Note: Descriptions are shown in the official language in which they were submitted.
The present invention relates to a process of producing novel
cephalosporin derivatives. More particularly, the invention
relates to a process of producing the novel cephalosporin deriva-
tives represented by the formula
..
R ~ OEI-CONH ~ ~ ~ (D)m~(E)n
NH N ~ CH2S- ~B ~ R
CO ~ COOH 4 ..
Rl R2 V
''
wherein R represents a hydrogen atom or a hydroxy group; (~ ;.` .
represents a 5- or 6-membered ring having 1-2 nitrogen atoms, a ~. .
quinoline ring, or a -thiopyran ring; R and R , which may be the .. :
15 same or different, each represents a hydrogen atom, a hydroxy ~ ;I
; group, a lower alkyl group, a lower alkoxy group, a lower alkyl- . -
thio group, or an oxo group; ~ represents a 5-membered ring
having 1-4 nitrogen atoms and/or 0-1 sulfur atom, a 6-membered .. ;
. ring having 2 nitrogen atoms, or a .:.
' 20 1,2,4-triazolo ~3,4-b~-1,3,4-thiadiazole ring, if R and R
!I can be present, which may be the same or different, each repre-
~i sents a hydrogen atom, an oxo group, a lower alkyl group, or an ` . .
~' .
amino group; D represents a sulfur atom or a -NHCO- group; E
~ represents an alkylene group or a phenylene group;.G represents .
:'~ 25 a carboxy group or a sulfo group; n is 0 or 1 when m is 0; and
x n is 1 when m is 1, and the non-toxic salts thereof.
f, As the compounds of this invention show antibacterial a~ti-
f ;vities in a wide range of gram positive bacteria and gram negative
:f: ~ .
t bacteria and show, in particular, excellent antibacte:rial ..
~ 30 activities to Pseudomonas and PrOteus strains, they can be appliecl ..
:' .
:
to human beings and animals as agents for treating various dis-
eases infected with these bacteria~
Cephalosporin derivatives effective to Pseudomonas strains
have not hitherto been known bu-t by the discovery of the novel
cephalosporin derivatives of this invention, it becomes possible
to treat the diseases caused by these strains.
Examples of the 5- or 6-membered ring having 1-2 nitrogen
atoms shown by ring ~ in this invention are a pyrrole ring, an ~ ~
imidazole ring, a pyrazole ring, a pyridine ring, a pyrimidine ~ ~-
...
10 ring~ a pyridiazine ring, a pyrazine ring, etc. These rings may -~
be partially saturated. Examples of the 5-membered ring having -
1-4 nitrogen atoms and/or 0-1 sulfur atom .shown by ring ~ are
a pyrrole ring, an imidazole ring, a pyrazole ring, a triazole
ring, a tetrazole ring, a thiazole ring, an isothiazole ring, a ;~
15 1,2,4-thiadiazole ring, a 1,3,4-thiadiazole ring, etc. Also,
examples of the 6-membered ring having 2 nitrogen atoms are a
pyrimidine ring, a pyrazine ring, a pyridazine ring, etc. These ;
1: .
rings may be partially saturated. Example of the lower alkyl
group in R -R or the lower alkyl group forming a lower alkoxy ;~
group or a lower alkylthio group in Rl-R4 are a methyl group, an
ethyl group, a propyl group, an isopropyl group, a butyl group,
etc. Furthermore, examples of an alkylene group in substituent
E are, ~or example, a methylene group, an ethylene group~ a ~ri- ~ -
~ : .
~ methylene group, an isopropylidene group, an ethylidene group, a
. ~: .. . .
propylene group, etc.
he compound~of this invention shown by general formula V
4~
can be;prepared by, for example, reacting the compound represented
by general ~ormula I
R- ~ CH-COMH ~ S
2 O~ ~ ~ CH2OCOCH3
COOH -
-: '' :
j 2
wherein R has the same meaning as in general formula V and the
heterocyclic carboxylic acid represented by general formula II
Rl ~ . :
R2 II ~ ~
.
.:
. wherein ~ , R , and R have the same meani.ng as in general ~ .
~ormula V or a reactive derivative thereof to produce the com~
pound shown by general formula III
:-., ~. ,
R ~ CH-CONE~ S~ III . .. ;
;1 IH O N ~ CM2OCOcH3 .. ~ .
COOEI
~, and then reacting the product with the mercapto-substituted ~ ~ .
.iJ , ~ . .....
heterocyclic ring compound represented by general ~ormula IV
.3~ ~ . ...
~ (D)m-(E)n-G
:~ M-S - ~ R3
.j ~ '. .
:J ~ : wherein M represents a hydrogen atom or an alkali metal atom and
R , R , ~ , D, E, G, m and n have the same meaning as in general -:~
formula V or, more precisely, a heterocyclic compound substituted
by a carboxyl group or a sulfo group, and a mercapto group.
~ ~t
~,; The reaction are illustrated by the ~ollowing reaction
formulae;
R ~ 7H-coNH r~
~: ~ 2 O ~ CH20COCH3 .'.'!~ ' '',
: COOH . ;
~ 3 ;.
; :'. ~ '' '.'
., ~. . .
Step 1 ¦ ~ - COOH II
R-- ~ CH-CONH- r s ~
NH O N \ ~ -cH2OCOCH3 III
1 COOH
C~
(D)m-(E)n-G
Step 2 ~ ~R rv
~ R
R ~ CIH CONH ~ S ~(D)m-(E)n-G
~H O N ~ CH2S ~ R~3 V
2 R
.
wherein ~ ~ D, E, G, R, R , R, R , R , m and n have
the same meaning as in the above formulae.
.
l~ The reaction of step 1 is carried out by reacting the com~
! `
pound of formula I and an almost equimolar or excessive amount
of the compound of formula II or ~he reactive derivative thereof.
'.. :: ::
I ~ 25 When the compound of formula II is used in a free state or
the salt thereof, the reaction is carried out preferably in the .: : .
presence of a condensing agent such as ~ dicyclohexylcarbodi- `,;.
imide; a phosphoric acid trialkyl ester, e.g., triethyl phosphate;
~ ,
a phosphorus oxyhalide, e.g., phosphorus oxychloride; a phosphorus ;
trihalide, e.g., phosphorus trichloride; a thionyl halide, e.g~
. . ',
thionyl chloride; an oxa~olyl chloride; and the like
Examples of the preferred reactive derivatives of the com-
pounds of formula ~I are, for example, acid halides, acid azides;
acid anhydrides; mixed acid anhydrides such as an alkylcarboxylic
acid mixed acid anhydride, an alkyl phosphoric acid mixed acid
anhydride, a dialkyl phosphorous acid mixed acid anhydride, a
sulfuric acid mixed acid anhydride, etc., as well as an active
amide with imidazole, an active ester, such as a p-nitrophenyl
ester, etc.
When an alkylcarboxylic acid mixed acid anhydride, an acid
halide, etc., is used as the reactive derivative of the compound ;
of formula II, the reaction is ordinarily carried out in an
organic solvent such as acetone, tetrahydrofuran, dimethyl for-
mamide, chloroform, dichloromethane, hexamethyl phosphoramide
(hexametapol~, etc., or a mixed solvent thereof in the presence
of a base such as triethylamine, dimethylaniline, etc., under
cooling or at room temperature.
, The compound of formula III formed is usually isolated and
'Ji purified by an ordinary chemical procedure such as extraction,
j 20 recrystallization, etc., and as the case may be, the reaction
mixture containing the compound of formula III is applicable
., .
for the reaction with the compound of formula rv without further
~;~ isolation or purification~
The reaction of step 2 is carried out by reacting the com-
q 25 pound of formula III and an almost equimolar or excessive amounk
of the compound of formula IV. The reaction is ordinarily carried
`1 :
ouk in aqueous solution, in an inert organic solvent such as
acetone, ether, chloroform, nitrobenzene, dimethylsulfoxide,
dimethylformamide, methanol, ethanol, etc., or a miXture thereof.
I'he r~action is also carried out in a neutral state or a slightly
:"
:', , , ,, ,, , , ,, . , ,'
alkaline state and in the cass of using the compound of formula
IV wherein M is a hydrogen atom, the reaction is carried out in
the presence of a basic material such as, for example, an alkali
metal hydroxide, an alkali metal carbonate, triethylamine, etc.
5 Ths reaction may be carried out at room temperature but there is
no particular limitation about the reaction temperature, and ~or
example, the reaction system may be heated to a boiling point of
the solvent used in the reaction.
The order of the reaction of step 1 and step 2 may be rever-
10 sed. The aimed product of formula V may be isolated and purified
by an ordinary chemical operation such as extraction, recrystal-
lization, etc. Furthermore, the compound of formula V can be
converted into a pharmacologically allowable salt thereof, e.g.,
inorganlc salts thereof, such as an alkali metal salt, an
15 ammonium salt, etc., and organic salts thereof, such as a tri-
ethylamine, diethanolamine, lysine, ornithine, et~.
i There are many compounds of formula V thus prepared but in
., .
