~’HOSPHOKUS-SITIZOGE~ COMPOCNDS. VI1
September 1967
923
TABLE I l’I!uwiim~I,- .\NU ~ ’ I I V Y I ’ H I N Y L I D Y N E U R E . \ S , -.ZZIRIDINEC.\RBO
IDES, . \ N U
-SE!dlC.\RB.\ZIUES
l~2l’(O)NIIcol~’ R‘
It
NU.
I
CI
i~ I Ioc I-r
I1
Cl
111
CsH:,SH
I\-
CsHjNH
v
C~H~NII
N HC&B r-p
VI
EN
KHOCH,
\‘I1
CNCONH
J.1 I1 CI T:,ONI-ICOK1i IX CsH,NTISHCOKII X NH?h-FTCO?;H SI NH?NTI(CH~)COKH S I 1 (CIIr)?NNHCOSH SI11 CITLVHCSNHNHCONII \?I-SI11 at 4”/miii.
RIP, o c a
Formula
---Calcil, C H
xN
---FoiiniI,
C
t
I&--.
i
X
“3
91-92 dec 70 dec
C2Hd212NZOJ’ 1 1 . 6 2 . 4 1 3 . 5 11.7 2 . . i 13..i C3H,C12N202P 17.7 2 . 3 18.8 1 7 , s 2 . 7 13.6
,Vq
l.jO-lt72
C~~ITI~NIOJ’ 5 7 . 0 5 . 4 1 7 . 7 56.7 3 . 3 17.4
295-297
Ci,H17SsOaP
=(-=+O -*
N3 N€ IOC If:{ SFINIjC6II, XHSH:! S (C Hr)SH,
NIIS(CII,,), NIINHCSNHCII,
52.4 4 . 2 20.4 5 2 . 5 4 . 2 20.2
206-208
C19HlsBrN402P51.3 4 . 1 1 2 . 6 5 1 . 6 3 . 8 12..5
99-101
C~H~~NT~O 32.7 ~ P 6 . 0 2 5 . 4 3 2 . 7 6 . 0 2*5.*5
283-287dec HI-182dec 211-212dec 188-140dec 126-129 dec 1ti0-161 dec 203 dec
C8HI:,rU’,O3P CsI{I>S&7P C21T124N&4P C3H12N!,04P CBH],?U’!,OIP C9H24KiIOIP C!,II,IS1204P&
33.8 22.!1 X.7 1X 4 22. 1 3 0 . ti 22.1
5 . 0 2 7 . 8 33.8 5 . 3 2 7 . 6
4.S 26.7 2 2 . 8 4 . 7 2 6 . 4 4.9 25.3 4 . 5 46 8 5 , s 4 0 ,5 6.8 33.7 4 . 3 34.4
50.6 4 . 9 1:3.:3 4 . 6 2 3 . 0 .5. 8 3 0 . 3 6.7 2.21 4 . 2
23.1 46.2 40.3 3 i .6 34.2
‘1
turn cleave yielding hydroxylamines, credence being lent to this hypothesis by the work of Boyland, et uL.,~; which indicated that the alkylating hydroxamic acid, X-hydroxyurethan, is the active metabolite formed from urethan which is responsible for chromosome breaks. These authors concluded that these radiomimetic effects may be caused by some simple hydroxylamine derivative released in the tissue. In an earlier study of ~ the possibility urethan, Berenblum, et ~ l . , ’mentioned that the carcinogenic activity of this drug is due to formation of an effective metabolite through some oxidative mechanism. Hydrazides IX-XI11 may, therefore, allow for an appropriate rat.e of autoxidation and incorporation in tumor cells prior to bioconversion to cytotoxic products. This rationale is essentially that offcred for hydrazide 1IAO inhibitors whereby these acyl deriva,tives serve as “carrier” moieties to produce peripherally inactive agents relying on metabolic: degradation to release the active compounds in the brain.” In consideration of these data, an estimation of the degree of M A 0 inhibition by certain of the semicarbazide derivatives was considered worthy of determination. None of the compounds produced reversal of reserpine-induced blepharoptosis; however, central stimulation of some nature occurred upon administration of XI as indicated by the convulsion induced by this compound. Semicarbazides of certain hydrazines are kriown to possess potent RIA0 inhibition;18 however, phosphorylated semicarbazides may not provide for tissues selectivity and, in view of their lack of stability toward oxidation in d r o , it is likely that they may undergo oxidative biodegradation with cleavage to an active moiety prior to distribution to the target organ. Preliminary antitumor screening indicates that VI may be the most promising of the series, giving n (15) 1,:. Hoylaiitl, I t , X w y , l i I’cygie, a i d l i . \\‘illianis, B i o l , l r r n r .J . , 89, I l 3 p (1968). (16) I . Herenblum, 1). Ben-Ishai, N. Ilaran-Glirra, .4. I.ai)idot, I:. Simon, and N . Trainin, Bwchem. Phormacol., 2, 168 (1959). ( 1 7 ) J. H . Diel, A. Horita, and A . E . Drukker in “Psychopharmaeo-
logical Agents,“ RI. Gordon, Ed.. Academic Press Inc., New York. N . Y . , 1964, p 359. (18) Reference 17, p 414.
