July 1969
70 1
NOTES
2-Benzyl-5-(N,N-dimethylamino)y-valerolactone, an Inhibitor of Plasma Cholinesterasela SHCRREL C. SMITH'^
AND
FLOYD W. D U N N ' ~
Department of Biochemistry, Cniversity of Tennessee College of Basic Medical Sciences, Memphis, Tennessee 38103 Received February 20, 1969
2-Benzyl-5-amino-~-valerolactone (I) was prepared as a possible intermediate for the synthesis of 2-benzyl5-aminolevulinic acid.2 It was recognized that the N,N-dimethyl derivative of I might possess acetylcholine-like a ~ t i v i t y ; ~ therefore, 2-benzyl-5-(IYJN-dimethylamino)-y-valerolactone (11) was prepared and tested against isolated human plasma pseudocholinesterase system. I1 was found to have an 150 of 2.14 X in a previously described assay p r o c e d ~ r e . ~ CH,C,H,
R , N C H 2 ~ 0
I,R=H 11, R = CH3 Experimental Sections
2-Benzyl-2~carboxy-5-bromo--pvalerolactone.-A suspension of 34.3 g (0.193 mole) of N-bromosuccinimide in 200 ml of H20 was added to a solution of 45 g (0.192 mole) of allylbenzylmalonic acid6 in 200 ml of EtzO. After being stirred for 12 hr the Et20 layer was removed and the HnOlayer was extracted twice with 25 ml of EtzO. The combined ethereal extract was concentrated to a heavy oil; this was dissolved in 75 ml of EtpO, and petroleum ether (bp 37-60') was slowly added until precipitation was complete. The crystals were removed by filtration and recrystallized twice from EtpO-petroleum ether (bp 37-60'); yield 27.3 g. Concentration of the mother liquor produced additional product which weighed 12.5 g after three recrystallizations. The total yield was 40.8 g (67.87,), mp 135" dec. Anal. (C13HlaBr0,)C, H. 2-BenzyI-5-phthalimido-~~-valerolactone.-2-Benzyl-~carboxy5-bromo-~-valerolactone (20 g, 0.064 mole) was heated at 130" for 48 hr t o effect decarboxylation. The resulting liquid showed ir absorption bands as expected. It was used in a modified Gabriel synthesis,? conducted a t 90" for 24 hr. Isolation by the reported procedure and crystallization from C&-petroleum ether yielded 14.7 g (69%) of product, mp 130-135". Five recrystallizations gave a product with mp 135137". Anal. (CdIleNO,) H, X ; C: calcd, 71.85; found, 71.42. 2-BenzyI-5-amino-~-valerolactone Hydrochloride (1).-*4 3.65-g was added to sample of 2-benzyl-5-phthalimido-~-valerolactone 200 ml of 6 V ; HC1. The mixture was alternately refluxed and (1) (a) This investigation x a s supported b y Public Health Service Grant N o . AI03710 from t h e Xational Institutes of Health and Biochemistry
Training Grant GM-203. (b) Taken in part from t h e M.S. Thesis of Sherrel C. Smith, University of Tennessee, J u n e 1965. (c) T o whom requests for reprints should be directed: Department of Chemistry, hbilene Christian College, dbilene, Texas 79601. (2) S.S.Smith and F. W. D u n n , J. Med. C h e m . , 12, 699 (1969). (3) -1.Enders and U. Schmidt, Arzneimittel-Forsch., 4 , 331 (1954). (4) (a) .A. Lasslo, P. D. \Taller, A. L. Meyer, and B. V. R a m a Sastry, J . .Wed. P h a r m . Chem., 2, 617 (1960); (b) A . Lasslo, J. G. Beasley. G. G. Nelms. and G. J. Epperson, ibid.,6, 811 (1963). ( 5 ) -411 melting points were determined on a Fisher-Johns melting point apparatus and are uncorrected. T h e elemental analyses were performed by Galbraith Laboratories, Knoxville, Tenn.. a n d &!I-H-W Laboratories, Garden City, hlich. Where analyses are indicated only b y symbols of t h e elements. analytical results obtained for those elements were within 10.40/, of t h e theoretical values. T h e cholinesterase inhibition study was kindly performed b y Dr. James G. Heaslry, Department of Medicinal Chemisiry, University of Tennessee, Alemphis. ( 6 ) R . T. Arnold, X D e M . Campos, a n d K. L. Lindsay, J . A m . Chem. Soc., 75, I044 (1953). (7) J. C. Sheehan a n d W. A. Bolhofer, i b i d . , '72, 2786 (1950).
