Professional Documents
Culture Documents
Harold Kadin
1. Description 80
1.1 Name, Formula, Molecular Weight 80
1.2 Appearance, Color, Odor 80
2. History 80
3. Synthesis 84
4. Physical Properties 84
4.1 Spectral Properties 84
4.2 Solid State Properties 97
4.3 Solution Data 106
5. Stability 107
5.1 Solid State Stability 107
5.2 Solution Stability 108
6. Analytical Tests and Methods 112
6.1 Elemental Analysis 112
6.2 Spectrophotometric Methods 112
6.3 Chromatographic Methods 113
6.4 Titrimetric Methods 120
7 Analysis in Biologic Fluids and Tissues and in Animal Rations 120
7.1 Thin Layer Radiochromatography (TLRC) 120
7.2 Gas Chromatography-Mass Spectroscopy (GC-MS) 122
7.3 Gas Chromatography-Flame Photometric Detection (GF-FPD) 123
7.4 High Performance Liquid Chromatography with UV Detection (HPLC-UVD) 124
7.5 Spectrofluorometry 124
7.6 Radioimmunoassay (RIA) 124
7.7 Semiautomated Ellman Colorimetry 125
7.8 High Performance Liquid Chromatography with 126
Electrochemical Detection (HPLC-ECD)
7.9 High Performance Liquid Chromatography with 128
Fluorescence Detection (HPLC-FD)
8. Drug Metabolism-Pharmacokinetics 128
8.1 Blood Level Studies 128
8.2 Urinary Excretion Studies 129
8.3 Miscellaneous Distribution Studies 134
9. Acknowledgments 131
10. References 131
11. Review Coverage Dates 137
1. Description
1.1 Name, Formula, Molecular Weight
Captopril, Capoten@, or Lopirins
is l-(3-mercapto-2-D-methyl-l-oxopropyl)-L-proline
(S,S) with -
Chem. -
Abstr. Registry Number
62571-86-2.
RELATIVE
ENZYME INHIBITOR IN VlTRO INHIBITION
CAABOXYPEPTIDASE A
D-2-EENZYLSUCCINIC ACID
-7 FH2 ?-
0 = CCH,-CH-C -C = 0
ANGIOTENSIN-CONVERTING ENZYME
SUCC1NYL.L-PROLINE 1
D-2-METHYLSUCCINYL.L-PAOLI" 15
CAPTOPRIL 14000
F i g . 1. Key steps i n t h e d e s i g n o f a s p e c i f i c i n h i b i t o r of
t h e a n g i o t e n s i n c o n v e r t i n g enzyme.
CAPTOPRIL 83
3. Synthesis
A process (14) is presented (Figure 2) in a
chemical reaction sequence which follows this
brief description.
Methacrylic acid (I) is condensed with thio-
lacetic acid (11) to give racemic 2-methyl-3-
acetylthiopropionic acid (111). L-proline is then
acylated with the acid chloride (IV) of the thio-
ester (111). The resulting proline thioester (VI)
is resolved from its - - isomer by aqueous cry-
R,S
stallization. Saponification of compound VI with
sodium hydroxide affords the sodium salt of
.
captopril which after acidification yields
captopril (VII)
4. Physical Properties
4.1 Spectral Properties
4.11 Infrared Spectra
The infrared spectrum of captopril in
chloroform is presented in Figure 3 and as a KBr
pellet in Figure 4. The infrared spectrum in the
latter indicates the presence of the following
frequencies and their structural assignments (15).
cm-1 Assignment
1750 C = 0 (COOH group)
1725
1640 C = 0 (amide)
2560 S - H
DiffereQfes in the fingerprint regions
(1350-900 cm ) of the mineral oil mull infrared
spectra of batches 3 and 4 (Figure 5 and 6,
respectively) indicate that the low melting batch
3 and the high melting batch 4 are polymorphs (see
Section 4.21) .
CAPTOPRIL 85
Figure 2
CH2 = CCOOH
MW = 86.01
AcSH
MW 76.11
+ AcSCH2CHCOOH
MW = 162.20
Methacrylic Thiolacetic Acetylthioisobutyric
IV
AcSCH2CHC
\ COOH
c1 COOH
MW = 180.65 MW = 115.13 MW = 259.32
Acid Chloride" L-Proline Proline Thioester
"'?
