-7.12 (m, 2H), 8.0 (m, 2H); ELMS 204.
Solution of 3-bromopropyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II (1200 mg, 3.69 mmol), A solution of (2S)-N-(4-hydroxy-5-mβthoxy-2- nitrobenzoyl)pyrrolidine-2-carboxaldehyde diethylthioacetal of formula III (1472 mg, 3.69 mmol) and K2CO3 (1530 mg, 11.07 mmol) in dry acetone (20 mL) was refluxed for 48 h. After the completion of reaction as indicated by TLC, EtOAc, the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crude product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (1:3) gave the pure (2S)-N- [{4-[6'-(4"-fluoroρhenyl)-2'- methylpyrimidine-4'-yloxy]-propoxy}-5-methoxy-2-nitro benzoyl] pyrrolidine-2- carboxaldehyde diethyl thioacetal of formula IV. 1H NMR: (CDC13) δ 1.2-1.4 (m, 6H), 1.7- 2.42 (m, 6H), 2.65 (s, 3H), 2.65-2.8 (m, 4H), 3.2- 3.3 (m, 2H), 3.95 (s, 3H), 4.3 (m, 2H), 4.6 (m, 2H), 4.65-4.8 (m, 1H), 4.85 (d, 1H J = 4.2), 6.78 (s, 1H), 6.9 (s, 1H), 7.1-7.2 (m, 2H), 7.7 (s, 1H), 8.0-8.1 (m, 2H); FABMS 939 (M+H)+\
(2S)-N-[{4-[6 ' -(4"-fluorophenyl)-2 ' -methylpyrimidine-4 ' -yloxy]propoxy } -5-methoxy -2-nitrobenzoyl]pyrrolidine-2-carboxaldehyde diethyl thioacetal of formula IV (500 mg, 0.78 mmol) was dissolved in methanol (10 mL) and added SnCl2.2H2O (873 mg 3.88 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 (3x30 mL). The combined organic phase was dried over Na2S04 and evaporated under vacuum to afford the crude (2S)- N-[{4-[6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'-yloxy]propoxy}-5-methoxy-2- aminobenzoyl]pyrrolidine-2-carbo xaldehyde diethyl thioacetal of formula V.
.A solution of formula V (300 mg, 0.50 mmol), HgCl2 (300 mg, 1.10 mmol) and CaC03 (120 mg, 1.20 mmol) in CH3CN/H2O (3:1, 15 mL) was stirred at room temperature for 12 h until TLC (EtOAc), indicates complete loss of starting material. Then organic layer is evaporated in vacuum and the residue is diluted with EtOAc. To this saturated NaHCO3 was added slowly at room temperature and the mixture is filtered through celite and washed with ethyl acetate. The filtrate is evaporated in vacuum to get crude 7-methoxy-8-[6'-(4"- fluorophenyl)-2'-methylpyrimidine-4'-yloxy]propoxy-(l laS)-l,2,3,l la-tetrahydro-5H- pyrrolo[2,l,c ][1 ,4]benzodiazepin-5-one of formula VI, which was further purified by column chromatography on silica gel eluting first with ethyl acetate to remove traces of mercuric salts and further eluted with EtOAc-methanol (9:1). 1H NMR: (CDC13) δ 1.9-2.15 (m, 4H), 2.25-2.4 (m, 2H), 2.65 (s, 3H), 3.5-3.8 (m, 3H), 3.95 (s, 1H), 4.0-4.2 (m, 2H), 4.5 (m, 2H), 6.78 (s, 1H), 6.8 (s, 1H), 7.1-7.2 (m, 2H), 7.5 (s, 1H), 7.5 (d, 1H, J = 4.3 Hz), 7.95- 8.2 (m, 2H); FABMS: 631 (M+H)+\
Example 2
A solution of 6-(4-fluorophenyl)-2-methyl-4-pyridinol of formula I (1000 mg 4.90 mmol), 1, 4-dibromobutane (2650 mg, 12.25 mmol) and K2C03 (2030 mg, 14.70 mmol) in dry acetone (40 mL) was refluxed for 48 h. After the completion of reaction as indicated by TLC, EtOAc-hexane (2:8), the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crude product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (4:96) gave the pure 4-bromobutyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II. 1HNMR: (CDC13) δ 1.9-2.1 (m, 4H), 2.6-2.7 (s, 3H), 3.45-3.55 (t, 2H), 4.4-4.5 (t, 2H), 6.8 (s, 1H), 7.1-7.2 (m, 2H), 8.0-8.1 (m, 2H); ELMS 204.
Solution of 4-bromobutyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II (1300 mg, 3.83 mmol), A solution of (2o>N-(4-hydroxy-5-methoxy-2- nitrobenzoyl)pyrrolidine-2-carbox-aldehyde diethylthioacetal of formula III (1532 mg, 3.83 mmol) and K2CO3 (1585 mg, 11.47 mmol) in dry acetone (20 ml) was refluxed for 48 h. After the completion of reaction as indicated by TLC, EtOAc, the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crude product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (30:70) gave the pure (2S)-N-[{4-[6'-(4"-fluorophenyl)-2'- methylpyrimidine-4'-yloxy]butoxy}-5-methoxy-2-nitrobenzoyl]pyrrolidine-2- carboxaldehyde diethyl thioacetal of formula IV. 1H NMR: (CDC13) δ 1.9-2.15 (m, 6H), 2.25- 2.40 (m, 2H), 2.65 (s, 3H), 3.5-3.8 (m, 3H), 3.95 (s, 1H), 4.0-4.2 (m, 2H), 4.50 (m, 2H), 6.78 (s, 1H), 6.80 (s, 1H), 7.1-7.2 (m, 2H), 7.5 (s, 1H), 7.65 (d, 1H, J = 4.4 Hz) 7.95-8.10 (m, 2H); FABMS 939 (M+H)+>.
(2S)-N-[ {4-[6 ' -(4' '-fluorophenyl)-2 ' -methylpyrimidine-4 ' -yloxy]butoxy} -5-methoxy- 2-nitrobenzoyl]pyrrolidine-2-carboxaldehyde diethyl thioacetal of formula IV (500 mg, 0.76 mmol) was dissolved in methanol (10 mL) and added SnCl2.2H2O (855 mg 3.80 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 (3x30 mL). The combined organic phase was dried over Na2SO4 and evaporated under vacuum to afford the crude (2S)-N-[{4- [6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'-yloxy]butoxy}-5-methoxy-2- aminobenzoyl]pyrrohdine-2-carbox aldehyde diethyl thioacetal of formula V.
