July, 1963
hhTHYLGLYOXAL
BIS(CCASYLHYDRAZONE) ANALOGS.
11
43 1
Studies on Methylglyoxal Bis(guany1hydrazone)l Analogs. 11. Structural Variations on Methylglyoxal Bis(guanylhydrazone)2 FREDBAIOCCHI, C. C. CHESG,W. J. HAGCERTY, JR.,LELASDR. LEWIS,T. IC=N-NH-C(=SH)NHdifferent classes. These modifications include the substitution of the methyl group, substitution of the hydrogen atom attached to the aldimine carbon, separation of the two guanidoimino and guanylhydrazonyl groups by various aliphatic moieties as well as by aromatic and alicyclic ring systems, substitution of the terminal hydrogen atoms of the guanylhydrazonyl portion, and monoguanylhydrazonyl derivatives. Several substituted benzophenone guanylhydrazones have shown confirmed activity in cell culture cytotoxicity tests.
group (111: R1 = CH,, Rz = alkyl). 3. Substitution of the hydrogen attached to the aldimine carbon in I by an aryl group (111: R1 = CH3, Rz = aryl). 4. Attachment of the two guanidoimino (=?JKHC(=?;H)SH*) moieties to a cyclic ring (IV). 5 . Separation of the two guanidoimino moieties by more than 2 CH&=SSHC(=NH)SHz carbon atoms (V). 6. Attachment of the two guaniI HC=N?;HC(=NH)NHz doimino moieties to a quinoid ring system (VI). 7. I Attachment of the two guanylhydrazonyl (>C= NKHC(=NH)NHJ groups to an aromatic ring (VII: The present investigation involves the preparation R = H or alkyl). 8. Substitution of the hydrogen of compounds with the fundamental >C=XNHCatoms a t the terminal nitrogen atoms in I by groups (=NH)NHmoiety in an attempt to correlate the other than alkyl groups (VIII). 9. Compounds constructure-activity relationship of this still unclassified taining only one guanylhydrazonyl function (IX). carcinostatic agent. Guanylhydrazones have been Substitution of the methyl group in I by alkyl groups, found to possess interesting biological properties: the in vitro growth inhibition of Mycobacterium t u b e r c ~ l o s i s , ~ as well as the substitution of the hydrogen atoms at the terminal nitrogen atoms in I by alkyl groups has been effective against tuberculosis of warm-blooded animals6: studied. prevention of the growth of mold in argicultural products’; bacteriostatic activitiess; they affect the activity of monoamine oxidase, diamine oxidase and histidine decarboxylaseg; and are effective in preventing the epidemic typhus infection in animals caused by rickettsiae. Compounds of the following types have been studied: I1 I11 11VI 1. Substitution of the methyl group in I by an aryl group (11: R1 = aryl). 2. Substitution of the hydroT1 gen attached t o the aldimine carbon in I by an alkyl ,C=X The discovery that methylglyoxal bis(guany1hydrazone) (I) possesses the first significant remission in adult acute myelocytic leukemia3 has initiated a systematic search in this field. A study in the homolog series of I has already been r e p ~ r t e d . ~
(1) The “Chemical Abstracts” name for this compound is 1,l’-[(methyl)ethanediylidenedinitriloldiguanidine. An acronym, “methyl GAG,” was suggested by Dr. Emil Frei, 111, of the National Cancer Institute. (2) This investigation was supported by the Cancer Chemotherapy National Service Center. National Cancer Institute of the National Institutes of Health, Public Health Service (Contract SA-43-ph-3025). (3) B. L. Freedlander and F. A. French, Cancer Rea., 18, 360 (1958). (4) E. G. Podrebarac, W.H. Nyberg, F. A. French, and C. C . Chenp, J . M e d . Chem., 6 , 283 (1963). ( 5 ) M. Naito, A. Shihoda, M. Ohta, F . Fujikawa, K. Nakajima. H. Fujii, A. Tokuoka, and Y. Hitosa, J . Pharm. Soc. (Tokyo), 74, 1047 (1952). (6) R. Behnisch, F. Mietzsch, and H. Schmidt, German Patent, 859,011 (Dec. 11. 