Cytotoxic compounds. Part 21. Chloro-, methoxy-, and methoxy-carbonyl-derivatives of (bis-2-chloroethylamino)-phenols and -anilines

摘要

2258 J.C.S. Perkin ICytotoxic Compounds. Part 21 2 Chloro-, Methoxy-, and Methoxy-carbonyl-derivatives of (Bis-2-chloroethylamino)-phenols and -anilinesBy Anthony J. Abela Medici. Leonard N. Owen,* and Constantine Sflomos, Department of Chemistry,Imperial College, London SW7 2AYNew or improved syntheses are described of some NN-bis-2-chloroethylanilines carrying both a free phenolicgroup and a methoxycarbonyl ring substituent.A study has been made of the hydroxyethylation, with ethylene oxide, of a variety of chloro-, methoxy-, andmethoxycarbonyl-nitroanilines, and of methoxy- and methoxycarbonyl-N-acylphenylenediamines. Bishydroxy-ethylation was inhibited by an o-methoxycarbonyl group and by an o- or p-nitro-group, but otherwise the NN-b is- 2 - hydroxyet hyl derivatives were obtained and subsequently converted in to the NN- bis- 2 - chloroet h yl com-pounds.Reduction of the nitro-group, or hydrolysis of the acylamino-group, in these dichlorides led t o NN-bis-2-chloroethylanilines carrying both a free amino-group and also a methoxycarbonyl-, methoxy-, or chloro-group asring substituents.The ring-substituted (bis-2-chloroethylamino) -phenols or -anilines are precursors of mustard urethanes havingpotential importance as anti-tumour agents.IN Part 17 attention was drawn to the particularchemotherapeutic advantage shown by aromatic nitrogenmustards of types (1) and (2), the most favourable anti-tumour activity (chemotherapeutic indices in the range50-150) being shown3 by those carbaniates in whichthe substituent in the non-mustard ring is chloro,methoxy, or methoxycarbonyl.7 No mustard carba-mates are known in which an additional substituent ispresent in the mustard aryl ring, and our objective wasto synthesise suitably functionalised aryl mustards fromwhich such urethanes could be prepared. Many NN-bis-2-chloroethylanilines, carrying two additional ringsubstituents, have been described,: but very few includea phenolic 6-8 or amino-group 9?10 through which, byreaction with an isocyanate or a chloroformate re-spectively, the carbamate function could be generated.In the original synthesis 2*6 of the phenolic mustard(4) the yield from the corresponding triol (3), by selectivereaction of the hydroxyethyl group with thionyl chloride,was poor. Reinvestigation of this stage has now shownthat a major by-product is the benzoxazine (27), whichevidently arises by an intramolecular cyclisation of thetype previously studied by Knorr.ll Recently, catalysisby zinc chloride has been reported to be very effectivefor the conversion of alcohols into chlorides by thionylchloride,12 and by the use of this method a much im-proved yield (58) of the mustard (4) was obtained.Reaction of the triol (5) (prepared by hydroxyethylationof methyl 5-aminosalicylate) with this reagent likewisegave an acceptable yield (52) of the mustard (6).t.Simple nitrogen mustards show very low values because oftheir high toxicity, and even the clinically useful Chlorambucil hasan index of only 12 when measured under the same condition^.^$ A comprehensive list has been ~ o m p i l e d .~1 Part 20, A. Behzadi and L. N. Owen, J.C.S. Perkin I , 1974,2 P. D. Edwards, D. L. D. Foster, L. N. Owen, and M. J .3 T. J . Bardos, 2. F. Chmielewicz, and P. Hebborn, A n n . NcwT. J. Bardos, N. Datta-Gupta, P. Hebborn, and D. J .5 A. J. Abela Medici, Ph.D. Thesis, University of London,6 B. J. Johnson, Ph.D. Thesis, University of London, 1963.7 B. R. Baker, W. W. Lee, A. P. Martinez, L. 0. Ross, and L.2287.Pringle, J.C.S. Perkin I, 1973, 2397.York Acad. Sci., 1969, 163, 1006; personal communication.Triggle, J . Medicin. Chem., 1965, 8, 167.1976.Goodman, J . Org. Chrm., 1962, 27, 3283.In their synthesis of the mustard ($9, Baker and hisassociates failed to achieve selective reaction of theliydroxyethyl groups in the triol (7) and they found itnecessary to effect temporary protection of the phenolicgroup; the overall yield of the phenolic mustard wasvery low.In view of our successful reactions withthionyl chloride-zinc chloride on the triols (3) and (5)we therefore treated the triol (7) in the same way. Theproduct, however, had a high chlorine content and thelH n.m.r. spectrum (which included resonances forphenolic hydroxy-groups at 7 -1.55 and -0.72)indicated that it consisted essentially of the chlorinatedcompounds (28) and (29) in equal proportions. Reportsof nuclear halogenation by thionyl chloride are rare,13but the orientation of the substituents in the triol (7) ismore f avourable for electrophilic substitution than inthe isomers (3) and (5).Selective formation of thedesired mustard (8) was eventually achieved in 65yield by use of the pyridine-methanesulphonyl chloridereagent .I4Substituted nitroanilines are potential startingmaterials for the synthesis of amino-mustards byeventual reduction of the nitro-group, but the suscepti-bility of the aniline to bishydroxyethylation is verydependent on its base strength and therefore on thenature and orientation of the s u b ~ t i t u e n t s . ~ ~ ~ ~ ~ Theweakly basic methyl 5-amino-2-nitrobenzoate, whentreated at ambient temperature with ethylene oxide inM. Articoand W. C. J . Koss, Biochem. Pharmacol., 1968, 17,883.W. C. J . Ross, G. P. Warwick, and J , J .Roberts, J . Chem.Soc., 1955, 3110.lo P. Kristian, A. Hulka, K. Antos, P. Nemec, and L. Drobnica,Chem. Zuesti, 1959, 13, 103; V. Bieksa, D. Rurduliene, and J.Degutis, Liet. T.S.R. Mokslu Acad. Darb., Ser. B, 1971, 64(1),133; 1973, 77(4), 55, 67; 0. Rarauskaite, V. Bieksa, and J .Degutis, ibid., 1971, 66(3), 139; J. Degutis and V. Bieksa, Liet.T.S.R. Acad. Darb., Sev. B, 1965,42(3), 71.l 1 L. Knorr, R e v . , 1889, 22, 2081.l2 T. G. Squires, W. W. Schmidt, and C. S. McCandlish, J . Org.Chepn.., 1975, 40, 134.13 H. Meyer, Monatsh., 1915, 36, 723; H. S. Mosher and M.Look, J . Ovg. Chcrn, 1955, 20, 283; cf. S. D. Saraf, Canad. J .Chem., 1969, 47, 2803.l4 M. Szekerke, R. Wade, and F. Bergel, J . Chem. Soc., 1965,1907.l5 W.C . J. Koss, J . Chem. Soc., 1949, 183.l6 M. Freifelder and K. R. Stone, J . Org. Chem., 1961, 26, 14771977aqueous acetic acid, gave only a low yield of the diol(9) , the main product being the AT-monohydroxyethylderivative. When the latter was treated with ethyleneoxide at 140 "C, a small amount of the diol was formed,( I ) X = 0 , Y = N H( 2 ) X NH,Y = OR ' R 2( 3 ) 2-OH( 4 ) 2 - O H( 5 ) 4-OH( 6 ) 4-OH( 7 ) 3 - O H( 8 ) 3 - O H( 9 ) 4 - N 0 2( 11 ) 3-NHAc( 1 2 ) 3-NHAc( 1 3 ) 3 - N H 2( 1 4 ) 2-NO2( 1 5 ) 5-NO2( 1 6 ) 5-NO2(I 0 ) 4-NO2( 1 7 ) 5-NH2( 1 8 ) 3-NHAc( 1 9 ) 3-NHAc(20) 3 - N H 2( 2 1 ) 5-NO;!( 2 2 ) 5-NO2( 2 3 ) 5-NH2( 2 4 ) 3-NO2( 2 5 ) 3-NO2( 2 6 ) 3-NH2co-0,5-CO2Me5- C02Me3-C02Me3 - C02Me4-C02Me4- C02Me3- C02Me3-C02Me4 - C02Me4 - C02Me4-C02Me4-OMe2- OMe2- OMe2- OMe4-OMe4- OMe4-OMe2-CL2- C l2 - c 14-CL4 - c L4-CLXOHCLOHClOHC lOHC lOHC lCLOHOHCLCLOHC IC lOHCIc1OHClCIOMeC0,MeC l( 2 9 )together with the lactone (30), identified analyticallyand by lH n.m.r.and mass spectra. Reaction of thediol with phosphoryl chloride gave the mustard (10) butthe overall yield was so low that the final reduction wasnot practicable.Hydroxyethylation of methyl 2-amino-4-nitrobenzoateunder various conditions gave small yields of the N-hydroxyethyl derivative but no diol ; methyl 2-amino-4-phthaliniidobenzoate (prepared by selective acylationof methyl 2.4-diaminobenzoate) likewise gave onlv themono-N-substituted product.Similar low reactivityhas been reported l7 for methyl 2-aminobenzoate, thoughits basic strength (pK, 2.23) is close to that of m-nitro-aniline (pKa 2.47), which forms a bishydroxyethylderivative without difficulty.ls The difference in be-haviour of these anthranilate compounds may thereforebe due to steric hindrance,16 possibly in conjunction withhydrogen bonding between an amine proton and theneighbouring carbonyl group. In contrast, when methyl2-acetamido-4-nitrobenzoate was reduced to the 4-amino-compound, no difficulty was found in preparingthe diol (1 l ) , from which the mustard (12) was obtained.Solvolysis with methanolic hvdrogen chloride thenafforded the amino-mustard (13), characterised as the2-chlorophenyl carbamate.The basic strength of o-anisidine (pK, 4.52) is onlyslightly less than that of aniline, whilst that of $-anisidine(pK, 5.30) is greater, but the dominating deactivationby an o- or $-nitro-group was evident in the hydroxy-ethylation of 2-methoxy-4-nitroaniline and 4-methoxy-2-nitroaniline. In aqueous acetic acid, only the N-2-hydroxyethyl derivative was found in each case, thoughunder more drastic conditions the 4-methoxy-compoundgave a 3 yield of the diol (14) together with 5 of theisomeric N-2-(2-hydroxyethoxy)ethyl derivative (31),separated by t.1.c.and distinguished by their lH n.m.r.spectra. 