Solution of formula V (300 mg, 0.50 mmol), HgCl2 (300 mg, 1.10 mmol) and CaCO3 (120 mg, 1.20 mmol) in CH3CN/H2O (3:1, 15 mL) was stirred at room temperature for 12 h until TLC (EtOAc), indicates complete loss of starting material. Then organic layer is evaporated in vacuum and the residue is diluted with EtOAc. To this saturated NaHC03 was added slowly at room temperature and the mixture is filtered through celite and washed with ethyl acetate. The filtrate is evaporated in vacuum to get crude 7-methoxy-8-[6'-(4"- fluorophenyl)-2'-methylpyrimidine-4 '-yloxy]butoxy-(l laS)- 1 ,2,3, 11 a-tetrahydro-5H- pyrrolo[2,l,c][l,4]benzodiazepin-5-one of formula VI, which was further purified by column chromatography on silica gel eluting first with ethyl acetate to remove traces of mercuric salts and further eluted with EtOAc-methanol (97:3). 1HNMR (CDC13) δ 1.92-2.42 (m, 8H), 2.60-2.95 (m, 12H), 3.2-3.88 (m, 6H), 3.92 (s, 6H), 4.14-4.28 (m, 4H), 6.76 (s, 2H), 7.5 (s, 2H), 7.66 (d, 2H); FABMS: 631 (M+H)+\
Example 3
Solution of 6-(4-fluorophenyl)-2-methyl-4-pyridinol of formula I (1000 mg 4.90 mmol), 1, 5-dibromoρentane (2820 mg, 12.25 mmol) and K2CO3 (2030 mg, 14.70 mmol) in dry acetone (40 mL) was refluxed for 48 h. After the completion of reaction as indicated by TLC, EtOAc-hexane (2:8), the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crude product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (3:97) gave the pure 5-bromopentyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II. 1H NMR: (CDC13) 1.5-1.6 (q, 2H), 1.7-1.8 (q, 2H), 1.9-2.0 (q, 2H), 2.6 (s, 3H), 3.4 (t, 2H), 4.3-4.4 (t, 3H), 6.7 (s, 1H), 7.1-7.12 (m, 2H), 7.95-8.05 (m, 2H); ELMS 204.
Solution of 5-bromopentyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II (1400 mg, 3.97 mmol), A solution of (2S)-N-(4-hydroxy-5-methoxy-2- nitrobenzoyl)pyrrohdine-2-carbox-aldehydediethylthioacetal of formula III (1590 mg, 3. 97 mmol) and K2CO3 (1640 mg, 11.898 mmol) in dry acetone (20 mL) was refluxed for 48 h. After the completion of reaction as indicated by TLC, EtOAc, the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crude product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (27:73) gave the pure (2S)-N-[{4[6'-(4"-fluorophenyl)-2'- methylpyrimidine-4 ' -yloxyjpentoxy } -5 -methoxy-2 D -nitrobenzoyl]pyrrolidine-2- carboxaldehyde diethyl thioacetal of formula IV. 1H NMR (CDC13) δ 1.2-1.5 (m, 8H), 1.60- 2.4 (m, 8H), 2.64 (s, 3H), 2.7-2.82 (m, 4H), 3.2-3.3 (m, 2H), 3.95 (s, 3H), 4.2 (m, 2H), 4.35- 4.48 (t, 2H, J = 6.5 Hz), 4.62-4.75 (m, H), 4.85 (d, 1H, J = 4.2 Hz), 6.78 (s, 1H), 6.82 (s, 1H), 7.05-7.20 (m, 2H), 7.65 (s, 1H), 7.95-8.1 (m, 2H); FABMS 939 (M+H)+ \
(2ιS)-N-[{4-[6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'-yloxy]pentoxy}-5- methoxy-2-nitrobenzoyl]pyrrohdine-2-carboxaldehyde diethyl thioacetal of formula IV (500 mg, 0.74 mmol) was dissolved in methanol (10 mL) and added SnCl2.2H20 (840 mg 3.72 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 (3x30 mL). The combined organic phase was dried over Na2SO and evaporated under vacuum to afford the crude (2S)- N-[{4-[6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'-yloxy]pentoxy}-5-methoxy-2- aminobenzoyl]pyrrolidine-2-carbo xaldehyde diethyl thioacetal of formula V.
Solution of formula V (300 mg, 0.47 mmol), HgCl2 (280 mg, 1.03 mmol) and CaCO3 (110 mg, 1.10 mmol) in CH3CN/H2O (3:1, 15 mL) was stirred at room temperature for 12 h until TLC (EtOAc), indicates complete loss of starting material. Then organic layer is evaporated in vacuum and the residue is diluted with EtOAc. To this saturated NaHCO3 was added slowly at room temperature and the mixture is filtered through celite and washed with ethyl acetate. The filtrate is evaporated in vacuum to get crude 7-methoxy-8-[6'-(4"- fluorophenyl)-2'-methylpyrimidine-4'-yloxy]pentoxy-(l laS)-l,2,3, 1 la-tetrahydro-5H- pyrrolo[2,l,c][l,4]benzodiazepin-5-on e of formula VI, which was further purified by column chromatography on silica gel eluting first with ethyl acetate to remove traces of mercuric salts and further eluted with EtOAc. 'HNMR (CDC13) δ 1.6-2.2 (m, 10H), 2.65 (s, 3H), 3.6- 3.8 (m, 2H), 3.95 (s, 1H), 4.1-4.2 (m, 2H) 4.45 (m, 2H), 6.84 (s, 1H), 6.86 (s, H), 7.1-7.22 (m, 2H), 7.65 (s, 1H), 7.68-7.71 (d, 1H, J = 4.4 Hz), 8.0-8.1 (m, 2H); FABMS: 631 (M+H)+\
Example 4
Solution of 6-(4-fluorophenyl)-2-methyl-4-pyridinol of formula I (1000 mg 4.90 mmol), 1, 3-dibromoρroρane (2475 mg, 12.25 mmol) and K2CO3 (2030 mg, 14.70 mmol) in dry acetone (40 mL) was refluxed for 48 h. After the completion of reaction as indicated by TLC, EtOAc-hexane (2:8), the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crude product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (5:95) gave the pure 3-bromopropyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II.