VI VI I
H3C 0
AcSCH2CHCN d)
COOH COOH
Proline Thioester MW = 217.28
Captopril
0
Ac = CH3C-
II
(a) Reflux (b) SOCl , DMF, Distillation
(c) H 0 + NaHCO Ci C12 Wash, HC1,
ci!ystallizaiG.on 41.
(d) H20 + NaOH, HC1, CH2C12 extraction
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WAVELENGTH (MICRONS)
F i g u r e 5. I n f r a r e d S p e c t r u m of C a p t o p r i l , B a t c h 3 , Mineral O i l M u l l
F R E w € w (CM-')
Table 1
Proton-NMR Data for Captopril
Proton Chemical Shift ( 6 )'PPM from TMS (ext.
COOH 9.81 (s)
CL-CH 4.60 (m)
B-CH2 2.03 (m)i 2.25 (m)
Y-CH,L 2.07 (m)
6 -CH2 3.63 (m)
9
-CH-C 2.44 (d,q) J=6,9
-STHA 2.82 (m)
-CH3 L 1.17 (d) J=6
SH 1.53 (ad) J=9,8
' multiplicities: d=doublet; q=quartet;
m=multiplet. J=proton-proton coupling
constants in Hertz.
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92
CAPTOPRIL 93
Table 2
Carbon # Chemical S h i f t ( 6 )
1 ppm from TMS
175.69
59.84
30.03
25.49
48.24
174.91
43.1
17.07
28.1
R e f e r e n c e d from c e n t e r peak o f t h e C D 3 0 D
m u l t i p l e t a t 4 9 . 0 ppm
S p e c t r a of c a p t o p r i l i n a q u e o u s m e t h a n o l ,
( F i g . 9 ) w a t e r , 0.1M sodium h y d r o x i d e and 0 . 1 M
h y d r o c h l o r i c a c i d ( F i g . 101, a r e p r e s e n t e d (17).
These s p e c t r a d e p i c t a n end a b s o r p t i o n s l o p e
w i t h o u t peak o r s h o u l d e r . The s l o p e s p e c t r u m i n
0 . 1 M sodium h y d r o x i d e was s h i f t e d c o n s i d e r a b l y
towards h i g h e r wavelengths. S i n c e weak s u l f h y d r y l
a b s o r p t i o n i s r e p o r t e d ( 1 8 ) i n t h e 220-230 nm
r e g i o n , t h i s a b s o r p t i o n s h i f t may b e d u e t o
i o n i z a t i o n o f t h e s u l f h y d r y l f u n c t i o n by t h e
a l k a l i . This s h i f t towards higher wavelengths
w i t h i n c r e a s e i n pH h a s been u s e d by O n d e t t i ( 1 9 )
t o d e t e r m i n e t h e pK o f t h e s u l f h y d r y l i n
captopril (Section 8.32).
HAROLD KADIN
. . . . . .
I ; 0 1 . . . . . . . . . . .0,
. !. !. ..
I '
. . ./ i ! .a,* , , , 4 , -
Fig. 9. U l t r a v i o l e t absorption s p e c t r a of C a p t o p r i l i n
10%aqueous methanol s o l u t i o n .
Instrument: Cary 15.
170 103
J
172+m/z
H
m/z 70
F
m/z 199 + H20
m/z 202 + CH3
M+
m/z 2 1 7 , 1 m / z 1{3 + C02 r m / z 140 + SH
I
L m / z 171
b m / z 126 + CH2SH
+ S C H 2 4 m/z 127 + C02
96 HAROLD KADlN
90.-
80-
>
F
U
II)
78.-
7
Z
W
t-
Z
H
50
W
>
H
t-
a
_I
w
L l
4.25 Hygroscopidity
Under ordinary conditions captopril is not
hygroscopic. Equilibrium moisture studies (23)
indicate no moisture pickup by captopril up to 50%
relative humidity at room temperature. Above 50%
R.H. it shows a tendency to cake after one to two
days.
Captopril did not exhibit any visual physical
changes and remained dry from 0 to 67% R.H. on
exposure for 14 days. Samples exposed to 81% R.H.
for 14 days appeared moist (24).
