Synthesis of potentially Cy-toac ti, e Amino A4iciclAmide Mus tarcis“b
Methods have beeii of ;imino aoid inoiio- anti 1~is~S-c~liloroethyl)amide::.These have been applied to hloroethv1)amide~of ur.-alanine, or.-pheiiy~alaiiiiie, and IILleucine, and t o the 2 et,hylarnides of glycine, ~.-alaiiine,ur,-alanine, ~~-pheiiylalanine, mrleucine, asparagine, and 01- arid utaniic acids. 111 most case‘: the products t o which compounds of this type readily rearrange iii witer, the c,orresporidiiig amirioeth.lcarbosylic esters. have heen iuolated and characterized. Wheii Iesi ed agaiiiat inells in tissue rulture, the “oiie-armed” Inu,tard ivatives were uniformly rytotoxic., whereits t Ire corresponding “two-arnied” derivat ivei were either in:ictivt= i howvrii t)orderline 1-yiotoxicity.
__
.
.-.
. .
(1) (a) Supporteil iiy a research grant, (C:I 02130) f r o m the Sational Cancer Institute, Kational Institutes of Ilealth, U. 3 . Public IIealtli Servire. (b) To whom inquiries should bt. addressed. ( 2 ) J. Ilo00 pg./ml. for the corresponding bis(2-chloroethy1)aiiiides of Lleucine, glycine, DL-alanine, and DL-phenylalanine. These results indicate a rather striking difference in behavior between the “one-armed” mustard derivatives which are uniformly cytotoxic aiid the “two-armed” derivatives which show only borderline cytotoxicity or are inactive in this system. The reason for this difference is riot too clearly understood and may simply reflect the difference in the rates of rearrangement to the corresponding esters that the two classes of conipourids undergo; or it may be related to the fact that the “twoarmed” mustards, lacking an aniide hydrogen, are less susceptible to enzymatic hydrolysis that would be required for “activation.” Of particular interest, however, is the fact that the “one-arnied” conipounds are substantially more cytotoxic than the parent mustard, 2-chloroethylamine, from which they are derived. Possibly used as ail amino acid amide, this simple alkylating agent reaches some more critical, intracellular kite where it is released, than it would administered in its native form. Experimental Section The compounds just described were prepared generally by procedures closely aiialogous to those in the specific cases reported in detail below. N-Carbobenzoxy-~~-phenyIalan~ne Cyanomethyl Ester.-Ncarbobenzoxy-DL-phenylalanine (1.5 g.) wa.5 treated with 1.05 ml. of triethylamine, and 0.95 nil. of chloroacetonitrile was added to the viscous mix, with a rise of temperature. The reaction mixture was left overnight at room temperature, and the excess chloroacetoiiitrile was removed under reduced pressure. The residue was taken up in ethyl acetate, arid the solution was washed with dilute HCI, then with saturated iXaHC03, dried (Na?S04), aiid evaporated. The white residue (2.0 9.) was crystallized from CCI,; yield 1.5 g., m.p. 96-97.5’. N-Carbobenzoxy-DL-phenylalanine 2-Hydroxyethylamide (Method A).-To a solution of 1.0 g. of S-carbobenzoxy-DLphenylalanine cyanomethyl ester in 7 ml. of ethyl acetate 0.17 ml. of 2-hydroxyethylamine was added, and the mixture was stirred overnight at room temperature. The solvent was removed under reduced prehsure aiid the re-idue was diaholved in CBC1,. The CHC1, solution waa washed with a small volume of cold, saturated S a c 1 coiitaiiiirig a few drops of dilute IICI, then with a small volume of cold, saturated NaHC03, dried (SazSOa), filtered, and evaporated. The residue (1.1 g.) was crystallized from benzene; m.p. 116-117.5” (sintering). ~~
(8) These studies have been carried orit under the direction of Dr. George E. Foley. We are grateful to Dr. Sidney Faiber, Director, Children’s Cancer Hesearch Foundation Inc., Boston, for permission t o refer to these preliminary results.
\
86
I
01.