2-Methoxy-5-nitroaniline, however, gave thediol (15) in good yield; conversion into the mustard (16)followed by hydrogenation over palladium gave theamino-mustard (17).Similarly, a high yield of the diol(18) was obtained from 3-acetamido-4-methoxyaniline ;conversion into the mustard (19) was best effected withthe pyridine-methanesulphonyl chloride reagent l4 andsubsequent hydrolysis with hydrochloric acid gave theamino-mustard (20). Both amines (17) and (20) rapidlydeteriorated, and therefore were immediately treatedwith phenyl chloroformate to give the stable phenylcarbamates.The behaviour of four chloronitroanilines on hydroxy-ethylation followed what had now become a familiarpattern. Diols could not be prepared from 2-chloro-4-nitroaniline or from 4-chloro-2-nitroaniline , onlymono-N-substitution being achieved, but the diols (21)and (24) were obtained from 2-chloro-nitroaniline and4-chloro-3-nitroaniline, respectively, and were subse-quently converted into the mustards (22) and (25);catalytic hydrogenation then afforded the amino-mustards (23) and (26).Hydroxyethylation of arylamines with ethylene oxidein aqueous acetic acid has never been investigated indetail. From the results which we have accumulated itis clear that the course of the reaction is very dependenton the nature and orientation of the substituents, but,even when a good yield of the bishydroxyethyl derivativeis obtained, t.1.c.shows that a complex mixture isformed. lH N.m.r. evidence indicates that acetatesSOC., 1953, 2386.17 J . L. Everett, J . J .Roberts, and W. C. J . Ross, J . Chem.18 P. D. Edwards, Ph.D. Thesis, ITniversity of T.ondon, 1972J.C.S. Perkin Iand chain-extension products such as (31) are frequentlypresent. When for electronic or steric reasons bis-hydroxyethylation is slow, the predominance of by-products makes the isolation of the required diolextremely difficult.EXPERIMENTAL'H N.m.r. spectra were recorded for solutions in deuterio-chloroform (Varian T-60 instrument), and hydroxy- oramino-functions were identified by deuterium exchange ;resonances for aromatic protons are omitted. 1.r. spectrawere recorded for all products, and were used to assist inidentifications and comparisons, but the absorptions wereunexceptional and are not reported. Kieselgel GF,,,(Merck) was used for t.l.c., and grade 3 neutral alumina(prepared from R.D.H.grade 1 alumina) for columnchromatography. Extracts were dried over magnesiumsulphate, and solvents were removed under reducedpressure below 50 "C. Petroleum refers to the fraction ofGeneral Procedure for Hydroxyethylation in A cetic Acid .-The amine, acetic acid, water (or dioxan), and freshlydistilled ethylene oxide were mixed a t 0 "C and stirred in asealed Aask a t ambient temperature for the time specified.The solution was then concentrated under reduced pressureand the residue was taken up in ether, dichloroniethane, orethyl acetate, and washed with aqueous sodium hydrogencarbonate. Evaporation of the dried extract then gave thecrude product, which was examined as individuallydescribed.Methyl 3-(Bis-2-chEoroethylamino)-4-hydvoxyben~oate (4).-(u) Thionyl chloride (12 ml) was added to a solution ofmethyl 3-(bis-2-hydroxyethylamino)-4-hydroxybenzoate(3.4 g) in dry benzene (25 ml).The mixture was boiledunder reflux for 20 min and then cooled. A precipitate wasformed ; this was collected and partitioned betweenchloroform and aqueous ammonia to give (from the washedand dried organic layer) the dichloride (4) (0.9 g), whichafter t.1.c. (chloroform) and recrystallisation from acetone-petroleum had m.p. 75-78" (lit.,, 76"), T 2.22br ( 1 H, OH),6.10 (3 H, 5 , CO,Me), and 6.57 (8 H, t, 4 x CH,). Evapor-ation of the benzene solution and purification of the residueby t.1.c. (CHC1,) gave methyl 4- (2-chloroethyl) -3,4-dihydro-2H- 1,4-benzoxazine-6-carboxylate (27), m.p.74-75", vmnx.1 702 cni-l, T 5.7 (2 H, t, OCH,), 6.10 (3 H, s, <:O,Me), 6.27(4 H, s, N.CH,.CH,Cl), and 6.50 (2 H, t, ring CH,*N) (Found:C, 56.3; H, 5.5; C1, 14.1; N, 5.5. Cl,H14C1N04 requiresC, 56.4; H, 5 . 5 ; C1, 13.9; N, 5.5). The n1.p. of amixture with the mustard (4) was 50-69". Other, un-identified fractions were also isolated by t.1.c.