Solution of 3-bromopropyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II (1200 mg, 3.69 mmol), A solution of (4R)-hydroxy-(2S)-N- (4-hydroxy-5- methoxy-2-nitrobenzoyl) pyrrohdine-2-carboxaldehyde diethylthioacetal of formula III (1472 mg, 3.69 mmol) and K2CO3 (1530 mg, 11.07 mmol) in dry acetone (20 mL) was refluxed for 48 h. After the completion of reaction as indicated by TLC, EtOAc, the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crude product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (1:3) gave the pure (4R)-hydroxy-(2S)-N-[{4-[6'-(4"- fluorophenyl)-2'-methylpyrimidine-4'-yloxy] propoxy}-5-methoxy-2-nitrobenzoyl] pyrrolidine-2-carboxaldehyde diethyl thioacetal of formula IV.
(4R)-hydroxy-(2S)-N-[{4-[6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'-yloxy] propoxy}-5-methoxy-2-nitrobenzoyl]pyrrolidine-2-carboxaldehyde diethyl thioacetal IV (500 mg, 0.78 mmol) was dissolved in methanol (10 mL) and added SnCl2.2H2O (873 mg 3.88 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 (3x30 mL). The combined organic phase was dried over Na2S04 and evaporated under vacuum to afford crude (4R)- hydroxy-(2S)-N-[{4-[6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'-yloxy]propoxy}-5- methoxy-2-nitrobenzoyl]pyrrolidine-2-carboxaldehyde diethyl thioacetal of formula V.
Solution of formula V (300 mg, 0.5 mmol), HgCl2 (300 mg, 1.10 mmol) and CaCO3 (120 mg, 1.20 mmol) in CH3CN/H20 (3:1, 15 mL) was stirred at room temperature for 12 h until TLC (EtOAc), indicates complete loss of starting material. Then organic layer is evaporated in vacuum and the residue is diluted with EtOAc. To this saturated NaHC03 was added slowly at room temperature and the mixture is filtered through celite and washed with ethyl acetate. The filtrate is evaporated in vacuum to get crude 7-methoxy-8-[6'-(4"- fluorophenyl)-2'-methylpyrimidine-4'-yloxy]propoxy-(4R)-hydroxy-(l laS)-l,2,3, 11a- tetrahydro-5H-pyrrolo[2,l,c][l,4]benzodiazepin-5-one of formula VI, which was further purified by column chromatography on silica gel eluting first with ethyl acetate to remove traces of mercuric salts and further eluted with EtoAc-methanol (9:1).
Example 5 Solution of 6-(4-fluorophenyl)-2-methyl-4-pyridinol of formula I (1000 mg 4.90 mmol), 1, 3-dibromoproρane (2475 mg, 12.25 mmol) and K2CO3 (2030 mg, 14.7 mmol) in dry acetone (40 mL) was refluxed for 48 h. After the completion of reaction as indicated by TLC, EtOAc-hexane (2:8), the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crade product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (5:95) gave the pure 3-bromopropyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II.
Solution of 3-bromopropyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II (1200 mg, 3.69 mmol), A solution of (4R)-hydroxy-(2S)-N- (4-hydroxy-5- methoxy-2-nitrobenzoyl)pyrroUdine-2-carboxaldehyde diethylthioacetal of formula III (1472 mg, 3.69 mmol) and K2CO3 (1530 mg, 11.07 mmol) in dry acetone (20 mL) was refluxed for 48 h. After the completion of reaction as indicated by TLC, EtOAc, the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crude product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (1:3) gave the pure (4R)-hydroxy-(2S)-N-[{4-[6'-(4"- fluorophenyl)-2'-methylpyrimidine-4' -yloxy] butoxy}-5-methoxy-2-nitrobenzoyl] pyrrolidine-2-carboxaldehyde diethyl thioacetal of formula IV.
(4R)-hydroxy-(2S)-N-[{4-[6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'-yloxy] butoxy}-5-methoxy-2-nitrobenzoyl]pyrrohdine-2-carboxaldehyde diethyl thioacetal of formula IV (500 mg, 0.78 mmol) was dissolved in methanol (10 mL) and added SnCl2.2H2O (873mg 3.88 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 (3x30 mL). The combined organic phase was dried over Na2SO4 and evaporated under vacuum to afford crude (4R)-hydroxy-(2S)-N-[ {4-[6 ' -(4" -fluorophenyl)-2' -methylpyrimidine-4' -yloxyjpentoxy } -5- methoxy-2-nitrobenzoyl] pyrrolidine-2-carboxaldehyde diethyl thioacetal of formula V.
Solution of formula V (300 mg, 0.5 mmol), HgCl2 (300 mg, 1.10 mmol) and CaCO3 (120 mg, 1.20 mmol) in CH3CN/H2O (3:1, 15 mL) was stirred at room temperature for 12 h until TLC (EtOAc), indicates complete loss of starting material. Then organic layer is evaporated in vacuum and the residue is diluted with EtOAc. To this saturated NaHC03 was added slowly at room temperature and the mixture is filtered through celite and washed with ethyl acetate. The filtrate was evaporated in vacuum to get crude 7-methoxy-8-[6'-(4"- fluorophenyl)-2'-methylpyrimidine-4'-yloxy]butoxy-(4R)-hydroxy-(l laS)-l,2,3,l la- tetrahydro-5H-pyrrolo[2,l,c][l,4]ben zodiazepin-5-one of formula VI, which was further purified by column chromatography on silica gel eluting first with ethyl acetate to remove traces of mercuric salts and further eluted with EtOAc-methanol (9: 1).
Example 6
Solution of 6-(4-fluorophenyl)-2-methyl-4-pyridinol of formula I (1000 mg, 4.90 mmol), 1, 3-dibromoproρane (2475 mg, 12.25 mmol) and K2CO3 (2030 mg, 14.70 mmol) in dry acetone (40 mL) was refluxed for 48 h. After the completion of reaction as indicated by TLC, EtOAc-hexane (2:8), the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crade product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (5.95) gave the pure 3-bromopropyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II.
