Eur J Med Chem (1994) 29,33 l-338 331

0 Elsevier, Paris

Synthesis and biological activity of 17-azasteroidal

neuromuscular-blocking agents

AK Vermal, CY Lee2, S Habtemariamz, AL Harveyz, DP Jindall*

IDepartment of Pharmaceutical Sciences, Panjab University, Chandigarh, 160 014, India;
2Department of Physiology and Pharmacology and Strathclyde Institute for Drug Research,
Universiry of Strathclyde, Glasgow Gl IXW, UK
(Received 10 September 1993; accepted 17 January 1994)

Summary - The new analogues, containing nitrogen at the 17-position instead of the 17a-position, of chandonium iodide 2 have
been prepared and evaluated for potential neuromuscular-blocking activity in in vitro studies using chick-biventer cervicis prepara-
tions. There is a slight increase in the interonium distance between the 2 quatematy heads of 13(DPJ-240). All the bisquatemary
compounds 13 (DPJ-240), 14 (DPJ-252), 15 (DPJ-262), 16 (DPJ-260) and 17 (DPJ-261) showed potent neuromuscular-blocking
activity but slightly less than chandonium iodide 2. Monoquaternary ammonium derivatives 12 (DPJ-246), 21 (DPJ-245), 23 (DPJ-
246) and 24 (DPJ-243) of 17-azasteroids were also synthesized and pharmacologically testled. They exhibited weak neuromuscular-
blocking activity as compared to the bisquatemary compounds and chandonium iodide 2.

bisquaternary 17-azasteroid / monoquaternary 17-azasteroid / neuromuscular-blocking activity / acetylcholine / carbachol /

chick biventer

Introduction between the 2 is more or less fixed. A large number

of azasteroidal neuromuscular-blocking agents with a
Neuromuscular-blocking agents cause interruption short duration of action and lesser side effects have
of transmission of nerve impulses at the skeletal been reported. Reviews on rational elements in the
neuromuscular junction. These are used clinically in development of superior neuromuscular-blocking
anaesthesiology to provide skeletal muscle relaxation agents and the aspects of the pharmacology of
for surgical procedures, as an adjunct to electrocon- aminosteroids including an account of azasteroids
vulsive therapy, to prevent shock-induced dislocations have been published [ 3-61.
and fractures, and to control muscle spasm in tetanus Pancuronium bromide 1 is in widespread clinical use
[l, 21. due to its non-depolarizing mechanism, medium dur-
The only short-acting neuromuscular-blocking drug ation of action, high potency and minimal side effects
in clinical use at present is suxamethonium, which
[7-91. Likewise, chandonium iodide 2 [ 10-131 17-
possesses various undesirable side effects due to
its persistent depolarizing mechanism of action.
Therefore, a number of research laboratories started a
programme of development of short-acting muscle
relaxants with non-depolarizing mechanisms of
action. In azasteroidal neuromuscular-blocking agents,
the steroid nucleus provides a supporting moiety for
the 2 quaternary heads, and the spatial relationship

