J O U R N A L OF T H E AMERICAN CHEMICAL SOCIETY VOLUME 70
SEPTEMBER 30, 1948
[CONTRIBUTION FROM
NUMBER 9
THELILLYRESEARCH LABORATORIES]
Biosynthesis of Penicillins. V.l Substituted Phenylacetic Acid Derivatives as Penicillin Precursors BY JOSEPH W. CORSE,~ REUBEN G. JONES, QUENTIN F. SOPER,CALVERT W. WHITEHEAD AND OTTOK. BEHRENS
In the previous papers of this series it has been shown that certain precursor substances when added to the culture media may profoundly influence both the quantity and kind of penicillins formed by penicillin-producing molds.' It has been established that the mold is able to incorporate the phenylacetyl portion of N-phenylacetylDL-valine into benzylpenicillin (Penicillin G 1. With these facts in mind it was of interest to determine whether phenylacetic acid derivatives containing substituents, R, on the benzene nucleus also could be utilized by the mold to form new penicillins (I). That such new penicillins are
compounds were selected on the basis of earlier work with a large number of phenylacetic acid derivatives (penicillin G precursors). These earlier experiments indicate that N-phenylacetyl-mvaline and N-2-hydroxyethylphenylacetamide were among those precursors best utilized by the mold. Also included in this paper is a series of derivatives of dithiophenylacetic acid. These compounds are the amides obtained from the acid and DL-valine, D- and L-penicillamine, DL-isoleucine, P,P-diethoxyalanine and methyl-n-propylamine. No significant stimulation was obtained from any of these products. CHs In the last two columns of Tables I and I1 are I recorded the results of tests carried out in shake flask cultures using P. notatum, strain NRRL 1976. A number of the compounds in Tables I R=+~-NH-CH-CH 0 CI N-CH and I1 were also tested with P.chrysogmum Q176 and in almost all cases the results were similar. /I I 0 COOH The methods of carrying out these tests and the formed, frequently to the exclusion of other peni- interpretation and significance of the results have cillins, has been demonstrated. The preceding been fully discussed elsewhere.6 In general, an paper1 of this series described the isolation and increase in penicillin yield indicates that a new characterization of a number of these new peni- penicillin has been formed. Additional evidence cillins. for the presence or absence of a new penicillin has The purpose of this communicationis to present been obtained in a number of cases by partition of the series of substituted phenylacetic acid deriva- the penicillin mixture in the Craig m a ~ h i n e . ~ tives which have been tested as precursor sub- However, final proof of the existence of a new stances, In Table I are listed the DL-valine de- penicillin rests with its isolation and characterizarivatives, and in Table I1 the N-2-hydroxyethyl tion. amides. The valine and 2-hydroxyethylamine One of the acids of this group, namely, p (1) For the preceding paper of this series see: Behrens, Corse, hydroxyphenylacetic acid, is the precursor of the Edwards, Garrison,Jones, Soper, Van Abeele and Whitehead, J . Bioi. fi-hydroxybenzylpenicillin naturally-occurring Chcm., 176, 793 (1948).
B-r
(2) Present address, University of California at Los Angeles, Los Angeles, California. (3) Behrens, Corse, Jones, Kleiderer, Soper, Van Abeele, Larson, Sylvester, Haines and Carter, J . Biol. Chsm.. 1711,766 (1948).
(4) Behrens, Corse, Jones, Manu, Soper, Van Abeele and Chiang, ibid., 176, 751 (1948). (5) Behrens, Corse, Huff, Jones, Soper and Whitehead, i b i d . . 178, 771 (1948).
2837
2835
J . W. CORSE,R. G. JONES, Q. F. SOPEK, C. W. WHITEHEADAND 0 . K. BEHRENS TABLE I
e z ~
SUBSTITUTED PHENYLACETYL-DL-VALINE DERIVATIVES
Solvent b
RO
1701. 70
I CH(C&)?
Analyses, % ’ N Calcd. Found
M. p.. OC.
Stimulation C
o-Aminod A 238-241 11.19 11.29 1.37 220-227 11.19 11.21 1.0 p-Aminod A 4.10 4.19 144-145 1.0 p-Benzyloxy A 125-127 4.33 4.50 1.0 B p-Carbethox yhydroxy 5.19 4.80 1.0 o-ChloroO 122-124 C 144-145 5.19 5.00 1.33 p-Chloro7 D 1.24 10.76 10.89 A 138-140 +Cyano8 148-150 3.88 3.94 1.0 A p-1odo9 5.04 4.84 114-115 1.0 p-Isopropyl’@ E 1.52 5.27 5.21 129 F p-Methoxyll 10.00 9.77 173-175 1.o o-Nitroi2 C 10.00 10.10 0.88 153-158 A m-Nitroi3 1.49 10.00 10.15 134-135 D p-Nitro“ 1.29 113-115 9.65 9.65 D p-Nitroe 5.04 4.87 1.0 130-132 A 2,4,6-Trimethyli6 Solvents References indicate the procedure in literature which has been followed for the preparation of the acid. used for recrystallization were: A. ethanol-ether-petroleum ether, B. ethanol-ether, C. methanol, D. ethanol, E. etherCompounds were tested a t 0.0008 molar concentration. The petroleum ether, F. methyl acetate-petroleum ether. value recorded represents the ratio: units in test container/units in control container. We acknowledge with thanks the numerous assays performed by Dr. J. M. McGuire. Prepared by catalytic reduction of the corresponding nitro compound with Hdams catalyst. e Isoleucine derivative.