cooled to room temperature four times. H,O and HCl were removed in a rotary evaporator. Absolute EtOH was repeatedly added in 20-ml portions and evaporated to remove excess HC1. The residue was taken up in absolute EtOH, and Eta0 was added until precipitation was complete; yield 2.2 g (83.3%), mp 196199" dec. Compound I gave a purple color with ninhydrin. Rf values were, respectively, 0.38 in BuOH saturated with Hp0, 0.77 in MeOH-KH3 (19: l), and 0.89 in pyridine-HpO (65:35). Anal. (CipHi6ClNOp) C, H, N. 2-Benzyl-5-(N,N-dimethylamino)-r-valerolactoneHydrochloride (II).-A solution of 2.0 g of I, 9.0 ml of 1 S NaOH, 5 ml of HpO,2 ml of 88% formic acid, and 2 ml of 407, CHZOwas heated for 6 hr a t 100". The mixture was concentrated t o a heavy oil under reduced pressure. Absolute EtOH in 20411 portions was added and was then evaporated four times. The oil was dissolved in 20 ml of anhydrous EtOH, and the S a c 1 was filtered off. Concentrated HC1 (40 ml) x a s added t o the filtrat'e and the solution was then concentrated to an oil. Absolute EtOH was added and evaporated four times. The dry crystals were thoroughly triturated with anhydrous Et20 and filtered; yield 1.9 g (8,5.0%), mp li5-1iio. Anal. (Cl,H20C1S0,) C, H, N.
N4,N4'-Decamethylenebis-4-aminopyridine and N g,Ng'-Decamethylenebis-9-aminoacridine JOHK
P. PBOLIKI, LOUISJ. LEXDVAY, A N D FRANK P. PALOPOLI The Sational Drug Company, Research Laboratories, Division of Richardson-Merrell Inc., Philadelphia, Pennsylvania 19144 Received December 31, 1968
The title compounds (see Table I) are essentially the "ring removal" and "ring addition" compounds of S4,~4'-decamethylenebis-4-an~inoquinaldine (l), a known antimicrobial agent, The bisdecamethylene derivatives of 2- and 3-aminopyridine were also prepared. The compounds were tested in vitro according to methods previously described.2 Table I1 contains the screening data for the amines, compound 1, and 9aminoacridine (9), a compound whose antimicrobial activity is well d ~ c u m e n t e d . ~ Experimental Section
Ng,Ng'-Decamethylenebis-9-aminoacridine (2) was prepared from 9-chloroacridine (25 g, 0.1 1 mol) and 1,lO-diaminodecane (8.6 g, 0.05 mol) according to the method of Strauss and Rosenstock.' N,N'-Bis(pyridy1)sebacyIamides (6-8).--Sebacyl chloride (30 g, 0.125 mol) was added dropwise t o a stirred mixture of the appropriate aminopyridine (23.5 g, 0.25 mol) and Et3X (25 ml) in T H F (250 ml). After about 10 ml of the chloride had been added, the mixt,ure began to boil, and addition was continued at such a rate that, reflux was maintained. ilfter the addition was complete (about 10 min) stirring was continued for an additional 1.7 min. The solvent was then removed by evaporation in oacuo. The residue was tritnrat,ed m-it,h500 ml of 10% &C03 solution, filtered, washed with HzO, and purified. N,N'-Bis(pyridy1)decamethyleneamines (3-5).--..1 slurry of the appropriate diamide 6-8 (12.3 g, 0.033 mol) in warm T H F (200 ml) was added, with stirring, to LAH (2.3 g, 0.06 mol) in T H F (30 ml). The resulting mixture was heated under reflux for 1.5 hr. The reaction mixture was cooled in an ice bath and stirred during the successive dropp-ise additions of H20 ( 3 ml), 15ccNaOH solution ( 3 ml), and H?O (20 ml). The mixtiire was
r.
(1) R. R. Straiiss and P. n. Rosenstock. S. Patent 8,362,875 (1968). (2) R . Strauss, A. McBurney, S . Romisher, and J. 31. Ueiler, Aritimicrobiirl Agents Chemotherapy, 5% (1963). ( 3 ) -1.Albert, S.D. Rubbo, and 31. I. Burvill. Brit. J . Ezptl. Patlrol.. 30, 159 (1949).
E v Iter icii iri SO.
coli
I !I
1"; 25
2
s
li
?;
I
x a
1'1 =
s 100 partial inhibition, S
= iio
artivity a t 500 pg 'nil.
filtered :ifid worked iip iii the iisiial niaiiiier. The resilltirig coiiip ~ ~ i i t i(free ~ l s base or 2HCl salt) were purified by crystalhatioil.