*Correspondence and reprints; this author previously wrote Paul D

Jindal. 1
332
methyl-3P-pyrrolidino-17a-aza-D-homo-5-androstene
dimethiodide exhibited a potent non-depolarizing
neuromuscular-blocking action of relatively short
duration and rapid onset in anaesthetized cat, being
4-5-fold more active than (+)-tubocurarine and only
slightly less active than pancuronium bromide 1.
Chandonium iodide 2 is undergoing clinical trials and
is being developed as a drug. Several modifications of
chandonium iodide have been reported [14-171.
It is widely held that the optimal interonium distance
between 2 quaternary heads for good neuromuscular-
blocking activity is 1.0-1.2 nm. As the interonium
distance falls below 1.0 nm, the neuromuscular-block-
ing activity diminishes with the result that ganglion-
blocking activity becomes prominent [ 181. In chando-
nium iodide 2, the nitrogen is located at the 17a-
position and in a conformational perspective it
2 compound 9. The vibrational stretches at 17 10 and
showed an interonium distance of 1.029 nm [19],
while in pancuronium 1 this value is 1.108 nm [20].
A number of new neuromuscular blocking agents, eg,
pipecuronium [21, 221, atracurium [23, 241 and miva-
curium [25], do not fall within these parameters. The
reason for these exceptions is uncertain.
We attempted to develop azasteroidal quaternary
ammonium compounds in which the interonium
distance is somewhat extended, as compared to chan-
donium iodide, to optimize the neuromuscular-block-
ing activity. In this paper we report the synthesis and
biological evaluation of bis- and monoquatemary
17-azasteroidal compounds. Shifting the nitrogen
atom from the 17a-position to the 17-position results
in a slight increase in the interonium distance
(1.10 nm) in compound 13 (DPJ-240) as measured by
Dreiding Models. This has one of the quaternary func-
Me Me
13
tions (3P-equatorial) well exposed and a double bond
at the 5,6-position, which will affect the geometry of
ring B as well as that of ring A.
Chemistry
17-Oxo-5-androsten-3P-ol 3 was subjected to oxima-
tion using the potassium tert-butoxidelisoamyl nitrite
system to give the 16-oximino derivative 4 [26, 271.
The oxime 4 was subjected to Beckmann rearrange-
ment in refluxing glacial acetic acid and acetic
anhydride to obtain 16,17a-dioxo-17-aza-D-homo-5-
androsten-3P-yl acetate 5 [28]. Its infrared spectrum
showed 2 bands at 1725 (C=O stretching) and
1695 cm-r (imide function). The 18-methyl singlet
had shifted downfield to 6 1.20 ppm. The imide 5
gave 6 on alkaline hydrolysis (scheme 1).
Oppenauer oxidation of 6 using a cyclohexanone/
toluene/dioxane system afforded 17-aza-D-homo-4
androstene-3,16,17a-trione 7. The NMR spectrum
showed signals at 6 1.23 (s, 3H, 18-CH,), 1.26 (s, 3H,
19-(X,), and 5.80 (s, lH, 4-H). On treatment with
pyrrolidine in methanol, the a, P-unsaturated ketone 7
gave enamine 8, which, on sodium borohydride re-
duction in methanol at room temperature, yielded
1690 cm-r were absent, while an intense band at 1660
cm-r was present, indicating the presence of an amide
function. Moreover, the NMR spectrum showed a
singlet at 6 1.00 (6H, 18-U!, and 19-CH,), that is, an
upfield shift of the 18-methyl signal indicating the
presence of a methylene function at position 17a. It
appears that there is a transformation of imide into its
corresponding amide due to regioselective reduction
[29-3 l] with sodium borohydride. The configuration
at position 3 may be assigned as in analogy with
earlier reports [32-341. Sodium/pentanol reduction of
compound 9 yielded diamine 10, which, on methyla-
tion in the formaldehyde/sodium borohydride system,
gave tertiary amine 11 (scheme 2).
The amines 10 and 11 were quatemized to give
monoquatemary compound 12 (DPJ-246) and bisquat-
emary 13 (DPJ-240), respectively. The bromide and
chloride analogues 14 and 15 were prepared by
passing bisquatemary iodide 13 through the corre-
sponding columns of IRA00 ion-exchange resin
[35, 361. Bulkier groups were also incorporated in
compound 13 to assess the effect of this kind of
change on the biological activity. With this aim,
diethiodide 16 (DPJ-260) and dibutiodide 17 (DPJ-
261) were synthesized from amine 11 (scheme 3).
It has been postulated [37] that neuromuscular-
blocking drugs bind to the acetylcholine receptor at
only one of the cationic heads and that the rest of the
molecule shields the receptor from the naturally re-
 333

lsoomyl nitrite
Pot. t- butoxide

5
6
Scheme 1.

leased acetylcholine. Therefore, it was considered of 19 in the cyclohexanone/toluene system gave an

worthwhile to prepare the monoquatemary derivatives oily unsaturated ketone 20 (scheme 4). The amine 19
in this series. The imide 5, on sodium/pentanol was acetylated to give 17-methyl- 17-aza-D-homo-5
reduction, yielded amine 18 1381 which on methyl- androsten-3P-yl acetate 22. Quatemization of amines
ation afforded tertiary amine 19. Oppenauer oxidation 19, 20 and 22 was carried out with methyl iodide in