4.26 Single Crystal X-ray Diffraction
Single crystal X-ray analyses have been
completed (25) for both the low (melting range
86-87OC) and high (melting range 105-106O) melting
polymorphs. Both forms are orthorhombic with the
following crystal data:
A. High melting polymorph -
a = 6.834(2), -b =
0
" 3 space group
8.821(2), c = 17.982(4)A; V = 1084A,
P2 2 2 wiFh four molecules3per unit cell; cal-
culakeh density = 1.33 gcm- .Refined to R = 0.04
for the 745 observed single crystal intensities.
B. Low melting polymorph - -=
a = 9.496(3), b
0
03 space group
12.304(3), c = 19.282(5)A; V = 2253A;
P2 2121 witF eight molecules egr unit cell;
calculated density = 1.28 gcm .
Refined to R =
0.06 for the 1093 observed single crystal inten-
sities.
The structure in both has the S,S absolute
configuration with a 2 ( T r a n s ) conformation about
the N-C(0) amide bond (the O-C-N-C(2) dihedral
angles vary from -4 to +6O). The molecular
conformation differ in detail, most notably in the
conformation about the (S)C-C(C0) bond.
Atomic coordinates relative to orthogonal
axes for the high melting form are:
CAPTOPRIL 99
C a p t o p r i l i n aqueous s o l u t i o n undergoes an
oxygen f a c i l i t a t e d , f i r s t o r d e r , f r e e r a d i c a l
oxidation a t its t h i o l to yield captopril
d i s u l f i d e ( 2 8 ) . H y d r o l y s i s a t t h e amide l i n k a g e
o c c u r s o n l y u n d e r f o r c i n g c o n d i t i o n s (see S e c t i o n
5 . 2 5 ) . O x i d a t i o n w a s d e l a y e d by a d j u s t m e n t t o
lower pH, a d d i t i o n o f c h e l a t i n g a g e n t s , i n c r e a s i n g
captopril concentration, u t i l i z a t i o n of nitrogen
o r l o w oxygen h e a d s p a c e , and i n c o r p o r a t i o n o f
a n t i o x i d a n t s . O x i d a t i o n seems t o o c c u r l e s s
r e a d i l y i n methanol ( 3 6 ) . N o d e g r a d a t i o n o f
c a p t o p r i l w a s o b s e r v e d ( 4 0 mcg/ml) i n t h i s s o l v e n t
f o r up t o 2 weeks a t 5OC.
5.21 S t a b i l i t y and S o l u t i o n pH
O x i d a t i o n r a t e c o n s t a n t s a t v a r i o u s pli v a l u e s
( 2 8 ) i n Table 5 , s u g g e s t t h a t c a p t o p r i l i s
o p t i m a l l y s t a b l e below pH 3.5, t h e o x i d a t i o n r a t e
b e i n g e s s e n t i a l l y c o n s t a n t from pH 2 t o 3. The
r a t e c o n s t a n t s i n c r e a s e r a p i d l y above pH 4 . Using
HPLC and c o l o r i m e t r y ( 3 8 ) , c a p t o p r i l a q u e o u s
s t a b i l i t y w a s s t u d i e d , a t 50 mcg/ml, i n a r o t a t i n g
b a s k e t d i s s o l u t i o n a p p a r a t u s f o r up t o 1 8 0 m i n u t e s
a t 37OC i n d i s t i l l e d water, and a t pH 1, 2 and 3 .
E x c e l l e n t s t a b i l i t y a t pH 1 and 2 b u t a p p r e c i a b l e
d e g r a d a t i o n a t pH 3 , and i n d i s t i l l e d w a t e r w a s
observed. S u r p r i s i n g l y , t h e r a t e of d e g r a d a t i o n
a t pH 3 exceeded t h a t i n d i s t i l l e d water. The
more r a p i d o x i d a t i o n a t pH 3 w a s a t t r i b u t e d t o
c a t a l y s i s v i a g r e a t e r t r a c e m e t a l s o l u t i o n from
the dissolution baskets.
