WO1994005641A1 - 4-SUBSTITUTED 6,9,-BIS (SUBSTITUTED-AMINO) BENZO [g] ISOQUINOLINE-5,10,DIONES - Google Patents

4-SUBSTITUTED 6,9,-BIS (SUBSTITUTED-AMINO) BENZO [g] ISOQUINOLINE-5,10,DIONES Download PDF

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Publication number
WO1994005641A1
WO1994005641A1 PCT/US1993/008242 US9308242W WO9405641A1 WO 1994005641 A1 WO1994005641 A1 WO 1994005641A1 US 9308242 W US9308242 W US 9308242W WO 9405641 A1 WO9405641 A1 WO 9405641A1
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Prior art keywords
amino
hydroxy
dione
bis
benzo
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PCT/US1993/008242
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English (en)
French (fr)
Inventor
A. Paul Krapcho
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The University Of Vermont
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Priority to AU50997/93A priority Critical patent/AU5099793A/en
Publication of WO1994005641A1 publication Critical patent/WO1994005641A1/en

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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D221/00—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00
    • C07D221/02—Heterocyclic compounds containing six-membered rings having one nitrogen atom as the only ring hetero atom, not provided for by groups C07D211/00 - C07D219/00 condensed with carbocyclic rings or ring systems
    • C07D221/04—Ortho- or peri-condensed ring systems
    • C07D221/06—Ring systems of three rings
    • C07D221/08—Aza-anthracenes