~'~ the preferred compound of formula V, ring ~ is a pyridine
' ring; substituent R is a hydrogen atom; R' is a hydroxy group or
an oxo group; ring ~ is a thiazole ring, a thiadiazole ring,
J or a tetrazole ring; D is a sulfur atom; E is an alkylene group
.~ .~:,, .
` having 1-2 carbon atoms; G is a carboxy group; m is 0 or 1; and
.~ - .
n is 1.
In more pre~erable compound of formula V of this in~ention,
I 25 ring ~ has a hydroxy group or an oxo group at the 4-position
-1 thereof; the alXylene group is a methylene group, G is a carboxy
group; D is a sulfur atom; the thiadiazole ring of ring ~ is -
a 1~2,4-thiadiazole ring or 1,3,4-thiadiazole ring cmd m is 1.
Practical examples of the compounds of formula V are as
30 follo~s: `
:., ': ~:
7-lD-c~ -Hydroxynico-tinoylamido)~t-phenylacetamido~-3-(5- ~
carboxymethylthio)-1,3,4-thiadiazole-~-yl)thiomethyl-~ -cephem- ~ .
4-carboxylic acid,
7-~D-~-(4-Hydroxynicotinoylamido)~-phenylacetamido~-3-t3- :.
5 carboxymethylthio-1,2,4-thiadiazol-5-yl)thiomethyl-~ -cephem-4- ~.
carboxylic acid, :; .
7- LD-~-(4-Hydroxynicotinoylamido)-~-phenylacetamido~3-(4- :
methyl-5-carboxythiazol-2-yl)thiomethyl-~ -cephem-4-carboxylic .
acid, . -
7- ~D-~-(4-~ydroxynicotinoylamido)-~-phenylacetamido~--3-(1-
carboxymethyl-lH-tetrazol-5-yl)thiomethyl-~ -cephem-4-carboxylic
acid,
7-[D-~-(4-Hydroxynicotinoylamido)-~-phenylacetamidoJ-3-(5-
carboxymethyl-4-methylthiazol-2-yl)thiomethyl-~ -cephem-4-
carboxylic acid,
7-~D-~-(4-Hydroxynicotinoylamido)~-phenylacetamido~-3-(5-
! carboxymethylthio-4-methylthiazol-2-yl)thiomethyl-~ -cephem-4- .. carboxylic acid, ~ ::
7-~D-~-(4-Hydroxynicotinoylamido)-~-phenylacetamido~-3-(5-
20 carboxymethyl-1,3,4-thiadiazo1-2-yl)thiomethy1-~3-cephem-4-
;, . . .
carboxylic acid, :. :
~'~ 7-~D-~-(4-Hydroxy-3-methylpyridin-5-ylcarboxamido)-~-phenyl- .~.
acetamido~-3-(5-carboxymethylthio-1,3,4,-thiadiazol-2-yl)thio-
~f~ methyl-~3-cephem-4-carboxylic acid,
7~[D-~-(4-~Iydroxy-6-methylnicot.inoylamido~-~-ph~nylacetamido~- -
3-(~5-carboxymethylthio-1,3,4-thiadiazol-2-yl)thiomethyl-~3-cepham- .
~; 4-carboxylic acid, and the like.
.1~ Now, ~or illustrating the excellent pharmacological effects
of the compounds of this invention, the values o:E MIC (.~/ml.) of .
i 30 the compounds against various bacteria are shown in I~able 1, in ^ .
which the compounds of -this invention used are shown by example
numbers. ~ ~
~ .
.
'~. 10 , ~ ~ ,
"~
,~ ' " ~. .
Il 15
i' '' ~''., .:"
', ' '~ ': , ,:
.;~i ~, ' ;": `
:~ ' .': ~ ',
j 20
. ~
.: ~
j ~: ~ '',"' '' " '
i 25
~: .:
'. , :, :', '
, ~ '~ . .''
! ;
' `'
8 ..
3 : ~
T A B_L E I
MIC (,~/ml)
Bacteria
Known compound
Cephalexin30 30 - ~100 - - 10 3
Cephaloglycin 25 100 100 ~100 >L00 >100 100 25
Cephazolin3.13 25 6.25?100 ~100 ~100 1.56 1.56
Cephalotin - 12.5 - ~100 ~100 - 3.13 0.78
Compound of0.190.19 0.18 25 50 25 25 6.25
Ex.
4< 0.09 0.190.39 12.5 6.25 6.25 3.13 3.13
5< 0.09 0.190.78 12.5 6.25 6.25 6.25 6.25
6< 0.09 ~0.090O39 12.5 6.25 6.25 3~13 6.25
9 0.19 0.390.39 12.5 12.5 12.5 3.13 3.13
10 <0.09 <0.090.39 25 50 50 6.25 3.13
11 0.39 6.250.56 25 12.5 12.5 25 3.13
12 0.19 0.190.39 25 12.5 25 6.25 0.39
13< 0.09 0.780.39 12.5 6.25 12.5 3.13 3.13
14 0.19 0.780.19 25 12.5 25 3.13 ~.25
0.19 0.39~ 0.09 25 6.25 12.5 6.25 ~.25
16 <0.09 < 0.090.19 25 25 25 6.25 12.5
21 <0.09 0.190.39 12.5 12.5 12.5 3.13 6.25
22 0.19 1.560.39 12.5 6.25 12.5 3.13 6.25
23< 0~09 < 0.090.19 12.5 6.25 12.5 1.56 6.25
~,~ 24 <0.09 < n.oso.ls 12.5 6.25 12.5 3.13 6.25
25< 0.09 1.560.39 12.5 6.25 12.5 3.13 6.25
' 26 0.19 0.390.78 12.5 12~5 25 6.25 12.5
27 <0.09 0.780.78 12.5 6.25 25 3.13 1.56
23 <0.09 12.50.78 12.5 6.25 12~5 3.13 6.~5
30~< ~.09 0.780.78 25 12.5 25 6~25 1~.5
32 <0.09 3.130.78 50 25 50 3.I3 6.25
,~ 33 <0.09 12.50.39 ~5 25 12.5 3.13 25
3; : 34 <0.09 0.780.78 25 12.5 25 12.5 12.5
': :
~L0~ 3
T A B L E I cont'd
MIC ( y/ml)
Bacteria
~ ~ ~ C) ~3)
Known compound
Cephalexin6.25 6.25 6.256.25 0.78
Cephaloglycir 3 3 3 10
Cephazolin 1056 1.56 1.561.56 0.19
Cephalotin 1.56 0.39 6.256.25
Compound of 1.56 0.78 6.25 25 0.78
4 1.56 0.39 0.783.13 0.19
1.56 0.39 1.566.25 0.78
6 1-56 0.39 0.783.13 0.39
9 1.56 0.39 1.566.25 0.39
1.56 0.19 1.566.25 0,39
11 0.78 0.39 6.25 25 0~39
~2 0.19 0.19 1.563 13 0 39
13 1.56 0,39 1.563.13 0.39
14 1.56 0.78 1.563.13 0,39
3.13 0.78 3.133.13 0.39
16 1.56 0.78 3~136.25 0.39
21 1.56 0.39 1.563.13 0.39
22 3.13 0.78 1.563013 0.39
23 1.56 0.39 1.563.13 0.39
24 1.56 0.78 1.563.13 0.78
0.39 0.19 1.566.25 0.39
1 26 6.25 1.56 3.136.25 0.78
f~ 27 0.78 0.39 0.783.13 0.78
f~ 29 1.56 0.78 0.783.13 0.78
f~ 30 6.25 1.56 6.2512.5 0.78
~ 32 1.56 1.56 0.783.13 0.78
3~; ~ 33 3.13 1.56 3.13 3-13
34 6.25 1.56 6.2512.5 0.78
. i , .
ga
3~
; ~ - .:. ., ~,
4:~.483
TABLE I cont ' d .
MIC ( ~/ml) . .
Bacteria
6)
Known compound
Cephalexin
0.78 1.56 1.56 6.25 12.5
Cephalog lycin .
`' 0.3 0.3 1 3 3 . : .
;. Cephazolin
0.19 0.19 0.19 0.78 1.56
; Cephalotin
<0 09 - 0.19 0.39 0,39
Compound of
Ex. 10.393.13 0.78 3.13 6.25
0.19 3.13 0.78 3.13 6.25
50.393.13 1.56 6.25 6.25
- 60,396.25 1.56 6.25 12.5
; 90.393.13 0.78 3.13 6.25 ';
,~ 100.193.13 0.39 3.13 3.13
11O. 396.250.393 . 13 6.25
123.133.13 0.78 3.13 6.25
13o 396.25 0.78 6.25 6.25
140.786.25 1.56 12.5 12.5
150.39 25 3.13 12,5 25
160.193.13 0.78 6.25 6.25 ~
210.393.13 1.56 6.25 6.25 `~;
220.786.25 1.56 12.5 12.5
230.7812.5 3.13 12.5 12.5 ~ ,
2~0,396.25 0.78 6.25 6.25
250.396.25 0.78 3.13 6.25
260 786.25 1.56 6.25 12.5 ;
. ~ . . .