(b) Thionyl chloride (12 ml) in benzene (10 ml) wasslowly added (30 niin) to a boiling solution of the same triol(3.4 g) in benzene (25 ml) containing powdered anhydrouszinc chloride (ca. 0.25 g). After being heated for a further20 min, the mixture (a precipitate had formed) was cooledand filtered. The solid was dissolved in dichloroniethane,and washed with aqueous ammonia and with water, and thissolution was then concentrated ; purification of the residueby t.1.c.gave the mustard (4) (2.24 g), m.p. 72-75",identified by the characteristic n.m.r. spectrum. Reactionwith phenyl isocyanate in the presence of triethylamine, for48 h a t 66-68', gave 2-(bis-2-chloroethylamino)-4-methoxy-carbonylphenyl N-phenylcarbamate, 1n.p. 89-90" (fromether-petroleum) (Found: C, 5 5 . 5 5 ; H, 4.8: C1, 17.0;b.p. 40-60".N, 6.8. Cl,H,oCl,N,04 requires C, 55.5; H, 4.9; C1, 17.2;N, 6.8).Methyl 5- (Bis-2-hydroxyethylamino) -2-hydroxybenzoate (5).-Methyl 5-amino-2-hydroxybenzoate l 9 (3.35 g) , water( 15 ml) , acetic acid (50 ml), and ethylene oxide (24 ml) after24 h gave a solid (4.7 g) which on recrystallisation fromchloroform-petroleum (b.p.60-80 "C) afforded the triol,m.p. 113-115", T -0.28 ( 1 H, s, phenolic OH), 6.05 (3 H,s, CO,Me), 6.20 (4 H, t , 0-CH,), 6.55 (4 H, t , N-CH,), and6.87br (2 H, aliphatic OH) (Found: C, 56.3; H, 6.7; N,5.3. C1,Hl,NO, requires C, 56.5; H, 6.7; N, 5.5).Methyl 5- (Bis-2-chloroethylamino) -2-hydroxybenzoate (6) .-Thionyl chloride (3.6 ml) in chloroform (3 ml) was slowlyadded (45 min) to a stirred solution of the triol (5) (1.0 g)in chloroform (12 ml) containing zinc chloride (cn. 0.1 g).The mixture was boiled under reflux for 15 min, and wasthen concentrated, neutralised with aqueous ammonia,and extracted with dichloromethane to give an oil. T.1.c.gave the dichloride (0.6 g), m.p.83-84' (from acetone-petroleum), z -0.28 (1 H, s, OH), 6.07 (3 H, s, CO,Me),and 6.12 (8 H, s, 4 x CH,) (Found: C, 49.2; H, 5.2; C1,24.2; N, 4.6. C,,H,,Cl,NO, requires C, 49.3; H, 5.2;C1, 24.3; N, 4.8).MethyZ 4-( Bis-2-hydroxyethylamino) -2-hydroxybenzoate (7).-Methyl 4-amino-2-hydroxybenzoate 2O (18.9 g), water(50 ml), acetic acid (200 ml), and ethylene oxide (25 ml),after 48 li gave, after recrystallisation from benzene, thetriol (15.3 g), m.p. 97-100" (raised by further recrystallis-ation to 101-103"; lit.,' 97-98'), T -0.94 (1 H, s,phenolic OH), 6.10 (3 H, s, CO,Me), 6.12 (4 H, t, OCH,),6.39 (4 H, t, N-CH,), and 6.57br (2 H, aliphatic OH);tribenzoute, m.p. 115-116" (Found: C, 69.9; H, 5.1; N,2.25. C,,H,,NO, requires C, 69.8; H, 5.15; K, 2.50/,).On reaction with thionyl chloride and zinc chloride inbenzene, as described for the preparation of the dichloride(4), the triol (1.2 g) gave an oil, which by t.1.c.(dichloro-methane) furnished as a main fraction (0.53 g) a 1 : 1mixture of methyl 3- and 5-chloro-4-(bis-2-chloroethyl-amino)-2-hydroxybenzoates (28) and (29), T - 1.55 (0.5 H,s, OH), -0.72 (0.5 H, s, OH), 6.05 (3 H, s, CO,Me), and6.37 (8 H, s, 4 x CH,) (Found: C, 44.0; H, 4.3; C1, 30.7;N, 4.5. Calc. for C,,H,,Cl,NO,: C, 44.0; H, 4.3; C1, 32.6;Methyl 4- (Bis-2-chloroethylanzino) -2-hydroxybenzoate (8) .-Methanesulphonyl chloride (5.3 nil) in pyridine ( 5 ml) wasadded to a solution of the triol (7) (4 g) in pyridine (25 nil).When the exothermic reaction had subsided the mixturewas heated on a steam-bath for 20 min, then cooled, andfiltered to remove pyridine methanesulphonate (5.8 g),n1.p.174", which was washed with dichloromethane andwith petroleum. The combined filtrate and washings werewashed successively with dilute hydrochloric acid, aqueoussodium carbonate, and water ; evaporation of the driedsolution then gave an oil, which by t.1.c. gave the dichloride(8) (2.95 g), m.p. 61-62" (from ether-petroleum) (lit.,'60.5-61"), z -0.98 (1 H, s, OH), 6.10 (3 H, s, CO,Me),and 6.28 (8 H, s, 4 x CH,).Methyl 5-A wino-2-nitrobenzoate .-Commercial 5-arnino-2-nitrobenzoic acid (30.4 g) was dissolved in dry methanol(200 ml) saturated with hydrogen chloride. The solutionwas boiled under reflux for 40 h, then evaporated to dry-l9 L.Gattermann, B e y . , 1894, 27, 1927.20 D. J. Drain, D. D. Martin, B. W. Mitchell, D. E. Seymour,and F. S. Spring, J . Chem. SOC., 1949, 1498; J . A. McCubbin,13. Y. Moir, and G. A . Neville, Canad. 1. Chem., 1970, 48, 942.N, 4.376)1977 226 1ness. 