Definitions

  • This invention is directed to 4-hydroxy-, alkoxy- or aralkoxy- 6,9-bis(substituted-a ino)benzo [g]isoquinoline-5,10-diones, and more particularly, to 4-hydroxy derivatives with 6,9-substituents which are (aminoalkyl)amino substituents. These compounds have been shown to have antitumor activity in vitro and in vivo.
  • Mitoxantrone is a broad spectrum oncolytic agent, whose activity is similar to that of the anthracycline antibiotic doxorubicin. Clinical trials have demonstrated that mitoxantrone has particularly promising activity in the treatment of advanced breast cancer, acute leukemia and lymphoma [Legha, Drugs of Today. 20. 629 (1984)].
  • SUBSTITUTE SHEET Ametantrone has been reported to be, in animals, about 10-fold less potent and cardiotoxic than mitoxan ⁇ trone. Because a delayed toxicity is observed only with mitoxantrone after administration of the two drugs by the i.p. route to non-tumor bearing rats at equieffective antitumor dosages, it is suggested that the presence of the 5,8-dihydroxy substitution in mitoxantrone might be implicated in the delayed deaths (Corbett et al. , Cancer Chemother. Pharmacol.. 6, 161 (1981)).
  • both mitoxantrone and ametantrone have a remarkable myelodepressive toxicity and both compounds show cross-resistance to cell histotypes developing resistance against doxorubicin mediated by overexpression of glycoprotein P.
  • Such a resistance which is named multidrug resistance, involves a number of antitumor antibiotics, among which amsacrine and podophyllotoxinic derivatives, and it is one of the main reasons for therapeutical failures in the treatment of solid tumors with said antibiotics.
  • novel anthracenedione antitumor agents having a higher therapeutical index than mitoxantrone and being effective both in inhibiting or delaying the growth of those solid tumors which are more refractory to chemotherapy (such as lung, breast, and colon tumors) and against tumor histotypes developing multidrug resistance.
  • SUBSTITUTE be related to the antitumor l,4bis[ (aminoalkyl)amino] anthracene-9, 10-diones; however, no antitumor activity data was reported for any of the above compounds.
  • l-[ (aminoalkyl)amino] derivatives (4) of 6,9-dihydroxy- benzo[g] isoquinoline-5, 10-dione related to mitoxantrone have been disclosed as DNA intercalators but they were completely devoid of any in vitro or in vivo antitumor activity (Croisy-Delcey et al. , Eur. J. Med. Chem. , 23. (1988), 101-106).
  • the compounds 5a and 5b are less cytotoxic than the analogues 6a and 6b.
  • the compound . 5a which is poorly cytotoxic in vitro, is inactive in vivo and it is both less active and less potent than the carbocyclic analogue 6a.
  • R CH 2 CH 2 N (Et) 2 6 a
  • R CH 2 CH 2 N (CH 3 ) 2
  • R CH 2 CH 2 CH 2 N (CH 3 ) 2 6b
  • R CH 2 CH 2 N (Et) 2 5d
  • R CH 2 CH 2 NHCOCH 3
  • the compounds of the invention have the formula
  • P is hydrogen, C_,-C 10 alkyl, C ⁇ -C. Q phenalkyl
  • R is ⁇ -C. 0 alkyl or C 7 -C 10 phenalkyl; C 2 -C 10 alkyl having one or two substituents selected from the group consisting of OR 1 and -NR 2 R 3 ; a dialkyl ether of the formula -(CH 2 ) n -0-(CH 2 ) m H or a dialkyl amine of the formula -(CH 2 ) n -NR 4 -(CH 2 ) m H wherein m+n equals an integer from 2 through 10, and may be unsubstituted or substituted by one or two hydroxy (OH) or -NR 2 R 3 groups;
  • R. is selected from the group consisting of hydrogen, C,-C 6 alkyl, C ⁇ - ' i o phenalkyl, C 2 -C 6 alkyl unsubstituted or substituted by -NR 2 R 3 ;
  • R 2 and R 3 may be the same or different and are selected from the group consisting of hydrogen, ⁇ ] -C w alkyl, C 7 -C 10 phenalkyl, phenyl, C 2 -C 10 alkyl substituted with one or two hydroxy (OH) groups; or one of R 2 or R 3 is H and the other is C(CH 3 ) (CH 2 OH) 2 ; or R 2 and R 3 taken
  • SUBSTITUTESHEET together with the nitrogen atom to which they are bound form an ethyleneimine ring or a 5- or 6-membered aromatic or nonaromatic heterocyclic ring which might contain another heteroatom such as sulfur, oxygen or nitrogen;
  • R 4 is selected from the group consisting of hydrogen, C ⁇ C ⁇ alkyl, C 2 -C 10 hydroxyalkyl, C 7 -C 10 aralkyl; as free bases and their salts with pharmaceutically acceptable acids.
  • the present invention also concerns the tautomeric forms, the single enantiomers and diastereoisomers of the compounds of formula (I) , as well as mixtures thereof.
  • the present invention also concerns the non-toxic salts of the compounds of formula (I) with acids acceptable for pharmaceutical and veterinary use such as those obtained by addition of inorganic acids like hydrochloric, hydrobromic, sulfuric, phosphoric, pyrophosphoric acid and/or of organic acids such as acetic, propionic, citric, benzoic, lactic, maleic, fumaric, succinic, tartaric, glutamic, aspartic, gluconic, ascorbic acids and the like.
  • acids acceptable for pharmaceutical and veterinary use such as those obtained by addition of inorganic acids like hydrochloric, hydrobromic, sulfuric, phosphoric, pyrophosphoric acid and/or of organic acids such as acetic, propionic, citric, benzoic, lactic, maleic, fumaric, succinic, tartaric, glutamic, aspartic, gluconic, ascorbic acids and the like.