!I
87 TABLEI11 C!Y.INOMETHYL ESTEILS OF N-CARBOBENZOXYAYINO L4~Ius NHCOOCH2C6Hs
I
R-CHCOOCH2CN [aIzQD, deg. ( c onc n. in acetone)
Tield,
c/o i4 84
bI.p., 'C.
CH2C6H6(DL-Phe) CH,CH( CH3)2 (DL-Leu) (~--.,-Bz-Glu) CH2CH2C02CH2C6H,
96 70
96-97
...
87-89
...
68
88-89
-20.5 (3.52)
CH2CH2CONH2( L - G ~ u - N H ~ ) CH2CONH2( DL-ASP-NH~)
63 30
133-134 108-110
70
9'3-101
R (amino acid)
CHI (DL-Ala) CH3 (L-Ala)
...
50-51 40-42
-35.6
%--
----Calcd., C H
Formula
C13H14N,0a C13H14Xj204
%--
---Found, C
N
59.51 5.38 10.69 59.4 59.51 5.38 10.69 59.6
H
K
5.4 5.4
10.9 10.8
(2.98)
CII,c€r,cI12-K=c(NIICOOC1
, , , ,
..
C16H17N3Oj
8.3 9.39 6.8
8.28 67.4 5 . 3 9.21 63.31 6.68 6.83 64.2 5 . 4
Ci~H18x204 67.44 5.36 C16H2&204 63.14 6.62 C22H22N2O6 64.38 5 . 4 0
56.42 5.37 13.16 56.51 5.46 12.96
CI4Hl6K3On 55.08 4 . 9 5 13.57 54.91 5.10 13.66
I,CJ 1~1,
(L-Ard
, , ,
Csnl1331;60, 62.4:; 5 . 4 0 11.38 62.33 5 . 5 1 11.27
TABLE IT~ - I ~ Y L ) ~ ~ U X Y.\NU E P HBIS( Y L~-- H Y D ~ ~ O . Y TYHEY L ) . I V I D E ~ OF NHCOOCH&e,H,
S - C . ~ ~ ~ B ~ B E ~ ZACIDY OXY.~MINO
I
R-CHCOX deg. (concn. in AIeOH)
[..ID,
Yield.
1%(Gly) H(G~Y) CH3 (DL-Ala) CH, (L-Ala)
oc.
X
%
N.p.,
N ( CHzCHZOH),
58 82 64 80
82-84 114-1 15. 113-115 113-114
HO
116-117 120-1 22
R (amino acid)
NHCHzCHgOH NHCHgCHZOH NHCHZCH2OH
...
Formula
--Calcd., C
CllHzON20B 56.7
-.-Found, %-N C H
%H
N
6.8
9.5
56.6
6.8
9.6
...
... - 13.5
CI3H18N204 CI3H&zO,
58 63 6 . 8 1 10.52 58,63 6.81 10.52
58.5 6 . 8 10.6 58.4 6 . 6 10.7
ClgH22S2Od C&i26Nd&
66.65 6.48 63.i.5 6.32
67.0 63.9
(2.29) CH2Cf,Hj (DL-Phe) NHCHzCHzOH C H X H ~ C O ~ C H Z C ~ H KHCH,CH,OIX , ( -.,-Bz-~-Glii)
82
...
