KOTES
September, 1964 pounds (VI, VII, and X) were also virustatic. Conipounds V and VI were virucidal against vaccinia in tissue culture; TI,X, aiid XIV showed activity against the hepatitis viral strain in &e. These results support the soundness of the assuinptioii on which this research mas planned, that ketoaldehydes derived from steroidal components would be antiviral. Experimental5 All a-ketoaldehydes discussed in this note have been described in the literature. R e prepared them in good yield by oxidation of the corresponding ketols with oxygen in aqueous methanol solution. Chemical and physical characteristics (especially infrared frequencies and rotatory indices) were in accwrd with literature data. Preparation of Schiff's Bases and N,N-Diaceta1s.-A mixture of the ketoaldehyde (1 mmole) and the respective primary amine (1-2 rnmoles) in 10 nil. of ethanol was stirred a t 20" for 24 hr., and the solution was concentrated until crystallization took place. ( 5 ) All melting points are corrected.
The Preparation of Penicilloyl-Polylysines, Skin Test Reagents for the Clinical Evaluation of Penicillin Hypersensitivity BERNARD B. LEVIXE Department of Jledaczne, .I e w York Unave?szty School oJ' Jledzczne, A jew l'ork, .\ e w Yo70 IZeceived d p r i l 8, IUS4
The benzylpenicilloyl (BPO) group has been denionstrated to be the major haptenic antigenic determinant of benzylpenicilliii (PG) hyper~ensitivity.'-~ Multivalent benzylpeiiicilloyl-polylysine (BPO-PLL) conjugates have been found to be effective elicitors of allergic skin reactions of the wheal-and-flare reactions in a significant percentage of patients with histories of allergy to PG.2S3 These materials are accordingly promising skin test reagents for the clinical evaluation of penicillin hypersensitivity. Penicilloyl-PLL conjugates have been prepared previously by reaction of penicillenic acids with p~lylysiiie.'-~ This procedure is tedious and results in conjugates contaniinated with peiiicilleiiic acid groupings and with other inipurities. This paper reports a new aiid simple iiiethod for the preparation of comparatively clean succinoylated iiiultivaleiit penicilloyl-PLL(S) conjugates of diff ereiit extents of conjugation directly froin penicillins. This method is based on the known reaction of penicillins with aliphatic amines a t high p H to form penicill o y l a i n i i i e ~ . ~Parker ~~ aiid Thiel have recently published a inethod of preparation of inaxiiiially coupled, ( 1 ) B.
(2) C. (1962). (3) A.
H. Levine a n d Z. Ovary, J . E z p t l . M r d . , 114, 875 (1961). W.Parker, J . Shapiro, 11. Kern, a n d H. N. Eisen, ibzd.,
116, 821
R. Levine, a n d V. H. Price, Immunology, in press. (1) 13. 13. Levine, J . Ezpi/. . l f e d . . 117, ltil (1963). ( 3 ) "Cliemistry of penicilli^^," El. T.Clarke, J. R. J o h n s o n . and I typical data are giveii iii Table 11. -4llergic skin reactions elicited by the RI'O-I'LI,(S) conjugates could he completely aiid specifically inhibited by thr uiiivaleiit hapten, E-UI'O-aiiiiiiocaproatc. ' confimiiiig the HI'O haptenic specificity of these alleigic reactions I'ully succinoylated peiiicilloyl coii*jugates of poly-1,lysiiie and of poly-I~-lysiiir 11W P fouitd t o bc I t o i i antigenic in guiiiea pigs12 a i d . accordingly, iiiay h c s $
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1 OS 11s IOSI 1011 1211s 126 1.iS J 149 '' a - B P 0 5 ~ - I ' L L ~) ~~~ coniajiis ( a-diitstereciisotiieiic beiizylpeni~ i , , ~ S ) ,from benzylc.illoyl BPO groups. l ) ~ l - f ~ P O ~ ~ - ~ ' I , L prepared penicillenic: wid cviritains :I diasterec,isoriierit, mixture of BP( ) groups. Suhsrripts refer t(i :ir.erage numbers of residues per inole of (>onjugate. Average diameters ( i n i n . ) of wheal :Rare. Ihplicate tests were done: i i i e i t n deviation for wheal, f l .O r i i n i . -1verage diameters (rnrn.) a i d ccrlor intensities of reatticins i n 3 individual guinea pigs sensitized with a given rabbit antipenicilliri serrini; 8 = strong, A W = tliirtler:ite strrrng, SI = rnoderntc,.
iioiiaiitigeiiic 111 iiiait. 1 urther atudics 011 tlw u 4 1 1 1 ness of these coiij ugates a? clinical reageiitb to dctc,ct pcwicilliii hypeiseiisitivity i t i iiiaii are iii p ~ ~ ) g r w b .