  • C.-. Q alkyl groups are methyl, ethyl, n-propyl, sec-propyl, n-butyl, sec-butyl, tert-butyl, n-pentyl, n-hexyl.
  • C 7 - C 10 phenalkyl are benzyl and 4-methoxy benzyl.
  • R is a dialkyl ether of the formula (CH 2 ) n -0-(CH 2 ) ra -H or a dialkyl amine of the formula (CH 2 ) n -NR 4 -(CH 2 ) (n -H and optionally substituted by one or two hydroxy or -NR 2 R 3 groups, at least two carbon atoms are preferably interposed between said oxygen atom and/or the -NR ⁇ - and -NR 2 R 3 groups.
  • the -NR 2 R 3 sub- stituent is a 5-6 member aromatic or not aromatic heterocyclic ring which may contain another heteroatom such as sulfur, oxygen and nitrogen, preferred examples of
  • SUBSTITUTE SHEET said heterocyclic rings are 1-imidazolyl, 1- pyrrolyl, l-tetrahydropyrrolyl, 1-pyrazolyl,
  • R is a C 2 -C 10 alkyl selected from the group consisting of:
  • the evaluation of the in vitro cytotoxic acti ⁇ vity of the compounds of the invention was performed using a human colon adenocarcinoma cell line (Lovo) isolated from a metastatic nodule and a subline with acquired resistance to a number of antitumor agents, among which are doxorubicin, VP-16 and vincristine.
  • This subline (named Lovo/DX) shows reduced accumulation of doxorubicin and overexpression of a protein (Grandi, M. , Geroni, C, Giuliani, F.C., British J. Cancer, (1986), 54 , 515).
  • the compounds were tested according to the MTT assay (Mos an, T.
  • a resistance index R.I. (defined as the ratio of the IC 50 for the resistant cell line to the IC 50 for the sensitive cell line) as high as 148 and 30 respectively were found, showing that this subline did have an acquired resistance to mitoxantrone and ametantrone.
  • R.I. defined as the ratio of the IC 50 for the resistant cell line to the IC 50 for the sensitive cell line
  • representative compounds of this invention were tested in the same resistant subline, a resistance index significantly lower occurred than with mitoxantrone and ametantrone.
  • the in vitro cytotoxic evaluation was also performed on L 1210 murine leukemia cells using the above mentioned cells maintained in suspension cultures (McCoy's 5A medium supplemented with 10% horse serum, glutamine, penicillin and streptomycin) grown in an humidified environment of 10% carbon dioxide and 90% air at 37°C.
  • the compounds were dissolved in dimethylsulfoxide (DMSO) and added to the suspended cells in appropriate concentrations. After 72 hours of continuous exposure, the cell concentration was counted using a Coulter Counter
  • growth inhibition 1- [cell number treated/cell number DMSO alone] x 100.
  • the ID 50 was calculated from the growth inhibition data and they are reported in Table IV in comparison with mitoxantrone aznd with the prior art compound 5_a (see Table 1 for structure of 5a) .
  • a representative compound of the invention is significantly more cytotoxic than mitoxantrone and dramatically more active than compound 5a.
  • the compounds of the present invention when tested against the adherent murine sarcoma cell line S180 and its subline S180/A10 made resistant to doxorubicin and 100 fold resistant to mitoxantrone, are more cytotoxic than mitoxantrane both in the sensitive and in the resistant cell lines and are not cross resistant to mitoxantrone.
  • Table V reports the data for a representative compound of the invention, 4-hydroxy-6,9- bis ⁇ [2-(dimethylaminb)ethyl]amino)benzo[g] isoquinoline- 5,10-dione, which shows a resistance index of 2 while mitoxantrone shows an RI of 98.
  • the compounds are effective in the treatment of tumors susceptible to azaanthracenedione therapy such as breast, ovary, lung, and leukemia.
  • a preferred dosage regimen would be from about 1 mg to about 50 mg per kilogram of body weight per day.
  • Unit dosage may be employed so that from about 70 mg to about 3.5 g of the active compound for a subject of about 70 kg of body weight are administered in a 24 hour period.
  • the dosage may be adjusted to be compatible to other treatment regimens, such as radiation therapy.
  • the pharmaceutical composition may be in the form of tablets, capsules, gel capsules, suppositories, lyophilized powders and solutions for intravenous administration.
  • the invention is illustrated by the following nonlimiting examples, and variations which are readily apparent to those skilled in the art.
  • MTT assay on human colon adenocarcinoma Lovo cells was performed according to
  • the compound and standards were dissolved in sterile water. All of the compounds were further diluted in culture medium for the in vitro test.
  • L1210 murine leukemia L1210 Murine leukemia cells were routinely main ⁇ tained in suspension cultures in McCoy's 5A medium supplemented with 10% horse serum, glutamine, penicillin and streptomycin and grown in a humidified environment of 10% carbon dioxide and 90% air at 37 ⁇ C.
  • each compound was dissolved in dimethylsulfoxide and added to 1 ml of L1210 cells (10 5 cells/tube) to attain final concentrations of 0.01, 0.1 and 1 ⁇ g of drug/ml of culture. After 72 hours of continuous exposure to the drug, the cell concentration was determined with a Coulter Counter. Growth inhibition was calculated for each drug using the following formula:
