September 1967
877
SCHISTOSOMICIDAL 4 - H Y U K O X Y 1 \ I E T H Y L - ~ - C H L O I ~ O A S I L I S E S
Preparation and Schistosomicidal Activity of Some 4-Hydroxymethyl-3-chloroanilines 1).RON, T.
13. LEXVIS, R . LORENZ, H. FREELE,D. A. BERBERIAN, AND
s. r h i C H E H
Steding-l.17znthrop Research Institute, Rensselaer, S e w York
Recezved M a r c h 4, 1967 N-(3-Chloro-4-hydrox~methylphenyl)-K’,~’-diethylethylenediamine (I\’) and N-(3-chloro-4-hydroxyniethylpheny1)piperazine (IT)were prepared by chemical and microbiological methods. They were found to be more active schistosomicidal agents than the corresponding 4-methyl derivatives (I11 and V). The p-pentylphenoxyhexyl analog of TI was prepared chemically and found to be more active than the 4-methyl derivative (VII). Both were far weaker than the structurally simpler compoiuids, IY and VI.
The discovery by Kikuth and Gonnert’ that lucanthone (I) was an orally effective schistosomicidal agent led to a great deal of structural modification of the parent nucleus in an attempt to develop better agents.* One of the simplest modifications which was active was a mirasan (111) in which the diethylaminoethylamino side chain and the p-methyl group are retained but the other features of the molecule are represented by a chlorine atom. This drug was quite active in mice, but apparently was ineffective in monkeys and man.* The related piperazine (V) is also an active schistosomicidal drug in mice but it is believed to be ineffective in higher species.* The latest entry in this class is the piperazine VII.4 The substance was reported to be an effective schistosomicidal agent in Schistosoma nzansoni infected mice. It was claimed to be active in monkeys also. 0
I/
NHCH,CH,N(C,H,), I
I R I. R = C H , 11, R = CHLOH
methyl was a necessary prelude for schistosomicidal action. In our laboratory it was found that I11 and V were quite active in mice but markedly less so in hamsters. Rosi, et ~ l . found , ~ that mouse liver microsomes efficiently hydroxylated I11 to IV, and V to VI but that hamster liver microsomes carried out this conversion very ineffectively if at all. It was also demonstrated that after oral administration of 111 to mice a substantial amount of IV was detected in the urine, while in a similar experiment with hamsters no I V could be seen on thin layer chromatograms. I t was concluded that as in the case of lucanthone, I11 and V were being metabolically converted by the animal host to therapeutically active agents. I n the present paper we describe the microbiological and chemical synthesis of the aniline derivatives IV, VI, and VI11 and discuss their schistosomicidal activity in mice and hamsters. The benz.yl alcohol (IV) was prepared according to Scheme I. Catalytic reduction of ethyl 2-chloro-4SCHEME I
111. R = CH3 IV. R = CH,OH
qcl COOC2H,
c1
IV
NHCH~CH,N(C~H~),
x
v. R = C H , ; R = H VI, R = CH,OH; R = H VII, R = CHj;R’= p-(CH,),OC,H,C(CH,),c,Hj VIII, R = CHiOH; R = p-(CH,),OC,H,C(CH,),C,Hj Recently it has been demonstrated that a microbial transformation product of lucanthone, the 4-hydroxymethyl derivative (11),is also a metabolite of I and is indeed the therapeutically active form of 1ucanthone.j It was also found that the biological activity of the related thiochromones and xanthen-4ones is greatljenhanced when the 4-methyl group was replaced by the 4-hydroxymethyl group. It was thought that in this group of compounds metabolic hydroxylation of the 4(1) W. Kikuth and R. Gonnert, Ann. T r o p . M e d . Parasitol., 48, 259 (1949). (2) R . Gorinert. BUZZ. World Health Organ., 46, 702 (1961). (3) H. Mauss, H. Kolling, and R. Gonnert. M e d . C h e m . , A b h a n d l . M e d . C h e m . Forschungsstaetten I . G. Farbenind., 6 , 185 (1956). (4) A. 0. Geisnler, P. hl. Bauman. A. Alter, P. .\I. Bsuman, A. 0. Geiszler, and G. F. Otto, Reports given a t the Annual Meeting of the American Society of Tropical Medicine and Hygiene, New Tork, N. Y., Nov 7, 1964. ( 5 ) D. Rosi, G . Peruzzotti, E. \V. Dennis, D. I\. IlerLerian, H . Freele, 13. 1’. Tullitr, and S. .\rclier, J . M e d . Chem., 10, 867 (19ti7).