6 Oppenouer
oxidotion

Sodiu m-pentanol

Me
0 Me

Scheme 2.
334

Me Neuromuscular-blocking activity
@P
Mel N-Me
Me
10 b The neuromuscular-blocking activity of all the
Q compounds was investigated on chick biventer cervicis
21
0& \ preparations. All compounds could abolish the twitch
C N
\
Me 12
response by indirect stimulation. Responses to exogen-
ous acetylcholine and carbachol were also reduced but
responses to KC1 were not significantly affected by
the test compounds. Twitches recovered on wash-out
of the compounds. All compounds caused concen-
tration-dependent parallel shifts of the dose-response
curve to carbachol to the right. The maximum
RX
11 responses to carbachol were not reduced in the
Resin lCl,Brl - presence of the compounds. Plots of (dose ratio - 1)
against negative log molar concentration of antagonist
gave straight lines with slopes close to 1, which is
consistent with competitive antagonism (table I).
It is interesting to note that the location of the nitro-
13 R , R’ , R* =Me; X=1 gen atom at the 17-position had a significant effect on
the biological activity of the newly synthesized
14 R, R’, R’ =Me; X = Br
compounds in the in vitro studies. All the bisquater-
15 R, R’, It’= Me; X= Cl nary compounds were found to possess potent neuro-
muscular-blocking activity, but were slightly less
16 R, R’nEt, R2= Me; X=1 active as compared to the prototype chandonium
17 R,R’=n-But, R’=Me; X=X iodide 2. It appears from the results that the chloride
form 15 (DPJ-262) is more active than the iodide
Scheme 3. 13 (DPJ-240) and bromide 14 (DPJ-252) forms of
bisquatemary compounds. A correlation between the
size of the N-substituent and antagonistic activity
ethanol to afford 24 (DPJ-243), 21 (DPJ-245) and 23 was also observed. Increasing the size of the sub-
(DPJ-248) respectively (schemes 4 and 5). An stituent from methyl 13 (DPJ-240) through ethyl
alternative procedure [39] to 13 and related 16 (DPJ-260) to butyl 17 (DPJ-261) groups resulted in
compounds [40] has recently been reported. a reduction in the activity.