  • % growth inhibition l-[cell number treated/ cell number DMSO alone] x 100.
  • the growth inhibition data was then used to calculate the IC 50 value (the calculated drug concentration required to inhibit cell growth by 50% of control.
  • P388 Murine Leukemia cells were maintained in vivo by serial intraperitoneal (i.p.) injections of 10° cells in CD2F1 mice. For test purposes, mice were
  • the best mode to prepare the compounds of the present invention is the process depicted in Scheme 1.
  • the starting material for the process is dimethyl-5-methoxy-cinchomeronate or 3-methoxy-4,5- dicarbomethyoxy-pyridine (II) [Korytuyk, N. and Angelino, N.J., J. Med. Chem., (1977), 23, 745].
  • Compound (II) according to reaction (a) , is hydrolyzed to the corresponding 5-methoxy-cinchomeronic acid (III) by alkaline hydrolysis which can be conducted in water, ethanol, methanol or mixtures thereof in presence of an alkaline hydroxy salt such as sodium hydroxide. The reaction is conducted at 95°C for one hour (reaction a).
  • the anhydride IV is then condensed with 1,4-difluorobenzene in presence of sec-butylthium in THF at -78°C, leading to 3-methoxy-4-(2' ,5'-difluorobenzoyl)- pyridine-5-carboxylic acid (V) (reaction c) .
  • the ketoacid V can then be cyclized leading to the 4-hydroxy- 6,9-difluorobenzo[g]isoquinoline-5,10-dione (VI) (reaction d) by heating in fuming sulfuric acid [30% S0 3 ] at a temperature of 130-135°C for 30 minutes.
  • the benzo[g]isoquinoline-5, 10-dione (VI) can then be transformed into the intermediates of general formula VII where ⁇ >. is c _ ⁇ c _ 0 alkyl, benzyl, p-methoxy-benzyl (reaction e) .
  • the reaction can be performed by reacting compound VI with an ethereal solution of a diazomethane, phenyl-diazo- methane or p-methoxyphenyldiazomethane in a solvent mixture such as methanol-THF at a temperature ranging from 0°C to room temperature.
  • the intermediates (VII) by reaction with an amine of formula (IX) :
  • R has the same meanings as defined in formula (I) for R or it is a group which may be converted to R, lead to compounds of general formula (VIII) , which then
  • SUBSTITUTE SHEET optionally can be transformed into compounds (I) performing one or more of the following steps:
  • the diester II (1.71 g, 0.0076 mol) was added to a magnetically stirred solution of sodium hydroxide (1.28 g, 0.032 mol) in water (5.6 mL) .
  • the mixture was heated to 95"C for 1 h and then cooled in an ice bath. Coned HCl was added dropwise to the stirred mixture and the white solid which separated was collected by filtration and washed with ice water (2 mL) .
  • a second crop of III was obtained by further acidification of the
  • the THF was removed under a slow stream of nitrogen and the residual brown solid was placed in water (2 L) and stirred in an ice bath.
  • the amber solution was acidified with coned HCl and the resulting precipitate was collected by filtration.
  • the precipitate was washed with ice water (5 mL) and ether (10 L) .
  • the off-white solid was dissolved in hot chloroform and allowed to separate out slowly.
  • the white solid was collected by filtration to yield V, 251 mg (78%) , mp 210- 211°C.
  • the keto acid V (48 mg, 0.104 mmol) was added to fuming sulfuric acid [30% S0 3 ] (0.20 mL) and the mixture was heated rapidly in an oil bath to 130-135°C for 1/2 h. The resultant golden-yellow solution was cooled and poured over ice-water (4 L) . The mixture was extracted with CH 2 C1 2 (6 x 10 mL) and the yellow extracts were dried over Na 2 S0 4 . Removal of the solvent by rotary evaporation led to VI (40 mg, 94%) mp 160°C (dec).
  • SUBSTITUTESHEET 49 h The solvent and excess reagent were removed by a slow stream of nitrogen. After ice (5 g) was added to the tacky blue solid, the mixture was extracted with dichloromethane (6 x 5 mL) . The blue solution was dried with Na 2 S0 4 , and the solvent was removed by rotoevaporation. The blue solid was chromatographed on a silica gel column (1.25 x 25 cm).
  • the gradient eluants were: CHC1 3 (100 L) ; 1% MeOH/CHCl 3 (100 L) ; 5% MeOH/CHCl 3 (400 L) ; 10% MeOH/CHCl 3 (100 mL) ; 35% MeOH/CHCl 3 (400 mL) . Removal of the solvents led to the blue product 30 mg (68%) mp 204-
  • the gradient eluents were: chloroform; 10% MeOH/chloroform (elutes pale rose band) ; 30% MeOH/chloroform (150 L) . Removal of the solvents led to the blue product (25 mg, 61%) . mp 227- 229°C.
  • SUBSTITUTESHEET 4-Benzyloxy-6.9-Bisf ( 2-rN-(t-ButoxycarbonyI amino]ethyl) amino]benzo[g]isoquinoline-5.10-dione (VIII ,P ⁇ CH-.Ph.
  • the gradient eluents were: chloroform (100 mL) ; 1% MeOH/chloroform (100 mL) ; 2% MeOH/chloroform (200 mL) [elutes rose mono-substituted compound, 6 mg] ; 5% MeOH/chloroform (200 mL) . Removal of the last eluent led to product (221 mg, 87%), mp 192-4 ⁇ C.
  • the gradient eluents were: chloroform (50 mL) ; 1% MeOH/chloroform (100 mL) ; 2% MeOH/chloroform (100 L) [elutes slight red band]; 4% MeOH/chloroform (100 L) ; 10%