270.786.251.566.2S 6.25
290~786.25 1.56 6.25 12.5 ~ ;~
300.7812.5 1.56 12.5 25
; J0.78 3.13 1.56 3.13 6.25
i'.., ~
330.783.13 3.13 12.5 25
341.5612.5 3.13 12.5 25
. ~
:- ~ ~ . ,: ,
: j: : : :
, ~ - :
' '~ ~ . .~ . ' '
: -
Proteus vulgaris OXK US
Proteus vulgaris OX l9US
Proteus mirabilis IFM OM-l9
Pseudomonas aeruyinosa ATCC 8689
Pseudomonas aeruginosa 99 ~Gentam~ycin resistant~
Pseudomonas ovalis IMA 1002 .
Escherichia coli kauffmann 0-1
Klebsiella pn~umoniae ATCC 10031
Salmonella typhi H901 W ~ :
Salmonella enteritidis
Shigella ~lexneri 2a 1675 .:
Shigella sonnei II 37148 :~
Bacillus megatherium 10778
Bacillus subtilis ATCC 6633 ~;
~ . . .
~ Micrococcus ~lavus AT~C 10240
.
Staphylococcus aureus 209P ~ -
Staphylococcus Shimanishi
Staphylococcus 3numa ~ -
... ..
.
. ... .
.: .,
.: . ..
'":": '~
, ............................. . . . ..
.... . .. .
. .. .
..
3 ~ ~
Other merit of the compounds of this invention is that the
salts, for example, sodium salts of these compounds are easily
dissolved in water at pH o~ living bodies, which makes them
suitable for injections at high concentration. The solubility -
: ,
5 of these compoundc~ are illustrated in Table 2 together with a ~ ;~
control example.
Table 2
..
Test sample
~Exa~nple no.)
4 - b The disodium salt of the compound was dissolved
in water in an ~nount of more than 2~/o ghowing
p~ 7.4.
6 " ;
13
16
23
24 "
. .
26 - b
.,: '
Control sample* The sodium salt of the compound was dissolved
' clearly at 0.5%, the solution of the salt
I became slight turbid at 1%, became turbid at i~
i 5%, bec~ne thickly turbid at l~/o~ and was
gelled at 20%. .. ;
(*): 7-~D~ 4-hydroxynicotinoylamido~ phenylacetamido~-3-t5-
methyl-1,3,4-thiadiazol-2-yl)thiomethyl-~ -cephem-4-
carboxylic acid.
~ Still other merit of the compounds of this invention is that `~ -
,~ 25 they are resistant to ~-lactamase produced by gram negative
`;~ bacteria.
For example, the relation between the ~nount (~/ml) o~ a
unaltered test sample at an initial concentration of the sample `~ ;
~ 100 ~/ml against ~-lact~nase isolated from Pseudomonas aeruginosa
``~ 30 Zl at various dilutions of the enzyme were investigated.
~: .
1 1 ~: , .
; ~ , '~' ' ' '
The results of the investigation is shown in Table 3.
Table 3
Test sample Dilution (magnification) of ~-lactamase
x ~ x~ x8 x 16 x 32 x 6~ x 128 x 25~
Compound o 0 29~6 61,0 100 100 100 100 100 ~ `
Example 4 - b
Cephalothin O O O 0 2.6 11.8 46.8 81.6
Cefazolin O O O O 0 9.2 21.5 84.6
Table 3 (cont.
Test sample Dilution
x 512 x 1024
Compound o~ 100 100
Example 4 - b ;
15 Cephalothin 100 100
Cefazolin 96.7 100
: ~ '.', " '' ' '
Other merit of the compounds of this invention is that they
are not inactivated in the internal organs in bodies. For -~
.'' .: ':.,:
example, when the compounds of this invention and cephalotin are
each homogenated with the internal organs, the compounds of this
invention are not decomposed while cephalotin is decomposed. -;
The relation between the reaction period of time and the ~ -
anti-bacterial activity (residual activity, %) when these com~
: .~:
pounds are incubated with the plasma and the liver homogenate of
a male Sprague-Dowley rat is shown in Table 4. The residual
antibacterial activity was measured by a thin-layer cup me-thod
: ~ .
according to Bacillus subtilis ATCC 6633.
12
, :.,, :.
~ 3
Table 4
A*(%) B*(%~
Time(minute)rrime(minute~
0 15 30 90 0 15 30 90
Compound of 100 97.0 93.9 103.0 100 93.5 91.3 102.2
Example 4
Compound of 100 92.0 102.0 102.0 100 95.7 8~.1 91.3 ~-
Example 6
Compound o~ 100 103.0 92.0 100.0 100 100.0 100.0 102.5
Example 9
'''; ~' '
Cephalothin 100 87~0 66.0 36.0 100 17.2 12.2 12.6
Cephalorid- 100 97.2 89.5 72.2 100 82.3 86.0 91.1
ine
", ' .
A*: Residual activity l%) when incubated with 9~/O plasma.
B*: Residual activity (%) when incubated with l~/o liver homo-
genate. -
~; The compounds of this invention are usually administered by
an intravenous injection or an intramuscular injection~ the
dosage is ordinarily 1 g. psr day per adult, and the amount can
be increased up to 5 g. according to the condition of diseases
and the patient. The dosage for a child is 20-40 mg./kg. and the
amount can be increased up to 100 mg./kg. according to the con-
dition of disease and the patient~
Exam~le l-a
; 25 To 100 ml. of dichloromethane were added 4,2 g. of cephalo-
glycin monohydrate, 3.0 g. of anhydrous magnesium sulfate, and
~ ~ 2,1 ml. of triethylamine and af~er stirring the mixture for one
;~ ~ hour at room temperature, magnesium sul~ate was ~ilterecl off to
provide a dichloromethane solution of cephaloglycin triethylamine
salt. The solution was cooled to -30 C. and then 1~75 CJ. of 4-
13
oxo-4H-thiopyran-3-carbonyl chloride and 20 1 ml. of triethylamine
were added to the solution followed by stirring for one hour.
~ hen, the temperature of the mixture was returned to room
temperature and after stirring for one hour, dichloromethane was
- 5 distilled off under reduced pressure. Wa~er was then added to -~
the residue formed and after adjusting the pH thereo~ to 2 by ~1
adding dropwise 4~/0 phosphoric acid to the mixture, the reaction
mixkure was extracted with 400 ml. of a mixture of butanol and
,~ :
ethyl acetate in 1 : 1 by volume ratio. The extract was washed
with wa~er, washed twice with a saturated aqueous sodium chloride
:~ :,:.:, .
! solution, and after filtering off insoluble matters, the filtrate
~ was dried over anhydrous magnesium sulfate. Then, when a butanol ~ i
'! solution of 30~/0 sodium 2-ethylhexanoate, precipitates formed,
`I which was recovered by filtration, washed with ethyl acetate and
i 15 ether, and then re-precipitated from a mixkure of methanol and
ether to provide 2.8 g. of sodium 7-D-L~-(4-oxo-4H-thiopyran-3- -
~t
carbonylamino)-~-phenylacetamido3cephalosporanate having a
melting point of 185-195C. (decomp.) ;
.'' :,'~,,
3~ Infrared absorption spectra: ~ ~
~, . .
~K cm. : 3450 (NH~, 1770 (~-lactam), 1660 (amide),
~ 1610 (carboxylate), and 1740 (ester).
t Nuclear magnetic resonance spectra: (D6DMS0 -~ CD30D).
2.00 (3M, s, CH3C0), 3.30 (2H, q(AB type), ~ ~ ),
, H ~ -;
25 4~84 (lH, d, ~ ), 4.93 (2H, q(AB type),/~CH2-0-),
~ ~ O
5.60 (lH, d, ~ ), 5.89 ~lH, s, ~ -CH
7,40 ~6H, ~ ~ ), 8.39 ~lH, q,
9.37 (lH, d, ~ ).
14
,3~
Example l-b
In 50 ml . of water were suspended 820 mg. of sodium 7-tD-~-
(4-oxo-4H-thiopyran-3-ylcarboxamido)-~-phenylacetamidolcephalo-
sporanate and 350 mg. of (5-mercapto-1,3,4-thiadiazol-2-yl)-succi -
5 namic acid and then after adding 3 ml. of an aqueous 10% sodium
bicarbonate to the suspension, the mixture was stirr~d for 23
hours while heating to 55 C. After the reaction was over, in
soluble matters were filtered o~f from ~he reaction mixture and
then the pH of the filtrate was adjusted to 2 by adding thereto 1
N hydrochloric acid. The precipitates formed were recovered by
filtration, washed thoroughly with water, and then with ether, and
dried over phosphorus pentoxide under reduced pressuxe. The pro-
duct thus obtained was dissolved in 12 ml of dimethyl sulfoxide
and aeter adding to the solution 0.9 ml of a 30/O n-butanol solu-
tion of sodium 2-ethylhexanoate, the mixture was stirred for 5
minutes. ~hen, 120 ml of ethyl acetate was added to the reac~ion
mixture and the resultant mixture was stirred for 10 minutes. The
precipitates thus formed were recovered by filtration, washed
thoroughly with ethyl acetate and then ether, and dried over
phosphorus pentoxide under reduced pressure to provide 480 mg
the disodium salt Oe 7-LD-~ ~4-oxo-4H-thiopyran-3-ylcarboxamido)-
~-phenylacetamido~-3-[5-(3-carboxypropionylamino)-1,3,4-thiadiazol-
2-yl~thiomethyl-~ -cephem-4-carboxylic acid.