Acknowledgment.--The authors \\-ish t o th:mI; 1 h . Robert' Strauss, Miss Ariiie JIcBurney, and A h . Beverly Slowicki for the microbial assays.
3 l - Sulfato-N2-isonicotinylhydrazine.
A Potential Metabolite of Isoniazid'
During studies on the metabolism of the antituberculous isoniazid (INH), in animals3 and we considered the possibility that, one of the group of acidlabile urinary metabolites was an ^\'-sulfate conjugate of I K H . This was suggested by earlier in v i u 0 ~ 7and ~ i.n vitro7 studies, using various laboratory animals, in which arylamines such as aniline and 1- and 2-naphthylamine were conjugated with sulfate to form highly acidlabile sulfamates. Since 1K.H is acetylated by thc same pigeon liver enzyme that acetylates arylamines,b :L similar parallel activity seemed possible for sulfate conjugation. If highly acid labile, an INH-sulfate (1) Supported in part b y U. S. I'ulilic
H e a l t h Service Grant .21-03682. ( 2 ) Komedical Research, Life Sciences Division, Btanford Researall Institiire, XIenlo P a r k , Calif. 94026. (:O , I . I I . Peters and V. E. lIayes, A r , , l t , l ! t l e r , i . l ' t i i i r m o c o d y n , , 169, 340 lIU6li). (1) (a) .I. H. l'eters, li. Y. Xiller, anLl 1'. 1 3 r o \ ~ nA , i d . B i o c h e n . , 12, 37il (1$165): ( b ) .I. Phurmarol. L'zptl. T h e r a p . , 160, 298 (1965). (5) 1'. 13o,vland, r). hlanson, and S. F. I). Orr, Bioriiem. J . , 66, 117 ) I). \'. I'arke. i 6 i d . , 7 7 , 4!i3 l I 9 6 0 ) . ) ( a ) .\. 13. Roy. Biorhim. ~ ' 3 i o p h y . q ..Ictii, 30, 1!13 (1'358): i l l ) B ./., 74, 49 (1960). ( 8 ) .J. \ \ . .lenne ani1 1'. 1 ) . 13oycr, Wiochiin. B?opl,U,>..
coiljugate would contribute t o the incompletrly itlciit Ified acid-labile fraction of urinary I S H nietabolitr.;. Therefore, \\e synthesized the I S H analog of :iryl sulfamates, Si-sulfato-S2-i~o~ii~otiriylhydr:tzir~e, l)y procedures employed previou4y.i It was found to t)e atable to the mild acidic conditiolis known to hydrolyzr the pyruvic and a-lietoglut :uic acid hydrazone.: of I S H 3 and the aryl sulfaniatehj (pH 1.0, room Icmper:iture). However, it could be bplit quantitatively t o I N H by heating at 43" in 1 N HCl for 24 hr, conditioiiy developed for the hydrolysih of :icetylisoniazid t o ISH \pithout the decomposition of ISH.4a When thii liytlrolysis procedure wa5 applicd t o urine collectrd froni dogs receiving I S H in itudies reported previously, no increase of I N H content could be detected. Tliew results indicated that the sulfate conjugate n-:is not :I metabolite of I X H ill the (log. Thereforr W P tc>stctl the vi~oit:ihility of t h c iulia1 e compound iii h u h r quent experiment-. For preliminary te on tho toxicity of thii c.oi11single doses of 43.1 m g 1.g pound, we ndmini.t iv (equivaleiit to 25 mg of ISH'lig) to six 26-day-old Ho1tzm:tn female r:& Similar groups of untrc:ittd id n i t * receiving 2%5nip of I S H 'kg served :I- COIISo iinto\wrd rcxction- were noted in t lie rai iiig either I S H O r thc ISH-sulfate con,iug:itc. The ruts J v c w weighed inimdintely before irijrctioii :tiid daily for t h e next 9 days. Necropsiea \ w r r p ~ r formed aftcr killing the aiiimuls with CHCld, \v1th bpecial atteiition being given t o the liver, kidney,, :mI \pleen, which were' cxamirid internally a i w ~ l :is l Yternally. So gros;.; chaiigc~from the untreutrd r:itw r e iiotcd ill tlw t rcJiitctl groups. In utlditioii, t i o toxic effects of the conipouiitl> \yere noted in the growth uf the animal-, :t.z cvideiicecl by nearly identical weight gains in the trc:itr(l :itid i u i t reated groups cluriiig lh(. btudy period T o determine t h e 211 i > i wyt:ibility of the IXH--uhat(~ cwijugate, iiijcci et1 four yourig adult female b(a:Igltu clogs (weight riiiige. 5 . 2 cJ.7 kg) with doseh of 9 . ( j
-
(1
