,July 1969
SCHISTOSOMICIDAL XASTHENONES ASD CHLOROTOLUIDIXES
large mass of polymeric pot residue. H, N.
Anal. (Cl6H&1N3O) C,
N-(6-Chloro-2-methoxy-9-acridinyl)-N[ 3-( diethy1amino)propyllacetamide (XX).-A solution of 46.3 g (0.1 mole) of 6chloro-9-( [ 3-(diethylamino)propyl]amino]-2-methoxyacridine dihydrochloride hydrateI6 in warm HzO was made basic with N H 4 0 H and extracted with CHCla. The combined CHC13 extracts were dried (AlgS01) and treated with 25 ml of AcC1. The mixture was boiled under reflux for 48 hr, filtered hot, and cooled to give 11.9 g of yellow solid, mp 266-274". This material w s presumed to be starting material from its lack of C=O absorption in the ir. The filt.rate was concentrated to dryness to give a yellow semisolid. Crystallization from 95% EtOH gave 6.0 g of a solid of indeterminate melting point which was discarded. The EtOH filtrate was concentrated to dryness, and the residue was dissolved in HZO, filtered, and made basic with NaOII. The gummy material which formed solidified on standing and was crystallized twice from heptane. The bright yellow crystals thus obtained, mp 113-114.5', weighed 11.2 g (27Yc) and showed strong C=O absorption in the ir a t C, H,.N. 1665 cm-l. Anal. (C23H2sClNaO~) 1-[ 5-[(7-Chloro-4-quinolyl)amino]salicyl) -4-piperidin01 Dihydroch1oride.-To a mixture of 4'-hydroxyacetanilide (45.3 g, 0.3 mole) and 30.4 g (0.3 mole) of 4-piperidinol in 200 ml of i-PrOH was added dropwise during 1 hr 22.5 ml of 407, CH20. The mixture was heated under reflux for 5 hr, the solvent was removed in vacuo, and 100 ml of H20 was added. The mixture was heated on a steam bath for 3 hr, cooled, and neutralized until just acid to congo red. 4,7-Dichloroquinoline (59.4 g, 0.3 mole) and 50 ml of EtOH were added and the resulting mixture was heated on a steam bath for 3 hr. The pasty mass was stirred with 2 1. of H2O and filtered. The filtrate was diluted to 4 1. and made alkaline R-ith NH40H. The solid was collected by filtration, digested with a mixture of boiling MeOH-EtOH, and filtered. The residue was suspended in hot EtOH and treated with concentrated HC1. The suspension of the hydrochloric acid salt was diluted with MezCO and filtered to yield 97.0 g (70c0) of product,, mp 298-300" dec. Anal. (C21Hz2C1?TaO:!.2HC1.0.25H?O)C, H, N, H20.
607
N-(2-Cyanoethyl)-N-ethylacetamide.-To a mixture of 147 g of HOAc and 225 g of AczO was added dropwise 236 g (2.4 moles) of 3-(ethylamino)pr0pionitrile.~~Heat was evolved and the temperature was maintained a t 50-60" by controlling the rate of addition. The solvent was removed in vacuo and the residue was distilled to give 278 g (83%) of product, bp 95' (0.3 mm), nZ6D1.4640. Anal. (C7H12N20) C, H, N. The ir spectrum contained a carbonyl band a t 1647 cm-l. N-( 3-Aminopropyl)-N -ethylacetamide.-N-( 2-Cyanoethyll-Nethylacetamide (272 g, 1.94 moles) was hydrogenated in 500 ml of toluene over 60 g of Raney cobalt in the presence of 60 ml of Et3N a t 100" and an initial hydrogen pressure of 140.6 kg/cm2. The solvent was removed in vacuo and the residue was distilled to give 170 g of product, bp 90-99" (0.2 mm). The ir spectrum showed a strong carbonyl peak a t 1635 and a shoulder at 1670 cm-I; in CC14 a split peak a t 1678, 1652 cm-l was present. The material was redistilled through a 30-cm T'igreux column and yielded 42 g (l5y0)of fraction A, bp 73" (0.2 mm), n Z 6 ~ 1.4725, and 77 g (27%) of fraction B, bp 83' (0.2 mm), n Z 5 ~ 1.4668. The ir spectrum of fraction A showed C=O a t 1640 and a split primary amine band a t 3300, 3370 cm-I and was presumed to be the desired X-(3-aminopropyl)-X-ethylacetamide. ,4nal. (C7HIcN20) C , H, N. Fraction B exhibited a C=O peak a t 1650, a strong N H absorption a t 3300, and an amide I1 band a t 1.560 em-' and was presumed to be the rearranged terminal amide S-[3-(ethylamino)N H, ~ ~X. ) propyllacetamide. Anal. ( C ~ I I I ~ C,