Solution of 3-bromopropyl- 6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II (1200 mg, 3.69 mmol), A solution of (4R)-hydroxy-(2S)-N-(4-hydroxy-5- methoxy-2-nitrobenzoyl)pyrroUdine-2-carboxaldehyde diethylthioacetal of formula III (1472 mg, 3.69 mmol) and K2CO3 (1530 mg, 11.07 mmol) in dry acetone (20 mL) was refluxed for 8 h. After the completion of reaction as indicated by TLC, EtOAc, the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crude product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (1 :3) gave pure (4R)-hydroxy-(2S)-N-[{4-[6'-(4"-fluorophenyl)- 2'-methylpyrimidine-4'-yloxy] pentoxy}-5-methoxy-2-nitrobenzoyl] pyrrolidine-2- carboxaldehyde diethyl thioacetal of formula IV.
(4R)-hydroxy-(2S)-N-[{4-[6'-(4"-fluoroρhenyl)-2'-methylpyrimidine-4'-yloxy] pentoxy}-5-methoxy-2-nitrobenzoyl]pyrrolidine-2-carboxaldehyde diethyl thioacetal of formula IV (500 mg, 0.78 mmol) was dissolved in methanol (10 mL) and added SnCl2.2H20 (873 mg 3.88 mmol) was refluxed for 1.5 h. Reaction mixture was then carefully adjusted to pH 8 with saturated NaHCO3 solution and then extracted with ethyl acetate (3x30 mL). Combined organic phase was dried over Na2SO4 and evaporated under vacuum to afford crude (4R)-hydroxy-(2S)-N-[ {4-[6' -(4' ' -fluorophenyι)-2 ' -methylpyrimidine-4' -yloxy] pentoxy}-5-methoxy-2-aminobenzoyl]pyrrolidine-2-carboxaldehyde diethyl thioacetal of formula V.
Solution of formula V (300 mg, 0.50 mmol), HgCl2 (300 mg, 1.10 mmol) and CaCO3 (120 mg, 1.20 mmol) in CH3CN H2O (3:1, 15 mL) was stirred at room temperature for 12 h until TLC (EtOAc), indicates complete loss of starting material. Then organic layer is evaporated in vacuum and the residue is diluted with EtOAc. To this saturated NaHC03 was added slowly at room temperature and the mixture is filtered through celite and washed with ethyl acetate. The filtrate is evaporated in vacuum to get crade 7-methoxy-8-[6'-(4"- fluorophenyl)-2'-methylpyrimidine-4'-yloxy]pentoxy-(4R)-hydroxy-(l laS)-l,2,3,l la- tetrahydro-5H-pyrrolo[2,l,c][l,4]benzodiazepin-5-one of formula VI, which was further purified by column chromatography on silica gel eluting first with ethyl acetate to remove traces of mercuric salts and further eluted with EtOAc-methanol (9:1). Example 7
Solution of 6-(4-fluorophenyl)-2-methyl-4-pyridinol of formula I (1000 mg, 4.90 mmol), 1, 3-dibromopropane (2475 mg, 12.25 mmol) and K2CO3 (2030 mg, 14.70 mmol) in dry acetone (40 mL) was refluxed for 48 h. After the completion of reaction as indicated by TLC, EtOAc-hexane (2:8), the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crude product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (5:95) gave the pure3-bromopropyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II.
Solution of 3-bromopropyl-6-(4-fluorophenyl)-2-methyl-4-pyrimidyl ether of formula II (1200 mg, 3.69 mmol), A solution of (4R)-hydroxy-(2S)-N-(4-hydroxy-5- methoxy-2-nitrobenzoyl)pyrrohdine-2-carboxaldehyde diethylthioacetal of formula III (1472 mg, 3.69 mmol) and K2CO3 (1530 mg, 11.07 mmol) in dry acetone (20 mL) was refluxed for 48 h. After the completion of reaction as indicated by TLC, EtOAc, the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crade product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (1 :3) gave pure (4R)-hydroxy-(2S)-N-[{4-[6'-(4"-fluorophenyl)- 2'-methylpyrimidine-4'-yloxy] propoxy}-5-methoxy-2-nitrobenzoyl] pyrrolidine-2- carboxaldehyde diethyl thioacetal of formula IV.
To a stirred solution of (4R)-hydroxy-(2S)-N-[{4-[6'-(4"-fluoroρhenyl)-2'- methylpyrimidine-4'-yloxy]propoxy}-5-methoxy-2-nitrobenzoyl]pyrrolidine-2- carboxaldehyde diethyl thioacetal of formula IV (660 mg, 1.00 mmol) in dichloromethane (10 mL) triethylamine (125 mg, 1.20 mmol) was added under N2 atmosphere at 0°C. After stirring for 5 min. acetylchloride (100 mg, 1.20 mmol) was added drop wise at same temperature and reaction mixture was allowed to stir at room temperature and reaction mixture was allowed to stir at room temperature for overnight and then poured into water, extracted with dichloromethane and dried over Na2SO4 and evaporated in vacuo to afforded the corresponding compound (4R)-acetyloxy-(2iS)-N-[{4-[6'-(4"-fluorophenyl)-2'- methylpyrimidine-4'-yloxy]propoxy}-5-methoxy-2-nitrobenzoyl]pyrrolidine-2- carboxaldehyde diethyl thioacetal.
(4R)-acetyloxy-(2S)-N-[{4-[6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'-yloxy] propoxy}-5-methoxy-2-nitrobenzoyl]pyrroUdine-2-carboxaldehyde diethyl thioacetal (600 mg, 0.81 mmol) was dissolved in methanol (10 mL) and added SnCl2.2H2O (880 mg 3.90 mmol) was refluxed for 1.5 h. Reaction mixture was then carefully adjusted to pH 8 with saturated NaHCO3 solution and then extracted with ethyl acetate (3x30 mL). Cσrribmed organic phase was dried over Na2SO and evaporated under vacuum to afford the crade (4R)- acetyloxy-(2<S)-N-[ {4- [6 ' -(4 " -fluorophenyl)-2 ' -methylpyrimidine-4 ' -yloxyjpropoxy } -5 - methoxy-2-aminobenzoyl]pyrrohdine-2-carboxaldehyde diethyl thioacetal of formula V.