COOC2H5 IX
IV
QC1 NHCOCH,N( C,H Xa
)L
1iitr~bellZOate gave a mixture of IX and ethyl p aminobenzoate. The Kaney nicliel-hydrazine method’ gave much better results. Alkylation of the ester IX with diethylaminoethyl chloride afforded X in very poor yield. Reduction (LiA1H4) to IV completed the synthesis. Some hydrogenolysis apparently took place since the presence of I11 could be detected on tlc. A somewhat better synthesis from the amino ester I X consisted of chloroacetylation of the amine to furnish the X-chloroacetyl derivative which in turn was treated with diethylamine to give the amino amide ( 6 ) D. Rosi, .\. .J Rlerola, and S. .\rclier, L i f e S e i . , in press. ( 7 ) D. llalcom and A . Furst. J . A m . C h e m . Soc., 76, 4334 (llc.53).
September 1967
SCHISTOSOMICIDAL 4-HYDROXYMETHYL-3-CHLOROASILIXES
879
was refluxed using a Dean-Stark apparatus t o collect water. I n 2 hr 9 ml of water was collected. T h e solution was coiiceiitrated and the residue was distilled. The fraction, bp 153-164" (0.1 mm), was collected as the product, 86.0 g (96.5%). This crude material was used in the next step. P 0 C l 3 (20.3 g, 0.13 mole) was added dropwise with stirring t o 7 5 ml of I I J I F as t h e temperatiire rose to 40'. I t was st,irred at this temperatlire for 0.5 hr, cooled, and then added with efficient stirring to a soliitiori of 22..5 g (0.1 mole) of l-(m-chlorophenyl)-4formylpiperazine ( X I ) i n 50 ml of DAIF which was being heated on the steam bath. After the addition was complete the reaction mixtiire W ~ stirred P a t 95" for 1 hi. ilboilt 60 ml of I I h I F was removed in u a m o aiid the residiie was poiired ont,o a slurry of ice R I ' ~water. The cold rolutioii was made alkaline with 35%; Ka01-I and the extracted ICHCI,). The combined extracts were washed Kith water aiid then conceiitrated. The dark residue was heated for 0..5 hr with 100 nil of 2..5 .\- HCI on the steam bath. After r d i r i g overnight, the crystalline precipitate was collected (111 a filter aiid washed with isopropyl alcohol. The yield of mp 247' dec. hydi,ochloride was 14.2 g (,5r5cy)j A n a l . Calcd for CIIHI~CIIY?O.TIC~l: X, 10.73: CI, 27.1.5. N-(3-Chloro-4-hydroxymethylphenyl)-N',N'-diethylethylene- Foillid: K,10.80: C1, 2 7 . 3 5 . diamine ( I V ) from Xa.-A solutioii of 23.4 g (0.177 mole) of AlClr 2-Chloro-4-(1-piperaziny1)benzyl Alcohol (VI).--A slurry of i i i :NO ml of tetrahydrofuran ( T H F ) was prepared b>- cautious 28.5 g (0.109 mole) of the hydrochloride XI11 in 200 ml of methaaddition of t h e salt to the solvent. T h e solution was added i i i irol was treated with 9.0 ml of 35Cc NaOH solution and cooled to oiie portioii to a suspension of 52 g (1.37 moles) of LiAIH4 ill 0'. T o this mixture there was added a solution of 4.5 g of NaBH4 2.0 1. of T H F at -12". The free base from 117 g (0.374 mole) i l l 40 ml of water. The methaiiol wa8 removed in vacuo; care of the eyter ( S a ) i n 200 ml of T H F was added over a 45-miii was takeii not to overheat the reaction mixture. T h e residue was period to the LiA1H4-A1C1, solution with stirring while the temtreated with 50 ml of IC; S a O H sollition aiid the mixtiire was experature of the latter was kept betweeu -8 and -12". The tracted (CT-ICI,). The combined extracts were decolorized with mixture was stirred for 6 hr at - 6 to -12" arid then hydrolyzed charcwal and dried illrierite). After evaporation, there remained b>- the carefiil addition of 163 ml of 35% aqiieoiis NaOH. The 23.2 g (9.iT) of the desired piperazine (\-I), m p 118-120" resiiltiiig emiilsioii was broken with 20 g of powdered KOH. (Lincor), identical iii all respects with material prepared by the T h e mixture wa.q filtered aiid the filter cake was washed with microbiological roiite. T H F . The filtrate was concentrated t o dryness in UUCZLO leaving Microbiological Oxidation of 1-(3-Chloro-4-methylphenyl)a residiie which weighed 84 g. It was dissolved in 75 ml of warm piperazine Hydrochloride.