ti23
May 19tis TABLE
1
N-CHLOROETHYL DERIVAT~VE~ ZXCH2CH2Cl NO. ZN nrp, oc Eluent Yield, % Formulaf U 47 C14HiiClzNS' 118-120 1 2-Chlorophenothiazinyl~~3 b 50 C1SH1IC1F3XSh 80-81 2 2-(Trifluoromethyl)phenothiaziriyld a 35 CigHisClNS 88-90 3 9-Methylbenzo [a]phenothiazinyl a 45 Ci~Hi6C1~s 87-88 4 10-Methylbenzo [a]phenothiazinyl ... 37 CnH14ClNS 163-164 5 Benzo [b]phenothiazinyle Kindly supplied by lh. c Woelm neutral alumina, activity grade 1. a 3: 1 ligroin (bp 70-9O0)-C6H6. b Ligroin (bp 60-90"). Harry L. Yale of the Squibb Institute for Medical Research, New Brunswick, N. J. e Recrystallized from CHC13. f All compounds showed proper analytical values for C, H, N unless otherwise noted. 0 Not analyzed. h C: calcd, 54.6'2; found, 55.11.
TABLE I1 2-[BlS(2-HYDROXYETHYL)AMINO]ETHYL DERIVATIVES ZliCHzCH2N( CH2CHzOH)z No.
ZN
LIP, o c
Eluent
lield. %
Formulad
2-Chloropheno thiazinyl Oil U 82 CiaH2iClN202S Oil b 79 GdLiF3NzOzS 2-( Trifluoromethy1)phenothiazinyl 9-Methylbenzo [a]phenothiazinyl Oil a 97 C~JH&;~OAS 10-Methylbenzo [a]phenothiazinyl Oil a 90 C?~HZE.N&S Benzo [b] phenothiazinyl 94-95 C 72 CzzHzaNz02S 4: 1 CBHB->I~ZCO.9: 1 C&6-?rlezC0. 7: 3 CtiHti-?rIezCO. Difficulty was experienced in securing proper elementary aiialytical values on the bis(2-hydroxyethy1)amiiioethyl derivatives. However, the bis(2-chloroethy1)aminoethyl derivatives and the hydrochlorides gave proper analytical values.
1 2 3 4 5
the corresponding hydroxyethyl compound a t ice temperature. The mixture was allowed to warm slowly to room temperature '2- [BIS(2-CHLOROETHYL)AMIN 01ETHYL DERIVATIVES0 and then heated on a steam bath for 1 hr. Excess POCl3 was ZNCHSCHnN(CH,CH,Cl)z removed under reduced pressure and the residual oil was disYield, solved in acetone. The solution was poured over crushed ice and No. ZN % Formulab neutralized (Na&O3). The resulting solution was extracted 1 2-Chlorophenothiazinyl 43 CiaHigCIjSzS several times (CHCI,). The combined extracts were washed 2 2-(Trifluoromethyl)phenothiazinyl 60 C19H1gC12F3N2S (HzO), concentrated, and placed on a chromatographic column of The product was eluted with C6H6 to yield 0.5 g (19%) 3 9-Methylbenzo [a]phenothiazinyl 33 C Z ~ H ~ ~ C ~ Z N Z Florisil. S of oil. Conversion to the hydrochloride gave, after crystalliza4 10-Methylbenzo[a]phenothiazinyl 67 C13H2&12NzS tion fiom CHCl3-Et20, 94Yc yield of colorless crystals, mp 1085 Benzo [blphenothiazinyl 59 CzzHzzClzNZS 110". Anal. (CiQH23C13NnOS)C, H, X. All compounds in this table were oils. * C, H, N analyses. TABLE
111
Q
HYDROCHLORIDES ZNCHzCHzN(CH,CHzC1)2. HC1
Z-[~lS(%CHLORoETIIYL)AMiNO] ETHYL
NO.
1 2 3 4 5 a
ZN 2-Chloropl~enothiazinyl 2-(Trifluoromethyl)phenothiazinyl Y-hleth~lt~enzo[a]phenothiazinyl 10-hlethyibenzo[a]phenothiazinyl Benzo[Dlphenothiazinyl
Acknowledgment.-This investigation was supported by Public Health Research Grant No. GA-040068 from the Sational Cancer Institute.
hlp, Yield, "C 70 Formulaa 140-142 83 CisHnoClrNIS 140-141 97 CIOHZ~C~F~NZS 176-177 94 CaHaClaNzS 160-161 96 C2sHzClaNzS 151-153 88 CzzHd2laNnS
C, H, X analyses.
Cysteine Analogs as Potential Amino Acid Antagonists in Bacteria WALTER A. ZYGNUNTA N D TELLIS A. MARTIN
&lead Johnson Research Center, Evansville, Indiana ,$77,91 stirred a t ice-bath temperature for 1 hr and at room temperature for 18 hr. An almost colorless precipitate formed in the course of the reaction. CtiHs and HzO were added to the reaction mixture. The C & . - E ~ Z Olayer was separated and concentrated by evaporation. Ligroin (bp 70-90") was added to produce a 3: 1 mixture by volume of ligroin and benzene. Chromatographic separation of the solution over Alcoa grade F-20 alumina, using a 3: 1 mixture of ligroin (bp 70-90") and benzene as eluent, yielded 2.5 g (677,) of white solid, mp 82-83'. IO-( 2- [Bis(2-hydroxyethyl)amino]ethyl ) -3-methoxyphenothiazine.-A solution of 2.0 g (6.9 mmoles) of 10-(2-~hloroethyl)-3methoxyphenothiazine in 30 ml of diethanolamine was stirred at 140-150° for 30 hr and cooled to room temperature, and 50 ml of cold H20 wm added. The suspension was extracted (CHC1,) and the extract was washed (H20). The CHCl3 was evaporated and the resulting oil was dissolved in CsH,. The solution was chromatographed ovei 60-100 mesh Florid. Elution with C& yielded a small amount of lo-( 2-chloroethyl)-3-methoxyphenothiazine. The product, an oil, was eluted with a 1:4 mixture of acetone and benzene. The yield was 1.8 g (73y0). Anal. ((319H24NzO&S) H, N ; C: calcd, 63.30; found, 63.77, 63.80. lo-( 2- [Bis(2-~hloroethyl)amino] ethyl } -3-methoxyphenothiazine.-POCla (10 nil) was added slowly to 2.0 g (6.8 mnioles) of
Received December 21, 1967
Relatively few compounds have been reported to be cysteine antagonists in microorganisms. Allylglycine' inhibits in part utilization of cysteine in bacteria and yeast. Klubes and Schultze2 found that S-(l,2dichloroviny1)cysteine inhibited growth of Escherichia coli and showed that the L enantiomer was the more active isomer. Cysteine thioethers3 from chloroethylenes have also been reported to inhibit growth of fungi and algae. I n the present studies, 21 cysteine or cystine analogs were tested as inhibitors of cysteine or cystine utiliza(1) K. Dittmer. H. L. Goering, I. Goodman, and S. J . Cristol, J . Am. C h e n . Soc., 70, 2499 (1948) (2) P. Klubes a n d M. 0. Schultze, Bzocliim. B i o p h y s . Bcla, 78, 114 (1963). ( 3 ) L. L. Moliinnry, 1. C . Uldridgu, atid J. C . Cowan, J . d m Chem. Soc., 81, 1423 (lQ5g).
