July, 1962
-4NTITCBERCULOUS AQENTS.
11
823
Antituberculous Agents. 11.’ N,N’-Diisopropylethylenediamine and Analogs R. G. SHEPHERD AND R. G. WILKINSON Organic Chemical Research Section, Lederle Laboratories Division, American Cyanamid Company, Pearl River, New York Received .Youember P, 1R61 The preparation and antituberculous activity of sonie forty homologous alkylenediamines [(C,H2,+1NH)2(CH~)m] is given along with a discussion of the structural features required for high activity. N,N’-Diisopropylethylenediamine (I) and its secondary (IV) and tertiary (V) butyl analogs displayed in mice about onehalf thr antituherculous activity of streptomycin.
In the course of general screening of compounds for antituberculous activity in mice, K,N’-diisopropylethylenediamine (I) was foundza to possess a high level of activity comparable to that of streptomycin. I n vitro testingzb demonstrated a selective activity against mycobacteria with no appreciable activity against various Gram-negative and Gram-positive bacterin or against fungi. No activity \vas observed against various bacterial infections in mice. Aliphatic polyamines such as spermine and spermidid or substituted polymethylene-a,w-diamines4 and ethylenediamine$ have been reported to hare antimycobavterial activity but only in vitro. High toxicity but no antituberculous activity mas observed in animals, which me have confirmed.?“ I n the earlier series of diamines, maximum activity was obtained when an unbranched alkyl or alkylene group of 12 to 18 carbons was present, giving a detergent type of structure. Detergency probably accounts for this activity in vitro which is non-specific and is reduced by the presence of protein. The authors5 mncluded that “thr actirr compounds behaved as nonsprcific microbic poisons.” PtLper 1. €2. 0.Wilkinson. R. G . Shepherd, .J. P. Thomas a n d C. Baughn, J . A m (1) Chem. Soc., 88, 2212 (1961). (2a) Personal communication from J. P. Thomas and G. S. Redin of these Laboratories, whom we thank for permission to quote their unpublished data, determined b y the method of hf. Baker, M. Schlosser a n d H. J. White, Ann. N . Y . Acad. Sci., 52, 678 (1949). (2b) Personal communication from M. Hauck and A. C. Dornbush of these Laboratories, whom we thank for permission to quote their unpublished data. (3) J. G . Hirsch, “Ciba Foundation Symposium on Experimental Tuberculosis,” Little Brown & Co., Boston, hlassitohusetts, 1955,p. 117. (4) D. E . Ames and R . E. Bowman, J. Chem. Soc., 1057 (1952). (5) F.A. Barkley. G. W. Mast, G . F. Grail, L. E. Tenenbaum. F. E . Anderson, F. Leonard and I>. M . Green, Antibiotics and CkemotherapU, 6, 554-560 (1956).
Thc slrikingly different structural requirements for w t i i ity i i i thc present series, the very marked antimycobacterial specificity arid thc high activity in vivo demonstrated that the active structures discussed herein are of a new type. Analogs of S,;"\T'-diisopropylethylrnediamine were synthesized with variation of the length niid branchiti& of the alkylene chain and variation i ) f the size, the tmnching and number of K-alkyl substituents. Symmetrically disubstituted diamiim I\ err generally prepared by catalytic reductive alkylation6 with the appropriate ketone or aldehyde, or by condensation of amines with an alkylene dihalidc. Sodium borohydride gave satisfactory reductive alkylation of ethylenediamine with valeraldehyde whereas catalytic reduction with platinum oxide mas u~~successful.Previous mention7 of reductive alkylation nit h sodium borohydride has involvcd only aromatic Schiff bases and the scopv of this reductioii is beiiig csltminrd. Condensation of an alkyl hxlide with ethylenediamine was iiot as generally uscful method as those mentioned above. S-AIethylation of N,S'-dialkylrthylenediamines by the Eschweiler-Clarke variation14 of the Lrrickart reaction proceeded rapidly :tnd in high yield The unsymmetrically substituted cthylciiediamiries~ e made w eithcr hy reductive alkylation of S-substitutcd cthglenediamincs or t)\amination of S-substitiited 2--cahloroethyl:imine hydrochlorides. Antituberculous atc*tii-itiesarc indicated in Tables I and 11. l'hcl testing results on a large number of c>ommcrciullyavailable substituted alkylenediamines arc not incliidcd sinw t hcy Showd no :tppreciahlr nntimycobacterial activity. S,N'-Diisopropylethylenediamine and its S,S'-di-sec-butyl ;ind ditert-butyl analogs were equally active against the otherwise fatal infection with Mycobacterium tuberculosis H37R,. in niicc. Thv median effective dosesZa required for s:urviv:d for at least 60 dzys wcrv 180-200 mg./kg./day when giren orally oilre n day, orally by drrigdiet, subcutaneously or intraperitoncally By the last two rou of administration, the activities w c r ~ahoiit one-half the activity of streptomycin. Thc diisopropyl compound TVW about one-half :I< toxic in mice :LS the di-wc-hiityl and tli-tcrt-htyl analogs. judging bv body weight loss.2a The following generalixatioiis can be made ahout the autitubcrciilous activity of Y,N'-dialkylethylencdiamine~in mice Highest :irti\.it,v mas ohserved when the alkyl groups wcrc isi)propyl (I), L~
t c h s
U S Lmerson O i ~ a n i cReactions, T 01 TV 174-23: (1948) 17) J. H.Billman and 8. C. Diesing, J 0 1 0 Cliein 22, 1008 ( 1 9 i 7 1 , 4 \romp .I 0 1 g ~ h r m 26, 482 ilwn \I))
(,
N \l.ilher
a i d
\I
July, 1962
-&&-TITUBERCULOUS AOEHTS.