Me
Sodium-
pentonol

Me
OMe
N
Me Mel
b

0 LYF ’
20 21
Scheme 4.
 335

Me
/Me
Me N Me1 ~

0
ii \ II
Me-C-O J3F Me-C-O

23

HO

Scheme 5. 24

Monoquaternary compounds, 12 (DPJ-246), 21 (MeOH) 238 nm (log E 3.96) (0.1 N NaOHiMeOH) 288 nm
(DPJ-248), 23 (DPJ-248) and 24 (DPJ-243) carrying (log E 4.29); IR (KBr) v 3375, 3195, 1730, 1630, 1455, 1300,
polar functionalities at the 3-position, more or less at 1145, 1045,945 and 865 cm-l; tH-NMR (CDCl,/DMSO-d,): 6
the same distance from the 17-cationic head, showed 1.0 (s, 3H), 1.13 (s, 3H), 5.46 (m, lH), and 12.4 (s, lH,
exch/D,O).
weak neuromuscular-blocking action in the in vitro
tests as compared to 13 (DPJ-240). 16,17a-Dioxo-17-aza-D-homo-S-androsten-Spy1 acetate 5
Thus, the present study indicates that 2 cationic A mixture of 16-oximino- 17-oxo-5-androsten-3P-ol 4 (1 .O g),
heads at an appropriate distance, and other structural acetic anhydride (15 ml), and glacial acetic acid (10 ml) was
features, are required for a favorable neuromuscular- refluxed for 18 h under anhydrous conditions. The solid resi-
blocking activity. due, obtained after removing the solvent under reduced press-
ure, was washed with petroleum ether (60-80”) and water. The
material obtained was crystallized from ethanol to afford 5,
Experimental protocols
Chemistry Table I. PA, values for the test compounds.
__-_
Compound P& Slope
Melting points are uncorrected. NMR spectra were recorded on
an EMP-390, 90 MHz NMR instrument using tetramethylsil-
ane (TMS) as the internal standard. IR spectra were obtained 12 (DPJ-246) 6.42 kO.07 I.1
with a Perkin-Elmer 882 soectronhotometer in KBr oellets. 13 (DPJ-240) 7.15 + 0.03 1.0
The purity of the compounds was established by thin-layer
chromatography and by elemental analysis (C, H, N). Anhy- 14 (DPJ-252) 6.99 kO.07 0.95
drous sodium sulfate was used as drying agent for organic 15 (DPJ-262) 7.16 0.87
solvents.
16 (DPJ-260) 6.85 f 0.06 0.83
16.Oximino-I 7-oxo-5-androsten-3P-014 17 (DPJ-261) 6.11 f 0.04 1.03
To a stirred solution of 17-oxo-5-androsten-3P-ol 3 (2.0 g) in
potassium tert-butoxide (prepared by dissolving potassium 21 (DPJ-245) 6.1 +0.2 ndb
metal (1 .O g) in tert-butanol (17 ml)) was added isoamyl nitrite
(1.8 ml) at room temperature under a nitrogen atmosphere. The 23 (DPJ-248) 5.03 kO.16 ndb
reaction mixture, after being stirred overnight, was diluted with 24 (DPJ-243) 5.6 I!I 0.2 ndb
water (100 ml), acidified, and the yellow aqueous suspension
was extracted with chloroform (4 x 100 ml). The combined ChandoniumC 8.1 1.0
chloroform extract was washed with sodium bicarbonate sol- TubocurarineC 6.0 1.0
ution (10%) and extracted with sodium hydroxide solution
(lo%, 4 x 100 ml). The combined alkaline extract was washed apA, values were estimated from the Gaddum equation:
with chloroform (100 ml) and acidified while cold. The precipi- dose ratio - 1 = [Al/K,, where K, is the antagonist dissocia-
tated product was filtered, washed, and dried. The product tion constant; bnd: slopes were not calculated because only
obtained was crystallized from methanol to afford 4, 1.0 g 2 sets of dose-response curves were obtained; Cvalues are
(45.0%); mp: 255-257°C (lit [26, 271 250-251°C); UV,,, from identical experiments by Gandiha et al 1975 [12].
336
0.45 g (40.0%); mp: 252-254’C (lit [28] 257-259°C);
IR (KBr) v 3300, 1725,1695,1440, 1370, 1260,1025 and 785
