N o,rES CHjOCH,C(CHz)LEHj
N
I
N CHd&COOCJL I1 CH,OCH,C( CH~)IOCHJ
I1
N I
N 11
CHjOCHX(CH2)jCHj V
discontinuance of Walker 236 testing system in the general screening.
T ~ B LI E --Antitumor Compounds
mg/k:!
I
230 100
acti\it> against \\ A-25b\Tt did, Sur1 I \ ors T-C
6 16 6 6 6 6
- 27 - 14 -9
T/C
(Yo)
9 8 63
Experimental Section
1,4-Dimethoxy-2-butanone Azine (I) and 1,4-Dimethoxy-Zbutanone Hydrazone (III).--I solution of 32 g (1 mol) of aiihydrous NIHa in 250 ml of anhydrous AIeOH vias stirred in an ire 6 6 -lh 18 I1 .0) bath while 33 g (0.25 mol) of l,ldirnethoxy-%-biitaiioiie (pre6 6 -7 72 103 pared by the method of Heiiriion and Kupiecki12 from 1 , P b u t y n diol dimethyl ether13) in 50 ml of hIeOH was added dropwise 6 6 - 16 41 I11 105 over a period of 30 min. The reaction mixt,ure was then stirred 6 6 -1i 25 70 a t room temperature for 16 hr. Removal of hIeOH and excess 6 '6 -7 42 45 NzHl followed by fractional distillation gave 22 g (60yc yield) 6 6 -3 89 30 ~ and 11.0 of t,he hydrazone (111),bp 91-92!' ( 5 . 3 mm), n Z 41.4671, g (17% yield) of the azine (I), bp 138-140" (2.6 mm), 7L2'D 1.4664. 1,4-1) imethoxyA n d . (CGHI~NZO~), C, H, X ; (CizHzJizO~),C, H, N. -2 60 4-butanone 400 6,6 Ethyl Pyruvate Azine with 1,4-Dimethoxy-2-butanone (II).A solution of 16.6 g (0.11 mol) of 1,4-dirnethoxy-2-butaiioiie -3 86 I IT 400 6 '6 hydrazone (111) in 50 m lof anhydrous MeOH was stirred at 0" while 13.2 g (0.11 mol) of ethyl pyruvate in 50 ml of VeOH was compounds were not reported. Wiley and coworkers4J added dropwise over a period of 30 min. The mixture w a d allowed to stir a t room temperature overnight, arid then distilled to prepared a series of substituted hydrazones of pyridoxal, give 19.0 g (70% yield) of the azine, bp 119-120' (0.4 mm), indole-3-carboxaldehyde and 2-methoxynaphthalden% 1.4708. Anal. (CnHzo3204),C, H, N. hyde, but only t w o of 35 compounds prepared were Ethyl acetoacetate azine with ethyl pyruvate (IV) was prefound t o have significant activity against sarcoma pared by a procedure analogous t,o the foregoing. The product, 183 in mice. mp 88-89", was purified by sublimation at 65' (0.1 mm). ilnal. (CIIHIINPOI), C, H, 3. A survey in the general area of biological activity 1-Methoxy-2-hexanone azine with 1,4-dimethoxy-2-butanone of azines again revealed little. Some synthetic azine ( V ) was prepared in an analogous fashion, bp 123-126" (2.3 mm), dyes, such as methyl violet and heliotrope, mere n Z 5 D 1.4628. d n U l . (CI~H&~O,), C, IT, 5 . chimed t o have good tuberculostatic effect against Ethyl acetoacetate azine with 1,4-dimethoxy-2-butanone ( V I ) human type tubercle bact'eria.6 The molluscacidal had bp 135-138' (0.35 mm), 12% 1.4938. Anal. (ClrHv&&a), activity displayed by azines of certain p-ben~oquinones~ C, H, N. 45