11
825
sec-butyl (IV) or tert-butyl (V). Without branching at the a-carbon, there was no appreciable activity as in the methyl, ethyl, n-propyl, n-butyl (11), isobutyl (111) or n-pentyl (VI) compounds. Activity dropped off rapidly when the secondary alkyl series (VII-IX, XIIXVI) was ascended but decreased more slowly in the tertiary alkyl series (V, X, XVII) . There was no activity when the two isopropyl groups were on the same nitrogen as in X,N-diisopropylethylenediamine or when only one such group was present as in N-isopropylethylenediamine. One compound with two different branched N-alkyl groups (XXVII) had high activity while the related compounds (XXVI, XXVIII, XXIX) were inactive. Increasing the number of S-alkyl groups to three or to four lowered the activity, in most cases drastically. Thus, addition to (I) of one X-methyl group (XX) reduced activity by one-half while the presence of two S-methyl groups (XXI) led to inactivity as also occurred with the tertiary nitrogen analog, 1,4-diisopropylpiperazine (XXV) . Diamines with three (XXII) or four (XXIII) isopropyl groups had only one-tenth the activity of I. Although activity mas present in tetraethyl- and tetra-n-pentylethylenediamine (XXXI), the latter more active compound showed toxicity near the minimal effective dose. Lengthening the alkylene chain to three or to four carbons gavc inactive trimethylene (XXXVI, XXXVII) and tetramethylene (XXXVIII) analogs of the highly active I and Y. Substitution of the ethylene chain with one methyl (XXXV) decreased activity threefold whereas substitution on both carbons (XXXIX, XXXXI) led to inactivity. N,N'-Diacetylation of I as in XXTV removed activity. N-Substitution with long alkyl groups (XIX, XXXITI, XXXIV), reported5 to be beneficial for in vitro antibacterial activity, produced no measurable activity in vivo. The very selective activity connected with steric hindrance around the two nitrogen atoms suggests that the activity of these compounds may be associated with ability to form a specific type of metal chelate. lbility of antituberculous drugs to complex with metal ions is well known* but proof is still lacking that chelation is the key to their mechanism of action. Although no evidence in support of a mechanism of action is available for these diamines, chelation was employed as a working hypothesis in guiding the synthesis of analogs. A different type of chelate is formed by some of the active compounds ( 8 ) h'l B Chenoaeth, Phaim. Ress (1961).
8, 57 (19563;
W 0 . Foye, .I Pharm SCI, 60, 93
826
R. G. SHEPHERD A N D K. C;. WILBIKSOS
Vol. 6
TABLE SYSTHESIS, PROPERTIES A S D ;INTIh.fYCOB.~CTERIAT,
Catalyst'' Tirue,
[HZ hlethod'
A
I'
Reactantsh 6 RzCO 1 ED
11'
A
2 RCHO 1 ED
IIIA
.\
2 RCHO 1 ED
IT
'
.A
2
nm
1 ED
premc ;kg./om.*) ] % redn. 3 Pt-C' 93 105%
I"
VI10
\.I11
IS
A
.I
A
135% 1 PtzO
-100 E t O H 21 250 sno Et011
:3
SI
L RCHO
i
1 ED 1 iKaRH4
80 3
2 RL'O 1 ET)
nm
2 1 Ed
2 R2CO
aono 1:tori 18 F't-C' 62 89 %
8
18 Pt-C:
72
4.5 20-65'
1147:
,400EtOII
I 8 Pt-C' 72 86 7
8..i
XIIIQ
A
STI
A
m-ioio
100
so=
A
n
27-723 100
in
H
1 1 7
I'tO.:
90%
~ 1 1 7 ~
29 250
7% .50-1005 10 H ~ O
1 ED
s
nnne I 250 -100 EtoH
n
f'
.50n
3 Pt02 3 40% 1 Pt.O? :i
92 74
V"
hr. Temp. Rnlvent" 1
.?Or 400 E t O H
I::
2 RBr I ED
2 RiCO 1 ED
160 E t O H 20 H?O 94
800
18 Pt-C' 62 90%
500 E t O I l zoo H ~ O 6 32-70' 400 Et013
2 . . i I'tO?