1952). (7) F. Fujikan-a, A. Tokuoka. AI. Takimura, and K. Miura, J . Pharm. SOC.(Tokyo), 74, 518 (1952). (8) (a) F. Fujikawa, A. Tokuoka, K. Miura, E. Kometani, 9. Nakazawa, T. Omasu, and T. Toyoda, ibid., 73, 20 (1953); (h) 6. Hayashi, Kumamoto Pharm. Bull.. 1. 93 (1954); (0) S. Petersen and G. Domagk. Naturwiss., 41, 10 (1954); (d) M. Torigoe, PhaTm. Bull. (Tokyo), 3, 337 (1955); (e) F . Mietzsch, German Patent, 958,832 (Feb. 28, 1957): ( f ) P. Montegazza, F. Pacchiano, and G. Cavallini, Antibiot. Chelhotherapy, 11, 405 (1961); (9) G. Cavallini, E. Massarani, D. Nardi, L. Mauri. and P. Rlontepazsa, J . M e d . Pharm. Chem., 4, 177 (1961). (9) E. Werle, A. Schauerand G. Hartung, Klin. Wocherhr., 33, 662 (1955). (10) T.I.. Chen, IIua Ilsiieh II~uehPao, 44, 349 (1958).
VI1
VI11
IX
The guanylhydrazone derivatives (see Table I) were prepared by the reaction of the aminoguanidine salts with the corresponding carbonyl compounds in an aqueous or aqueous-alcohol medium in the presence of a catalytic amount of acid. It is of interest to note that when more than one “true” carbonyl group was present in a molecule, the extent of condensation between the carbonyl groups and aminoguanidine did not depend on the amount of the latter used, but rather on the naturc and solubility of its salt. For instaim,
43%
,
_-
I
._
July, 1963
METHYLGLYOXAL BIS(GGANYLHYDRAZONE) AKALOGS. I1
433
when l-phenyl-l,2-propanedionewas condensed with excess aminoguanidine dihydrochloride, the desired 1,1'-(met hylphenylethanediylidenedinitrilo) - diguanidiiir (111, R, = CH3, R2 = CeH,) wab obtained. 011 thcfi other hand, when amiiioguanidine sulfate was used, only one carbonyl group reacted. Thc resulting riionoguanylhydrazone derivative was assigned the cu-acetylbcnzylideiieaiiiinoguaiiidille structure (X) rather than its p isomer (XI). This was, in part, duc to tlie hyperconjugation effect escrted by tlic methyl group of l-phenyl-l,2-propanedionewliich nould teiid to make tlic carbonyl group next to the methyl group
General Preparation of Guanylhydrazones.---~:~~ioiis dt,s I I ~ miinoguanidine v-ere prepared in situ from coinnic:rc~i:tlly:LV:II! :hie arriinoguariidine bicwbonat.e.2' l o :I solution of 0.1 1 mole (15 g,j of urninoguctnidine t)ic.irIu>nate in 125 ml. of water rras added slowly the desired x i i l (:I fcw drops of amyl alcohol were added in order t o prevent fo:tniiiiy) until the pH of the solution was less than 7 . l'he scilution \V:P filtered from trace :tmounts of insoluble solids. The approl)ri:itct carbonyl ctmipountl n m then added slowly to tlie slightly :icicl filtrate at cu. BO". T h e amount of cw-lwnyl compound usctl i1.25 surh that, the ratio of aniinoguanidine 1xlr carbonyl function LV:IF I : 1. If the carbonyl compound did not dissolve in the :iqui'ous mixture, ethanol was added until thc reaction ~nisturc. w ; i \ lioniogeneous. The svlution was then stirred at rooni toriil)c'r:ilure for 16 hr. If a precipitate was Eorrncd the solid proi1uc.t \\-:is isolated by filtration. Otherwise tlie renction mixturt) WM cl-:~pi rntcd until :I soliil resitluc was ciht:iinecl. I.leir> vents are listed in 7Y:il~lwI arid 11. 1-