Muclear magnetic resonance spectra (D2O):
~5 S: 2.66 (4~, -NHCOCH2C~ C02Na).
..
In~rared absorption spectra:
~ YmaX cm. : 1762 ~-lactam), 1660 (amide).
Exam~le 2-a
To a solution of 900 mg Oe c~phaloglycin tr:Lethylamine salt
and 17 ml of dichloromethane was added 550 mg of p-nikroE~henyl
3-hydroxypyridazine-4-carboxyl~te under ice-cooling and then after
adding 15 ml of dimethylformamide, the resultant mixture was
stirred for one hour.
Then, the mixture was further stirred for 17 hours at room
temperature. After the reaction was over, 23 ml. o dichloro
methane was added to the reaction mixture and the mixture was
extracted thrice with 40 ml. of an aqueous 1.5% sodium bicarbonate.
The aqueous layer recovered was washed with dichloromethane, the
pH of the aqueous solution was adjusted to 2 with 1 ~ hydrochloric
10 acid, and then the precipitates formed were extracted with 60 ml. ~
of a mixture of bu~anol and ethyl acetate in 1 : 1 by volume ratio. ~--
The extract was washed with water and ~hen a saturatecl aqueous
sodium chloride solution, and dried over anhydrous magnesium
sulfate. To the solution was added 1.3 ml. of a 30/O n-butanol
~' 15 solution of sodium 2-ethylhexanoate, -the precipitates formed were
;~l recovered by filtration, washed with ethyl acetate and then ether,
i~ and dried over phosphorus pentoxide under reduced pressure to
provide 600 mg~ of sodium 7-[D-~-(3-hydroxypyridazin-4-ylcarbox-
amido)-~-phenylacetamido]-cephalosporanate.
Nuclear magnetic resonance spectra tD6-DMSO)
8.16, 8.30 (hydrogen of the nucleus of pyridazine ring).
Infrared absorption spectra:
cm. 1 1760 (~-lactam), 1664 (amide).
Bxample 2-b
In 25 ml. of water were suspended 400 mg. of sodium 7- LD-~- :
(3-hydroxypyridazine-4-ylcarboxamido)-~-phenylacetamido]cephalo- ~;
sporanate and 170 mg. of (5-mercapto-1,3,4-thiadiazol -2--yl)-
~$ succinamic acid and then 1.5 ml. of an aqueous 1~/~ sodium bicar~
~; bonate solution was added to the suspension. ~he resu]tant mix-
~i 30 ture wa~ stirred for 20 hours at 60C. A~ter the reaction was
16
, ., ~ ,' ' ~ ' ':'
over, insoluble matters Eormed was filtered off from the reaction
mixture and the pH of the filtrate was adjusted to 2 by adding
thereto 1 ~ hydrochloric acid~ The precipitates thus formed
were recovered by filtration, washed thoroughly with water, 25 ml.
of a mixture o~ methanol and ether in 1 : 5 by volume ratio, and
then ether, and dried over phosphorus pentoxide under reduced
pressure.
The product obtained was dissolved in 4 ml. of dimethyl
sulfoxide, and a~ter adding to the solution 0.5 ml. of a 30~/0 n-
butanol solution of sodium 2-ethylhexanoate, the resultant mix-
ture was stirred for 5 minutes. To the reaction mixture was
added 50 ml. of ethyl acetate and the mixture was stirred for 10
minutes. q'he precipitates thus ~ormed were recovered by filtra-
tion, washed thoroughly with ethyl acetate and then ether, and
dried over phosphorus pentoxide to provide 260 mg. oE the disodium
salt of 7-tD-~-(3-hydroxypyridazin-4-ylcarboxamido)-~-phenylace-
tamido]-3-{5-~3-carboxypropionylamiino)-1,3,4-thiadiazol -2-yl} ;
thiomethyl-~3-cephem-4-carboxylic acid.
Nuclear magnetic resonance spectra (D20):
~: 2.60 (4H-NHCOC~I2CH2C02Na~
Infrared absorption spectra: ~ `
1 ~m cm. : 1762 (~-lactam), 1664 (amide).
Example 3 a
~¦ After cooling a solution of 1.55 g. of cephaloglycin triethyl-
25 amine salt and 25 ml. of dichlorome~hane to -30 C., 0.60 gO of
2,4-dihydroxypyrimidine-5-carboxylic acid chloride and the mixture
was stirred for 2 hours at temperatures of from 20 C. to -30 C.
and then for 2 hours at room temperature. After the reaction was
,; :. : .
over, the reaction mixture was concentrated under reduced press-
ure and then 30 ml. of water was added to the concentrate. Then,
:' ~ . .'
the p~ of the solu-tion was adjusted to 2 by adding 1 N hydro-
chloric acid and then the precipitates formed were extracted with
120 ml. of a mixture of butanol and ethyl acetate in 1 : 1 by
volume ratio.
The extract was washed with water and a saturated aqueous
sodium chloride solution successively and dried over anhydrous
magnesium sulfate and ~iltered. To the filtrate was added 1.8 ~;
ml. of a 30~/0 n-butanol solution o sodium 2-ethylhexanoate and `
the precipitates formed were recovered by filtration, washed with
..
ethyl acetate and then ether, and dried over phosphorus pentoxide
under reduced pressure to provide 0.72 g. of sodium 7-[D~ 2,4-
' dihydroxypyrimidin-5-ylcarboxamido)-~-phenylacetamido]cephalos~
.j '' '' "
poranate.
Nuclear magnetic resonance spectra (D6-DMS0):
~ 15 J 8.28 (hydrogen of the nucleus of pyrimidine)
s', Infrared absorption spectra:
~KBr cm. 1 1760 (~-lactam), 1676 (amide). ;
~ Example 3-b , ;
,~ In 25 ml. of water were suspended 400 mg. of sodium 7- ~-t~-
20 (2~4-dihydroxypyrimidin-5 ylcarboxamido)-~-phenylacetamido~- ;
~ cephalosporanate and 170 mg. of (5-mercapto-1,3,4-thiadiazol-
,~ 2~yl)succinamic acid and after adding 1.5 ml. of an at~ueous 10% -
sodium bicarbonate solution to the suspension, the mixture was
stirred for 22 hours under heating to 55 C. After the reaction ,~
was over, insoluble matters formed were filtered off from the
3;~ ~ reaction mixture and the pH of the ~iltr~te was adjusted to 2 by
adding thereto 1 N hydrochloric acid~ The precipitates formed
were recovered by filtration, washed thoroughly with water and
~', then ether, and dried over phosphorus pentoxide under reduced
'~ 30 pressure. By treating the product as in Example 2-b, 220 mg. of
.` .
18 ~
the sodium salt o~ 7-lD-~-(2,4-dihydroxypyrimidin-5-ylcarbOXamidO)-
~-phenylacetamido~-3- ¦5-(3-carboxypropionylamino)-1,3,4-thiadiazol-
2~yl) thiomethyl-L\3-cephem-4-carboxylic acid was obtained.
Nuclear magnetic resonance spectrum (D20)
~: 2.62 (4~, -~HCOCH C~I C0 Na)
Infrared absorption spectra:
cm. 1760 (~-lactam), 1664 ~amide).
Example 4-a
In 40 ml. of dichloromethane was suspended 1.4 g. o~ 4-
hydroxynicotinic acid and after adding 1.4 ml. of triethylamine
to the suspension, the mixture was stirred for 30 minutes at room ~-
.j .' :
' temperature to form an almost transparent solution. The solution
;~ was e~ooled to 0-5 C. and 10 ml. of dichloromethane containing
0.73 ml. of thionyl chloride was added dropwise to the solution
i 15 at the same temperature. Thereafter, the mixture was stirred ~or
one hour at room temperature to provide a suspension o~ 4-
hydroxynicotinoyl chloride.
,.~,.. ~, .
In 100 ml. o~ dichloromethane was suspended 4.25 g. o~
`l cephaloglycin monohydrate and then 2.8 ml. of triethylamine was
dissolved therein. Furthermore, 5 g. of anhydrous magnesium
sul~ate was added to the system and after stirring the mixture
for about 10 minutes at room temperature, the mixture was filtered
to provide a solution o~ cephaloglycin triethylamine salt. The
solution was cooled to -20 C. and the suspension of 4-hydroxy-
- -. ., 25 nicotinoyl chloride prepared above was added dropwise to the
solution with stirring. Thereafter, the cooling bath was removed
to raise the temperature up to room temperature and the mixture
was stirred ~or about one hour at room temperature. `
The reaction mixture obtained was concentrated at low
;~ :
~` 30 temperature under reduced pressure and the solicl residue formed
19 ~ .,: .
