Journal of ilfedicinal Chemistry, 1970, Val. 13, S o . 4 765
Norm Characteristics of the new l-phenyl-2-arylhydrazono-l,2,3butanetriones are summarized in Table 11. 2-Amino-4-methyI-6-phenylpyrimidine.-Guanidine nitrate ( 2 . 5 g, 0.02 mol) was added to l-phenyl-1,3-butanedione (3.2 g, 0.02 mol) containing 10 S NaOH (10.0 ml) and 3IeOH (20.0 mlj. The mixture was stirred for 10 hr at 50-60” and left for another 1% hr at room temperature. The precipitated 2-amino-4methyl-6-phenylpyrimidine was collected and washed with 1IeOH and hot H20. It was recrystallized from RIeOH (2.3 g, 6!jC;,),mp 171’. A n a l . ( C I ~ H ~ , S , )H, C , N. 2-Amino-4-methyl-6-phenyl-5- (2,4-dimethoxyphenylazo)pyrimidine. Method A.-The diazotized 2,4-dimethoxyaniline, free from HXO?was added to a well-cooled aiid stirred solution of 2-aniiiio-4-methyl-6-phenylpyrimidiae (3.7 g, 0.02 mol j in AcOH (43.0 ml) containing sufficient XaOAc so as to maintain the reaction mixtIire at pH 6-7. The mixture was st,irred for another 6 hr at 0-5’ and left for 24 hr at room temperature. The product thus precipitated was collected, washed well with HIO, and recryotallixed from DNF-EtOH (3.6 g, 6655) as golden yellow plates, nip 160”. A n a l . (ClgHl&502)C, H, N . Method B.-Guanidine nitrate (2.5 g, 0.02 mol) n-as added to l-phenyl-2-(2,4- dimethoxypheny1)hydrazono-1,2,3-butanetrione (6.5 g, 0.02 mol) which in turn was prepared by coupling diazotized 2,4-dimethoxyaiiiline with l-phenyl-1,3-butanedione, containing 10 S S a O H (10.0 mlj and MeOH (20.0 ml). The mixture wa3 stirred for 12 hr at, 60-70” and left for another 12 hr at room temperature. 2-Amino-4-methyl-6-phenyl-5-(2,4-dirnethoxypheny1azo)pyrimidine precipitated, was collected, and was washed successively with lIeOH aiid hot H?O. It was recrystallized from DlIF-EtOH (2.7 g., 517,) as golden yellow plates, mp leOo. BY similar procedures, several 2-amino-4-methyl-6-phenyllazopyrimidines were prepared; they are summarized in Table I.
Acknowledgment.-The authors are greatly indebted to Drs. H. B. Wood and H. W. Bond of the Cancer Chemotherapy Sat,ional Service Cent’er for t’heir cooperation and for making the screening data available. We are also thankful to Professor W.U. Malik, Head of the Chemistry Depart,ment, for providing t8henecessary facilities for this work and t,he C.S.I.R., Xew Delhi (India) for a Junior Research Fellowship (held by R.A.S.).
Synthesis of 5-Iodo-3-indolylphosphodiesters of 5-Fluorodeoxyuridine As Possible Chromogenic Cancer Chemotherapeutic Agents’ K. C. Tsorr, S. AOYAGI,A N D E. E. MILLER Harrison Dcpartment of Surgical Research, School of Jlediczne, C‘niwrsity of Pennsylvania, Philadelphia, Pennsylvania 19104 Rrceived A u g u s t 14, 1969
I n the design and synthesis of anticancer agents based on enzyme rationa,le,2 it would be desirable to have one of the enzymatically hydrolyzed products as a chromogen so that the fa,te of such enzyme-mediated drugs cen be followed. Previously, we reported the synthesis of several tetrezolium mustards which could be reduced in vivo to a more toxic, colored formazan mustard. The present paper reports the incorporation (1) This work was supported by AEC Research Grant dT(30-1)3784 and U. S.Public Health Service Research Grant Ch-07339. (2) K . C. Tsou, S. B. Damle, R . W.Crichlow, R. G . Ravdin, and H. TT. Blunt, J . Phaim. Sei.. 16, 484 (1967). (3) I could be observed only after long incubation.
Both 1 and 2 \vcrc c o m p r e d with I.'UI>R in Hel:i cell tiqsue culture. Compound 1 was found to be mow eflective than 2 against, tliii cell line, equal to ITJDII ai 10-j d/ concentration. Interestingly, only in Hela cell3 treated with the 3' derivative could the liberated 5,s'diiodoindigo be seen. The 3' derivative was noi 111drolyzed. It is thus our opinion that this ricw chiornogenic derivative of FUDR would provide a more direct means of btudying the mechanism of E'UDIZ in c ; ~ n c ( ~ r research. Such an objective ha. been a long-term g o d of developmental enzyme hiztochemistry in our h h ~ r : i tory :tnd elseir here. Experimental Section 5-Fluorodeoxyuridine 5'-(5-Iodo-3-indolyl) Phosphate (dFC.~'-IIP).-~'-O-AC-~FT-~ (1.06 g, 3.7 mmol) was treated i n dr) pyridine n i t h ,j-jodo-.\.-;\c-indox?-l pho..phodic.hloridate obtained from 606 mg ( 2 mniol) of 5-iodo-~~'--Ac-iiidoxyl fur 24 h r at r ~ m n temperature. The reartioil mixt,ure was evaporated in Z'UCLLO, treated with 20 ml of concentrated YHaOH, left for 24 hr at room temperature and evaporated below 30". The resiiltant oil was diluted with H20, treated with activated charcoal, and filtered. The filtrate was diluted to 1.6 1. and absorbed 011 ti DEAE-Sephadex column (2.2 X 38 em). The coliimii wab washed with 0.02 XI SH,HCOa (ca. 1.7) and connected l o a linear gradient produced from 0.02 -11(4 1.) and 0.3 JI NHIIICO,~ (4 1.). After the developer in the gradient apparatus wad used up, additional solution (0.4 M , 5 1.) was led to the column. The peak fractions were freeze-dried to yield t,he almost colorless product, ca,. 710 mg (61% ). After rechromat>ography, the analytically pure sample was obtained ca. 300 mg. Anal. (C,:II,gFIYdO8P): C, 34.94; €1, 3.27; K, 9.59; I, 21.72; P, 5.30. Found: C, 34.87; H, 3.47; K,9.63; I, 21.45; P, 5.20.
Journul of Jlrdicinal Chemistry, 1970, Vol. 13, S o . 4
767
TABLI: I1 PHYSICAL PROPERTIES OF PHOSPHOUIESTERS OF FUDR Rf values" Sols 1
Compounds
0.64 dT-3'-IIP 0.59 dT-5'-IIP 0.57 dFU-3'-IIP 0.60 dFU-j'-IIP I I P in solvent 1, 0.63; 2, 0.31; 3, 0.31.
uv spectrab
Soh 2
Sols 3
Amin ( m r )
(4
Amax ( m d
0.63 250(9200) 0.68 0.61 250(9100) 0.65 0.49 251(8300) 0.71 0.45 251(7800) 0.69 All spectra were recorded in dist,illed H20 (neutral).
(e)
268(12600) 268(12200) 271 (11800) 271(10900)
C o m p a r i s o n of E f f e c t of FUDR, F U D R - 5 ' - p - I 1 a n d FUDR-3'-p-I1 o n Hela Cell Growth