W M li~rgely att,ributed t,o the parent q u i n ~ n e s , ~ - ~ Acknowledgment.-The authors wish to thank 11r. :iud the d l known biological activity of nitrofuran John Gravatt' for his assist'ance in performing analytical derivatives could possibly be t'he reason for the antimeasurements. microbial activity exhibited by a number of J-nit,ro(12) G. F. Hennion and F. 1.' Iiapiecki, .I. Org. Chem., 18, 1601 (1953). ?-furfural aziries.loill (13) W.Reppe, J u ~ t u . Liebig, ; ArLri. Ctiem., 696, 38 (1Y35). The activity exhibited by compounds 1-111 is in a wag rather unique in that similar activit'y was not observed by et8hylacetoacetate azine with ethyl pyr/%Alanyl Thio Esters' uvate (117) nor by l,4-dimethoxy-2-butanoneJthe parerit compound of 1-111. Logically, other azine deI N D CHARLLS G . SICIIINLR GIL CLIFTOS~ rivatives of 1,4-dimethoxy-2-butanoiie, such as compounds V and VI, were prepared for a preliminary Department of Chemzstr?j,*Yorth Texas State Ytr.ucture-activit'y study. However, the study could L'niuerszty, Denton, Texas 76203 not be cont'inued in our laboratory because of the
Recezved October 10, 1969
G. Irick, J . Jfed. Phnrm. Chem., 5, 49 (1962). ( 5 ) R . H. W l e y a n d R. L. Clevenger, &id., 6, 1367 (1962). (6) 11.G. Good, Zantr. Bukteriol., Prrrasitenk. B b t . I. O r i g . . , 169, 99 (1957); (4) R . H. Wiley and
O r g . Chem., 26, 1614 (1960). (8) A . Halawani and N . Latif, J . E g y p t . M e d . Assoc., 37,957 (1954). (9) T. von Brand. 13. Mehlman, and hl. 0. Nolan, J . Pnrasitol., 36, 475 (1949). (10) J. D. Jolinston, U. S. P a t e n t , 3,099,663 (1963); Chem. Abstr., 60, 28U.l~ (1964). (11) J. 13. .Juiinrton, U. S.P a t e n t , 3,206,257 (l'Jti7); Ckem. d b a t r . , 67, 310011 (I'JHT,.
The introduction of chemically active centers in organic molecules which may serve as alkylating agents has been an area of recent interest in thesyntheses of (1) This invedtigation was supported in Vart lis Kational Institutes of IIealtlr Research Grant N u . C.\-08102. Sational Cancer Institute, and b y tlie Nortli Texas State University Vacnlty Researcli Fund. (2) Sational Defense Education .\et B'dlow, Title 11'.
7
9 I O I I 12 13 14 15 L E N G T H OF A L K Y L G R O U P
8
16
J o u i n a l of Jlctlzcmul Chcttiistr y j 1,970,l-ol. ii, -1o. .2 577 was filtered aud washed with dry EtZO. The resulting solids were recrystallized (;\IeOH-Et,,O) to yield hygroscopic compounds which were maintained in a desiccator. (See Table I). n-Decyl 3-Aminopropionate Hydrochloride.-The same procedwe as used for t,his compolnid as for the thio ester. The solid had nip 84-85" and analyzed satisfactorily for C, H, and Iu.
RIetal Complexes of 1-Substituted 3-Hydroxyureas
-1.0
8
IO 12 14 16 TOTAL CHAIN L E N G T H
18
20
Figiwe 2.--6 cornparisoil of the coilcentration of @-alaiiyl thioesters and long chain alkylamines required t o completely inhibit the growth of Luclobaci//us arabinosus. The legend iiidicates the total number of atoms in each of the chairid att'ached to the amiiio groiip.
cantly less toxic. These dat'a suggest' that the inhibitory properties of the thio esters are a function of their lipid character rather than of their alkylating ability.