24
:3
%->
July, 1962
ANTITUBERCULOUS AQENTS.
I1
827
I .4CTIVITY O F SUBSTITUTED
ETHYLESEDIAMINES Rel. act.@ in vivo/
B.p.O (mm.)d Yield, or m.p.' % 169-169.5 (760) 60
37
258-2 5Y 115-120 (17)
Purif.,' ml./g.
0 . 3 HnO 3 EtOH
9
-----hnalyees---Calcd. over C H 66.6 14.0 66.7 1 3 . 8 4 4 . 3 10.2 44.6 10.4 69.7 1 4 . 0 ti9.8 14 0
Inhib. Found
conon." in vitro
N C1 19.4 19.6 12.9 32.6 1 2 . 8 32 3 16.3 16.1
1.0 60 ca. 0 . 1 >250i ca. 0 . 1 >250i
94-97 (15)
70
69.7 69 7
14 0 13 9
16.3 16 2
94-96 ( 17)
89
7 9 0 9
14 0
2 EtOH 4 MenCO
69 69 49 48
14 0 10 7 10 7
10 3 15 9 11 4 11 2
28 9 29 0
1.0 125
0 3 Hn0 3 EtOH
48.1 48 4
10.7 IO 7
1 1 . 1 28.9 11 2 28 6
1.0 500
EtOH
52.7 53 1
11.1 11 0
26.0 25.8
250i
90
138-142 (15)
23
180.5-182.5
90
93-95 (1)
70
247-255,6
97
126-127.5 (16)
27
290-291 dec.
70
2 HnO 40 E t O H
53 5 53 3
9 7 9 8
4 MeOH 7 MepCO
55.8 55.6 73.6 73.6
11.4
11 MeOH
55.8
4 HpO 6 EtOH
10 4 10 4
55.9
Y 8 11.6 9 4 14.1 12.3 14.5 12.1 11.4 9.3 Il..i 9 3
55.8
11 4
55 8
11 7
23.5 23 5
250i
9 . 3 23.5 9 2 23 3
,500
250i
828
Ethyienedianiinr,
X\
N ,i S '-Bis(.l-Iieptyl) -
XVIIt
S,N'-Bis(2,1.1-triiiietliyl-Ipentyl 1
XVIII-A'
"
2 RKO 1 El?
N.~'-Bis(rr-n~ethylhrnevl,-
XVIII R
SIX% YX''
XXI
XXII
SXIII
LFtters refer t o pruceduras described u n d u EslJoriiiir,ntaI. " Reactants. given in moltratios, are abbreviated in a manner which, from the structure of the product a n d the synthetir method, should make their structure obvious. E D is ethylenediamine; R'CI, HCl is the alkylor dialkyl-aminoethyi chloride hydrochloride. ' Amount of catalyst (g.) and volume of solvent (nil.) correspond to 1 mole of diamine. P t / C is .l. T. Baker R- Co. 10% Pton-carbon. TI desired t o convert kg./cni.* t o atmospheres multiply b y 0.97. Boiling points (uncorr.) 3 t pressure indicated. Melting points below 270° are corrected. e Yields of bases represent distillate of boiling point indicated; occasionally where hydrochloride salts were recovered from other fractions they are included. Yields of hydrochlorides [include second crops of high purity) Recrystallization from solvent pairs by dissolving in thP are for conversion of base to salt. more polar solvent a n d adding the less polar solvent, using the volumes given per g. of solut?. Except in those cases involving fractional crystallization of isomers, two recrystallizations were sufficient t o a t t a i n a constant m. p. A single solvent denotes recrystallization by heating and cooling. Relative antiinycobacterial activityZa (upper figure) against a lethal infection with Mycobacterium tUberCdOsi8 H37Rv in mice is based on dosages in the drug diet giving significant, (4 days or more) prolongation of survival time, with (I) taken as the standard. Ratios based on preliminary evaluation d a t a are labeled "ca. 0.2.'' Inactivity a t the highest dose tested i R indicated hg < O . l i , r f c . hMinimal conrentration (suhjert t o f a - f o l d \-ariation) in mre./ml.
