TABLEI THEEFFECT OF CHOYEMPERINE O N EHRLICH ASCITES TUMOR-BEARING AXICE
The compounds were obtained by condensation of the appropriate haloacyl chlorides or bromides with piperazine hexahydrate or 2-methylpiperazine in chloroNo. of form in which almost all derivatives of haloacylpiperaDose, daily Mean PCVa zines are readily soluble. The acid chlorides give higher (Test/Control) T/C ing/k:: doves Survivors yields than bromides. The piperazine derivatives were 400 1 0/4 ... ... obtained in good yield as solids which can be easily 1 4/6 2.4510.4/3.410.6 0.72 ’LO0 100 1 6/6 2.46&0.5/3 4 k 0 . 6 0.72 purified by recrystallization. The new derivatives 50 0 0/5 ... ... of 2-methylpiperazine described in this paper, exclud25 3 4/5 1.0 1 0 . 3 / 2 . 2 f O . 4 0.45 ing XVI and X I X obtained in the solid state, are (as a Packed cell volume in milliliters 1 atandard deviation. crude products) oils or resins, crystallizing slowly and decomposing during distillation under reduced preset a1.j The latter compound was not lethal at 200 sure. Among them we obtained XI1 and XVImg/kg administered daily for 11 days. If biological XTX in a pure crystalline form. We could not obtain activity depends upon opening of the cyclic diester ring in a pure state the products of condensation of 2-chlorostructure, these data indicate that crosemperine may be propionyl, 2-bromopropionyl, 3-chloropropionyl, and more susceptible to enzymic attack than monocro3-chlorobutylryl chlorides with 2-methylpiperazine. taline. Alternatively, the methylated nitrogen of the Therefore, these compounds were not submitted to present compound may alter its distribution properties, biological investigations. resulting in increased toxic effects to the host but reThe derivatives of 2-methylpiperazine and the derivaduced toxic effects to the tumor. tives of piperazine previously describedlag3were exCrosemperine was tested up to 1600 pg/ml concenamined for cytostatic activity by the test of Constantration against Staphylococcus aureus, Escherichia coli, tinescu and co-w0rkers.j The method is based upon Bacillus cereus, Saccharomyces cerevisiae, Candida the determination of the smallest amount of compound albicans, and Aspergillus niger, using a serial dilution which, dissolved in a 1.0 m N caffein solution (0.0212 method’ arid lvas found inactive. wt o/G), will, after 24 hr at 25 f l o ,induce the characteristic alteration of all the mitotic figures of cario(7) R. F. Smith, D. E. Shay, and N. J. Doorenbos, J . Bacterzol., 8 6 , 1295 (1963). kinesis in the wheat radicular meristemas introduced into the solution. The alterations are identical with those produced by the same agent in healthy and canCytostatic Bis(haloacy1) Derivatives of Piperazine cerous animal tissues with an intense proliferative activity. These “chromatoc1azic’16 changes consist a n d 2-Methylpiperazine mainly of mitosis protraction, chromosome fragmentation, the formation of ana- and telophasic bridges, STEFAN G ~ o s z r c oSKI, ~ JUL~USZ SIENKIEWICZ, LILIANAN.IJM.IN, 1t001c.iOTELEANU, A N D ~ I A R IRETEZEANU A the appearance of micronuclei, and the grouping of chromatin in clusters of various sizes. Under the Llepartment of Pharmaceutical Chemistry, Medical Academy, same conditions, but without the addition of caff ein, LMi, Poland, and Institute of Pharmaceutical Research, only a slight inhibition of mitosis with no “chromatoBucharest, Rumania clazic” effects can be observed. On the other hand, Received November 18, 1967 the caffeine solution alone, depending on concentration, induces a weak “~tatmodieretic”~~’ activity, Disubstituted haloacylpiperazines were of interest to which consists of the formation of polynuclear cells. us as intermediates which can be used to obtain amine I t should also be emphasized that cytostatic substances, derivatives.’ Carbon and co-workersZa discovered which shorn a mechanism of action different from alantitumor activity for N,N’-bis(3-bromopropionyl)kylation, do not induce any of the previously described piperazine, introduced recently under the name pipochanges in the cariokinesis pictures. They produce, bromanlZb and N,~’-bis(3-chloropropionyl)piperazine at most, an inhibition of mitosis. was obtained some years ago by one of us.’& We Among 16 substances examined, pipobroman (111) decided to synthesize a series of derivatives3 and to showed (Table I) highest alkylating activity, followed examine their cytostatic activity. Also the mechanism by XI1 and 11. Substances I, X, XIX, and XX did of action of these compounds has not yet been elucinot show cytostatic activity. The remainder indated.4 In order to explain the relation between creased the statmodieretic activity of caff ein and had chemical structure and cytostatic activity of the comweak antimitotic or alkylating properties. pounds, we varied both the acyl substituent, introducing On the basis of experimental data, it can be concluded alkyl groups and a halogen atom in positions 2 or 3, that the bromine derivatives show a higher activity i~ndthe amine part, replacing piperazine by 2-methylthan chloro analogs. Position 3 is much more favorable piperazine. than 2. The introduction of a methyl radical or branched acyl group into the ring reduces the alkylating (1) (a) S. Groszkowski, Roczniki Chem., 34, 707 (1960); 38, 229 (1964); (h) S. Groszkowski and J. Sienkiewicz, Ann. Pharm. Franc., in press. activity. The absence of the halogen atom or too (2) (a) R. J. Stein, J. A. Carbon, J. Langdon, and R. K. Richards, J . Lab. small or too large acyl radicals abolish activity. Cliri, .Wed.. 56, 949 (1960); (b) VeroyteB. ( 3 ) (a) S. Groszkowski, J. Sienkiewicz, and L. Najman, Farmacia (Buoharest), 16, 263 (1967): (h) S.Groszkowski, L. Korzycka, and A. Wesolomski, f i o c z n i k i Chem., in press. (4) (a) J. Louis, R . J. Rohn, and R. W.Monto, Proc. A m . Assoc. Cancer Res., 3 , 246 (1961); (b) T. J. McNair, E. A. Wibin, E. T. Hoppe, J. L. Schniidt. and F. ,\. de Peyster, .I. Sury. Res., 3, 130 (1963); ( 0 ) C. E. Sasjleti, J. A I . JVslden, and H. H. Spencer, Cuncer. Res., 26, 275 (1865).
( 5 ) D. G. Constantinescu, h l . Constantinescu, AI. Retezeanu, R. Oteleanu, and V. Stoenesou., Compt. R e n d . , 263, 176, 1061 (1961); 264, 1665 (1962). ( 6 ) P. Dustin, Jr., “Expos& Annuels de Biologie Cellulaire,” Masson and Cie., Editeurs, Paris, 1956, p p 189-240. (7) .%. Gosselin Compl. Rend., 210, 544 (lY40).