A considerable amount of work hab been done to indicate the existence of a definite correlation between the metal binding properties of therapeutically active compounds and their a ~ t i v i t y . ~ - For ~ instance, therapeutically active analogs of tetracycline form 2: 1 complexeswith Cu (II),S i (11),and Zn(I1j ions whereas the inactive analogs appear to form only 1:1 complexes.: Hydroxyurea (HUj6 and ethylhydroxyurea (EHVj' have been found to be very active against 1,1210 lymphoid leukemia. When patients with myelogenous leukemia were placed on HU therapy, acetohydroxamic acid was identified in their blood. To explain the mode of action of HU in biological systems, two theories have been proposed. According to Fishbein and Carbone,sHU is hydrolyzed to hydroxylamine, which cleaves thio esters such as acetyl-coenzyme -4, as shown in eq 1. h second theoryg suggested that "*OH
Experimental Sectionla Biological Testing Procedures.-The organisms used in this study were E. coli 9723, L. arabinosus 17-5, L. casei 7469, and P. cerwisiae 8042. An inorganic salts medium was used for E. coli,ll and a previously reported amino acid medium was utilized for I,. arabinosus and L. cusei12 except that the vit.amin supplement contained 0.2 gg/ml of calcium pantothenate, and 500 pg of glutamic acid was added per tube for assays with L. casei. An acid-hydrolyzed casein medium13 was used for P. cerevisiae with the Salts A concentration increased fourfold. Growth assay times and temperatures were 16 hr at 37" for E. coli, 24 hr at 37" for L . casei, 20 hr at 30" for L . arabinosus, and 21 hr a t 30" for P. cerevisiae. The amount of growth was determined on a Bausch and Lomb Model 20 spectrometer as per cent transmission a t 600 rnp.l4 p-Alanyl Thio Ester Hydrochlorides.-A 20-ml sample of the appropriate thiol \Tas cooled to appoximately 5", and 3 g of @-alanylchloride. HC18 was added. The react,ion mixture, protected frcm moisture, was stirred for 15 min at ice bath temperature and then allowed to come to room temperature. Stirring was continued overnight. The mixture was again cooled to 5", aiid EtsO (50 ml) was added to give the desired product' which (10) 3Ielting points \\.ere determined iritli a Thomas-Hooi-er Capillary Melting Point Apparatus. T h e authors are indebted t o Mrs. Sarah R . Bryant for assistance with the microbial testing procedures. 32, 120 (1945). (11) E. H. .\nderson, Proc. S a t . A c n d . S c i . , V.S., (12) .J. M. Ravel. L. \Voods, B. Felsing, and I\-. Shire. J . B i d . Chem., 206, 391 11954). 113) E. 31. I,anrforil. .Jr., IT-. 11, Ilartliny. and \V. Sltive, a i r c h . B l ' o r h r m . B i o p h y s . . 73, 180 (1958). (14) R . E. hlasinyale. Y. R. liryant, C. G . Skinner, J . Xasli, and P. F. liruae. ,Jr., J . - U e , / . C h e m . , 12, 152 (1Y6Y).
+ 2 H D +3 H l O H + NH4+ + HCOa+ CHdCOSCoA +CH3COSHOH + HS-Co4
KHiCOXHOH
fl)
hydroxyurea caused fragmentation of isolated DSA and induced chromosomal abnormalities in mammalian cells. The proposed mechanism is shown in eq 2 . oxidation
KHICOSHOH
-+ 2( HOX :) +Ha?rT?02 - Hi0
(2)
Hyponitrous acid (H2S2O2)like hydroxylamine and other compounds, causes cleavage of the main chain of cellular DSA. Our results based on the decomposition of Fe(II1)-EHU complex suggest that hydroxyureas may be involved in a n oxidation-reduction reaction, producing K O . I n this paper, we also wish t o describe the synthesis, metal-binding properties, and antitumor activity of several 1-substituted 3-hydroxyureas. The synthesis of 1-substituted 3-hydroxyureas la-g was achieved by a slight modification of the method (1) J. C. Dabrowiak, 31. .I, Thesis. \Vestern llicliiyan University, Iialamazoo, hfich., 1 9 6 i . (2) J. T. Doluisio and .I.S . XIartin, J . .\fed. Chern.. 6 , 20 (1963). ( 3 ) R. C. \Varner and I . JVeber, J . d m e r . Chem. Soc., 76, 5094 (1953). (-1) K. \\-. Iiohn, S u t u r e , 191, 1156 (1961). ( 5 ) J . T . Doluisio and -1.N . Martin, J . M e d . Phnrm. Chem., 6, 16 (1963). (6) 13. dreams. K.Losee, and .J. Hernstein. i h i d . . 6 , ( 2 ) . 201 (1963). (i) Privarp communication, C a n r r r Cliemotherap?. National Srrvice Crnter, I3etliesda. 11~1. (8) \V. Fislll~einand P. Carbone. S c i e n c e , 1 4 2 , 106Y (1963). (Y) .i, 13endicli. E. Horenfreund. G . Iiornguld, and 31. liriln. J . . Y , I ~ . Cuncer I d . , 82, 667 (1964).