-The stock culture of Bspergdlus henzetie and the solution was diliited with 100 ml of pentane. sclerotiorzcm was grown initially at 26" for 10-14 days on slants 011staiidirig 44..5 g ( 4 6 5 ) of IT separated, mp 64-67". Upon in a nutrierit medium consisting of Difco maltose (40 gll.), recrystallizatioii from benzene-pentane with the aid of charcoal, Difco proteose peptone S o . 3 (10 g/l.), aiid Racto agar (15 g/l.). piire white I\. was obtained which melted at 66-68". The5e slants were used to prepare seed for the fermentations in 1-(p-f-Pentylphenoxyhexyl)-4-(m-ehlorophenyl)piperazine.the followiiig way. Sterile distilled water (10 ml) was added t o A solutioii of 81.5 g of 6-(p-t-peiitylphenoxj-)hexyl bromide,lZ the slant and the spores and some vegetation growth were shaken 40 g of ni-chloropheiiylpiperaziriej 50.5 g of triethylamine, and loose with a sterile hooked needle to form a iiniform siispensiori. ,500 ml of toluene wan heated under reflux for 6 hr. T h e mixture This was added to a 2-1. flask containing 700 ml of a hterile soywas left overnight and then filtered to remove the triethylamine dextrose mediiim of the following composition: 15 g of soybean hydrohromide. T h e residue was concentrated to a small volume g of salt, 5 g of ILHPO:, meal, 20 g of dextrose, 5 g of yeast, .i mid p u t on a 200-g silica gel column and eluted with 2Yc isopropylaiid erioiigh t a p water to make the filial voliime 1 1. The pH was amine-ether. Thi.q operation removed some color. T h e eluate adjiisted t,o 6.5 with HC1 jiist prior to sterilization a t 121' for wa3 evaporated to dryness arid converted to the hydrochloride 15 min. These cotton-pliigged flasks were placed on a rotary i i i ether with alcoholic HCI. There was obtaiued a tc shaker (2.5-cm throw at 240 rpm) aiid iiiciibated for aboiit S g of the hydrochloride, m p 116-119O. A sample was days a t 26". lized from ethatiol-ether for anal? Ten liters of a soy-dextrose mediirm of the following composi:Inal. Calcd for C2iH;,ClXr0 .F tion was iised for the fermentation itself: 1 kg of cerelose, 1.50 g 2-Chloro-4- [4-(p-t-penty1phenoxy)hexyl-1-piperazinyl]benzyl of soyhean meal, .iO g of yeast, .50 g of salt, 2.5 g of lIgSO4. Alcohol (VIII).-To a solution of 29 g of POCI, in 44 g of dimethyl7H20, 13.8 g of N a H 2 P 0 4 . H 2 0 301 , g of Na2HPO*.12H?O,and formamide (IIJIF) there was added a solution of 50 g (0.104 mole) t a p water to make the final volume 10 1. The solution with p H of l-(p-f-peritylpheiiox~hexyl)-4-(m-chlorophetiyl~piperaziiie hy7.3 was sterilized as ahove. One seed flask was added to the drochloride in 150 ml of 1 ) J t F over a period of 15 miri with the ahove solution which was stirred i n a fermenter kept in a water temperature held at 15". -4fter the addition was complete, bath a t 28". After 24 hr the addition of 1-was begiin. About the mixture w-as heated at 120-130° for 35 mill aiid then poured 10 g/day was added i n tLvo portions over a period of 3 days. ciiito 1 kg of ice. The solid was filtered, washed with water and The t,otal amoiint of sitbstrate consumed was 53 g. Foiir such diliite NH,, atid then taken rip iii ether. The extracts were fermentations were carried oiit simiiltarieoiisly so t h a t 212 g washed ( H 2 0 ,3: NaOH, saturated NaC1). The et,herlayer was of 1-(3-chloro-4-methylphenyl)piperaziiie hydrochloride 10.86 dried (JIgSO,) mid concentrated t o leave a viscoiis oil, 48 g. This