Solution of formula V (350 mg, 0.50 mmol), HgCl2 (300 mg, 1.10 mmol) and CaC03 (120 mg, 1.20 mmol) in CH3CN/H20 (3:1, 15 mL) was stirred at room temperature for 12 h until TLC (EtOAc), indicates complete loss of starting material. Then organic layer is evaporated in vacuum and the residue is diluted with EtOAc. To this saturated NaHC03 was added slowly at room temperature and the mixture is filtered through celite and washed with ethyl acetate. The filtrate is evaporated in vacuum to get crade 7-methoxy-8-[6'-(4"- fluorophenyl)-2'-methylpyrimidine-4,-yloxy]propoxy-(4R)-acetyloxy-(l laS)-l,2,3, 11a- tetrahydro-5H-pyrrolo[2,l,c][l,4]benzodiazepin-5-one of formula VI, which was further purified by column chromatography on silica gel eluting first with ethyl acetate to remove traces of mercuric salts and further eluted with EtOAc-methanol (9:1).
Example 8
Solution of 6-(4-fluorophenyl)-2-methyl-4-pyridinol of formula I (1000 mg, 4.90 mmol), 1, 3-dibromopropane (2475 mg, 12.25 mmol) and K2CO3 (2030 mg, 14.70 mmol) in dry acetone (40 mL) was refluxed for 48 h. After the completion of reaction as indicated by TLC, EtOAc-hexane (2:8), the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crude product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (5:95) gave the pure3-bromopropyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II.
Solution of 3-bromopropyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II (1200 mg, 3.69 mmol), A solution of (4R)-hydroxy-(2S)-N-(4-hydroxy-5- methoxy-2-nitrobenzoyl)pyrrolidine-2-carboxaldehyde diethylthioacetal of formula III (1472 mg, 3.69 mmol) and K2CO3 (1530 mg, 11.07 mmol) in dry acetone (20 mL) was refluxed for 48 h. After the completion of reaction as indicated by TLC, EtOAc, the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crude product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (1:3) gave pure (4R)-hydroxy-(2S)-N-[{4-[6'-(4"-fluorophenyl)- 2 ' -methylpyrimidine-4 ' -yloxy] butoxy } -5 -methoxy-2-nitrobenzoyl] pyrrolidine-2- carboxaldehyde diethyl thioacetal of formula IV. To a stirred solution of (4R)-hydroxy-(2S)-N-[{4-[6'-(4"-fluorophenyl)-2'- methylpyrimidine-4'-yloxy]butoxy}-5-methoxy-2-nitrobenzoyl]pyrrolidine-2- carboxaldehyde diethyl thioacetal of formula IV (674 mg, 1.00 mmol) in dichloromethane (10 mL) triethylamine (125 mg, 1.2 mmol) was added under N2 atmosphere at 0.°C. After stirring for 5 min. acetylchloride (100 mg, 1.20 mmol) was added drop wise at same temperature and reaction mixture was allowed to stir at room temperature for overnight and then poured into water, extracted with dichloromethane and dried over Na2SO4 and evaporated in vacuum to afforded the corresponding compound (4R)-acetyloxy-(2S)-N-[{4- [6 ' -(4 " -fluorophenyl)-2 ' -methylpyrimidine-4 ' -yloxy]butoxy } -5 -methoxy-2-nitrobenzoyl] pyrrohdine-2-carboxaldehyde diethyl thioacetal.
(4R)-acetyloxy-(2S)-N-[{4-[6'-(4"-fluorophenyl)-2'-methylpyrimidine-4'-yloxy] butoxy}-5-methoxy-2-nitrobenzoyl]pyrrolidine-2-carboxaldehyde diethyl thioacetal (600 mg, 0.79 mmol) was dissolved in methanol (10 mL) and added SnCl2.2H2O (880 mg, 3.95 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 (3x30 mL). Combined organic phase was dried over Na2SO4 and evaporated under vacuum to afford crude (4R)- acetyloxy-(2S)-N-[ {4-[6 ' -(4" -fluorophenyl)-2 ' -methylpyrimidine-4 ' -yloxyjbutoxy } -5- methoxy-2-aminobenzoyl]pyrrohdine-2-carboxaldehyde diethyl thioacetal of formula V.
Solution of formula V (360 mg, 0.50 mmol), HgCl2 (300 mg, 1.10 mmol) and CaCO3 (120 mg, 1.20 mmol) in CH3CN/H2O (3:1, 15 mL) was stirred at room temperature for 12 h until TLC (EtOAc), indicates complete loss of starting material. Then organic layer is evaporated in vacuum and the residue is diluted with EtOAc. To this saturated NaHC03 was added slowly at room temperature and the mixture is filtered through celite and washed with ethyl acetate. The filtrate is evaporated in vacuum to get crade 7-methoxy-8-[6'-(4"- fluorophenyl)-2'-methylpyrimidine-4'-yloxy]butoxy-(4R)-acetyloxy-(l laS)-l,2,3,l la- tetrahydro-5H-pyrrolo[2,l,c][l,4]benzodiazepin-5-one of formula VI, which was further purified by column chromatography on silica gel eluting first with ethyl acetate to remove traces of mercuric salts and further eluted with EtOAc-methanol (9:1).
Example 9
Solution of 6-(4-fluorophenyl)-2-methyl-4-pyridinol of formula I (1000 mg, 4.90 mmol), 1, 3-dibromopropane (2475 mg, 12.25 mmol) and K2CO3 (2030 mg, 14.70 mmol) in dry acetone (40 mL) was refluxed for 8 h. After the completion of reaction as indicated by TLC, EtOAc-hexane (2:8), the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crade product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (5:95) gave the pure 3-bromopropyl-6-(4-fluoro phenyl)-2-methyl-4-pyrimidyl ether of formula II.
