246 Journal of Medicinal Chemistry, 1971, Vol. 1.4,N o . 3
TABLE I1 Gastric an tisecretory activity EDw, m g / k Shay ratb
Toxicity
LD50,mg/kg No.
1
2 3 4 5
6 7 8 9 10 11 12 13
14 15 16 17 18 19 20 21 22 23
per os (mice)
570d (545-595) 618 (573-668) 673 (576-787) 628 (576-686) >1.000 470 (398-554) >1.000 >1.000 575 (532-623) 70 (58.6-83.6)
1.42 2.30
>10- 4
3.5.10-5
5.10
>io-'
4.05
>io-'
1.90 2.64
>io-'
11.50 3.70 3.25
1.25.10-4
>io-' >io-'
>10-4
NA (20)
>1.000 >1 .ooo 830 (686-1004) 924 (775-1101) >1.000 >1.000 >1.000 >1.000 >1.000 >1.000 653 (568-751) >1.000 >1.000
Reference products
Anticholinergic activity pAiC
NA (100) 16.2 4.28
>10-4 >10-4
4.32
>10-4
7.90 1.50 7.82 NA (100) 50 82 NA (50) 4.60 NA (100) LDwa
NOTES
vacuum. The solid residue was recrystd from CeHe to give 6 g (60%)) mp 108-109°. Anal. (CsH12NS)N, S. The hydrochloride was prepared by adding etheral 4 N HC1 (6 ml) to 3a (4 g) in EtOH (150 ml). The solvents werc removed under vacuum and the residue was recrpstdfrom 90 ml of EtOAcEtOH (50:40); yield 3.9 g (8070), mp 180-181' dec. A n d . (CsHizNS.HC1) N, S. Gastric Antisecretory Activity in the Rat.-Gastric antisecretory activity was evaluated in the 4 hr polyrus-ligated rat, using the technique of Shay? The compds were suspended in 20'/;, gum syrup, and were administered intraduodenally immediately after pyloric ligation to groups of 6 male Sprague Dawley/CD rat.s weighing 221 f 1.77 g.t Free acid output was calcd for each rat and expressed as pequiv/4 hr per 100 g of body weight. Data of the whole test series have been pooled for the control group (183 rats),$ and the mean value of each group receiving drugs was compared to the mean value of this control group using t.he Student's "t" test. Per cent inhibition whs calcd in comparison with the control values representing loo%, and plotted on semilogarithmic paper vs. mg/kg of dose. EDbowas read from the graph. Three to five doses were used for each compound. After completion of this manuscript, some pharmacological results on 2-pyridyl thioacetamide were described by J. Borsy, el al., at the 4th World Congress of Gast,roenterology i n Copenhagen. These results are in full agreement with ours. ~
>10- 4
>io-' >10-4
(5) H.G.Shay, S. A. Komarow, S. 9.Felds. I). Meranse, h l . Gruenstein, and H. Siplet, Gastroenterology, 5 , 43 (1945). t m zk standard error of the mean. Control values for these experiments with 185 rats were: vol: 3.09 i 0.06 m1/4 hr per 100 g . Free acid concn: 85.8 f 1.2 mequiv/l. Free acid output: 271.8 zk 7.9 pequiv/4 hr per 100 g. Total acid concn: 111.0 zk 1.0 mequiv/l. Total acid output: 348 ct 8.6 pequiv/4 hr per 100 E .
Antimicrobial Compounds. 1. Synthesis and Antimicrobial Activity of Some Alkylidene, Cycloalkylidene, and Arylidene Derivatives of 3-Hydrazinopyridazine
>io-'
ED% Shay-rat
Atropine sulfatee
207 ip 0.462 ip (182-227) PPT * HClf 592 PO 5.406 >10 - 4 (526-667) Thioacetamideo ~ 2 2 ip 0 35.0 ip a Acute toxicity was determined orally in male albino mice CJI strain. LDm calcd by C. S. Weil's method [Biometrics, 8, 249 (1962)l. Numbers in parentheses are fiduciary limits of LDw. Compounds are given by intraduodenal route. NA = inactive compound. Numbers in parentheses are the maximum dose assayed. Anticholinergic activity was established on perfused guinea pig ileum by pA2 technique [E. Bulbring, A. Grema, and 0. It. Saxby, Brit. J . Pharmacol., 13, 440 (1958)l. Drugs were Van Tamelen and Baran (footnote dissolved in 0.97, saline. a, Table I ) have LDu mice as 750 mg/kg sc. Atropine sulfate from FLUKA AG, Buchs (Switzerland). f 2-Phenyl-2-(2-pyridy1)thioacetamide .HCl synthesized in our laboratories. 0 Thioacetamide from Schuchardt, Munchen (Germany).
this position, were inactive (18) or showed only mild activity (19). Experimental Section
2 4 2-Pyridy1)butanothioamide (5a).--2( 2-Pyridy1)butanonitrile (8.2 g, 0.056 mole) was dissolved into a mixture of EtaN (5.6 g, 0.056 mole) and pyridine (8 9). The soln was satd with dry HzS a t room temp and treated in a sealed tube to 100' and maintained for 15 hr. After cooling, the mixture was poured into H20 (100 ml). The suspension was extracted with CHCla, the extracts were washed (HzO), dried (NazSOd), and concd under
31. LIKAR,B. DRINOVEC,
Institute of Microbiology, University of Ljubljana, Ljubljana, Yugoslavia
M.