'
July, 1962
.\NTITUBERCI-LOUS
AQEZITY. I1
829
(continued) B.g.0
Formula CieH36Nz 2HC1
(mm.)d Yield.e Purif.f or m.p.O % ml./g. 126-129(1 5) 62 193 5-194.5
87 1 E t O H 2 MeCO
113-116(0 3)
78
190-195
96
CLEH~PNZ '2HCl
118-120(1 2 ) 234-235 5
41 98
ClaH24NZ (+).2HCl
204-207 (12) 247-248 5
8 51
meno 2HC1
295-295 5
(i)CioHisNz '2HC1
127-135 (12) 257.5-260
CleHseNz ,2HC1
60.5 60.7
11.8 11.9
46
2 EtOH 10 MenCO 25 E t O H
63.3 63.1 63.3 63.7
7.7 7.9 7.7 7.9
70 97
25 MeOH
50.6
3G E t O H
50.6 66.5 66.8
249-253
26
CgHzzNz
174-179 (760)
53
164-168
66
CioHz4Nz '2HCI
228-229 98s
96
CiiHneNz
201-202.5 (760) 81
.2HC1
118.5-120
. --.\naly~es- . Calcd. over Found C H N 74.9 14.2 10.9 74.6 1 4 . 1 1 0 . 8 5 8 . 3 11.6 8 . 5 58.7 11.7 8 . 4 74.9 1 4 . 2 1 0 . 9 75.1 14.3 10.8 5 8 . 3 11.6 8.5 58.6 11.5 8 7
3 MeOH
CisHasNz 2HC1
2HC1 "/rHzO
4 MeOH 4 MezCO
-
100
CIIH~NZ
110-115 (16)
16
'2HC1
210.5-212.5
82
7.9 8.0
--c1
2 1 . 5 ca. 0 . 2 21.1 60
21.5 21.4
< O . li 30
1 9 . 9 ca. 0.5 30 19.6
8 . 3 20.8 8 . 2 20.6 8.2 20.8 8.2 20.8
7.7 11.8 7 . 5 11.8 12.4 6.0 12.4 6 . 2
Rel. act.O i n visa/ Inhib. c0ncn.h in vitw
29.9 29.9 15.1 15.0
1OOOi
2 EtOH 4 AiezCO
55.8 55.6
11.4 11.4
9.3 23.5 9.3 23.1
ca. 0 . 1
>lOOOi
giving 100% inhibitionzb of Mycobacterium smegmatis (ATCC607). Inactivity a t the highest W. R. Boon, J . Chem. SOC.,307 (1947), reconcentration tested is indicated b y >250i, etc. ported b.p. 169-171°, m.p. 250'. i F. B. Zienty, J . An. Chern.. Soc., 68, 1388 (19413, reported 'Lit.' b.p. 210°, m.p. 187'. Lit.' b.p. b.p. l l O - l l l o (8 mm.). It Lit.< b.p. 212-214'. J . A. King and F. H. McMillan, J . Am. Chem. Sot., 68, 1774 196-198°. m.p. 275-280' dec. (1946), reported b.p. 165-175' (90 mm.j and b.p. 149O (26 mm.). G. N. Vyas and 9. G. DhoR. A. Donia, J. A. Schotpate, Current Sci. (India),26, 356-7 (19561, report m.p. 305-310". ton, L. 0. Benta and G. F. P. Smith, J . O r g . Chem., 14, 946 (1949), report b.p. 86-87' (2 mm.). A lower boiling fraction (75-100°, 15 mm.) probably impure monoalkylated product was also J. L. Szabo and W. F. Bruce, U. S. Patent 2,739,981, March isolated in about 30% yield. 27, 1956, reported b.p. 95-97'. ' No reduction of the mixture of ketone and ethylenediamine occurred until the Schiff base was formed by the benzene azeotrope method. a Lit.g b.p. 125127' (2 mm.). Lit.," 92% yield, b.p. 125-127O (2 mm.). " J. L. Szabo and W. F. Bruce, U. S. Patent 2,709 700, May 31, 1955, reported a superior reduction of the Schiff base in glacial acetic acid; howevcr, they reported no yield or properties. Linsker and Evans11 reported 97% yield, m.p. 246-24So by a procedure we have found unsuccessful. to The crude intermediate &(N-methyl-N-iso xopy1amino)ethyl chloride hydrochloride was prepared by the method of J .H. Biel, J . A m . Chem. Soc.. 71, 1308 (1949).