A solution of 3-bromopropyl-6-(4-ftuoro phenyl)-2-methyl-4-pyrimidyl ether of formula II (1200 mg, 3.69 mmol), A solution of (4R)-hydroxy-(2S)-N-(4-hydroxy-5- methoxy-2-nitrobenzoyl)pyrrolidine-2-carboxaldehyde diethylthioacetal of formula III (1472 mg, 3.69 mmol) and K2CO3 (1530 mg, 11.07 mmol) in dry acetone (20 mL) was refluxed for 48h. After the completion of reaction as indicated by TLC, EtOAc, the reaction mixture was poured on to the water and then extracted with ethyl acetate. Evaporation of the organic layer gave the crade product, which was further purified by column chromatography on silica gel eluting with EtOAc-hexane (1:3) gave pure (4R)-hydroxy-(2S)-N-[{4-[6'-(4"-fluorophenyl)- 2 '-methylpyrimidine-4 '-yloxy] pentoxy}-5-methoxy-2-nitrobenzoyl] pyrrolidine-2- carboxaldehyde diethyl thioacetal of formula IV.
To a stirred solution of (4R)-hydroxy-(2S)-N-[{4-[6'-(4"-fluoroρhenyl)-2'- methylpyrimidine-4'-yloxy]pentoxy}-5-methoxy-2-nitrobenzoyl] pyrrolidine-2- carboxaldehyde diethyl thioacetal of formula IV (688 mg, 1 Ommol) in dichloromethane (lOmL) triethylamine (125 mg, 1.20 mmol) was added under N2 atmosphere at 0°C. After stirring for 5 min. acetylchloride (100 mg, 1.20 mmol) was added drop wise at same temperature and reaction mixture was allowed to stir at room temperature overnight and then poured into water, extracted with dichloromethane and dried over Na2SO4 and evaporated in vacuum to afforded the corresponding compound (4R)-acetyloxy- (2S)-N-[{4-[6'-(4"- fluorophenyl)-2'-methylpyrimidine-4'-yloxy]pentoxy}-5-methoxy-2-nitrobenzoyl] pyrrolidine-2-carboxaldehyde diethyl thioacetal.
(4R)-acetyloxy-(2S)-N-[{4-[6 ' -(4" -fluorophenyl)-2' -methylpyrimidine-4' -yloxy] pentoxy}-5-methoxy-2-nitrobenzoyl]pyrrohdine-2-carboxaldehyde diethyl thioacetal (600 mg, 0.78 mmol) was dissolved in methanol (10 mL) and added SnCl2.2H2O (880 mg 3.90 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 (3x30 mL). Combined organic phase was dried over Na2SO4 and evaporated under vacuum to afford crude (4R)- acetyloxy-(2S)-N-[ {4-[6 ' -(4 ' ' -fluorophenyl)-2 ' -methylpyrimidine-4 ' -yloxy]pentoxy } -5- methoxy-2-aminobenzoyl]py rroUdine-2-carboxaldehyde diethyl thioacetal of formula V.
Solution of formula V (365 mg, 0.50 mmol), HgCl2 (300 mg, 1.10 mmol) and CaC03 (120 mg, 1.20 mmol) in CH3CN H2O (3:1, 15 mL) was stirred at room temperature for 12 h until TLC (EtOAc), indicates complete loss of starting material. Then organic layer is evaporated in vacuum and the residue is diluted with EtOAc. To this saturated NaHC03 was added slowly at room temperature and the mixture is filtered through celite and washed with ethyl acetate. The filtrate is evaporated in vacuum to get crude 7-methoxy-8-[6'-(4"- fluorophenyl)-25-methylpyrimidine-4'-yloxy]pentoxy-(4R)-acetyloxy-(l laS)-l,2,3, 1 la- tetralιydro-5H-pyrrolo[2,l,c][l,4]benzodiazepin-5-one of formula VI, which was further purified by column chromatography on silica gel eluting first with ethyl acetate to remove traces of mercuric salts and further eluted with EtOAc-methanol (9:1). Biological Activity: In vitro biological activity studies were carried out at National Cancer Institute (USA).
Cytotoxicity: Compounds Via and VIb were evaluated in vitro against sixty human tumour cells derived from nine cancer types (leukemia, non-small-cell lung, colon, CNS, melanoma, ovarian, prostate, and breast cancer). For each compound, dose response curves for each cell line were measured at a minimum of five concentrations at 10 fold dilutions. A protocol of 48 h continuous drug exposure was used and a sulforhodamine B (SRB) protein assay was used to estimate cell viability or growth. The concentration causing 50% cell growth inhibition (GI50), total cell growth inhibition (TGI 0% growth) and 50% cell death (LC50, -50% growth) compared with the control was calculated. The mean graph midpoint values of log10TGI and log10LC50 as well as log10 GI50 for Via and VIb are listed in Table 1. As demonstrated by mean graph pattern, compound VIb exhibits an interesting profile of activity and selectivity for various cell lines. The mean graph mid point of log10 TGI and log10 LC50 showed similar pattern to the log10 GI50 mean graph mid points.
Table 1. log10 GI50 log10 TGI and log10 LC50 mean graphs midpoints(MG_MLD) of in vitro cytotoxicity data for the compounds Via and VI against human tumour cell lines.
Compound Logιo GI50 LogioTGI LogιoLC50
" via 73 ^34 ^4A0
Vib -6.41 -5.57 -4.69
The comparison of the data of Table 2 reveals the importance of the alkane spacer. As the alkane spacer increased from 3-4 the cytotoxic activity has moderately enhanced. The 4 carbon spacer of compound VIb confers a suitable fit in the minor groove of double helix DNA and shows slightly higher activity in this series of compounds Via and VIb. Table 2. Log LC50 (concentration in mol/L causing 50% lethality) Values for Compounds Via and VIb Cancer Compound Compound Via VIb
Leukaemia -4 03 -5 41 non-small-cell lung -4 07 -4 61
Colon -4 12 _4 67
CNS -4 10 -4 40
Melanoma -4 14 -5 22
Ovarian -4 05 -4 33
Renal -4 18 -4 39
Prostate -4 00 -4 45
.orβeiSt -4 09 -4 52
Each cancer type represents the average of six to eight different cancer cell lines.