-4.5 (2.88)
8.18 6.i6
6.6 6.3
8.0 6.8
R--CHCOX
56 45 65 57
115-117 115-117 1C2-104 102-104a 124-126 125-127 125-127 119-121 130-1 31
Calcd., %-----Found, Formula C H C I N C H C CnHlaCINzOs 53.23 5 . 5 9 13.10 10.35 53.0 5 . 6 C12Hi&INz03 53.23 5.59 1 3 . 1 0 1 0 . 3 5 5 3 . 3 5 . 5 Ci3HiiClNzOa 54.83 6 . 0 2 12.46 9 . 8 4 54.9 6 . 2 CiaHi.CINz03 55.1 6 . 1 C ~ ~ H I ? C ~ N 54 ~ O83~ ~6 . 0 2 1 2 . 4 6 9 . 8 4 62.4 5 . 3 Ci8HnClNzOa 63.24 5 . 8 7 9 . 8 3 7.77 Ci9HnClNz03 63.56.0 6 3 . 2 4 5 . 8 i 9.83 7.77 59.0 7 . 2 CisHzaClNzOa 58.80 7.0Y 1 0 . 8 5 8 . 5 7 61.2 6 . 0 8.19 6.47 61.02 5 . 8 2 CizHzrC1N 208
1 2 . 3 10.1 10.0 7.9 9.6 7.8 10.9 8.6 8.1 6.7
H
29
180-182
CiaHzoClXaOa
6.1
10.4
12.2
C
58
182-184d
CisHzaClSsOa
C D
50 18
192-194 192-1'd3e
51.3 5 . 5
10.7
1P.i
B
68
5.8
5.5
11 0
B A
71
B
68 75
Yield, No. 1 2 3 4 5 6 7 8 9 10 11 12
13 14
K (amino acid) H (G~Y)
X A I etliod XHCHKHzCl .1 C NHCHzCHzCl '4 NHCHzCHzCI B NHCHzCHzC1 K H C HzCHzC1 h . I NHCHzCHzCI B NHCHzCHzCI B NHCHzCHzCI h NHCHzCHzCI
H (Gly) CHa (DL-Ala) CHs (DL-Ala) CHa (L-Ala) CHzCeHa (DL-Phe) C H ~ C E (DL-Phe) H~ C HICH (C Ha)z (DL-Leu) CHzCHzCOzCHzCeHa [L-~-BZ-GIU) CHzCHzCONHz NHCHzCHzCI (L-G~u-NHz) CHICH~CONHZ NHCHzCHiCl (L-GIu-NHz) CHzCONHz ( D L - . ~ s ~ - N H ??;HCHzCH&l ) C HzCON H? NHCHzCHCl ( DL-ASP-NHz) CHzCHzCHzNHCHzCHKl
I
N=C(NHCOzCHzCaHb)z (L-Arg) H (Gly) 16 H (Gly) 17 CHa (DL-Ala) 18 CHZCEHE (DL-Phe) 19 CHzCsHa (DL-Phe) 20 CHiCH (CHs)a (DL-Leu) 21 CHzCH2SCHa (DL-hICt) 15
N(CHzCHzCI)? N(CHzCHzC1)z N(CHzCHzC1)z N(CHzCHzC1)z N(CHzCHzCl)% N(CHzCHzC1)z N(CH~CH?CI)?
B
55 IT
56 65 84
4 B
n
.1I.p., "C.
5%
68 54
7--
5.90
10.37
CiaHisClNsO~ CnHiaC1NsOi
51.30
5.54
10.81 12 82
165-166
Ca?Ha6ClNsO.
60 23
5 69
5.56
10 9T
87-89 87-89' 102-104 127-129 126-128n 101-103 99-101
Ci~HisClzNzOa CiaH~aCkNzOa CisHzaCIzNzOa C~iHzaCIzNzOa CnHzaClzNzOa CiaHz~ClzNzOs Ci:HzaCI?Nz035
50.47
5.45
21.28
8.41
50.4 5.4
21.1
8.4
51.87 5 . 7 6 20.46 59.58 5.72 16.75
8.07 6.62
51.9 60.1
20.5 16.0
7.9 6.5
7.20 6 88
5 5 . 6 6 . 8 18.3 50.3 5 . 8 17.3
( 5 , 7.87)
52.9
I N 13.1 10.3 13.0 10.3 12.5 9.9
92.iI
55.55 6.73 18.22 50.12 5 . 9 4 1 7 . 3 9
12.29
%--
60.2
5.9 5.9
7.3 7.1 (S, 7.7)
Undepressed when admixed with 3. [ C L ] ~ ~-35.6" U (c 2.98, acetone). e [ . i j z O ~-17.8" ( c 1.5'2>methanol). Uiidepressed when admixed with 10. e Uiidepressed when admixed with 12. Uiidepressed when admixed with 15. u Uiidepressed wheii admixed with 18.
'