: ~:
' ,:
was dissolved in 100 ml. of cold water. when the pH of the solu-
tion was adjusted to 2 with hydrochloric acid, white crystals
were precipitated, which were recovered by Eiltration~ washed with
water and a small amount o~ acetone, and then dried.
The crystals thus recovered wPre dissolved in 30 ml. of .
dimethylformamide and insoluble matters formed were filtered off.
. Then, a n-butanol solution of 3~/o potassium 2-ethylhexanoate was .
.~ added to the filtrate until the precipitation ceased. Further-
more, 100 ml. of ether was added to the suspension and after
10 allowing to stand the mixture for a while, the precipitates were ~ .
recovered by filtration and washed with acetone. Then, by puri- .
fying ths precipitates by dissolving them in water~containing .:
dimethylformamide followed by ~iltration and reprscipitating with ;;
the addition of acetone, 1.35 g. of potassium 7-D- ~ -(4-hydroxy- ~ .
pyridine 3-carbonylamino)-~-phenylacetamino]cephalosporanate was
obtained as the yellowish brown powdery crystals. Melting point
195-202 C, (decomp.) .
.,~ ~ . ~, . . .
In~rared absorption spectra: ~ ,
a cm. : 3400 (NH, OH), 1760 (.~-lactam~, 1740 (acetate),
1655 (amide), 1600 (carboxylats).
Nuclear magnetic resonance spectra (CD30D ~ D6-DMSO):
1.97 (3H, CH3COO-), 3.13 (2H, ~ ), 4-8 (2H, CH20),
i ~ 5.01 (1H, ~ t ), 6.6 (lH, ~ ), 5.82 (lH, :~
~: 25 : t ~S- ) :
O ~ `"
~' 6.38 (lH, ~ CH-), 7.4-7.6 (5H, ~ ), . .: ;
` ' ~ NH , .
8.SO (lH, ~ ), 7.86 (lH
l 30
-:- ~
i( ,,` : ~ " ~.:
.:
xample 4-b
To 32 ml. o~ water were added 500 mg. of sodium 7- tD-~-(4-
hydroxynicotinoylamido)-~-phenylacetam,ido~cephalosporanate, 209
mg. of (5-mercapto-1,3,4-thia,diazol -2-yl)thioacetic acid, and
203 mg. of sodium bicarbonate and the mixture was stirred for 22
hours at 55C. Then, the pH 0~ the reaction mixture was adjusted
to about 2 by adding 1 N hydrochloric acid under ice-cooling and
the white precipitates formed were recovered by filtration) washed
thoroughly with water and then ether, and dried to provide 400 `
mg. of 7-CD-O,~4-hydroxynicotinoylamido)-~-phenylacetamidO]-3-(5-
..... . .
carboxymethylthio-1,3,4-thiadiazol -2-yl(thiomethyl-,,~ cephem-4-
carboxylic acid.
Melting point about 250 C. (decomposed gradually).
Infrared absorption spectrum:
1 15 ~ma cm. : 1775 (~-lactam).
Z Nuclear magnetic resonance spectra (D6-DMSO): !' '
ppm: 3.58 (2H), 4.13 (2H), 4.32 (2H), 5.02 (~I), 5.77 (lH),
5.79 (lH), 6.43 (lH), 7.36 (5H), 7.80 (lH), 8,.43 ~lH),
and 9.42 ~lH).
1 20 Ex~mple 5
J To 44 ml. o~ water were added 700 mg. of sodium 7-~D-~-(4-
hydroxynicotinoylamido)-~phenylacetamido]cephalosporanate, 312
mg. of 3-~5-mercapto-1,3,4-thiadiazol -2-ylthio)propionic acid,
and 283 mg. of sodium bicarbonate and then the mixture was stirred
for 2 hours at 55 C. Then, by treating the product as in Example
4-b, 52C mg. of 7-[D- * (4-hydroxynicokincylamido)-~-phenylaceta-
;' ~ mid~ -3-(5-carboxyethylkhio-1,3,4-thiadiazol ~2-yl)thiomethyl~
.. . . .
cephem-4-carboxylic acid was obtained. - `~
Melting point about 250 C. (decomp.)~
Infrared absorption spectrum:
:
21 ~` ~
r cm. 1 1775 (~-lactam).
max
Nuclear magnetic resonance spectra (D6-DMSO): ~
; ~ppm: 2.74 ~2H)~ 3.42 (2~I), 3.58 (2H), 4.32 (2H), 5.02 (lH), ~ -
5.77 (lH), 5.79 (lH), 6.43 (1~), 7.36 (5H), 7.80 (lH),
8.43 (lH), and 9.43 (lH).
Example 6
To 40 ml. of water were added 615 mg. of sodium 7-~D~ (4-
hydroxynico-tinoylamido(-~-phenylacetamido~-cephalosporanate, 235
mg. of 3-(5-mercapto-1,3,4-thiadiazol -2-yl)thiopropionic acid,
and 218 mg. of sodium bicarbonate and the mixture was stirred Eor
2 hours a-t 55 C. Then, by treating the product as in Example
, 4-b, 450 mg. of 7-LD-~-(4-hydroxynicotinoylamido)-~-phenylacet-
amido]-3-(5-carboxyethyl-1,3,4-thiadiazol -2-yl)thiomethyl-a -
cephem-4-carboxylic acid was obtained.
Melting point about 250 C. ~decomp.)
In~rared absorption spectrum:
~m x cm. : 1775 (~-lactam).
Nuclear magnetic resonance spectra (D6-DMSO)
~ppm: 2.73 (2H), 3.25 (2H), 3.58 (2H), 4.35 (2~I), 5.01 tlH~,
5~78 (lH), 5.80 (~ 6.44 (lH), 7.36 (5H), 7.81 (lH),
1 8.44 (lH), 9.43 (lH).
.~ .; . .
Example 7-a ;
' To a suspension consisting o~ 4.23 g. of cephaloglycin mono-
',~ hydrate, 3.0 g. of anhydrous magnesium sulfate, and 70 ml. of
25 methylene chloride was added 3.1 ml. of triethylamine and a~ter ~ -
stirring the mixture for one hour at room temperature, magnesium
i~ ... .
sul~ate was filtered off to provide a triethylamine salt solution
o~ cephaloglycin. To the solution was added dropwise a solution
~; . ,
of nicotinoyl chloride ~-oxide prepared by adding dropwise 0.73 ml.
o~ thionyl chloride to a solution consisting of :L.39 g. of
: - ,
- 22 -
nicotinic acid N-oxide, 1.4 ml. of triethylamine, and 30 ml. of
methylene chloride under ice-cooling followed by stirring for
1.5 hours. In this case, the pH of the reaction mixture was
maintained at 8-9 by adding triethylamine.
Thereafter, the reaction mixture was stirred for 3 hours at
a temperature of -20 + S C. and then allowed to stand overnight
at -20 C. To the residue formed by distilling o~f n butanol from
; the reaction mixture were added ice-water and a mixture of n- -
butanol and ethyl acetate in 2 : 3 by volume ratio and adjusted
the p~ thereo~ to 2.0 by th~ addition of dilute hydrochloric
acid.
rrhe organic layer formed was recovered, washed thrice with
water, dried over anhydrous magnesium sulfate, and was mixed with
a n-butanol solution of 30% sodium 2-ethylhexanoate, whereby
~ 15 precipitates formed. By recovering the precipitates by filtration
I and re-precipitating them with a mixture of methanol and ether,
~ 2.7 g. of sodium 7 ~D-~-(pyridine-N oxide-2-ylcarboxamid) ~
, ... ..
phenylacetamido~cephalosporanate was obtained.
' Melting point about 263 C. (decomp.)
I~rared absorption spectra:
~m cm. : 3300 (N~I), 1760 (~-lactam), 1735 lester), ;~
1665 (amide), and 1615 (carboxylate).
. .: . ,. . . .: .
Nuclear magnetic resonance spectra ~D6-DMS0)~
`~ ~ppm: 2.00 (s, 3H, -OC-CH3), 3.48 ~q, 2H, ~ H )~ ;
,~25 4.92 (d, lH, ~ ), 4.95 (q, 2H, ~ -CH2-O~
,~5.60 (q, lH, ~ 3, 5.91 (d, lH, ~ _C-CO-), ~ ;;
N ~ -~
7.42 (m, 7H, ~ ~ ~ C0_), 8.42 ~q, lH,
::. -
23
~' , '' ' .
