S OTES
May 1966
423
TABLE 11: SCREESING D \T.\" N(CH2CH2Cl),
I
I
CH,-p.T3 %I
-N3 Pyridine
3-rlcetylpyridiiie
Ethyl isoiiicotiriate
Quinoline
Lepidine
Isoquinoline
Test systemb
Dose, mdkg
AB AA AA \T A WA WA ITA WA ITA JVA4 WA KB KB AA Ah AA aA AA AA AA AA AX WA WA w.4 Wh KB AA A.4 Ah A.1 AA ilA AA WA
100 33 10 41 20.5 10.2 8.0 6.0 5.1 4.0 2.0
JTA WAA
Cures
6 6 0 0
0 0 0 0
Animal at dif (T - C)'
- 56 - 29 - 25 - 17 -6
- 17 c
-i
-2
---Turnor Test
Control
0 .0 0 .0 1.1 3 ,0 1 , r, 2.5 5.9 8.5
5.0 5.0 5 0 8.0 8.0 5.0 8.0 8.0
ut----.
T / C, %
--Cell ED&
culture--Slopee
0 0
22
.,-
r) I
56 50
-i *3,
106
4.3
-0.56 - 0.49
1 3
-0.33
1 .ti
33 10 3.0 50 10 3.0 33 10 3.0 13 6.5 3.25 1.6
33 10 3.0 100 33 10 3.0 41 20.5 10.2 5.1
- 13 2 6 3 8 20
6 4 0 0
- 59 - 40 - 19 - 15
0.0 0.9 3.0 6.9
0 10
--
33 l i
-9 1
2 0 0 0
- 29 - 10 - 1s -6
0.6 4.7 4.3 5.2
KB KB Tests carried out by the CCKSC according to their normal screening protocol. darcoma 256, KB = cell culture. T = test animal, C = control animal. d ED,, control growth. a Slope = difference in response for a tenfold difference in dose. 1- { p - [Bis(2-p-tolylsulfonyloxyethyl)amino] benzyl lpyridinium (IV).-p-[Bis(2-p-tolylsulfonyloxyeth~l)ap-Toluenesulfonate mino] benzaldehyde6 was reduced with NaBH4 as described for the preparation of I. The crude alcohol, mp 78-80', was treated with p-toluenesulfonyl chloride and pyridine as described for the preparation of I1 t o give a 667, yield of the hemihydrate of IT', mp 79-81' (from acetone). Anal. Calcd for C ~ ~ H ~ O X ~ O & . O . C, ~ H58.32; ~ O : H, 5.42; N, 3.68; S, 12.63. Found: C, 58.20; H, 5.31; N, 3.65; S, 12.64. 1- { p - IBis(2-p-tolylsulfonyloxyethyl)amino] benzyl ] quinolinium p-toluenesulfonate wab obtained ill 56Y0 yield ab its heniihydrate, nip 97-99' (from ac.etoiie-dimethylforriiaiiiide), in a procedure similar to the preparation of IV. Anal. Calcd for Ca~H,,N209S3.0.5H20:C, 60.52; H, 5.34; N, 3.45; S, 11.85. Foimd: C, 60.38; 60.24; H, 5.50, 5.57; N, 3.46; S, 11.55. (6) A Cohen and R . S. Tipson, J . M e d . Chcm., 6 , 822 (1963).
8.9 8.9 8.9 8.9
5.0 5.0 5.0 5.0
12 94 86 104
11.0 1.6 -0.46 * AA = toxicity test, WA = Walker carcino= dose (pg/ml) that inhibits growth to 50% of
2,4-Bis(arylamino)pyrimidines as Antimicrobial Agents DOLLY OHOSH~ Department of Biochemistry, Universify College of Scienre, Calcutta 19, India Received January 26, 1966
Although a large number of 2,4-diaminopyrimidines with hydrogen, alkyl, or aryl substitution in the 5 and 6 positions (type A) have been synthesized and screened (1) Officer, Scientists' Pool, Council of Scientific and Industrial Research, Government of India.
Ar HN
('ornpd
.\ r
Yield. r. 'C (crude)
Reaction time.
min
AIp, "C"
--I. Scott, and E. Young. Brit. J . Pharmacol.. 3, 156 (1948); 3, 160 (1948); (c) G. €1. Hitchings and J. J. Rurchall, Advan. Enzymol.. 27, 417 (1965).
13) (a) C. K. Banks, J . Sni. Chem. Soe.. 66, 1127 (1944); (b) {bid., 73, 3011 (1951); ( c ) T. B. Johnson and C . 0. Johns, A m . Chem. J . . 3 4 , J75 (1405); id) tI. I,. Wheeler and If. S. Iiristol, < b i d . , 33, 437 (1905). ( 4 ) 1). R o y , P. Ghosli, and T3. C. Giilia. J. O w . Chem., 26, 1909 (1960). ( 5 ) D. Roy, S. Ghosh, and '3. C . Guha, Arch. Biochem. B i o p k w . , 92, 366 il'36l).
B
A
Experimental Section
2,4-Bis(p-chloroanilino)pyrimidine ( I I ) . -'~2,,3-1 )i('hIOIY,~J) riiiiidine ( 1 . 5 g, 0.01 mole) was added to a solutinn of p-chlornaniliiic~ (2.6 g, 0.02 mole) in dilute HC1 (0.2 nil of conwiitrated HCI i i i 30 1111 of R-ater) and slo1~1,vheated to reflux on a miid bath Tvifli bshaking. Crystals began t,o appear within a few miiiutcs of rcfluxing. The refluxing was discontinued after 30 min, a n d t h v reaction mixture was kept overnight i n a refrigerator. Thc~ crj-stalline product was filtered off and washed with cold water. This compound could he readily recrystallized from 95% etti:iiiol, The yield was almost quantitative. The other compounds listed in Table I were synthesized bj. name general method as described for 11. As indicated in 'rablo I the time of refluxing had to be extended in certain cases :tiid some compounds u-eie recrystallized as the hydrochlorides sincci they could not be satisfactorily crystallized in their basic form. All compounds were dried in zraciio a t 100" for 20 hr heforc ~nalysis. Inhibition of Growth of Microorganisms.-All ~~~niplJUiltis ~t'ri, tested for their antimicrobial activity against Streptococriis ,faecalis, Escherichia coli B, and a pathogenic strain of ?.em!, Candida alhicans. The concentrations of synthetic compounds necessary for 50% inhibition of growth were determined turbidimetrically b y serial-dilution technique in test tube3 using liqiiid growth medium4 (shown in Table 11). t h ( 3
Acknowledgment.-Tlw author eqjresses her grat itude t o the Council of Scientific and Industrial l l ~ iearrh, Go\ erninent of India, for financial support. Thr author also wishc- t o tliank Profcssor Saile41 Chandra Roy for enc~ouragc~mcntand generous hrlp i l l caarryirig out tliis work.