'l'lie residue was partitioned between ether andaqueous sodium carbonate to give the crude ester (27 g),m.p. 86-94". This was dissolved in hot methanol andwater was gradually added to precipitate a red oil, whichwas removed ; more water was added, in small portions, toprecipitate further small amounts of red oil, until the colourof the solution had changed from orange to bright yellow.When the yellow solution was cooled in ice the requiredmethyl estev was obtained, m.p.98-100" (Found: C, 49.0;H, 4.3; N , 14.2. C,H,N,O, requires C, 49.0; H, 4.1; N,14.3q0) ; S-acetyl derivative had m.p. 109" (lit.,21 11 1-The red oil crystallised, and after purification by t.1.c.gave an isomeric ester, m.p. 194--195" (from methanol)(Found: (., 49.3; H, 4.2; N, 14.4) evidently derivedfrom an impurity in the original acid. The m.p. (sig-nificantly different from those of the known methyl amino-nitrobenzoates) suggests that i t is one of the three isomersnot yet d e ~ c r i b e d . ~Hydroxyethylation of Methyl 5-Amino-2-nitrobenzoate.-The amine (2.0 g), water (10 nil), acetic acid (30 nil), andethylene oxide ( 4 ml), after 48 h gave an oil, which bycolumn chromatography (ether) gave (i) unchanged amine(0.8 g) ; (ii) iizethyl 5-( 2-hydroxyethylanzino) -2-nitrobenzoate( 1 .1 g), an oil, T 4.5br ( 1 H, NH), 6.07 ( 3 H, s, CO,Me),6.22 ( 2 H, t, OCH,), 6.63 ( 2 H, t, NCH,), and 7.35br ( 1 H,OH) (Found: C, 49.8; H, 5.2; N, 11.4. CloHl,N,O,requires C, 50.0; H, 5.0; N , 11.7); and (iii) methyl5-(bis-2-hydroxyethylamino)-2-nitrobenzoate ( 9 ) (0.1 g ) ,an oil, T 6.4br ( 2 H, OH), 6.10 ( 3 H, s, CO,Me), and 6.2( 8 H, ni, 4 ? CH,).Fraction (ii) (1.0 g), ethylene oxide ( 3 ml), and dioxan( 3 ml) were heated in a sealed tube for 16 h a t 140 "C togive (t.1.c. in ether) starting material (0.3 g), the diol ( 9 )(0.1 g), and the lactone (30) of 5-(bis-2-hydroxyethylamino)-2-nitrobenzoic acid (0.1 g ) , m.p. 143-144" (from dichloro-methane-petroleum), T 5.03 ( 2 H, s, ring OCH,), 5.73( 2 H, t, aliphatic O*CH,), 6.03 ( 2 H, s, ring N-CH,), 6.08br( 1 H, OH), and 6.47 ( 2 H, t, aliphatic NCH,), M + 252(Found: C, 52.7; H, 4.7; N, 11.1.C1,H1,N,05 requiresC , 52.4; H, 4.8; N , 11.1; M , 252). When thisoxazocinewas treated for 1 h with boiling methanol saturated withhydrogen chloride, i t gave the diol ( 9 ) , identified by then.m.r. spectrum.Alethy1 5-( Bis-2-chloroethylamin0)-2-nitrobenzoate ( 10) .-Phosphoryl chloride (1.0 g) in benzene ( 3 ml) was added toa solution of the diol ( 9 ) (0.5 g) in benzene ( 3 ml). Themixture was boiled under reflux for 6 h, then cooled, washedwith aqueous ammonia, dried, and concentrated to an oil,which by t.1.c. (dichloromethane) gave the dichloride, m.p.89-91" (from dichloromethane-petroleum), T 6.06 ( 3 H,s , C@,Me) and 6.22 ( 8 H, q , 4 x CH,) (Found: C, 44.8;H, 4 .5 ; N , 8.45. Cl,H14Cl,N,04 requires C , 44.9; H, 4.4;N, 8.7).Hydroxyethylation of Methyl 2-Amino-4-nitrobenz0ate.-The amine (2.0 g), water (25 ml), acetic acid (50 ml), andethylene oxide (25 ml) after 48 h gave a solid, which wasseparated by column chromatography (ether) into un-changed aniine ( 1 .O g) and methyl 2-(2-hydroxyethylamino)-4-nitrobenzoate (0.6 g), m.p. 112-113", T 1.87br ( 1 H , NH),6.07 (2 H, m, 0-CH,), 6.10 ( 3 H, s, CO,Me), 6.57 ( 2 H, t,N-CH,), and 7.73br ( 1 H, OH) (Found: C, 50.2; H, 5.3;112").21 J . F. Runnett and M. M. Rauhut, J. Org. CheuPz., 1956, 21,944.N , 11.6. C1oH12N2O5 requires C, 50.0; H, 5.0; N , 11.7).Similar results were obtained when water was replaced bydioxan, and also when the amine (2.0 g), dioxan ( 5 ml),and ethylene oxide ( 2 nil) were heated in a sealed tube for16 h a t 150 "C.Hydroxyethylation of Methyl 2-Amino-4-phthalimidobenzo-ate.-A mixture of methyl 2,4-diaminobenzoate (1.7 g),phthalic anhydride (1.5 g), toluene (20 ml), and triethyl-amine (0.5 nd) was boiled in a Dean and Stark apparatusfor 2 h.When cooled, the solution deposited the 4-phtlzalimido-derivative (8.05 g), m.p. 207-208" (from chloro-form-petroleum) (Found: C, 64.65; H, 4.2; N, 9.4.C,,H,,N,O, requires C , 64.9; H, 4.1; N , 0 . 5 0 ~ ) . Thisproduct (0.5 g), dioxan (10 ml), acetic acid (10 nil), andethylene oxide ( 2 nil) after 48 h gave a solid; recrystallis-ation from dichloromethane-petroleum gave tisetlzyl 2- (2-lzydroxyethylamino)-4-phtlzalimidobenzoate, m.p.185-186",T 6.09 ( 3 H , s, CO,Me), 6.13 ( 2 H, t, OCH,), 6.57 ( 2 H, t ,NCH,), and 8.27br (2 H, XH, OH) (Found: C, 63.4; H,4.6; N, 8.1. C,,H1,F205 requires C , 63.5; H, 4.7; N,8.2y0).Methyl 2-A cetainido-4- (bis-2-hydroxyethylanzino) benzoate( 1 1 ) .