mole) was iised as a substrate. T o each fermentatioii vessel w a s di,.wlved i n 600 ml of methatiol, aiid 8.0 g of NaRH, was there was added 130 ml of 10 *YNaOH t o stop fiuther growth of added i i i 2.0-g poi,tioiis over a period of 1.5 hr. The methanol the organism. Each vessel wah extracted with two 20-1, portion* was removed aiid the residiie was allized from aboiit 1 I . of of CH,CI,, and the extracts were partiall>-coricelitrated in z w u o . hexaiie. A semisolid (30 g ) separa rid was chromatographed The residiies from foiir siich extractions were combined and theti oti 100 g of silica gel iisiiig 2C; isopropylamine aiid ether as the evaporated t o fiirnish a cryatalliiie residue weighing 150 g. eliitiiig solvelit. The talliziiig fractions were combined and This was recrystallized from 800 ml of ethyl acetate after an recrystallized from boi hexaiie to fririiish 20 g of piire \.III, insolitble fractioii (4.5 g, nip 180') was filtered off. On cooling mp 73-74". t a b of \Ideposited, the warm stiliition, 120 g of ci,eam-colored ( Anal. Calcd for CrrH~iClSyO2:S , 5.92, Foillid: N, 6.02. melting at 122" (cor). A secoiid slightly lesh piire crop weighing 2-Chloro-4-( 1 -piperazinyl)benzaldehydeHydrochloride (XIII). 23..i g was collected o i l coiiceiitratioii of the mother liqiiors. T h e --A solutioii of 78.6 g (0.40 mole) of I-( m-chloropheny1)pipertotal yield of conibitied crops was 143.5 g or 73% of the theoaziiie, 22.2 g (0.48mole) of formic acid, aiid 100-ml of beiizeiie retical. Anal. Calcd for CIIHI,C1N40:IY, 12.36: C1, 13.64. Foruid: E,12.43: C1, 15.44. T h e iIv *pectriim showed the followitig 211 nig ( e 24,300), 236 (l3,900), 291 sh (1SOO). The iinir spectriirn (%O(,; all the acid chloride was added the solution was refluxed for 1.5 hr, cooled, and diluted with 1 1. of pentane. T h e solution was placed in the refrigerator overnight and filtered. and the crystals were collected, washed with pentane, and dried; 135 g (83TC), mp 119-122". Anal. Calcd for CI1H,,C1?NO,: N , .?.(IT; C1, 25.68. Foiiiid: K, 5.16: C1, 23.95. Ethyl 2-Chloro-4-(~-diethylaminoacetamido)benzoate.A solution of 133 g (0.481 mole) of the above ester aiid 106 g (1.44 moles) of diethylamine in 1 1. of benzene was refluxed for 4 hr. The whole \?-as rediiced t o half-volume atid washed with water. The organic layer was extracted with diliite HC1. The acid extracts were washed with ether, made basic, and extracted with ether again. The second ether extract wa8 dried arid conceiitrated. The rebidiie was di,\solved iii isopropyl alrohol aiid treated with 1 eqiiiv of coiiceiitrated HC1. The mliitioii was treated with ether t o the point of turbidit,y and set aside and tals of the hydrochloride which separated were ied: 144 g (X6S;), m p 150-l.~lo. .\nul. C d r d for C l ~ I I ~ ~ C I N ~ O ~C1, ~ H20.30. C 1 : Foiiiid: C1, 20.09.
CDCb risiiig TMY its ail iiiteriial standard) was compatible with the assigned structure. Nmr sigiials were observed at 6 7.31 (1 TI doublet, J;,g = 8 H z , Cj-H), 6.82 and 6.73 (overlapping 13 doiiblet, I FI quartet,,J?,6= 2.8 112, C2-Hand Cs-H, respectively), 4.6'2 (2 €1 singlet, -CH,O-), 3.14 and 2.!17 (10 H, unresolved sillglets, OH, N H , arid (CFl.")4). There was no CHa signal. The misture melting points with naniples prepared by the other routes showed no depressioii. The ir spectra of these sample> were essentially superimposahle. Microbiological Oxidation of N-(3-Chloro-4-methylphenyl)N',N'-diethylethylenediamine Hydrochloride (HI).-Essentially the same procedure was used here. A total of 161 g (0..58 mole) of riubstrate was coiiverted over i t i i x-,l:t,~.period. T h e substance cted and coiic.eiitrateJ a- i i i the above procedure.