Claims

We Claim:
1. Novel pyrrolo[2,l-c][l,4]benzodiazepine of formula VI wherein R is H , OH, OAc and n is 3 to 5:
Figure imgf000020_0001
FORMULA VI Pyrrolobenzodiazepine as claimed in claim 1 of the stractural formula shown below:
Figure imgf000020_0002
3. Pyrrolobenzodiazepine as claimed in claim 1 of the stractural formula shown below:
Figure imgf000020_0003
Pyrrolobenzodiazepine as claimed in claim 1 of the structural formula shown below:
Figure imgf000020_0004
5. Pyrrolobenzodiazepine as claimed in claim 1 of the structural formula shown below:
Figure imgf000020_0005
6. Pyrrolobenzodiazepine as claimed in claim 1 of the stractural formula shown below:
Figure imgf000021_0001
7. Pyrrolobenzodiazepine as claimed in claim 1 of the structural formula shown below:
Figure imgf000021_0002
8. Pyrrolobenzodiazepine as claimed in claim 1 of the structural formula shown below:
Figure imgf000021_0003
9. Pyrrolobenzodiazepine as claimed in claim 1 of the stractural formula shown below:
Figure imgf000021_0004
10. Pyrrolobenzodiazepine as claimed in claim 1 of the stractural formula shown below:
Figure imgf000021_0005
11. A process for the preparation of novel pyrrolo[2,l-c][l,4]benzodiazepine of formula VI wherein R is H , OH, OAc and n is 3 to 5:
Figure imgf000022_0001
FORMULA VI the process comprising reacting 6-4-(4-fluorophenyl)-2-methyl-4-pyrimidinol of formula I
Figure imgf000022_0002
FORMULA I with a dibromoalkane in an aprotic water miscible organic solvent in the presence of a mild inorganic base up to refluxing temperature for a period up to 48 h, isolating n- bromoalkyl-6-(4-flurophenyl)-2-methyl-4-pyrimidinyl ether of formula II with (2S)- N-(4-hydroxy-5-methoxy-2-nitrobenzoyl)pyrrolidine-2-carboxaldehyde diethyl thioacetal of formula LH
Figure imgf000022_0003
in presence of a mild inorganic base in presence of aprotic water miscible organic solvent up to refluxing for a period of 48 h, isolating (2S) - N - [(4 - [6" - (4' - fluorophenyl) - 2' - methylpyrimidine - 4' - yloxy] alkoxy} - 5 - methoxy - 2 - nitrobenzoyl] pyrrolidine - 2 - carboxaldehyde diethyl thioacetal of formula IV where n is 3 to 5,
Figure imgf000023_0001
FOMMULA IV reducing the compound of formula IV with SnCl2.2H2O in presence of organic solvent up to a reflux temperature, isolating the (2S)-N-[{4 - [6' - (4" - fluorophenyl) - 2' - methylpyrimidine - 4' - yloxy] alkoxy} - 5 - methoxy - 2 - amonobenzoyl] pyrrolidine - 2 - carboxaldehyde diethyl thioacetal of formula V where n is 3 to 5
Figure imgf000023_0002
reacting the compound of formula V with a deprotecting agent to obtain the pyrrolo[2,l-c][l,4]benzodiazepines of formula VI.
12. A process as claimed in claim 11 wherein the aprotic water miscible organic solvent is selected from the group consisting of acetone, THF and DMF.
13. A process as claimed in claim 11 wherein the mild inorganic base is selected from the group consisting of K2CO3, CsCO3 and BaCO3.
14. A process as claimed in claim 11 wherein the isolated (2S) - N - [{4 - [6" - (4' - fluorophenyl) - 2' - methylpyrimidine - 4' - yloxy] alkoxy} - 5 - methoxy - 2 - nitrobenzoyl] pyrrolidine - 2 - carboxaldehyde diethyl thioacetal of formula IV is reduced using SnCl2.2H O in presence of an organic solvent.
15. Use of a novel pyrrolo[2,l-c][l,4]benzodiazepine of formula VI wherein R is H , OH, OAc and n is 3 to 5:
Figure imgf000023_0003
FORMULA VI in the treatment of tumours in a subject suffering from cancer.
16. Use as claimed in claim 15 wherein said cancer comprises leukemia, non-small-cell lung, colon, CNS, melanoma, ovarian, prostate and breast cancer.
17. A method for the treatment of cancer in a subject suffering therefrom comprising administering a pharmaceutically effective dose of a pyrrolo[2,l- c][l,4]benzodiazepine of formula VI wherein R is H , OH, OAc and n is 3 to 5:
Figure imgf000024_0001
FORMULA VI
18. A method as claimed in claim 17 wherein the cancer is selected from the group consisting of leukemia, non-small-cell lung, colon, CNS, melanoma, ovarian, prostate and breast cancer.
PCT/IN2003/000122 2003-03-31 2003-03-31 NEW PYRIMIDINE LINKED PYRROLO[2,1-c][1,4]BENZODIAZEPINES AS POTENTIAL ANTITUMOUR AGENTS WO2004087712A1 (en)

Priority Applications (2)

Application Number Priority Date Filing Date Title
PCT/IN2003/000122 WO2004087712A1 (en) 2003-03-31 2003-03-31 NEW PYRIMIDINE LINKED PYRROLO[2,1-c][1,4]BENZODIAZEPINES AS POTENTIAL ANTITUMOUR AGENTS
AU2003222444A AU2003222444A1 (en) 2003-03-31 2003-03-31 NEW PYRIMIDINE LINKED PYRROLO(2,1-c)(1,4)BENZODIAZEPINES AS POTENTIAL ANTITUMOUR AGENTS

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/IN2003/000122 WO2004087712A1 (en) 2003-03-31 2003-03-31 NEW PYRIMIDINE LINKED PYRROLO[2,1-c][1,4]BENZODIAZEPINES AS POTENTIAL ANTITUMOUR AGENTS

Publications (1)