T/C (yo)of GS against Walker 236 subcutaneous a t a dose of 1 mgjkg. Subsequent testing at higher dose levels and against other systems may afford greater activity. X gave the most favorable response, T/C (%) = 27, against Walker 265 subcutaneous a t a dose of 80 mgjkg. Five compounds were incidentally screened for antimalarial properties; none extended survival time sufficiently to be classified as being active under the test conditions. Phosphorylation of ureas and semicarbazides results in the formation of compounds containing a large number of donor atoms, particularly in the case of the phosphinylidynetrissemicarbazides(IX-XIII) and l,l’,l”-phosphinylidynetris-3-methoxyurea (VIIT) which may be considered to be tridecadentatc ligands. Upon potentiometric titration to determine their upproximate ability to chelate, three derivatives cwmplexed with cupric ion arid two with zinc in :i 2 : 1 ratio. Only X I was examined with vanadium ion which it sequestered i n a ratio of 2 : 1. Experimental Section Chemistry.-SIelting poiiits were taken on a FisherJohiis apparatus and are uncorrected. Coleman carbon, hydrogen, and nitrogen analyzers were used for the elemental analyses. Infrared spectra of all starting materials, intermediates, and products were taken in a Nujol mull on a Beckman IR-8 spectrophotometer and gave the expected absorptions. A Corning Model 12 pH meter was used in the potentiometric titration and a J‘arian A-60 spectrometer for the nmr spectra. The experimental procedures given below refer to the compounds listed in Table I. Synthesis. 1-Dichlorophosphinyl-2-methoxyurea(I) and NDichlorophosphinyl-1 -aziridinecarboxamide (II).--A solution of phosphorisoc\.Lttiatitiic dichloijdelg (SO nimoles) iii 100 ml of ether was cooled t o 5-10’, aiid a11equimo1:tr :mount of axiriditie or methoxyaminem in 50 ml of ether was added dropwise. Followiiig filtration the residues were wnshetl with ether and plxcecl in n vaciiiiiii desiwator. N-Dianilinophosphinyl-1 -aziridinecarboxamide ( III)..-Ci)mpouiid I1 ( 2 5 mmoles) wiis added i l l portioils t o aniliiie (100 (19) Z. 13. Papanastassious and T. J . Bardos, .I. .Wed. Pharm. C h e m . , 6 , 1000 (1962). (20) T. C . Bissot, R. W.Parry, and D. €1. Campbell. J. Am. Chem. SOC., 79,‘796 (1957).
(i(i
SI1
..i 4
( i,llll> were crystallized from ethanol-water iIY) or dioxatie-water (I-). 1 -Diaziridinylphosphinyl-2-methoxyurea(VI).--Aziridine ( 3 0 ininc~les)and triethylamiiie ( 5 0 mmole,\j i t i 100 rnl of wc*etcltir \vwe 11,r;itedwith I1 ( 2 2 mnic~le.)itr t h e same m a t i i i e r 11' tlesrt~ikic~tl f o r 111. Followiiig holvettt rernclvttl the i.esitiue \va> esti,nrtctl \ v i l l i elhcr :itit1 c,otrc'eiitr:itii)ii of the ether extract produced thc, \v:itei~-st~li~hle pi~otiuc*twhich gave >I Iiegtttive te.-t with 0.1 .\+ AgSO:,.