Adrenergic Neurone Blocking Agents. 11.' Some Dioxane- Substituted Derivatives of Guanoxan
$yilt heaes are t i e d b e d ( J f 2-rnethyl- atid Irans-S-methyl-a-griaiiidiiionlet h!.l-1,4-t)eiizodic)~:iri i t l i d of .syn- :I 11t1 c~nli-2'-giianidinospiro( 1,4-benzodiosaii-~,lf-c~yclopropa~ie~) (2a and b). The stereochemistry of the cornpounds was determined by iinir spectroscopy, aiid their adrenergic neiirone blockilig properties were compared with those of guanosaii.
111 u recent publication1 we described the synthesis mid adrenergic iieurone blockiiig properties of a series of guanidines related to guanoxan (1, R = H). =2lthough i n a r i y derivatives tvith substituents in the aromatic
4
3
CH, O*CH,
0 1
2a s y n
banti
ring were described, t'he oiily reference to a subst'it.ueiit i n the dioxane ring was to the &methyl derivative which was prepared and tested as a mixt'ure of cis and trans isomers. We have now prepared the pure tmns-3-metjh\,l derivat'ive (1, R = CH,) and t'he %-methylderivative of gu:inoxan. Following report,s of t8hc biological activit,y of phenoxycycloproI,vlAmiIies~,~j it, seemed reason:tblc to synthesize t'he spirocyclopropyl compounds (2). Accordingly, both epimers (2a and b) were prepared. Chemistry.-The 3-methyl derivat'ive (1, R = CH,) was approached via the mixture of esters 3 and 4 (1% = C2H5),obtained by reaction of catechol with ethyl 2,3-dibromobutyrate in the presence of K2C03and ( I ) Part I of this series: J. .lugstein. 8 . h1, Green, .\, 11. Blonro, G. \ \ . I f , Potter, C. R . Ivorthing, and T. I. \$-ripley, J . ,Wed. Chem., 8 , 446 (1985). ( 2 ) Hoffman-La Roche and Co., A. G . , Belgian Patent 613,910 (Aug 14, 1962): Smith Kline and French Laboratories, U. 8 . Patent 3,156,725 (NOT I O , 1964). ( 3 ) (a) C. Kaiser, 13. SI. Lester, C. I,. Zirkle, .\. liuryer, C. S. I l a y k , T. I,. 1)elia. and L. %irnpil)l,J . .Veri. l'/turm. Ciiem.. 6 , 1243 (1962); (I>) ( ' . I.. Xirklr, C ' . Kaiser, I). Ei. Tedesrlri, It. 1:. 'I'edesc,lii. ani1 .\, I3rlrger, ibiri.. 6 , 1285 (1962).
H
O$H
0
CO,R H
COLR
A
B
B
A
: i c e t o ~ i e . ~lleductioii of thc ester mixturc (I,i-iIII~j ielded the mixture of dcohols ( 5 , It = H) from which the trans isomer was isolated readily b\ crJ .tallixation; the cis isomer could not be isolated in a purr state from this reaction. The bans-alcohol (5a, It = H) was converted t o thc to\ylate (5a, It = Ts), which upoil treatment 1% ith gu:inid~ne~ yielded the desired priicluct . 1 (R = CH.,).
J
5
6
a = trans
b = cis
Initially, t.he tians series ( i f compounds wts cst:lt)lished as such by conversion of t,he ester mixture n.it,li lit'hium aluminum deuteride to the corresponding misture of deuterioalcohols 6. I"rom this mixt'ure :i pur(" crystalline alcohol h:Lviiig :L relativel!, h r g e vicili:tl I
1) . I . K O < ). I,. Or". ('hew, 26, : 4 ; 4 ! l \Iilrirli ('lieiiiiwl ( ' o .
;ilile f r o i i ;
I
I!ltil 1 ,
tliis
e>,i?r iiiisLiirr. i i
TION
ii(il~l-