JAPELJ,
* A, POLLAX,A.
POVSE, AND
P. JERMAN
Research and Development Institute, Krka, Pharmaceutical and Chemical Works, Novo Mesto, Yugoslavia Received June 1 1970 I
Various sulfonamidopyridazines1v2have been reported to have antibacterial activity with a low toxicity. In our experiments we have found that 3-thenoylamino6-chl~ropyridazine~ exhibits good antimicrobial activand a few ity. A number of hydrazin~pyridazines~ alkylidenehydrazinopyridazines5 have been reported to (1) (a) G.W. Anderson, H. E. Faith, H. W. Marson, P. 9. Winnek, and R. 0. Roblin, J . Amer. Chem. Soc.. 64, 2902 (1942); (b) G. Grundmann, Chem. Be?., 81, 1 (1948); (e) W. G , Overend and L. F. Wiggins, J. Chem. Soc., 239 (1947); C. Pedrali and A. Mantegani, J . O w . Chem., 28, 778 (1958); (d) T. Horie, K. Kinjo, and T.Ueda, Chem. Pharm. Butt,, 10,580 (1962); (e) I. Satoda, F. Kuaada, and K. Mori, Yakugaku Zasshi, 8% 233 (1962); (f) 8. Kukolja, Z. CeniC, and D. Kolbah, Tetrahedron, 19, 1153 (1963). (2) M. Kumagai and M. Bando, Nippon Kagaku Zasshi, 84,995 (1963). (3) M . Likar. P. Schauer, M. Japelj, M. Globokar, M. Oklobdtija, A. Povbe, and V. Sunjib, J . Med. Chem., 18, 159 (1970). (4) (a) E. Schlitter, J. Druey and A , Marxner, Foriechr. Arsneim.-Forsd. 4, 295 (1962); (b) J. Druey and A . Marxner, J . Med. Pharm. Chem., 1, 1 (1959); (c) D.Bargeton and J. Roquet, Arch. Ent. Pharmacodyn., 187,428 (1962). (5) (a) D. Libermann, BdZ. Soc. Chim. F r . , 1430 (1954); (b) Britislr Patent 856,409; Chem. Abstr., 65, 13459 (1961); (c) British Patent 788,502; Chem. Abstr., 66, 11967 (1961); (d) E.Bellasio, F,Parravioini, and E. Testa, Formoco Ed. Sci., 84, 919 (1869).
Journal of Medicinal Chemistry, 1971, Vol. 14, N o . 3 247
NOTES
TABLEI ALKYLIDENE,CYCLOALKYLIDENE, AND ARYLIDENE I)ERIVATIVES
OF
3-HYDRAZINOPYRIDAZINE
R2&NH-N=C==R4 RI "N
No. 1
RI
R2
c1 c1 c1
Ri
R4
Mp, OC
Recrystna aolvent
% yield*
H H CH, CHs CH3
H =C (Ph)CHa 175177 A 49.0 2 H =C(CH&OOCzHa)CHa 122-125 B 55.0 207-208 H =C(CHzCOOCzHs)CHa 3 B 49.8 4 c1 H =C(CHzCH(CHa)z)CHa 104-1 07 A 60.0 142-144 5 c1 H =C(CHs)z C 62.2 6 c1 156-159 D 50.0 H H =C(CHs)i 7 c1 H H =C(CHzCH(CHs)z)CHs 128-129 A 61.6 8 c1 CHs H =C(Ph)CHs 205-207 A 53.6 9 c1 H H =C6HlO 158-160 A 68.0 CHs 10 c1 H =C6HlO 160-162 A 55.7 11 c1 113-116 CHa CHa 1) 65.0 =CsHio 12 C1 CHs =C (Ph)CHa 120-122 CHI A 61 .O 13 c1 CH3 102-104 CHs =C(CHa)* B 56.0 14 c1 H CHs 150- 153 A 48.0 =C6Hio CHs 15 c1 H =C(CzHa)CHa 109-112 E 57.0 16 c1 H H =C (CzHr)CHs 99-103 E 48.0 17 c1 CH3 CHs =C(CzHa)CHa 187-189 A 35.0 a A, 100% EtOII; B, 100% MeOH; C, 67% MeOH; D, 67% EtOH; E, n-hexane. I,Yields are given for products. Compounds were analyzed for C, H, N. 13esults were within 0.47, of calculated values.
have good hypotensive properties, but no mention was made of their antimicrobial activities. Recently we have prepared a number of new alkylidene, cycloalkylidene, and arylidene derivatives of substituted 3-hydrazinopyridazines (Table I) in order to examine their antiviral, antibacterial, and antifungal activities. Results of Microbiological Assays.-New alkylidene, arylidene, and cycloalkylidene derivatives of 3-hydraeinopyridaeine have been tested for their antimicrobial effects using standard techniques as described previously.6 Representative strains of pathogenic and saprophytic bacteria, fungi, and viruses have been used as test organisms and the positive results are presented Table 11.