Acknowledgments.-The authors express their appreciation to Dr. Paul E. Thompson and conorkers for the antimalarial studies, Mrs. Eva Gold and Miss Maria Limson for assistance in the chemical work, Mr. William Pearlman for carrying out the hydrogenations, Ah. C. E. Childs and associates for the microanalyses, and Dr. J. 11. Vandenbelt and coworkers for determination of their and uv absorption spectra.
Comparison of Schistosomicidal Activity of Xanthenones and 4-Methyl-3-chloroanilines and Their Hydroxymethyl Analogs in Swiss Mice and Syrian Hamsters Infected with Schistosoma mansoni D. A. BERBERIAN, E. W. DENNIS,H . FREELE,D. ROSI,T. R. LEWIS,R. LORENZ, ASD S. ARCHER Ste,ling-Winthrop Research Institute, Rensselaer, S e w YO& 1214.4 Received March 86, 1969 The schistosomicidal activity of a number of xanthenones, 4-methyl-3-chioroanilines, and their hydroxylated derivatives were tested against Schistosoma mansonz. It was demonstrated that hydroxylation enhanced schistosomicidal activity one- to sixfold in the mouse and two- to 33-fold in the hamster.
A series of xanthenones initially synthesized in the thirties by Mauss' and designated as "miracils" were demonstrated by Kikuth and Gonnert2 to be orally effective against Schistosoma mansoni infection in white mice and monkeys. Of the several xanthenones, lucanthone ( U r a c i l D) was found to be effective not only in experimental animal infections but also in natural infections of humans. Although highly efficacious in the monkey, lucanthone was less effective in humans and mice. Of the hundreds of xanthenones synthesized during the past quarter of a century a few were more active than lucanthone in experimental infections in animals but less effective when field tested against schistosome infections of man. The erratic and unpredictable activity in different hosts was found (1) H. Mauss. Chem. Ber., 81, 19 (1948). (2) W. Kikuth and R. Gonnert, 2. Tropenmed. Parasztol., 1, 234 (1949).
to be related to the ability of the host to hydroxylate the 4-methyl group para to the alkylamino side chain. Hycanthone, the hydroxymethyl analog of lucanthone, was the most active of the several metabolites produced by each host species. It was also demonstrated that the variety and the proportion of lucanthone metabolites whether urinary excretion products or products obtained after incubation with liver microsomes were different for each host specie^-^ Swiss mice and Syrian hamsters infected experimentally with S . mansoni were treated with xanthenones, 4(3) D. Rosi, G. Peruzzotti, E. U '. Dennls, D. A. Berberian. H. Freele, and 5. Archer. Nature. 208, 1005 (1965). D. Rosi, G . Peruazotti. E. W. Dennis, D. A . Berberian, H. Freele. (4) and 5. Archer, J . Med. Chem., 10, 867 (1967). ( 5 ) D . Rosi, T. R. Lewis, R. Lorenz, H. Freele, D. A. Berberian, and S. Archer, ibid., 10, 877 (1967). ( 6 ) D. Rosi, A. J. Merola, and $. Archer, Life Scz., 6, 1433 (1967).