-A solution of methyl 2-acetamido-4-nitrobenzoate 23(12 g) in methanol (300 ml), with loo/ palladium-charcoal(1.2 g), was hydrogenated for 18 h a t 50 "C and 25 atm togive methyl 2-acetamido-4-av~zinobenzoate (9.3 g ) , m.p. 215-216" (from chloroform) (Found: C, 57.5; H, 5.0; N , 13.5.CloHl,N,O, requires C, 57.7; H, 5 . 8 ; N, 13.4596). Thisamine ( 1 .O g), dioxan (15 ml), acetic acid (15 ml), andethylene oxide ( 4 ml), after 100 11 gave an oil, which byt.1.c.(ethyl acetate) gave two main fractions: (i) methyl2-acetamido-4- (2-hydroxyethylapnino) benzoate (0.4 g) , m.p.130-131" (from dichloromethane-petroleum), T - 1.38br( 1 H, XH), 6.12 ( 3 H, s, CO,Me), 6.0-7.6 (7 H, i n ) , and7.77 ( 3 H, s, A c ) (Found: C, 55.3; H, 6 . 4 ; I-, 10.4.Cl,Hl,K,0,,0.5H,0 requires C , 55.15; H, 6.55; S, 10.7);and (ii) the diol ( 1 1 ) (0.85 g), m.p. 118-120" (from dichloro-methane-petroleum), T - 1.3br ( 1 H, XH), 5.9br ( 2 H, OH),6.14 ( 3 H, s , CO,Me), 6.3 ( 8 H, m, 4 x CH,), and 7.83 ( 3 H,s, Ac) (Found: C, 56.9; H, 6.6; N, 9.4. C14H2,,N205requires C, 56.75; H, 6.8; N, 9.45:;).Methyl 2-Acetamido-4-(bis-2-chloroethylatnino) benzoate ( 18).-Methanesulphonyl chloride (1.2 nil) was added to asolution of the diol (1 1 ) (1.6 g ) in pyridine ( 1 2 nil).Themixture was heated a t 100 "C for 20 min, then poured onice and extracted with dichloromethane. Concentration ofthe washed and dried extract and purification of the productby t.1.c. gave the diclzloride (12) (0.8 g), m.p. 73-75" (frompetroleum), T - 1.28br (1 H, NH), 6.12 ( 3 H, s , CO,Me),6.22 ( 8 H, t, 4 x CH,), and 7.78 ( 3 H, s, Ac) (Found: C,50.5; H, 5.35; C1, 21.6; S, 8.25. C,,H,,Cl,S,O, requiresC, 50.5; H, 5.4; C1, 21.3; N , 8.404). A slightly loweryield of the same product was obtained by the use ofthionyl chloride in benzene.Methyl 2-Amino-4- (bis-2-chloroethylanzino) benzoate ( 13) .-h solution of the acetamido-compound (12) (0.53 g) inmethanolic 50 hydrogen chloride (50 ml) was boiled underreflux for 1 h, the end of the condenser being fitted with aballoon to prevent loss of hydrogen chloride.Evaporationthen gave the crude amine hydrochloride (0.52 g), m.p.135-137". This appeared to be unstable and it wasdissolved in pyridine ( 1 ml) and benzene (10 ml) and22 J . J . Blanksma and D. Hoegen, Rec. Trav. chant., 1946, 65,333J.C.S. Perkin Itreated with 2-chlorophenyl chloroformate (0.4 g) to give2-chloro~henyl N-2-methoxycarbonyl-5- (bis-2-chloroethyl-amino)phenyZlcarbarte (0.5 g), m.p. 11 1-1 12" (fromether-petroleum), T -0.98br (1 H, NH), 6.13 (3 H, s,CO,Me), and 6.27 (8 H, t, 4 x CH,) (Found: C, 51.4; H,4.3; C1, 24.2; N, 6.0.C1,Hl,C1,N,O, requires C, 51.2; H,4.3; C1, 23.9; N, 6.3).Hydroxyethy lation of 2-Methoxy- 4-nitroaniline .-(a) Ke-action of the amine (2.0 g) with ethylene oxide (4 ml) inwater (8 ml) and acetic acid (26 ml) for 52 h gave a brownsolid, which was dissolved in dichloromethane. Thesolution was diluted with petroleum until no more dark oilwas precipitated. From the decanted solution, 4-(2-hydroxyethylamino) -3-methoxynitrobenzene (0.5 g) crystal-lised; m.p. 87-88', T 4.6br (1 H, NH), 6.06 (3 H, s,OMe), 6.09 (2 H, t, OCH,), 6.57 (2 H, t, N-CH,), and 7.9br(1 H, OH) (Found: C, 50.9; H, 5.8; N, 13.2. C,H,,N,O,requires C, 50.9; H, 5.7; N, 13.2). A similar result wasobtained in dioxan-acetic acid, and (at 150 "C) in dioxanalone.Starting material (ca. 60) was recovered in allexperiments.Hydroxyethylation of 4-Methoxy-2-nitroaniline.--Theamine (2.0 g), water (8 ml), acetic acid (26 ml), and ethyleneoxide (2 ml) after 60 h gave a solid, which on recrystallis-ation from dichloromethane-petroleum gave 1-( 2-hydroxy-ethylamino)-4-methoxy-2-nitrobenzene (2.25 g), m.p. 90-92",T 1.8br (1 H, NH), 6.03 (2 H, t, OCH,), 6.20 (3 H, s, OMe),6.43 (2H, t, N-CH,), and 7.95br (1 H, OH) (Found: C, 51.0;H, 5.9; N, 13.1. CgHl,hT@, requires C, 50.9; H, 5.7; K,13.27A).(b) The aniine (5.0 g), ethylene oxide ( 5 nil), and dioxan(5 ml) were heated in a sealed tube a t 160 "C for 16 h.Column chromatography (ether) of the product gave(i) unchanged amine (0.8 g) ; (ii) the N-2-hydroxyethylderivative ( 1.2 g), m.p.88-91', spectroscopically identicalwith that previously described ; (iii) 2-2-( 2-hydroxyethoxy)-ethylamino-5-methoxynitrobenzene (31) (0.4 g), an oil,T 1.87br (1 H, NH), 6.20 (3 H, s, OMe), 6.1-6.7 (8 H, m,4 x CH,), and 7.5br (1 H, OH) (Found: C, 51.4; H, 6.3;N, 11.0. Cl,H16N20, requires C, 51.55; H, 6.3; N,10.9) ; and (iv) 2-(bis-2-hydroxyethylamino)-5-methoxy-nitrobenzene (14) (0.2 g), an oil, T 6.13 (3 H, s, OMe), 6.38(4 H, t, OCH,), 6.77 (4 H, t, N-CH,), and 7.10br (2 H, OH)(Found: C, 51.2; €3, 5.95; N, 10.5. CllH16N205 requiresC, 51.55; H, 6.3; N, 10.9).3-(Bis-2-hydroxyethylam~no)-4-methoxynitrobenzene ( 15) .