susceptible toward niicleopliilic attack hy tlic basic arninoguanidine. Hence, the formation of XI is less likely and structure S is preferred. This postulation was further substantiated by the fact that the product gave a positive iodoform test. The possibility that XI might also be present as a minor product was ruled out by paper chromatographic studies. Only on(' spot !vas observed when the condensation product wa> developed in three different systems Tlic preparation of 1,l'-(iiiethylethaiiedi~lideiiediiiitl.ilo)-bis(3-aiiiiiioguaiiidine) (lrIII, T = S H J ~ta. accomplished by a 3-step syiithesis 3Iethylation of tliioscniicarbazide with methyl iodide yielded S(methy1thio)-semicarbazide. l1 Treatment of the latter with hydrazine gave S, S '-dianiiiiognanidine hydriodide in good yield. To facilitate isolation of the final product, tlie hydriodide was converted to the sulfate salt hy means of silver sulfate." Condensation of the resulting S ,S '-diamiiioguaiiidinc sulfate with inethyl glyoxal afforded the desired coinpound (1-111. T = STI,). I'relitninary antitumor screening results of the guanylhydrazones are listed in Tables I and 11.12--1'' With the information presently available it appears that in the bisguanylhydrazone group several generalizations can be made: (1) replacement of the methyl group in I by an alkyl or aryl group results in loss of the original activity, ( 2 ) the presence of the aldimiiie hydrogen is essential for activity, (3) the original activity is relinquished when the hydrogen atom a t the terminal nitrogcn atom in 1 is replaced by functional groups. In the iiionoguaiiylhydrazoiie series, the simple aliphatic analogs are totally inactive. Several substituted benzophenone guaiiylhydrazones have shown confirmed activity in cell culture cytotoxicity tests. 1mh
W.K i r r t e n a n d G. B. L. Smith, J. A m . Ctiem. Soc., 68, 800 (1936). (12) J. Thiele and E. Dralle, Ann., 302, 275 (1898). (13) H. Beyer and T. Pyl, Ann. Chem.. 606, 50 (1957). 114) P. Grammatical&. Bull. m c . c h i n . France, 446 (1952). (1.5) J. Thiele and W.Barlow. Ann., 302, 311 (1898). (16) S.Petereen and G. Domagk, German Patent 942,627 (May 3, 1956). (17) F. L. Scott, W. N. hlorrish, and J. Reillp. J. O r g . Clrem.. 22, 640 (1067). (18) I[. King and J. Wright, J. Chem. Sui... 2314 (1948). I,]!)) U. \Vedekind and S. Bronatein, .Inn.. 307, 293 (18YYl.
Found: C, :$1.5: I-I, i i . 3 ; T,58.2. N,N'-Diaminoguanidine Hydriodide.---To ii suspeiiyioii of 23.:{ g. io. 1 inulr, (.if 2 - i i t i 1 ~ t l i ~ ~ l t h i c ~ ' ~ ~ e 1 i i i ~ ~ ~ r ~ i~n n290 ~ i ~nil. e~i~drii~ditle l l f :LllSl)lutc 111~~t,llLL~~i~~ \\ :k,$ :Ldded :3,:3 g. (0.1 nlole o f -ellow precipitate formed. Recryst:illization of t,lie product f r o i n :I niixtiir'c. of ethanol :ind water g:Lve 20 g. (fX(;;,l of a diit,t: solid! 1 l l . p . L?Lt5" (v,itlieffetves~c.' 1.1 -i~~ethvlethanedir~liclenedinitriloi-bis(3-aminoguanidi~le
(11) G.
(20) All melting points (corrected) were taken o n a Thornos-IIoover m e l t ing point apparatus. T h e ultraviolet absorption spectra were determinril with 8 Beckman DK-2. a n d the infrared spectra were tnken with a Perkin Elmer Infracord. (21) Trojan Puwder (."ompi~n~~, .lllmtown, ['ennsyl~~ini:~. ' 2 ' 2 ) (a) I?. J'hillipp and J. 1,~. \Tilliarus, ./. .1m. Chem. .Sor., SO, 24%; .\. ITrnry. R . ( * . S I i ~ k o s k y an