~0 ~ ), 8.47 (q, lH, ~ ), 9.46 (d, lH,
C0- H
O -
-NH), and 12.14 (d, lH, -NH3
Example 7-b
To a solution of 400 mg. o~ sodium 7-[D~ (pyridine-M-oxide-
2-ylcarboxamido)-~-phenylacetamido3cephalosporanate in 25 ml. of
water was added 190 mg. of (5-mercapto-1,3,5-thiadiazol -2-yl)~
succinamic acid at room temperature and ~urther a saturated
10 aqueous sodium bicarbonate solution was added to the mixture to .
adju~t the pH of the solution to 6.8-7.2. The solution was
stirred for 26 hours at 55-60 C. and the pH thereo~ was then ;.
adjusted to 2.0 with diluted hydrochloric acid to form a suspen- ,. :
sion. The product thus obtained was extracted with a mixture of
15 n-butanol and ethyl acetate in 2 : 3 by volume ratio and the ` :
organic layer formed was washed thrice with water, dried, and
then mixed with a n-butanol solution of 30/0 sodium ~-ethylhexa-
noate and then ether to provide an oily material. The oily
material thus obtained was dissolved in methanol and ether was
20 added to the solution, whereby precipitates formed, which were .
recovered by ~iltration, washed with ether, and dried to provide ~ .
0.25 g. of sodium 7- CD-~-(pyridine-N-oxide-2-ylcarboxamido)-~
phenylacetamido-3- [5-~3-carboxypropionylamino3-1,3i4-thiadiazol-
2-y ~ thiomethyl-~ -cephem-4-carboxylate.
~ Melting point about 251 C. (decomp~
: . :
,. :
Infrared absorption spectra~
K~r cm. : 3300-3400 (broad, ~H~, 1760 (~-lact~n~
1665 (amide), 1590 (carboxylate3, ~
Nuclear magnetic resonance spectra (D20)~ -
30: ~ppm: 2.49 (m, 4H, -COCH2C~-I ), 3013 (q, 2H, ~ H
:: ,
24
,: :.
~3 : :
~CH2-S_ )~ 4-80 (d, lH, ~ - r~~
5.54 (d, lH, r~ ) 5 57 ( ~ C_<
: ,
7.30 (m, 7H, ~ -~ ~ H )~ 7.96 (q, lH, ~ ),
O O .: ' .~' .
and 8.18 (q, lH, H ~ )-
Exampla 8-a ;
After stirring 35 ml. of a methylene chloride solution con- `-
i taining 2.12 g. o~ cephaloglycin monohydrate, 1.5 g. of anhydrous
i~ magnesium sulfate, and l.4 ml. of triethylamine for one hour at
i~, room temperature "nagnesium sulfate was filtered oPf to provide
a solution of triethylamine salt of cephaloylycin. A~ter cooling
,i 15 the solution to -25c., 816 mg. of 2-imidazolidone-1-carboxylic
,lj: . ~. . .
acid chloride and urther 0.3 ml. of triethylamine were added
thereto.
sj . , ~,:
The mixture thus o~tained was stirred for 3 hours at -20 +
5 C and then allowed to stand overnight at -20 C. Then, the
solvent was distilled off from the reaction mixture under reduced
pressure and the residue obtained was dissolved in ice water.
The pH of the solution ~as ad~usted~ to 2 with diluted hydrochlori~
acid and the precipitates formed were recovered by filtration,
w~ashed with wa~er, and dried to give 2.1 g. o~ the precipitates. ~ ;
~The~precipitates were dissolved in 20 ml. of dimethylformamide and
then~3~m~1. of a n-butanol solution of 30/0 sodium 2-ethylhexanoate
was~added to the solution. Then, ethyl acetate was added to the
mlxture, whereby precipitates formed, which were recovered by
filtra~ion, washed with ether, and re-precipitated ~rom a mixture
0~ o~ mothanol and;ether to provide 1.5 g. of sodium 7-LD-~- (2~
. , . .:
imidazolidon-l ylcarboxamido)-~-phenylacetamido~cephalosporanate.
Melting point about 225 C. (decomp.)
Infrared absorption spectra:
~ma cm. : 3100-3500 (~H), 1700 (~-lac~am), 1720 (ester,
-N-C-N-), 1660 (amide), and 1605 (carboxylate).
Nuclear magnetic resonance spectra (D6-r)MSO).
J 1.98 ~s, 3H, -OCCH3), 3.28 (q, 2H, ~ H ), 3032,
3.69 (4H, H ~ H ~, 4.88 (d, lH, ~ , 4.91
o , . ..
(9.2H, ~ ) 5.55 (c~, lH, ~ ,_~, 5.66
(d, lH, ~ ), 7.34 (m, 5H, ~ ), 7.78
(lH, -NH), 9.13 (d, lH, -NH), 9.35 (d, lH, -NH).
Example 8-b
l To an aqueous solution of 800 mg. of sodium 7-LD_~_~2_0XO_
i imidazolin-l-ylcarboxamido)-Q~phenylacetamido]cephalosporanate
1 in 50 ml. of water was added 0.38 g. o~ (5-mercapto-1,3,4-thia-
diazol -2-yl)succinamic acid and then the pH of the solution was
adjusted to 7.2-7.5 by adding a saturated sodium bicarbonate
.j . ...
solution. The solution was stirred ~or 24 hours at 55-60 C. and
after cooling, the pH of the solution was adjusted to 2.0 by
diluted hydrochloric acid whereby precipitates formed. Ths pre~
cipitates were recovered by filtration, washed with water, dried,
and dissolved in 8 ml. of dimethylformamide. Then, 1 ml. of a
n-butanol solution of 3~/o sodium 2-ethylhexanoate was added to the
solution and then ethyl acetate was added thereto ~o ~orm pre-
cipitates, which were recovered by Piltration, washed wil:h ether, ~ `
.,~ ;
and re-precipitat~d from a mixture o~ methanol and ether to provide
26
. . :
. ~ ~ , , ,,, , , . , ~ :
750 mg. of disodium 7-lD-~'-(2-oxoimidazolin-1-ylcarboxamido)~
phenylacetamido~-3-l5-(3-carboxypropioIlylamino)-1,3,4-thiadiazol-
2-yl)thiomethyl-~ -cephem-4-carboxylate.
Melting point: about 246 C. (decomp.)
In~rared absorption spectra:
~ma cm. : 3200-3500 (NH), 1760 (~-lactam), 1720 if`
(NHCNH), 1675, 1660 (amide), 1600 (carboxylate).
Nuclear magnetic resonance spectra (D2O)
~ ppm: 2.56 (dd, 4H, -CO~H2CEI2COO-), 3.28, 3.60 (m, 4H,
H I I H ),
N N
~f :,. ,
4.82 (d, lH, ~ ~ ), 5.36 (s, lH, ~ ),
;... ...
~5 ~4 ~d, lH, ~ 1 ~ (, 3.92 (q, 2H, ~ CH ~;
ii 15
S ~ H ~ ~`
3.13 (q, 2H, ~ H )- -
'~Examp le 9 : :
To a mixture of 0.53 g. Oe sodium 7- ~D-~-(4-hydroxynicotinoyl-
amido)-~-phenylacetamido~cephalosporanate and 0.3 g. o e sodium
5-mercapto-1,3,4-thiadiazol -2-yl-thioethylsulfonate were added
35 ml. of water and 0.108 g. of sodium bicarbonate and the resul-
, tant mixture was stirred for 22 hours at about 57 C. Then, after
I ~ removing insoluble matters by filtration, the pH of the filtrate
was adjusted to about 1 with 1.5 N hydrochloric acid under ice-
'Z , .,
q 25 cooling and the precipitates thus formed were recovered by fil-
i tration. The preclpitates were washed with water and then ether
and dried to give 0.45 g. of 7-~D-~-(4-hydroxynicotinoyl~mido)~
phenylacetamido3-3-(5-sulfoethylthio-1,3,4-thiadiazol -2-yl)thio- ;
methyl-~ -cephem-4-carboxylic acid. :
Melting point: 200-220 C. (gradually decomposed).
27
, . . . ... .
., .~ '', ',
Infrared absorption spectrum:
KBr -1
~ cm. : 1774 (~-lactam~.
Nuclear magnetic resonance spectra (D6-DMS0):
~(ppm): 1.89 (2H), 3.47 (2H), 3.59 (2H), 4.34 (2H), 5.05
(lH), 5.79 (lH), 5.87 (lH), 6.52 (lH), 7.41 (5H),
7.89 (lH), 8.49 (lH), and 9.47 (1~3,
Example 10
To 45 ml. o~ water were added 700 mg. of svdium 7- tD-~-(4-
oxo-4H-thiopyran-3-ylcarboxamido~ ~-phenylacetamido~cephalospora-
nate, 284 mg. of (5-mercapto-1,3,4,-thiadiazol -2-yl)thioacetic
acid, and 260 mg. of sodium bicarbonate and the resultant mixture
was stirred for 20 hours at 50-54 C. When the pH of the reaction
mixture thus obtained was adjusted to about 2 with 1 N hydro~
chloric acid under ice-cooling and the precipitates formed were
recovered by filtration, 530 mg. of the aLmed product, 7- LD-~-(4-
oxo-4H-thiopyran-3-ylcarboxamido)-~-phenylacetamido]-3-(5-carbOxy-
methylthio-1,3,4-thiadiazole-2-yl)thiomethyl-~3-cephem-4-
carboxylic azid was obtained.
Melting point about 190 C. (decomp.)
: 20 Infrared absorption spectrum:
cm. 1 1778 (~-lactam).