-2-Methoxy-5-nitroaniline (20 g), water (80 ml) , acetic acid(380 ml), and ethylene oxide (30 ml) after 120 h gave thediol (24.5 g), m.p.74" (from dichloromethane-petroleum),T 6.05 (3 H, s, OMe), 6.35 (4 H, t, OCH,), 6.65 (4 H, t,N-CH,), and 7.5br (2 H, OH) (Found: C, 51.4; H, 6.2;N, 10.9. C,1Hl,N,05 requires C, 51.55; H, 6.3; N,( 16).-The diol (15) (0.51 g) was treated with phosphorylchloride (1 ml) in boiling benzene (3 ml) for 2 h. Thecooled solution was diluted with dichloromethane, washedwith aqueous sodium hydrogen carbonate and with water,dried, and evaporated to give the dichloride (0.48 g), map.55" (from ether-petroleum), T 6.0 (3 H, s, OMe) and 6.4(8 H, s, 4 x CH,) (Found: C, 45.2; H, 5.1; C1, 23.8; N,9.4. CllHl,C1,N,03 requires C, 45.1; H, 4.8; C1, 24.2;N, 9.6).23 F. H. Bergeim, K. Losee, and W. A. Lott, J .Amer. Chem.SOC., 1947, 69, 583.10.9).3- (Bis-2-chloroethylamino) -4-methoxy- l-nitrobenzene3-(Bis-2-chloroethylamino)-4-metlzoxyaniline ( 17) .-A solu-tion of the nitro-compound (16) (2.9 g) in methanol (70 ml),containing 10 palladium-charcoal (0.3 g) was hydrogen-ated at ambient temperature and atmospheric pressure for6 h. The solution was then filtered into methanolic 50hydrogen chloride (60 ml), and on evaporation the crudeamine hydrochloride was obtained as a solid (2.9 g) whichsoon became brown. It was immediately treated withbenzene (80 ml), pyridine (2 g), and phenyl chloroformate(2.0 g) ; purification of the oily product by t.1.c. (dichloro-methane), followed by crystallisation from ether-petroleum,gave phenyl N-3-(bis-2-chloroethylamino)-4-methoxyphe.~zy~-carbamate (1.0 g), m.p.63-66" (Found: C, 56.45; H, 5.3;C1, 18.3; N, 7.25. C1,H,,C1,N,O, requires C, 56.4; H,5.3; C1, 18.5; N, 7.3).( 18) .-5-Amino-2-methoxyacetanilide 23 ( 18.0 g) treated withethylene oxide (55 ml) in water (100 ml) and acetic acid(100 ml) for 44 h, gave (by extraction of the neutralisedsolution with ethyl acetate) the diol (18) (18.0 g), m.p. 90"(from ether), T 2.20br (1 H, NH), 6.15 (3 H, s, OMe), 6.19(4 H, t, 0-CH,), 6.50 (4 H, t, N-CH,), and 7.85 (3 H, s, Ac)(Found: C, 58.3; H, 7.6; N, 10.5. Cl3H,,N,O4 requiresC, 58.2; H, 7.5; N, 10.4).5-(Bis-2-chloroethylamino) -2-methoxyaniline (20) .-Meth-anesulphonyl chloride (0.7 ml) was added to a solutionof the diol (18) (0.5 g) in pyridine (3 ml) and the mixturewas set aside for 5 h.The precipitate (pyridine methane-sulphonate) was filtered off and the filtrate was concen-trated to remove most of the pyridine. The residue wasdissolved in dichloromethane and the solution was washed,dried, and evaporated to give 5- (bis-2-chloroethylamino)-2-methoxyacetanilide (19), an oil (0.33 g), purified by t.1.c.(ether), T 2.16br (1 H, NH), 6.17 (3 H, s, OMe), 6.32 (8 H,s, 4 x CH,), and 7.81 (3 H, s, Ac) (Found: C , 51.15; H,6.1; C1, 23.2; N, 9.0. Cl,H,,Cl,N20, requires C, 51.2; H,5.9; C1, 23.2; N, 9.2). This was dissolved in 50hydrogen chloride in methanol (10 ml), and the solutionwas boiled under reflux for 2 h, then evaporated to give thehydrochloride (0.23 g) of the amine (20).Treatment in theusual way with phenyl chloroformate in pyridine-benzenegave phenyl N-5-(bis-2-chloroethylamino)-2-methoxyphenyl-carbamate, m.p. 118-120" (from dichloromethane-petroleum) (Found: C, 56.65; H, 5.4; N, 7.2. C18H20C1,-N,O, requires C, 56.4; H, 5.3; N, 7.3).Hydroxyethylation of 2-Chloro-4-nitroaniline.--1'he amine(2.0 g), water (25 ml), acetic acid (35 ml), and ethylene oxide(10 ml) after 100 h gave a solid, which on fractional crystal-lisation from dichloromethane-petroleum gave unchangedamine ( 1 .O g) and 3-chloro-4-( 2-hydroxyethy1amino)- 1-nitrobenzene (0.45 g), m.p. 115-117" (lit.,24 120°) (Found:C, 44.5; H, 4.4; N, 12.7. Calc. for C,H,C1N2O,: C, 44.4;H, 4.2; N, 12.9). A similar result was obtained byheating the amine (5 g), clioxan ( 5 ml), and ethylene oxide( 5 ml) a t 140 "C for 16 h.Hydroxyethylation of 4-Chloro-2-n~troaniline.