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  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
PCT/US1993/008242 1992-09-08 1993-09-08 4-SUBSTITUTED 6,9,-BIS (SUBSTITUTED-AMINO) BENZO [g] ISOQUINOLINE-5,10,DIONES WO1994005641A1 (en)

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AU50997/93A AU5099793A (en) 1992-09-08 1993-09-08 4-substituted 6,9,-bis (substituted-amino) benzo {g} isoquinoline-5,10,diones

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US94183192A 1992-09-08 1992-09-08
US07/941,831 1992-09-08

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CN (1) CN1101038A (enrdf_load_stackoverflow)
AU (1) AU5099793A (enrdf_load_stackoverflow)
TW (1) TW276251B (enrdf_load_stackoverflow)
WO (1) WO1994005641A1 (enrdf_load_stackoverflow)
ZA (1) ZA936396B (enrdf_load_stackoverflow)

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4197249A (en) * 1977-08-15 1980-04-08 American Cyanamid Company 1,4-Bis(substituted-amino)-5,8-dihydroxyanthraquinones and leuco bases thereof
WO1992007558A1 (en) * 1990-11-02 1992-05-14 Spinx Pharmaceuticals Corporation 1,4-bis-(amino-hydroxyalkylamino)-anthraquinones for inhibiting protein kinase c
EP0503537A1 (en) * 1991-03-08 1992-09-16 The University Of Vermont 6,9 Bis(substituted-amino)benzo-[g]isoquinoline-5,10-diones

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4197249A (en) * 1977-08-15 1980-04-08 American Cyanamid Company 1,4-Bis(substituted-amino)-5,8-dihydroxyanthraquinones and leuco bases thereof
WO1992007558A1 (en) * 1990-11-02 1992-05-14 Spinx Pharmaceuticals Corporation 1,4-bis-(amino-hydroxyalkylamino)-anthraquinones for inhibiting protein kinase c
EP0503537A1 (en) * 1991-03-08 1992-09-16 The University Of Vermont 6,9 Bis(substituted-amino)benzo-[g]isoquinoline-5,10-diones

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
A.PAUL KRAPCHO ET AL: "Synthesis and antineoplastic evaluations of 5,8-bis[(aminoalkyl)amino]-1-azaanthracene-9,10-diones", JOURNAL OF MEDICINAL CHEMISTRY, vol. 28, no. 8, August 1985 (1985-08-01), WASHINGTON US, pages 1124 - 1126 *
M. CROISY-DELCEY ET AL: "Aminoalkylamino derivatives of dihydroxy-benzo[g!isoquinolinedione and of trihydroxy-naphtho [2,3-g]isoquinoline dione:synthesis and anti-tumor evaluation", EUROPEAN JOURNAL OF MEDICINAL CHEMISTRY.CHIMICA THERAPEUTICA, vol. 23, no. 1, January 1988 (1988-01-01), PARIS FR, pages 101 - 106 *

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AU5099793A (en) 1994-03-29
TW276251B (enrdf_load_stackoverflow) 1996-05-21
CN1101038A (zh) 1995-04-05
ZA936396B (en) 1994-06-13

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