cm-l; iH-NMR (CDCI,): 6 1.03 (s, 3H), 1.20 (s, 3H), 2.03 (s,
3H), 4.51 (m, lH), 5.36 (m, lH), and 8.33 (br, lH, exch/D,O).
16, 17a-Dioxo-I 7-aza-o-homo-5-androsten-3/%ol6
A mixture of 16, 17a-dioxo- 17-aza-o-homo-5- androsten-
3p-yl acetate 5 (2.0 g), potassium hydroxide (0.4 g), and
methanol (100 ml) was refluxed for 30 min. After acidification
with dilute hydrochloric acid, the reaction mixture was further
refluxed for 15 min. The solid residue obtained, after removing
solvent under reduced pressure, was washed with water, dried,
and crystallized from methanol to give 6, 1.2 g (68.0%); mp:
258-260°C; IR (KBr) v 3435, 3190, 1710, 1690, 1375, 1290
and 1060 cm-t; iH-NMR (CDCl,/DMSO-d,): 8 1.03 (s, 3H),
1.20 (s, 3H), 3.46 (m, lH), 4.48 (br, lH, exch/D,O), and 5.38
(m, 1H); MS: m/z 317 (M+). Anal for C,9H,,N0, (C, H, N).
I7-Aza-o-homo-4-androstene-3, 16,17a-trione 7 androstene 10 (0.5 g) in methanol (25 ml) was added form-
16, 17a-Dioxo-17-aza-o-homo-5-androsten-3P-ol 6 (2.Og) was
dissolved in a mixture of cyclohexanone (20 ml), toluene
(100 ml), and dry dioxane (3 ml) and the solution was sub-
jected to azeotropic distillation to remove traces of moisture.
The distillation was continued at a slow rate, till the dropwise
addition of a solution of aluminium isopropoxide (2.0 g) in dry
toluene (20 ml) was complete. The reaction mixture was
refluxed for 3 h and then allowed to stand at room temperature
for 12 h. The slurry was filtered and washed with toluene. The
combined washings and filtrate were steam-distilled until the
complete removal of organic solvents was effected. The re-
sidual aqueous suspension was allowed to stand at room tem-
perature to give a crystalline material which was recrystallized
from aqueous ethanol to give 7, 1.0 g (50.0%); mp:
168-170°C; UV,,, (MeOH): 241 nm (log E 4.28); IR (KBr):
v 3190, 3080, 1720, 1690, 1670, 1380, 1280, and 1110 cm-l;
‘H-NMR (CDClJ: 6 1.23 (s, 3H), 1.26 (s, 3H), 5.80 (s, lH),
and 8.58 (br, lH, exch/D,O); MS: m/z 315 (M+). Anal for
C&W% CC, H, N.
3-Pyrrolidino-I 7-aza-D-homo-3,5-androstadiene-16, I7a-dione 8
To a refluxing solution of 17-aza-o-homo-4-androstene-3, 16,
17a-trione 7 (1.5 g) in methanol (25 ml) was added freshly
distilled pyrrolidine (1 ml). Refluxing was continued further for
1 min to induce crystallization. The crystalline material was
filtered, washed and dried to give 8, 1.3 g (74.20%); mp:
292-294°C (decomp); UV,,, (MeOH): 276 nm (log E 4.28);
IR (KBr): v 3225, 1720, 1700, 1640, 1600, 1603, 1400, 1345,
1270, 1185, and 830 cm-l; lH-NMR (CDCl,): 6 1.04 (s, 3H),
1.24 (s, 3H), 3.21 (m, 4H), 4.90 (s, lH), and 5.16 (m, 1H).
Anal for CZIH,,N,O, (C, H, N).
3PPyrrolidino-I 7-aza-o-homo-5-androsten-16one 9
To a stirred susnension of 3-ovrrolidino-17-aza-o-homo-3.5-
androstadiene-16: 17a-dione 8 ii.5 g) in methanol (150 ml)
was added sodium borohydride (1.5 g) in small quantities at
room temperature. Stirring was continued for 2 h, the reaction
mixture was poured into ice-cold water (800 ml), and the
aqueous suspension was extracted with chloroform
(6 x 100 ml). The combined chloroform extract was washed
with water, dried, and solvent removed under reduced pressure
to give a solid residue which was crystallized from acetone to
give 9, 0.7 g (46.4%); mp: 234-236°C; IR (KBr): v 3328,
1660. 1630. 1480. 1375. 1056. and 890 cm-i: tH-NMR