-I2eactionof the amine (1.0 g) with ethylene oxide ( 5 ml) in aceticacid (20 ml) and water (10 ml) for 120 h gave a solid (1.1 g).A sample of this by t.1.c.was separated into unchangedamine (74) and 4-chloro- 1-(2-hydroxyethylamin0)-2-nitrobenzene (23y0), m.p. 107-108" (lit.,z4 107.5"), T 1.87br(1 H, NH), 6.10 (2 H, t, 0-CH,), 6.47 ( 2 H, t, NKH,), and8.05br ( 1 H, OH).24 C . B. Kremer and A. Bendich, J . Amer. Chem. SOC., 1939, 61,2658.5- (Bis-2-hydroxyethylamino) -2-methoxyacetanilid3-(Bis-2-hydroxyethylamino)-4-chloro- 1-nitrobenzene (21).-Reaction of 2-chloro-5-nitroaniline (20 g) with ethyleneoxide (90 ml) in water (80 ml) and acetic acid (380 ml) for100 h, followed by 70 h a t 50 "C, gave an oil (43 g) whichwas subjected to column chromatography (dichloromethane,followed by ether).The slow-running fraction whichcontained the required product (n.m.r. spectrum) wasfurther purified by t.1.c. (ether) t o give the diol (21) (2.6 g),m.p. 93- 95" (from dichloromethane-petroleum), T 6.30(4 H, t, O*CH,), 6.60 (4 H, t, NCH,), and 6.85br (2 H, OH)(Found: C, 46.3; H, 4.75; N, 10.6. CloHl,C1N,04requiresC, 46.1; H, 5.0; N, 10.75).3-(Bis-2-chloroethylamino)-4-chlorouniline (23) .---Reactionof the diol (21) (2.6 g) with phosphoryl chloride (5 ml) inboiling benzene (15 ml) for 5 h gave, after t.l.c., 3-(bis-2-chloroethylalnino)-4-chloro-l-nitrobenzene (22) (2.6 g), m.p.61-62" (from ether-petroleum) (Found: C, 40.5; H, 3.75;C1, 35.7; N, 9.5.CloHllCl,N,O, requires C, 40.4; H, 3.7;C1, 35.7; N, 9.47;). Hydrogenation of this compound(1.5 g) in methanol (35 ml) containing 10 palladium-charcoal (0.15 g) for 3 h, followed by filtration intomethanolic 15 hydrogen chloride (90 ml) and evapor-ation, gave the crude hydrochloride (1.5 g) of the amine(23). This was immediately treated, as previously des-cribed, with phenyl chloroformate in pyridine-benzene ;purification of the urethane by t.1.c. (dichloromethane-petroleum, 1 : 1) gave phenyl N-3-(bis-2-chloroethylumzno)-4-chlorophenylcarbamate (1.05 g), m.p. 64-66" (from di-chloromethane-petroleum) (Found: C, 52.95; H, 4.7; C1,27.2; N, 7.0.Cl,H,,C1,IL',O, requires C, 52.7; H, 4.4;C1, 27.4; N, 7.2).4-(Bis-2-hydroxyethylamino)- l-chloro-2-nitrobenzene (24).-4-Chloro-3-nitroaniline (2.0 g), water (10 ml), acetic acid(20 ml), and ethylene oxide (4 ml) after 120 h gave an oilwhich crystallised from dicliloromethane. Recrystallis-ation from acetone-petroleum gave the diol (24) (1.3 g),n1.p. 108", T (CD,),CO 5.9-6.6 (2 H, OH) and 6.40(8 H, t, 4 x CH,) (Found: C, 45.9; H, 4.9; N, 10.8.CloHl,C1N,04 requires C, 46.1; H, 5.0; N , 10.75). Afurther quantity of the diol (0.4 g) was obtained from themother liquor by t.l.c., which also furnished 4-( 2-acetoxy-ethyZamino)- l-chloro-2-nitrobenzene (77 mg), an oil, vmax,1 732 cm-l (OAc), 7 5.67 (2 H, t, OCH,), 5.90br ( 1 H, NH),6.57 (2 H, t, N-CH,), and 7.90 (3 H, s, Ac) (Found: C, 46.6;H, 4.2; N, 10.7. C,,HllC1N,04 requires C, 46.4; H, 4.3;N, 10.804).When 4-chloro-3-nitroaniline ( 1 .O g) was heated withethylene oxide (2 ml) in dioxan ( 3 ml) at 100 "C for 16 h,the products, separated by t.1.c. (dichloromethane), were(i) unchanged amine (0.45 g) ; (ii) l-chloro-4-(2-hydroxy-ethylarnino)-2-nitrobenzene (0.48 g), m.p. 74", T 5.3br ( 1 H,NH), 6.13 (2 H, t , OCH,), 6.70 (2 H, t, NCH,), and 7.lbr( 1 H, OH) (Found: C, 44.3; H, 4.2; C1, 16.7; N, 12.8.C,H,ClN,O, requires C, 44.4; H, 4.2; C1, 16.4; N, 12.9) ;and (iii) the diol (24) (0.26 g), m.p. 106-log", spectro-scopically identical with the authentic specimen.5-( Bis-2-chloroethylamino) -2-chloroaniline (26) .-Treat-ment of the diol (24) (5.75 g) with phosphoryl chloride (10ml) in boiling benzene (30 ml) in the usual way afforded4-(bis-2-chloroethylamino)- I-chloro-2-nitrobenzene (25) (4.0 g),n1.p. 144" (from methanol) (Found: C, 40.3; H , 3.75; C1,36.0; N, 9.6. CloHllC1,N,O, requires C, 40.4; H, 3.7;C1, 35.7; N, 9.4). A suspension of this nitro-compound(1.0 g) in methanol (60 ml) containing 10 palladium-charcoal (0.1 g) was hydrogenated under ambient con-ditions for 5 h, then filtered into methanolic 5006 hydrogenchloride (30 nil). The crude hydrochloride, obtained onevaporation, reverted to the free arnino-conzpound (26),m.p. 176-178", on recrystallisation from methanol-ether(Found: C, 44.7; H, 5.0; C1, 40.0; N, 10.3. CloH,,C1,N,requires C, 44.9; H, 4.9; C1, 39.8; N, 10.5).We thank the Commonwealth Scholarship Commissionfor a Scholarship (to A. J . A. M.) and the State ScholarshipFoundation of Greece for an award (to C. S.).7/783 Received, 6th May, 1977