4*
3*
. .1:Ih S
L1*
12*
a a
>lire-------
* 81.:.
rng,'kg ;(lay X 5
--Hamsters----EI)so I SE. rng 'kgiday X 5 I
.Act.
index
\c:. index
SCHIRTOSOMTCIDAL XAXTHENONER . 4 m CHT~OROTOLUIDISEP
,July 1969
GO9
TABLE I11 COMPlR.4TIVE SCHISTOSOMICIDSL ACTIVITYO F
-M , cie
EDso
13* (AIirnsnn) IC,*
CHI CHTOTT
li*
CH 3
.
* SE,
--Hamsters-EDso i S E , mg/kg/dag X 5
Z
mg/kg/day X 5
Act. index
NHCH?CH:N(CSHj)2.HCl r\;HCH?CH,N(CyH,), .HCI
12.1 12 . 1 1.5.0 f 2 . 8
1 1
5 3 . 0 1 13.4 7 . 0 1 0.9
1 7
FHJCH,\!H ,2HC,
5.2 fO . i
1
>100.0
1
2.1 f0.3
2
3 . 0 1 0.4
14.6 f 2 . 1
1
>400, 0
40 0 i 9 . 3
2
i X . O =k 1 5 . 3
Y
Compda
THEIR HYDROSYMETHYL AKALOGS
4-RIETHYL-3-CHLORO.~lrILISES AKD
-N
Act. index
\cH,H'
18*
19
>33
1
>.5
TABLE IV C O M P IR.\TIVE
SCHISTOSOMICIDIL ACTIVITY
O F 4-hIETHYL-3-CHLORO.4NILINES
\ND
THEIR HYDROXYMETHYL AXALOGS
-----.\Zice---
EDso
Act. mg/ka/day X 5 index
Z
P
Compda
----Hamsters--
* SE,
EDso
+ SE,
mg/kp/day X 5
Act. index
,,CH.CH,\ -S
21
CH,
22
CHgOH
23 **
CHa
24 *
CHSOH
2.2
CH a
26
CHLOH
NCH CH=CH,.
\CH,CH. / CH CH.
/
a
See footnote a, Table I.
-N
HCI
1
9 . 0 1 1.9
1
>400.0
1
\
'CH CH/
NCH CH=CH-. HCI
-NHCH?CH?N( CyHs)?* HCI
* Also reported
7.5 f 1.7
as Abbott A-16612.
methyl-3-chloroanilines, and their hydroxylated derivatives. It was found that hydroxylation caused a oneto six fold enhancement of schistosomicidal activity in the mouse and a two- to 33-fold enhancement in the hamster. The purpose of this publication is to present the findings relating to the schistosomicidal activity in
c
9 4 . 0 f 38.8
>4
>200.0
1
>400.0
1
3 6 . 2 3 ~7 . 2
B
2 i X . 0 1 63.7
?
4.0zk0.9
1
42.31 9 5
I
3.711.0
2
. i . l i 1.3
I(:
Also reported as Hoechst S-616.
mice and hamsters of a number of xanthenones, 4methyl-3-chloroanilines, and their hydroxylated derivatives. Experimental Section Procediire= iised for the infection of wails ( i i i t s t r a h h i s gln-
hratus Puerto Mean origin) and rodents (Swiss mice and Syrian hamsters) wit,h S. rnunsoni (Puert’o Hican strain) and for the testing and evaluating of schistosomicidal activity of compounds administered via the oral andlor parenteral routes havr. been previoiisly described by Berberian and Freel~.’