(CDCIJDMSO-d,):’ S 1.06 (s, 6H), 3.13 (s, 4H), and 5.33
(m, 1H). Anal for C*,H,,N,O.
3PPyrrolidino-I 7-aza-o-homo-5-androstene IO
Sodium metal (10 g) was slowly added to a refluxing solution
of 3p-pyrrolidino- 17-aza-o-homo-5-androsten- 16-one 9 (1 g)
in 1-pentanol (200 ml). The reaction mixture was refluxed till
the sodium metal had completely reacted. The reaction mixture
was steam-distilled to remove I-pentanol. The residual aqueous
suspension was cooled to room temperature and extracted with
chloroform (6 x 100 ml). The combined chloroform extract was
washed with water, dried and the solvent removed under
reduced pressure to give a solid residue which was crystallized
from acetone to gi;e 10, 0.25 g (27.0%); mp: 18&182”C;
IR (KBr): v 3260. 1440. 1365. 1280.1140. 890. and 805 cm-t:
1H-NMR (CDCl,): S 0.96 (s, 3H), 1100 (s,’ 3H); 2.63 (br, 5H);
and 5.33 (m, 1H); MS: m/z 342 (M+). Anal for &H,,N2
CC, H, N).
17-Methyl-3/%pyrrolidino-l7-aza-D-homo-5-androstene 11
To a stirred solution of 3P-pyrrolidino-17-aza-o-homo-5-
aldehyde solution (40% v/v, 1.2 ml) at room temperature and
stirring continued for further 1 h. Sodium borohydride (0.5 g)
was added in small quantities to the stirred solution at room
temperature. Stirring was continued for 2 h and the reaction
mixture was then poured into ice-cold water (250 ml). The
aqueous suspension was extracted with chloroform
(4 x 100 ml). The combined chloroform extract was washed
with water, dried, and the solvent was removed under reduced
pressure to give a solid residue which was crystallized from
acetone to give 11, 0.25 g (48.0 %); mp: 174-176°C; IR (KBr)
v 2780, 1445, 1370, 1280, 1175, 1140, 900 and 815 cm-t;
iH-NMR (CDCI,): S 1.03 (s, 6H), 2.23 (s, 3H), 2.66 (m, 5H),
and 5.40 (m, 1H); MS: m/z 356 (M+). Anal for C24H40N2
CC, H, NJ.
3L%Pyrrolidino-I7-aza-o-homo-5-androstene dimethiodide (DPJ-
246) 12
Methyl iodide (0.2 ml) was added to a solution of 3p-pyrroli-
dino-17-aza-o-homo-5-androstene 10 (0. I g) in absolute etha-
nol (20 ml) at room temperature and the solution was refluxed
for 1 h. The reaction mixture was cooled, filtered and concen-
trated to induce crystallization of 12, 0.14 g (76.5%); mp:
278-280°C. Anal for CZSH,,I,N2 (C, H, N).
17-Methyl-3@pyrrolidino-I 7-aza-D-homo-5-androstene dimeth-
iodide (DPJ-240) 13
Methyl iodide (0.2 ml) was added to a refluxing solution
of 17-methyl-3P-pyrrolidino-17-aza-D-homo-5-androstene 11
(0.1 g) in absolute ethanol (15 ml) and refluxing was continued
further for 30 min. The reaction mixture was cooled to room
temperature, filtered, and concentrated to induce crystallization
of 13, 0.15 g (83.5 %); mp: 290-292°C. Anal for C,,H,&N,
CC, H, N).
17-Methyl-3~-pyrrolidino-l7-aza-o-homo-5-androstene dimetho-
bromide (DPJ-252) 14
The preparation of 14 began with the preparation of ion-
exchange resin (bromide fo&). ‘Amberlite’ resin IR-400 (Cl)
(20 a) was taken UP in hvdrobromic acid solution (1N. 200 ml)
and ihe slurry was-heated on a boiling water bath ‘for’ 1 h with
stirring. The supernatant was decanted off and the resin was
repeatedly washed with distilled water until free from acid. The
resin so obtained was given similar treatment with sodium
hydroxide solution (lN, 200 ml) and finally with hydrobromic
acid solution (lN, 200 ml). The acid-free, damp resin was
taken up in aqueous methanol (50%) and the slurry was packed
in a column.