Nuclear magnetic resonance spectra (D6-DMSO)~
3.58 (2H), 4.14 (2H), 4.33 (2H), 5.02 (lH) r 5.77 (2H) ~ . . :
7.35 (6H), 8.~9 ~lM), 9~34 (lH), and 9.48 ( lH) .
~xam~le 11
To 50 ml. of water were added 0.8 g. of sodium 7-~D~-(4-
hydroxynicotinoylamido)-~-phenylacetamido~cephalosporanate, 0.281
~;~ g. of 5-mercapto-1,3,4 thiadiazol -2-carboxylic acid, and 0.296 g.
of sodium bicarbonate and the resultant mixture was stirred for
,:
~ 30 20 hours at 55 C. Then, the p~ of the reaction mixture ~hus
~ . .
~ 28 ~
:., ' ',`',' ' '. ~'
obtained was adjusted to 1-2 with 1 N hydrochloric acid under
ice~cooling and the white precipitates formed were recovered by
filtration, washed with water, and dried to provide 0.5 g. of
7-tD-~-(4-hydroxynicotinoylamido)-~-phenylacetamido~-3-(5-
S carboxy-1,3,4-thiadiazol ~2-yl)thiomethyl-~3-cephem-4-carboxylic
acid.
Infrared absorption spectrum:
cm. : 1770 (~-lactam). `~
max
Nuclear magnetic rasonance spectra (D6-DMSO)
~ppm: 3.59 (2H, q), 4.39 (2H, q), 5.02 (lH,d), 5.72 (lH),
5.84 (lH, d), 6.44 (lH, d), 7.37 (5H), 7.82 (lH, d),
8.45 (lE~, s), 9.44 (lH, d), 11.23 (lH, d), 1220 (lH,
broad).
Example 12
15To 70 ml. of water were added 700 mg. of 7- tD-~-~7-methyl- l~ -
thio-4-quinolone-3-carboxamido)~d-phenylacetamido~cephalosporanic
acid, 220 mg. o~ (5-mercapto-1,3,4-thiadiazol -2-yl)thioacetic
acid, and 315 mg. of sodium bicarbonate, and tha resultant mixture
was stirred for 20 hours at 55-60 C. After tha reaction was over,
20 the raaction mixture obtained was cooled, acidified with diluted ;;
hydrochloric acid, and the precipitates formed were recovered by ;:
filtration, washed well with water, and dried -to provide 400 mg.
of 7-~D-~-(7-methylthio-4-quinolon -3-ylcarboxamido)-~-phenyl-
. ,: .
acetamido~-3-(~-carboxymethylthio-1,3,4-thiadiazol -2--yl)thio-
methyl-~ -caphem-4-carboxylic acid.
Infrared absorption spectrum:
~ma cm. : 1780 (~-lactaim)~
~uclear magnetic resonance spectra (D6-DMSO):
ppm: 2.6Q (3H), 3.56 (2~I), 4.16 (2EI), 4.20 ~2~1), 5.05 (lH),
5.84 ~lH), 5~93 (lH), 7.40 (7H), 8.18 (lH) "3.70 (lH),
29
9.50 (lH), 11.00 (lH), and 12.60 (lH).
Example 13
In 50 ml. of water were suspended 0.80 g. o~ 7-[D-~(4-
hydroxynicotinoylamido ~-~-phenylacetamido~cephalosporanic acid
and 0.32 g. of (5-mercapto-1,2,4-thiadiazol -3-yl1thioacetic acid
and after adding further 0.45 g. of sodium bicarbonate to the
suspension, the mixture was stirred for 24 hours while heating
to 55 C. After the reaction was over, insoluble matters were
filtered off from the reaction mixture and the pH of the filtrate
was adjusted -to 2 by adding 5% hydrochloric acid to form precipi-
tates, which were recovered by filtration, washed wel:L with water
and then with water, and dried over phosphorus pentoxide under
: '
reduced pressure to provide 0.70 g. of the powder of 7- ~D-~-(4-
hhydroxy~icotino~ylamido)-~-phenylacetamido~-3-(3-carboxymeth
thio-1,2,4-thiadiaæol -5-yl)thiomethyl-~3-cephem-4-carboxylic
acid.
Melting point: 189-193 C~ (decomp.)
Infrared absorption spectra:
max cm. : 1770 (~-lactam) and 1650 (amide).
~ 20 Nuclear magnetic resonance spectrum (D6-DMSO~
f ; .
`~ d ppm: 4.04 (2H~ -SCH2C02H)
Example 14 - ;
1~ To 50 ml. of water were added 0.8 g. of sodium 7- LD-~-(4-
; hyd~o~y~icoti~oylamido)-~-phenylacetamido~cephalosporanate~
~,~ 25 0.293 g. of 2-mercapto-4-methylthiazole-5-carboxylic acid, and
0.296 g. o~ sodium bicarbonate and the resultan~ mi~ture was
stirred for 22 hours at 55C. Then, the pH of the reaction
mixture was adjusted to 1-2 with 1 N hydrochloric acid under ice-
cooling. The white precipita-tes thus formed were recovered by
. :. .
filtration, washed with water, and dried to provide 0.6 g. of
~' ,. .
7_CD_~_(4 hydroxynicotinoylamido )-~-phenylacetamido~-3~
methyl-5-carboxythiazol -2-yl) thiomethyl-L~3-cephem-4-carboxylic
acid.
Infrared absorption spectrum:
~max cm. : 1770 (~~lactam~
Nuclear maynetic resonance spectra ~D6~DMS0):
Jrppm: 2.56 (3H, S~, 3.56 (2H, q) 4.37 (2H, q) 5.01 (1H,
d), 5.74 (lH), 5.86 (lH, d), 6.44 (lH, d), 7.37 (5H),
7.82 (lH, d), 8.44 (lH, d), 9.44 (lH, d), 11.21 (lH,
d~, and 12.21 (lH, broad~. ;
EXAMPLE 15 -
To 40 ml. of water ~ere added 600 mg. of sodium 7-~D-a-(4-
hydroxynicotinoylamido)-a-phenylacetamido]cephalosporanate, '~
195 mg. of 5-mercapto-1-carboxymethyl tetrazole, and ]80 mg. of
sodium bicarbonate and the resultant mixture was stirred for 22
. .
hours at 55 C. The reaction mixture thus obtained was filtered
and the pH of the filtrate was adjusted to 1 with 1~ hydrochloric
~;~ acid under ice-cooling. The precipitates formed were recovered
~, . .; . . .
l; by filtration, washed with water and then with ether, and dried ~ ~
.. . . .
~, to provide 300 mg. f 7~ CD ~ ( 4~hydroxynicotinoylamido )-~-phenyl-
'~ 20 acetamido~-3-(1-carboxy~ethyltetrazol -5-yl)-thiomethyl-~3-cephem-
4-~carboxylic acid. ~ ;
Melting point: about 240 C. (gradually decomposed3
:.
Infrared absorption spec~rum: -
max cm. : 1775 (~-lactam3.
Nuclear magnetic resonance spectra (D6-DMS0)~
ppm: 3.60 (2H), 4.36 (2H3, 5.00 (1~), 5.34 (2H), 5.84 (2H),
6.48 (lH), 7.44 (5H), 7.88 (lH), 8.50 (lH), 9.52 (lH).
Example 16
l To 30 ml. of water were added 0.45 g. o~ sodium 7- LD-~-(4-
hydroxynicotinoyLamido)-~-phenylacetamido~cephalosporanate, 0.2 g.
of 6-carboxymethylthio-4~mercaptopyrimidine, and 0.14 g. o:E sodium
bicar~onate and the resultant mixture was stirred for 22 hours at
55 C. Af-ter Eilteriny the reaction mixture thus obtained, the
pH of the filtrate was adjusted to 1 with 1 N hydrochloric acid
under ice-cooling and the precipitates thus formed were recovered
by filtration, washed with water and then ether, and dried to
provide 0.42 g~ of 7 ~D~-(4-hydroxynicotinoylamido)-d~phenyl-
acetamidolZ-3-(6-carboxymethylthiopyr~midin -4-yl)thiomethyl-~ -
cephem-4-carboxylic acid.
Infrared absorption spectrum:
~m cm. : 1775
Nuclear magnetic resonance spectra (D6-DMSO):
~ppm: 3.50 (2H), 3.98 (3H~, 4.58 (lH), 5.00 (lH), 5.74 (2H),
6.42 (lH), 7.34 (6H), 7.78 (lH), 8.42 (lH), 9.40 (lH).
Example 17
To 37 ml. of water were added 0.5 g. of 7- CD-~-t4-hydroxy- ~
nicotinoylamido)-~-phenylacetiamido~cephalosporanic acid, 0.235 g. ~- -
of 6-carboxymethylthio-4-mercapto-2-methylpyrimidine, and 0.295 g.
of sodium bicarbonate and the resultant mixture was stirred for 22
hours at 50-55 C. The reaction mixture thus obtained was filtered
j and the pH of the filtrate was adjusted to about 2 with 1 N
; .:
hydrochloric acid under ice-cooling. The precipitates formed
were recovered by filtration, washed with water and then ether,
and dried under reduced pressure to provide 0.42 g. of 7- ~D-
~
(4-hydroxynicotinoylamido)-~-phenylacetamidolZ-3-(6-carboxymethyl-
~ 3
J thio-2-methylpyrimidin-4-yl)thiomethyl-a -cephem-4-carboxylic
~ . . ,
acid.