Formulac
CizHiiClN4 CioHiaClN~Oi CiiHi~ClN40z CiiHitClN~ CsHiiClN4 CiHoClNi CioHiaClN4 C1aHiaClN4 CioH~sClNi CiiHi5ClNt CIZHIIC~N~ CirHirClNi C9HiaCIN4 CiiHisClN4 CsHiaClN4 CsHiiClNi CioHisClNi the recrystallized
Neissevia catarrhalis, Listeria monocytogenes, Pasteurella pseudo f ubei-culosis, Corynebacterium diph t heriae, Bacillus antbacis, Salmonella typhi, Klebsiella, Pseudomoiias aeruginosa, Streptococcus hemolyticus, Candida albicans, Microsporum canis, and Aspeyyillus fumigatus. Compounds 6, 10, 11, 12, 13, 14, 15, and 16 markedly reduced the titer of influenza virus A, grown in allantoic cavities of embryonated eggs. Compounds 1, 2, and 11 were also active against influenza virus A,. Compounds 3 a t 0.0026 AI and 5 a t 0.0054 A4 protected 30% of mice infected with 100 I'DSOdose of Semliki forest virus (arbovirus group A). None of the compounds showed an inhibitory effect on the growth of herpesvirus and poliovirus type 1 grown in human embryonic kidney cell cultures.
TABLE I1 ANT~BACTERIAL AND ANTIFUNGAL ACTIVITY OF THE TEST COMPOUNDSO Microorganism Diplococcus pneumoniue Hemophylua influenrae Streptococcus viridans Salmonella paralyphi B Mycobacterium pholochromogenes M . acotochromopes M . nonphotochromogenenes M . rapid growers Nocardia aateroides Cryptococcus neoformans Truchophyton rubrum T . schoenleini 1'. interdigitale T . menlagrophites a
1
2
+ +
5
6
8
11 12 13 14
+ + + ++ + + ++ + + + + + + + + + + + + + + + + + + + +
+ meane a total inhibition of microbial growth.
Experimental Section' 15 16
+ +
+ + + ++ +
Chemistry.-Syntheses of the compounds listed in Table I were carried out by a general procedure, using the acid-catalyzed reactions of the substituted Bhydraziiiopyridaziries with aliphatic ketones, aliphatic-aromatic ketones, cyclopentanone, and cyclohexanone. Intermediary 3-hydrazino-6-chloropyridazine,~3-hydraxino-4-methyl-6-chloropyridazine, 3-hyd~azinod-methyl-6chloropyridazine,1° and 3-hydraziiio-4,.5-dimethyl-6-chloropyridazine11 were obtained according to the procedures described earlier. Preparation of Compounds l-l5.-The appropriate ketone (0.013 mole), the corresponding 3-hydrazinopyridazine (0.01 mole), HZS04 (0.0005 mole), and aliphatic alcohol (MeOH or EtOH) were heated under refliix for 2 hr, cooled, and filtered. The crude products were purified by recrystallizatioii (bee Table I).
(6) (a) P. Schauer and M. Likar, Pathol. Microbiol., ¶6, 371 (1965); (b) P. Schauer, M . Likar, M. Tiller, A. KrbavPiZ. and A. Polak, ibid., IS, 382 (1965): (c) P. Schauer. M . Likar, M. TiSler, and A. KrbavPiZ, ibid., 19, 506
(7) All melting points were determined on a 13otius hlikrolieistiscli apyarat u s and are uncorrected. AII compounds exhibited the expected ir spectra. (8) (a) W. P. Jencks, J . Amer. Chem. Soc., 61, 475 (1969); (h) G. J . Karabatsos, and R. A. Taller, ibid., 6S, 3268 (1963). (9) Roche Products Inc., British patent 711,756: Chem. Abslr., 49, 11724 (1955). (10) 8.Linholter and R . Rosenpn, Acta Chem. Scund., 16, 2389 (1962). (11) M. Japelj, B. Btanovnik, and M. Tiller, Monulsh. Chem., 100, 671
(1966).
(1969).
None of the compounds Were active against Shigella jlexneri, Staphylococcus aureus, Slreptococcus pyoyenes,