A solution of 17-methyl-3P-pyrrolidino-17-aza-o-homo-5-
androstene dimethiodide (DPJ-240) 13 (0.2 g), in aqueous
methanol (50%, 5 ml), was run through the column packed
with resin (bromide form) (25 ml damp solid). Elution with
aqueous methanol (50%) was continued till the eluate gave no
pale-yellow precipitate with silver nitrate solution. Combining
the eluates and removing the solvent under reduced pressure
gave a solid residue which was crystallized from
acetone/methanol to afford 14, 0.11 g, (64.0%); mp:
286282°C. Anal for C26H46Br2N2 (C, H, N).
I7-Methyl-3P-pyrrolidino-I 7-aza-D-homo-5-androstene dimetho-
chloride (DPJ-262) 15
A solution of 17-methyl-3P-pyrrolidino- 17-aza-D-homo-5-
androstene dimethiodide (DPJ-240) 13 (0.2 g), in aqueous
methanol (50%, 5 ml), was run through the column packed
with resin (chloride form, prepared in a similar fashion to the
bromide form) (25 ml damp solid). Elution with aqueous
methanol (50%) was continued till the eluate gave no white
precipitate with silver nitrate solution. Combining the eluate
and removing the solvent under reduced pressure gave a solid
residue which was crystallized from acetone/methanol to yield
15, 0.14 g (98.0%); mp: 318-320°C. Anal for C,,H,,Cl,N,
CC, H, N).
I7-Methyl-3P-pyrrolidino-I 7-aza-o-homo-5-androstene
diethiodide (DPJ-260) 16
Ethyl iodide (0.5 ml) was added to a refluxing solution
of 17-methvl-3~-ovrrolidino-17-aza-o-homo-5-androstene
(0.1 g) in abiolute’ethanol (15 ml) and refluxing was continued
for 7 h. The reaction mixture was cooled, filtered and concen-
trated to induce crystallization, The crystalline material was
filtered, washed and dried to afford 16, 0.02 g (10.7%); mp:
284-286°C. Anal for C2sHs012N2 (C, H, N).
17.Methyl-3/%pyrrolidino-I 7-aza-D-homo-5-androstene dihut-
iodide (DPJ-261) 17
n-Butyl iodide (0.5 ml) was added to a refluxing solution
of 17-methyl-3P-pyrrolidino-17-aza-o-homo-5-androstene
(0.1 g) in absolute ethanol (15 ml) and refluxing was continued
for 4 h. The reaction mixture was cooled, filtered, and concen-
trated to induce crystallization. The crystalline material was
filtered, washed, and dried to afford 17, 0.05 g (25.5%); mp:
258-260°C. Anal for C,,H,,I,N, (C, H, N).
17-Aza-D-homo-5-androsten-3/%ol I8
Sodium metal (10.0 g) was slowly added to a refluxing solution
of 16, 17a-dioxo-l7-aza-D-homo-5-androsten-3P-yl acetate 5
(1.0 g) in I-pentanol (200 ml). The reaction mixture was
refluxed till the sodium metal had completely reacted, and this
was followed by the removal of I-pentanol by steam distil-
lation. The aqueous suspension was cooled and extracted with
chloroform (6 x 100 ml). The combined chloroform extract was
washed with water, dried, and the solvent was removed under
reduced pressure to give a solid residue which was crystallized
from acetone to afFord 18, 0.4 g (50.0%); mp: 178-180°C
(lit 1331 191-193°C): IR (KBr): v 3396. 3320. 1458. 1362.
1059, 952 and 842 cm-i; tH-NMR (DMSd-d,): 6 0.93 is, 3H);
0.96 (s, 3H), and 5.29 (m. IH). Anal for C,,H,,NO (C, H, N).
17-Methyl-l7-aza-~-homo-5-androsten-3~-oll9
To a stirred solution of 17-aza-o-homo-5-androsten-3P-ol 18
(1.0 g) in methanol (30 ml) was added formaldehyde solution
(40% v/v, 1.5 ml) at room temperature and stirring was further
continued for 1 h. Sodium borohydride (1 .O g) was added in
337
small quantities to the stirred solution over a period of 2 h. The
reaction mixture was poured into ice-cold water (300 ml) and
the aqueous suspension was extracted with chloroform
(6 x 100 ml). The combined chloroform extract was washed
with water, dried, and the solvent was removed under reduced
pressure to give a fluffy material which was chromatographed
over a column of neutral alumina (40 g). Elution with petro-
leum ether/toluene (5:5) and removal of the solvent under re-
duced pressure afforded 19, 0.60 g (57.0%); mp: 98-100°C.
IR (KBr): v 3377, 2784, 1445, 1375, 1056, and 812 cm-i;
tH-NMR (CDCl,): 6 1.03 (s, 6H), 2.18 (s, 3H), 3.50 (m, lH),
and 5.37 (m, 1H). Anal for C20H31N0 (C, H, N).
I7-Methyl-17.aza-D-homo-4-androsten-3-one 20
17-Methyl-17-aza-D-homo-5-androsten-3~-ol 19 (0.5 g) was
dissolved in a mixture of cyclohexanone (7.5 ml) and toluene
(50 ml), and the solution was subjected to azeotropic distil-
lation to remove traces of moisture. Distillation was continued
at a slow rate till the dropwise addition of a solution of alu-
minium isopropoxide (0.6 g), in dry toluene (20 ml), was
complete. The reaction mixture was refluxed for 3 h and then
allowed to stand at room temperature for 12 h. The slurry was
filtered, washed, and the organic solvents were removed by
steam distillation. The residual aqueous supension was cooled
and extracted with chloroform (4 x 100 ml). The combined
chloroform extract was washed, dried, and the solvent was
removed under reduced pressure to give an oily residue of 20
which could not be crystallized, 0.22g (44.3%). This was used
11 as such in further reactions. UV,,, (MeOH): 242 nm. The
compound was further characterized by preparing its methio-
dide derivative.
17-Methyl-l7-aza-D-homo-5-androsten-3~-yl acetate 22
A mixture of 17-methyl-17-aza-D-homo-5-androsten-3P-ol 19
(0.25 g), acetic anhydride (5.0 ml), and pyridine (5 ml), was
heated on a water bath for 15 min. The reaction mixture was
poured into a mixture of crushed ice and potassium hydroxide
solution (lo%, 50 ml) and the aqueous suspension was ex-
11 tracted with chloroform (3 x 50 ml). The combined chloro-
form extract was washed, dried, and the solvent was removed
under reduced pressure to give an oily residue 22 which could
not be crystallized, 0.15 g (52.7%). Thin-layer chromatography
gave a single spot. The residue was characterised by synthe-
sizing its quaternary derivative.
17-Methyl-l7-aza-D-homo-5-androsten-3~-ol methiodide (DPJ-
243) 24
Methyl iodide (0.2 ml) was added to a solution of 17-methyl-
17-aza-D-homo-5-androsten-3P-ol 19 (0.1 g) at room temper-
ature and the solution was retluxed for 1 h. The reaction
mixture was cooled, filtered, and concentrated to induce
crystallization. The crystalline material was filtered, washed,
and dried to afford 24, 0.12 g (82%); mp: 280-282°C. Anal for
C,,H,,INO CC, H, N).
17-Methvl-l7-aza-D-homo-5-androsten-3~-vl , _ acetate methio-
dide (DFJ-248) 23
Methyl iodide (0.2 ml) was added to a solution of 17-methyl-
17-aza-D-homo-5-androsten-3b-vl a d
acetate 22 (0.15 a) in absol-
-:
ute ethanol (20 ml) at room temperature and the mixture was
refluxed for 1 h. The reaction mixture was cooled, filtered, and
concentrated to induce crystallization. The crystalline material
was filtered, washed, and dried to afford 23, 0.1 g (47.3%);
mp: 276-278°C; IR (KBr): v 2853, 1734, 1454, 1374, 1244,
1035 and 904 cm-t. Anal for C,,H3,1N02 (C, H, N).
338