of our observation:, on the schistosomicidal activity of :i number of hydroxylated derivative.: of xanthenones and 4-met hyl-X-c.hloroanilines. The findings have lent further ” :inti.cliistosom:tI agent rpInfection Controls.--A total of 686s live and seven gardleis of‘ t l i r route of dmiriistration. Airgentu, dead schistosomes were recovered at autopsy from 371 et al.,’? treated 211 patient. with single or multiple infection control mice utilized in 50 tests; the average intramuscular doseh of hycnnthone (2-3 number of live schistosomes per mouqe was lS.5. A concluded that the intramuscular route total of 4564 live and no dead hchistosomes nere found promising bec:iuce of the good tolerance a t autopsy from 189 infection control hamsters utilized cure.” .\[ore recently C l a r l q rt u1.,13 reported the rclin 39 tests; the average number of schistosomes per sults of their triitl- on :I total of 97 South African Ithohamster was 23.8. desiaii- n h o \\-ere foulid to be infected with either Schistosomicidal Activity.- Data on the schirtosoScliistosoim iiei71afob~umor S . t m / i s o ) z i or h t h . They micidal :Lctivity for xarit henones arid their hydroxyobtained :I highly wtisfartory cur(’ rate on both hemamethyl analogs are presented in Tables I and 11, and tobium :tnd nian~oiiiinfection-. They c o n d e r e d the for 4-methyl-3-chloroaniliiieh and their hydroxymethyl toxicity of hycaiit hone :is iinegligible” and concluded r ria logs in Tables I11 arid IV. that a single intramuscular injection \vas the preferred mode of :idminiitration. They stated that hyc:tnthone Discussion one drug which could be widely used i n m a s chemoSince the discovery in 1918 by Christopherson8 t8hat8 therapy and coiitrol of hematobium : i d mansoni infeetions. Both Brazi1i:in : i d South -Ifrican qtudies l)nbtart” emetic mas an eff ect’ive schistosomicidal agent,, lished hitherto :tnd thoze underway ha\ e :Ao conthe search for more active and less toxic compounds has firmed our 1:ibor:itory findings that a single intramuhcubeen relentlessly pursued at’ an ever increasing tempo lur dose of hycanthone i. as effective a s a &;-day courw wit,h the X.mansoni infection in Swiss mice being used of oral mc.tlic:ition. .llthough ineffective agaiu\t as a working model for routine in ~ J ~ Uscreening. O StanSchistosoma japoi! I ( ’ ~ C I ) Lhycanthone comes close to fulden9 estimated that’probably as many as 250,000 chemifilling the speculiitivc criteria of F:~irlev’~ aiitl S~Tcals have been test,ed to date in laboratory animals. wmc’’ for :jn itlc:il ~chi~toiomicide. Three nonmetallic schistosomicides of merit, have emerged from the extensive researches of the past three ‘11) \ Kat/ .I Pellegrino a n d C 1 01iLeira. i b s t r a c t s a n d Rerieiis, 8th International (‘ungresw- of Tropical lredicine anrl Malaria. Teheran decades : niridazole, lo lucant8hone hydrochloride, arid 1‘168 p 1101 hycnnt’hone, t’he hydroxyniethyl analog of lucanthone. (12) (‘, \ i r a e n i u r3 Garcia I< I’ 1 ) d i a i i x I L)a S i l r a , anrl I I t o < I D:rt,n presented in this paper constituted an extension rigries ( o i i ~ aref , 11 p 1102 1 7 ) 1). .\. 13erherian a n d H. Freele, J . Purtisitoi., SO, 438 (1964). 18) J. R. Christopherson, Lancet. 2 , 325 (1918). (9) 0. D. Standen. T r a m . Roy. SOC.7‘rop. .Mer/. Hyg.. 61, 563 (1987). (10) C. R . Lambert, 11. \Tilhelm, N. Rtriehel. F. Rradolfer. a n d P. Sclrmidt, Esperientia, 20, 452 (1964).
( 1 1) V r ) e V (‘lark? I ) \I. Rlair, a n d \ I C \ \ e b e r , Cenfriii 4 f r x r i n I \ f e d 18, 1 (1969) (14) S I1 Fairle\ Trari, R o y b o r . T r o p .\fed. Hyg., 46, 278 ( l q 5 l j ( 1 5 ) .I Ceirsonir ( ibs, Foundation bymposiiim on IEilharziasis, G 1, \Y I\ olstenholme and U O’( onnor Ed , L i t t l e Rroirn and C o I l o i t o n , \ l a w , 1962 p310