TABLE I1 0
I/
i”LT-(%HYDROXYETHYL)SUBSTITUTED PHENYLACETAMIDES ~ -__ -CHZC---NHCHZCH?OH
R R’L
p-Acetylamiiio’6 p-Allyloxy 4-Amino-3-nitro $-Amino p-t-Amyl p-Anisoylamino 4-Bromo-3-chloro o-Bromoi7 m-Bromol8 p-Bromoi1 Q-t-Butyl o-Chloroe m-ChloroitO p-Chloro’ 3,5-Diacei.omercuri-4-hydroxy 3,5-Dibromo-4-hydroxqP 3,4-Dibromo 2,4-Dichloro” 3,4-Dichloroh p-( 7-Dietliylaminopropoxy) p - ( p ,7-Dihydroxypropoxy) 3,5-Diiodo-4-hydroxy’
M. p., “ C .
145- 146 84-85 132 103-104 d
210-211 104-106 106-107 129-130 105-109 d
99-100 114-117 90-9 1
.....
200-202 125-127 118-119 113-1 14 d d
179-180
I. prakt. Chem., 62, 554 (1900) (7) Straus, Ann., 393, 317 (1912). (8)Jaeger and Robinson, J. Chem. SOC, 745 (1941). (9) Datta and Chatterjee, THIS JOURNAL, 41, 295 (1919). 110) Rossi, Ann. suppi., 1, 139 (1861). i. l l .) Cain. Simonsen and Smith. J. Chcm. Soc., 103. 1036 (1913). (13) Reissert, Bev., 80, 1041 (18971, ibid., 41, 3814; 3925 (1908). (13) Gabriel and Borgmann, Ber., 16, 2064 (1883). (14i Robertson, “Organic Syntheses,” Coll. Vol. I, John Wiley (6) hlehner,
Analyses, % N Calcd. Found
11.81 5.94
11.86 6.28
c
14.42 5.61
14.56 5.58
B
4.79 5.43 5.43 5.43 5.61 6.55 6.55 6.55 1.97’ 3.97 4.16 5.64 5.64 9.08 5.20 3.12
4.95 5.51 5.37 5.40 5.58 6.43 6.54 6.48 2.10 3.66 4.22 5.71 5.79 8.86 5.31 3.03
Stirnulationb
1.0 1.23 1.0 1.14 1.0 1.0 1.71 1.0 2.21 2.90 1.0 1.0 1.134 1.07 1.0 1.0 1.0 1.0 2.10 1.0 1.20 1.0
and Sons, Inc., New York, N. Y.,1941, p. 406. (15) The rnesitylacetic acid used was kindly furnished by nr. R. C. Fuson of the University of Illinois, cf. “Organic Syntheses,” ‘26, 65 (1945). (16) Ferber and Bendix, Bey., 72, 839 (1939).
(17) Shuttleworth, Rapson and Stewart, J . Chem. SOL,73 (1944). (18) Berger, J . prokl. Chem., 133, 331 (1932). (19) Wislicenus and Grtitener. Bcr.. UI. 1933 (1909). 1.20) Kenner and Morton, J. Chcm..Soc., 679 (1934)
PHENYLACETIC ACIDDERIVATIVES AS PENICILLIN PRECURSORS
Sept., 1948
2839
TABLE I1 (Cotttinued) Ra
M. p., "C.
Formula
Analyses, % ' N Calcd. Found
Stimulationb
2,3-DimethoxyZl 93 5.83 1.0 5.85 3,4-Dimethoxy21 96-98 5.81 1.0 5.85 1.27 3,4-Dimethyl 99-100 7.03 6.75 1.26 90-91 6.28 P-Ethoxy" 6.27 1.23 103-105 7.09 o-Fluoro 7.10 75-77 7.10 1.93 m-Fluoro 7.10 1.54 75 7.03 p-Fluoro 7.10 d 1.24 7.39 o-Hydroxyl .7.17 92-93 1.13 7.58 m-Hydroxyk 7.17 7.18 110-11'2 1.0 7.17 $-HydroxyaS 7.19 84-86 1.0 p-Hydroxy' 7.33 m 157-158 1.0 p-(N-2-Hydroxyethylcarbamyl)24 1.0 180-181.5 14.43 4-Hydroxy-3-phenylazo 14.04" 1.75 127-12!3 4.93 4.59 m-Iodo 1.83 112-113 4.32 p-10d09 4.59O d 6.20 1.0 5-Isopropyl-2-rnethyl" 5.95 d 1.33 6.18 p-I~opropyl~o*~~ 6.33 d 1.0 6.06 6.69 o-Methoxp 59 6.68 1.o 6.69 m-MethoxyZ6 6.64 1.22 86-88 fi-MethoxyQsr 6.69 6.42 1.o 99-100 6.28 3,4-Methylenedioxp 1.49 115-117 6.30 6.30 p-Methylmercapto 10.81 1.0 69 4-Methoxy-3-nitro 11.02 1.36 63-64 7.16 7.24 o-MethylP? d 1.39 7.21 7.24 m-Methyls 1.69 76-78 7.28 7.24 p-Methylzg 1.0 140-142 12.54 12.50 p-Nitr~l~