17-Methvl-I 7-aza-o-homo-4-androsten-j-one methiodide IDPJ- (Briggs MH, Christie G, eds) Academic, London, Vol3,39-65
245) 21. 7 Buckett WR, Hewett CL, Savage DS (1973) J Med Chem 16, 1116-l 124
Methyl iodide (0.2 ml) was added to a solution of 17-methyl- 8 Baird WLM, Reid AM (1967) Br J Anaesrh 39,775-780
17-aza-D-homo-4-androsten-3-one 20 (0.2 a) in absolute etha- 9 McDowell SA, Clarke RSJ (1969) Anaesthesia 24,581-590
no1 (20 ml) and processed as above to-afford 21, 0.28 g 10 Singh H, Paul D (1974) J Chem Sot Perkin Tram 1, 1475-1479
11 Gandiha A, Marshall IG, Paul D, Singh H (1974) J Pharm Pharmacol
(95.2%); mp: 278-280°C. Anal for C2,H341N0 (C, H, N). 26,871-877
Neuromuscular-blocking activity 12 Gaodiha A, Marshall IG, Paul D, Rodger IW, Scott W, Singh H (1975)
Clin Exp Pharmacol Physiol2, 159-170
Two chick biventer cervicis preparations [41] were mounted in 13 Harvey AL, Paul D, Rodger IW, Singh H (1976) J Pharm Pharmacol
lo-ml tissue baths containing physiological salt solution of the 28,617-619
followine comuosition (mM): NaCl. 118.4: KCl. 4.7: MaSO,. 14 Bhardwaj TR, Kapoor S, Shekhar CC, Jindal DP, Singh H (1988) Ind J Chem
1.2; KH2?04, i.2; CaCl;, 2.5; NaHdO,, 251 and glucbse,“ll.i: 27B, 209-212
15 Singh H, Gupta RK, Bardwaj TR (1988) Ind J Chem 27B, 508-512
The solution was maintained at 34°C and pH 7.3, and bubbled
16 Abraham J, Jindal DP, Singh H (1992) Ind J Chem 3 lB, 566569
with oxygen containing 5% C02. 17 Abraham J, Jindal DP, Singh H, Patnaik GK, Srimal RC (1993) Eur J Med
In preliminary experiments to determine the effective concen- Chem 28.231-234
trations of the neuromuscular-blocking compounds, twitches 18 Bowman WC, Webb SN (1972) J Pharm Pharmacol24,762-772
were evoked by stimulating the motor nerve every 10 s with 19 Palmer RA, Kalam MA, Singh H, Paul D (1980) J Cryst MO/ Srruct 10,
pulses of 0.2 ms duration and a voltage greater than that which 31-53
produced a maximum twitch. Contractures to exogenouslv 20 Savage DS, Cameron AF, Ferguson G, Hannaway C, Mackary IR (1971)
applied acetylcholine (1W M for 30 s), carbachol (2 x 10-S M J Chem Sot B, 410-415
for 60 s). and KC1 (4 x 10-Z M for 30 s) were obtained in the 21 Stenlake JB, Waigh RD, Dewar GH, Hughes R, Chapple DJ, Coker GG
absence’bf nerve stimulation prior to’ addition of the test (198l)EurJMedChem 16,515-524
compound and after block of twitches by the test compound. 22 Stenlake JB, Waigh RD, Urwin J, Dewar GH, Coker GG (1983)
In other experiments, concentration-effect curves to carbachol Br J Anaesth 55, 3S
were constructed in the absence and in the presence of 3 23 Tuba Z (1980) Arzneim Forsch 30, 342-346
concentrations of the test compounds. PA, values were calcu- 24 K@ati E, Bir6 K (1980) Arzneim Forsch 30,346-354
lated from plots of log (dose ratio - 1) against PA, [42]. 25 Savarese JJ, Ali HH, Basta SJ, Embree PB, Scott RPF, Sunder N, Weakly JN,
Wastila WB. El-Sayad HA (1988) Anesthesiology 68.723-732
26 Stodala FH, Kindal EC, Mckenzie BF (1941) J Org Chem 6,841-844
Acknowledgments 27 Hassner A, Pomerantz IH (1962) J Org Chem 27, 176&1766
28 Hassner A, Wentworth WA, Pomerantz IH (1963) J Org Chem 28,3W306
Authors DP Jindal and AK Verma are thankful to the 29 Bailey DM, Johnson RE (1970) J Org Chem 35,3574-3576
University Grants Commission, New Delhi for the financial 30 Brooks CJW, Ekhato IV (1982) J Chem Sot Chem Commun 943-944
31 Suginome H, Satoh G, Wang JB, Yamada S, Kobayashi K (1990) J Chem Sot
support and Panjab University Chandigarh, India, for providing
Perkin Tram 1 1239-1245
research facilities. 32 Majid MA, Palmer RA, Singh H, Paul D (1977) Acta Crystallogr 833,
364-3649
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