N,N',N"-Phosphinylidynetris-l-aziridinecarboxamide (VII), 1,1',1"-Phosphinylidynetris-3-methoxyurea(VIII), and Phos-
phinylidynetrissemicarbazides (IX~XIII).-Phrisphiti!.lid~ire triii-i~ry:iii:~le,,41fa Iiii1rg:uiii4sj I t r i 4 . j (:ii,ritig with d i l i i l t : 1 IC'I ;iiid with etliaitol I I I winovp the iiitreac~trds e i n i u i r h a z i t l e . SI q ~ v :ei t ) tinir pciik ~ l ) ~ I S O - ~6 /?.!It? l i ~ ,~X!C'll:~i.TI)2 . 0 g (6:t mniole,.) of Ihi. pridiirt i l l 100 1111 of \ v : i t v t ~ \Y:I* ntltlcd 2.12 g i s c ~ c ~ y i t t i a t e(phosphoryl
11 YI I
September 1067
~,~’-DICAHBETHOXY-~,2’-BIAZIHIDINE
927
activity by a procedure similar to that’ of Aceto and Harris21 as modified by Nematollahi, et a1.22 A dose of 1.1 mmoles/kg was administered orally to groups of ten mice (male, strain CZH 20 i= 1 g). Since I X and XI11 are insoluble iii water, a 3% subpension of acacia in distilled water was used as the vehicle in all cases except where otherwise noted. After 2 hr, five mice in each group were challenged with a 2.5-mglkg ip dose of reserpine; t,he other five being given 5 mg/kg ip of reserpine. The animals were left undisturbed for 3 hr then evaluat’ed as to degree of blepharoptosis by two individuals. Ten control mice received oiily vehicle aiid reserpine (2.5 arid 5 mg/kg for each half) as a coiitrol while isocarboxazide, administered in the same maiiiier as the test compounds at a dose of O X 5 mmole/kg, served as a standard. The following ptotic scoring system of A4cetoaiid Harris21 was used: 4 = complete, 3 = three-fourthb, 2 = onehalf, 1 = one-fourth closure of the eyelids and 0 = open eyelids. Since the administration of XI resulted in convulsive death of all animals at the l.l-mmoles/kg level, the test was repeated using O . 2 i 3 , 0.55, and 0.825-niole/kg doses (ayueoiis solutions;) iii ti groiip of five mice with a challengiiig dose of 6 mg/kg ip of‘ reserpiiie. The results of the testing are given in Table 111.
Anticancer Screening.-Derivatives containing aziridinyl moieties were tested against Walker 256 subcutaneous (WA); the remaining compounds against Walker 256 intramuscular ( N I L ) and/or lymphoid leukemia L1210 (LE).23 Screening results are presented in Table IV. Antimalarial Screening.-Five mice were infected with a lethal dose of P l a s m o d i u m berghei 3 days prior to administration of I, 11, VIC-X, and XI1 in doses of 40, 160, and 640 mg/kg sc in oiLY4 The mean siirvival time of infected control mice was 6.1 days. The changes in survival time are given as the mean survival time of the treated groiip miliris the meaii survival time of the control group. The re;iiilts of the rodent antimalarial tests are given in Table I\..
(21) A l . 1). .\cero and L. 1’. Ilarris, J . Ttixl‘col. A p p l . IJhurmactil,, 7, 329 (1952). (22) J. N e ~ n a t u l l a l ~I\.. i , Guess, and J . Autian, J . .Wed. Chem., 9 , 660 (1966).
(23) Test results furnirlied by the Cancer Clieiiiotliera~~yNationul Service Center, I3etliesda, J I d . (24) Test results furnished b y \\.alter Reed . h n y Nedical Center, n’ashington, D . C.
Acknowledgments.-The author wishes to thank 1\1. AI. Clay for his assistance in the JIAO inhibition study. The author is also indebted to D. A . Nontalvo, Sout’hwest Research Institute, Sari Antonio, Texas, 11. R . \Villcot,t, School of Chemistry, University of Houston, id J . E. Robbers, l’urdue Uiiiversity, for rcc“liiig arid interpreting the rimr spectra.
Alkylating Agents Related to 2,2’-Biaziridine. 11.’ N,N’-Dicarbethox~-2,2’-biaziridine 1’ETER
w.
F E I T .4SD O L E
TVAERJIOSE SIELbEX
Leo Pharmaceuitcal Products, Ballerup, D e n m u r k Received December 1 7 , 1966 Revised M a n u s c r i p t Received d p r i l 17, 1967 DL- and ( 2 I t :2’l