Publication Number Publication Date
WO2004087712A1 true WO2004087712A1 (en) 2004-10-14

Family

ID=33104971

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IN2003/000122 WO2004087712A1 (en) 2003-03-31 2003-03-31 NEW PYRIMIDINE LINKED PYRROLO[2,1-c][1,4]BENZODIAZEPINES AS POTENTIAL ANTITUMOUR AGENTS

Country Status (2)

Country Link
AU (1) AU2003222444A1 (en)
WO (1) WO2004087712A1 (en)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009109984A1 (en) * 2008-03-05 2009-09-11 Council Of Scientific & Industrial Research Quinazoline linked pyrrolo[2,1-c][1, 4]benzodiazepine hybrids as potential anticancer agents and process for the preparation thereof'
WO2012110840A1 (en) * 2011-02-15 2012-08-23 Council Of Scientific & Industrial Research Diaryl ether linked pyrrolo [2,1-c][1,4] benzodiazepine hybrids and process for the preparation thereof

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0101924A1 (en) * 1982-07-26 1984-03-07 Bristol-Myers Company Antitumor antibiotics
EP0781766A1 (en) * 1994-09-09 1997-07-02 Nippon Shinyaku Company, Limited Heterocyclic derivative and medicine

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0101924A1 (en) * 1982-07-26 1984-03-07 Bristol-Myers Company Antitumor antibiotics
EP0781766A1 (en) * 1994-09-09 1997-07-02 Nippon Shinyaku Company, Limited Heterocyclic derivative and medicine

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2009109984A1 (en) * 2008-03-05 2009-09-11 Council Of Scientific & Industrial Research Quinazoline linked pyrrolo[2,1-c][1, 4]benzodiazepine hybrids as potential anticancer agents and process for the preparation thereof'
JP2011513389A (en) * 2008-03-05 2011-04-28 カウンスィル オブ サイエンティフィック アンド インダストリアル リサーチ Quinazoline-binding pyrrolo [2,1-c] [1,4] benzodiazepine hybrid as an effective anticancer agent and method for producing the same
US8153627B2 (en) 2008-03-05 2012-04-10 Council Of Scientific & Industrial Research Quinazoline linked pyrrolo[2,1-C][1, 4]benzodiazepine hybrids as potential anticancer agents and process for the preparation thereof
WO2012110840A1 (en) * 2011-02-15 2012-08-23 Council Of Scientific & Industrial Research Diaryl ether linked pyrrolo [2,1-c][1,4] benzodiazepine hybrids and process for the preparation thereof
US8809321B2 (en) 2011-02-15 2014-08-19 Council Of Scientific And Industrial Research Diaryl ether linked pyrrolo [2,1-c][1,4] benzodiazepine hybrids and process for the preparation thereof

Also Published As

Publication number Publication date
AU2003222444A1 (en) 2004-10-25

Similar Documents

Publication Publication Date Title
EP1608664B1 (en) Non-cross-linking pyrrolo[2,1-c][1,4]benzodiazepines as potential antitumour agents and process thereof
EP2061795B1 (en) Pyrrolo[2,1-c][1,4]benzodiazepine hybrids and a process for the preparation thereof
US7189710B2 (en) C2-fluoro pyrrolo [2,1−c][1,4]benzodiazepine dimers
JP5657395B2 (en) Quinazoline-binding pyrrolo [2,1-c] [1,4] benzodiazepine hybrid as an effective anticancer agent and method for producing the same
EP1608663A1 (en) Pyrrolo (2,1-c)(1,4) benzodiazepines dimers as antitumour agents and process thereof
US6683073B1 (en) Pyrimidine linked pyrrolo[2,1-c][1,4]benzodiazepines as potential antitumour agents
EP1608650B1 (en) Pyrene-linked pyrrolo(2,1-c)(1,4)benzodiazepine derivatives useful as anticancer agents
US20050222133A1 (en) Process for preparing pyrrolo[2, 1-c] [1,4] benzodiazepine hybrids
US6800622B1 (en) Pyrene-linked pyrrolo[2,1-c][1,4]benzodiazepine hybrids useful as anti-cancer agents
RU2314309C2 (en) Pyrrolo[2.1-c][1.4]benzodiazepines, method for their preparing and pharmaceutical composition based on thereof
US7056913B2 (en) C8—linked pyrrolo[2,1-c][1,4]benzodiazepine-acridone/acridine hybrids
JP4520411B2 (en) Method for preparing pyrrolo [2,1-c] [1,4] benzodiazepine hybrid
WO2004087712A1 (en) NEW PYRIMIDINE LINKED PYRROLO[2,1-c][1,4]BENZODIAZEPINES AS POTENTIAL ANTITUMOUR AGENTS
JP4833663B2 (en) C2-fluoropyrrolo [2,1-c] [1,4] benzodiazepine dimer
EP2260039B1 (en) Chalcone linked pyrrolo[2,1-c[[1, 4]benzodiazepine hybrids as potential anticancer agents and process for the preparation thereof
JP4718328B2 (en) C8-linked pyrrolo [2,1-c] [1,4] benzodiazepine-acridone / acridine hybrid
EP2271648B1 (en) Isoxazoline linked pyrrolo [2,1-c][1,4]benzodiazepine hybrids as potential anticancer agents and the process for preparattion thereof
EP2373656A1 (en) Cinnamido-pvrrolor[2,1-c][1,4]benzodiazepines as potential anticancer agents and process for the preparation thereof
WO2010058416A1 (en) Quinazolinone linked pyrrolo[2,1 -c][1.4]benzodiazepine hybrids as potential anticancer agents and process forthe preparation thereof
EP2262809A1 (en) Benzophenone-piperazine linked pyrrolo[2,1-c][1,4]benzodiazepine hybrids as potential anticancer agents and process for the preparation thereof

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ OM PH PL PT RO RU SD SE SG SK SL TJ TM TN TR TT TZ UA UG UZ VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HU IE IT LU MC NL PT RO SE SI SK TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
122 Ep: pct application non-entry in european phase
NENP Non-entry into the national phase

Ref country code: JP

WWW Wipo information: withdrawn in national office

Country of ref document: JP