*
It. G . SHEPHERD A N D H. G. WIIXINSOS
830
\.()I. 5
TABLE SYNTHESES, PROPERTIES ASn
XXI\
Name N , N '-Ethylenebis(N-isopropylacetamide)
XXT
1.4-13ii~opropvl1)i1~rrazin~
Methodn Reactantsh E
AXT~XYCOR 1('TERT 41.
Zr d n .
Solvent"
or, I in0 no sol\ .
R
17
100O 8n 1r.o
XXVI"
XXT'II
S-I'ropyl-hT'-i~o~iropylrtli~lenediamine
n
S-sec-Butyl-N'-isoprol,yiethylenediamine
n
2 . 1 n-PrNH?
io 1 1 EtAIeCO 1.5 P t - r 1 RNHCnHdNHz 78 fi4r;
XXVIII*
N-(l,l-Diniethylproyyl)-N'-iaopropylethylenediamine
n
1 3 I-AniNH? 1 R'CI HCI
XXIS"
N-Isopropyl-S ' - 1 h n p l e t liylenediaminr
H
R PhSH 1 R ' r l HCI
N-Isopropyl-N', N ' - d i - n - ] ~ ~ n t v l pthylenediamine
n
SXX"
18 80C 700 E t O I i
I R'CI HCI
H
n
SXXII
:iO-Y0=
?no moil IS 80" 700 E ~ O I I .if) JfgO 18
80C
i n o I :toII
I 2 K?"" 1 R'C1 HCl
&8 80 5 7.50 E t 0 1 1 :in IT?O
1 NaOTT
XXXI
ir.~)
2 iRiNF? CmHdBr*
li
800 ,500 Et011 100 If.0 24 800
i n-Bu2NI-I I R'CI TTCI
ino SXXIII'
N , ~ , N ' - T r i i i i e t I i y l'-nonyl-~ r t hylenediamine
n
1 RAleNH I R'CI wi
I,;t(-)ir
.:I>
sno 700 E t O H 230 H20
SXXIVk
XXXV
S-Dodecgl- IU,N ', N '-t I I nieth>Iet hylenediaminr
B
SI,N2-Diisopropyl-1,2-propane-
A
3 hlezCO 1 R(NHz)z
8 Pt-(' 103 87 %
A
3 hIezCO 1 R(NHz)z
18 Pt-C 103
diamine
XXXt I
N , N'-Diisopropyl-trimethylenediamine
L R.\IeNH 1 R'CI HCI
140" 2000
118s
xylene 2.5 42-71' none 3.25 38-81' none
July, 1962
ANTITUBERCULOUS AQENTS.
I1
83 1
I1 . k C T I V I T T OF Fr.4RIOVTS . 4 L K Y L E N E D I A U I N E F
CioHzzNz '2HCI
B.p.' (mm.jd
Yield,'
or m.p.O
Purif.'
'73
ml./g.
107-109
73
3 MezCO
199-204 (760) 327
39 90
65-85 (12)
40
214-216
89
173-176 (760)
89
219-221.5
83
180-200 (760)
35
325-326
60
149-153 (15)
98
160 5-163
65
128-129 (5)
0 . 5 HnO 14 MeOH
49.4 49.4
10.0 1 1 . 5 2 9 . 2 10 0 1 1 . 3 28.9
2 MeOH 4 MenCO
3EtOH 4 MenCO
44.2 44.3 68.3 67.6 46.9 46.9
10.2 14.3 14.0 13.8 10.6 10.6
12.9 13.0 17.7 18.1 12.2 12.3
1H20 5EtOH
49.0 49.2
10.6 10.2
11.4 28.9 11.5 28.9
4 EtOH
52.6
52.5 74.3 73 8
8 . 0 11.2 2 8 . 2 8.1 1 1 . 5 28 4 14.1 11.6 1 3 . 8 11.7
63.9 64.1
12.2 12.5
17
150-153 (0.3)
87
160-165
85
195-205 (760)
75
204-205
93
135-139 (11)
47
267-267.6
97
128-138 (0.5)
31
263-264 5
73
172-176 (760)
67
169-179
85
191-195 (780)
69
299-302
96
-----Analyses--Calcd. over Found C H N CI 6 3 . 1 10.6 1 2 . 3 63.3 10.6 12.2
6 MenCO
32.6 32.4
30.8 30.6
Rel. activ.O i n mivo/ Inhib. concmh in uitvo 250i
lOOOi