CAPTOPRIL 109
Table 5
Oxidation Rate Constant for Captopril (5 mg/ml)
in Citrate-Phosphate Buffers at Various pH
Values at 5OoC
(day ) x 10f
PH Rate-Eonstan
2.13 8.38
2.59 9.01
2.89 8.22
3.13 8.31
3.53 9.92
3.88 9.13
4.23 12.94
4.67 19.43
5.16 28.93
5.63 42.03
5.22 Solution Stability, Metal Ions
and Chelating Agents
Transition metal ions most effectively cata-
lyze oxidation of captopril through a recycling of
oxygen free radicals (28). The most effective of
these catalysts are ubiquitous copper and iron, in
given order. As little as 1 ppm of copper has
been observed to catalyze captopril oxidation in
solution (28).
As has been demonstrated with cysteine (39)
lower levels of disodium edetate (EDTA Na2) may
enhance metal ion catalyzed thiol oxidation,
whereas higher levels retard oxidation.
Disodium edetate 0.1% (Na2EDTA 0.1%) best
stabilized 0.5 mg captopril per ml (of
citrate-phosphate buffer at pH 4, p = 0.5) in
Teflon-faced rubber sealed vials (37).
Analysis of urinary captopril was necessary
for dosage form bioavailability and dose titration
studies. The necessity for long term storage of
samples prior to analysis required development of
an acid-chelate stabilization (40). This
stabilization utilized diethylenetriamine
pentaacetic acid (DTPA) reputed (40) to be a more
effective metal chelator than Na2EDTA. A
110 HAROLD KADIN
116
Table 8
C a p t o p r i l HPLC S y s t e m s
cp COOH
TECHNICON AUTOANALYZER
FOR ELLMAN COLORIMETRY
P r
WASTE
RECORDER
IFLOW CELL I
COLOR I M ETER
WASTE
- FUNCTION NOMINAL
ML/MIN
~~
ELLMANS 0.8
WASTE
* = SOLVAFLEX SOLVENT TECHNICON
RESISTANT TUBING, SAMPLER I I
OTHER TUBING TYGON WITH 50 2/1 CAM
ELLMAN'S =
TEA-EDTA 0.08% ELLMAN'S
= 1.86% EDTA Nao 2H20 + IN 50% MEOH (COLD)
2Ooh TRIETHANOLAMINE + + 0.01 M
0.02% TWEEN 80 ACETATE pH 4.7
Figure 16
128 HAROLD KADIN
9. Acknowledgments
I would like to express my appreciation to
individuals who have been very helpful for the
contributions indicated: Drs. J. Fried and M.
Porubcan - NMR, Drs. Y.J. Wang and T. Prusik -
Stability, Mr. A. Restivo and Mr. D. Domina -
Synthesis and Solubility, Drs. A. Cohen and P.
Funke - MS and GC-MS, Ms. M. Malley and Dr. J.
Gougatas - Single Crystal X-Ray, Mr. F. Dondzila,
Mr. S. Perlman and Dr. J. Kirschbaum - HPLC, Mr.
H. Roberts - TLC, Mr. R. Poet and Dr. G. Brewer -
Proof Reading and Manuscript Clarity, Ms. D.
Walker - Word Processing, Dr. D. Cushman -
History, Drs. B. Migdalof and D. McKinstry - Drug
Metabolism - Pharmacokinetics.
10. References
1. Ondetti, M.A., Williams, N.J., Sabo, E.F.,
Pluscec, J., Weaver, E.R.; and Kocy, O.,
Biochemistry, -
10, 4033 (1971).
2. Gavras, H., Brunner, H.R., Laragh, J.H.,
Sealey, J.E. Gavras, I., and Vukovich, R.A.,
New Eng. J. Med., 291, 817 (1974).
3. Case, D.B., Atlas, S.A. Laragh, J.H., Sealey,
J.E., Sullivan, P.A., and McKinstry, D . N . ,
Progr. Cardiovasc. Dis.; 21, 195 (1978).
4. Cushman, D.W. and Cheung, H.S., Biochem.
Pharmac., -
20, 1637 (1971).
18. .
Karchmer , J H. , "Treatise on Anal. Chem. ' I ,
Part 11; Volume 13, Page 410 (1966), Edited
By Kolthoff, I.M. and Elving, P.J., John
Wiley Publishers.
19. Ondetti, M., Personal Communication, February
1976.
20. Puar, M.S. and Funke, P.T., Personal
Communication, February, 1976.
21. Kadin, H., Personal Communication, March,
1976.