Infrared absorption spectrum: ~
~ cm. : 1778 (~-lactam). ;;
Z~ 30 ~uclear magnetic resonance spectra (D6-DMSO):
32
` "'' '.' ~::: .-
7~
J ppm 2.50 (3H3, 3.52 (2H), 3.82 (lH), 3.98 (2H), 4.62 (lH),
5.02 (lH), about 5.73 (lH), 5.85 (lH), 6.45 (lH),
7.~3 (lH), 7.38 (5H~, 7.82 (lH), 8.46 (~I), and
9 44 (lH)-
~xample 18
TO 37 ml. of water were added 526 mg. f 7-LD d (4-hydroxy- ,~
nicotinoylamido)~-phenylacetamido~cephalosporanic acid, 220 mg.
-~ of l-carboxymethyl-3-mercapto-6(1H)-pyridaz:inone, and 300 mg. of
~- sodium bicarbonate and the resultant mixture was stirred for 22
hours at 50-55C. After the reaction was over, the reaction
mixture obtained was filtered and the pH of the filtrate was then
adjusted to about 2 with 1 N hydrochloric acid under ice-cooling.
The precipitates thus ~ormed were recovered by Eiltration, washed
j with water and then ether, and dried under reduced pressure to
provide 350 mg. of 7-LD-~-(4-hydroxynicotinoylamido(-d-phenyl-
acetamido]-3-(1-carboxymethyl-6(lH)-pyridazinon -3-yl)thiomethyl-
cephem-4-carboxylic acid. ~-
Infrared absorption spectrum: ~
cm. : 1775 (~-Iactam)- ~ ~;
;j 20 Nuclear magnetic re~onance spect~a (D6-DMS0):
~(ppm): 3.46 ~2H), 3.8~3 tlH), 4.33 (lH), 4.66 (2H), 4.93 (1
5.74 (2H), 6.40 ~lH), 6.88 (lH), 7.34 ~6H), 7.74 (lH), ~s ;~
8.40 (lH~, and 9.41 (lH).
xample l9
~o 30 ml. of water were added 420 mg. of 7- LD-~-(4-hydroxy- `-
niootinoylamido~-~-phenylacetamido~cephalosporanic acid, 220 mg.
of 7-amino-4-carboxymethylthio 6-mercaptopyr.imidine, and 240 mg.
o~ sodium bicarbonate and by treating the resultant mixture as
.. . .
. . "
;~ in Example 18, 310 mg. of 7-~D-~-(4-hydroxynicotinoylamido)J~-
30~ phenylacetamido]-3-(2-amino-4-carboxymethylthiopyrimidin-6-yl)-
~:
thiomethyl-~ -cephem-4-carboxylic acid was obtained.
nfrared absorption spectrum:
~KBr cm 1 1770
max
Nuclear m~gnetic resonance spectra (D6-DMS0) `-
~(ppm): 3.52 (2H), 3.92 (3~I), 4.46 (lH), 5.02 (lH), 5.76
(2H), 6.44 (2H), 6.68 (2H), 7~36 (5H), 7.82 (lH),
8.46 (lH), and 9.44 (lH). s
Example 20-a
To 20 ml. of methylene chloride were added 290 mg. oE 4-
hydroxy-2-methylpyrLm:idine-5-carboxylic acid and 0.3 ml. of tri-
:, . .
ethylamine and after cooling the resultant mixture to -10 C. with ~ -~
skirring, a solution of 148 mg. of ethyl chlorocarbonate in 3 ml.
of methylene chloride was added dropwise to the mixture. The
resultant mixture was stirred then for one hour at -10 C. and a
solukion of 500 mg. oE cephaloglycin and 0.2 ml. of triethylamine
in 10 ml. of methylene chloride was added to the mixture. The
resultant mixture was further stirred for one hour at 10 C. and
then for one hour at room temperature.
After the reaction was over, the solvent was distilled off
. . .
j 20 under reduced pressure and 5 ml. of water was added to the residue
After further adding thereto 30 ml. of a 1 : 1 mixture of n-
butanol and ethyl acetate, the pH of the mixture was adjusted to -~
3 with 1 N hydrochloric acid followed by stirring well. Then, an
,
organic solvent layer formed was recovered, washed with water and
then a saturated aqueous sodium chloride solution, and then dried
over anhydrous magnesium sulfate. The reaction mixture thus
~ obtained was concentrated at room temperature and then 20 ml. oE
i . .;~
ether was added to the xesidue. The precipitates formed were ~:
recovered by filtration, washed with water and then ether, and
,,: ... . ,,:
'~30 dried under reduced pressure to provide 300 mg. of 7-~D-d-(4
'~
~ '. , ' ':
hydroxy-2-methylpyrimidine-5-ylcarboxamide)-~-phenylacetamidol-
cephalosporanic acid.
Infrared absorption spectrum:
~ cm. : 1780 ~-lactam).
Nuclear magnetic resonance spectra (D6 DMSO~:
~ppm): 2.00 (3H), 2.41 t3H), 3.41 t2H), 4.64 (lH), 4.95
(lH), 5.04 tlH), 5.80 (lH), 5.84 (lH), 7.39 (5H),
8.55 tlH). k;~
Exi~mple 20-b
In 18 ml. of water were dissolved 300 mg. of 7-[D-~-t4-
hydroxy-2-methylpyrimidin -5-ylcarboxamido)~-phenylacetamido~-
cephalosporanic acid, 112 mg. of 2-mercapto-1,3,4-thiadiazol -5-yl-
thioacetic acid, and 183 mg. of sodium bicarbonate and by treating
the solution obtained as in Example 18, 240 mg. o~ 7-~D-~-(4-
~ 15 hydroxy-2-methylpyrimidin -5-ylcarboxamido)-~-phenylacetamido1-3- ~ -
1 (5-carboxymethylthio-1,3,4-thiadiazol -2-yl)thiomethyl-~ -cephem-
~ 4-carboxylic acid was obtained.
3 Infrared absorption spectrum:
K cm. : 1780 (~-lactc~m).
Nuclear magnetic resonance spectra tD6-DMSO): `
ppm): 2.36 t3H), 3.56 t2H), 4.12 t2~1), 4.20 tlH), 4.50
tlH), 5.02 (lH), 5.72 (lH), 5.84 tlH), 7.38 ~5H),
and 8.60 (lH).
~ Example 21
j 25 To a mixture of 0.5 g. of 7- LD-~-~4-hydroxynicotinoylamido)- ;
~-phenylacetamido]cephalosporanic acid and 275 mg. of 2-(1- ` ;
~ carboxyethylthio)-5-mercapto-1,3,4-thiadiazole were added 310 mg.
i ~ of sodium bicarbonate and 37 ml. of water and the resultant
mixture was stirred for 22 hours at 50-55 C. Insoluble matters
30 were filtered o~ ~rom the reaction mixture thus obtained and the ~ ~
. ~ .
' .' ., '''' '
.
pH of the filtrate was adjusted to about 2 with 1 N hydrochloric
acid under ice-cooling. The precipitates formed were recovered
by filtration, washed with water and then e-ther, and clried under ``
reduced pressure to provide 0.4 g. of 7-[Dv~-(4-hydroxynicotinoyl-
5amido)-O~phenylacetamido~-3-~5 (1-carboxyethylthio)-1,3~4-
thiadiazol -2-yl~thiomethyl-~3-cephem-4-carboxylic acid. ;
Infrared absorption spectrum:
~ K cm. : 1778 (~-lactam).
Nuclear magnetic resonance spectra (D6-DMSO):
~; 10 ~ppm): 1.54 (3~), 3.58 (2H), 4.20 (lH), 4.37 (lH), 4O45
tlH), 5002 (~ about 5.76 (lH), 5.85 (lH), 7.39
(5H), 7.82 (lH), 8.45 (lH), and 9.45 (lH).
le 22
~ To a mixture of 500 mg. of 7-LD-d-(4-hydroxynicotinoylamido)-
;, 15 ~-phenylacetamido~-cephalosporanic acid and 292 mg. of 2-(1-carboxy- `
l-methylethylthio)~5-mercapto-1,3,4-thiadiazole were added 320
i mg. of sodium bicarbonate and 37 ml. of water and by treating the
.! resultant mixture as in Example 21, 370 mg. of 7-~D-~-(4-hydroxy-
I nicotinoylamido)-~-phenylacetamido~-3-[5-(1-carboxy-1-methylethyl- ',~ 20 thio)-1,3,4-thiadiazol -2-yl~thiomethyl-~3-cephem-4-carboxylic
~ acid was obtained.
~,,~ .~ . . .
Infrared absorption spectrum~
,~ ~ cm. : 1777 (~-lactam).
i ~uclear magnetic resonance spectra (D6-DMS0) ,~
~ ppm): 1.57 (6H), 3.59 (2H~, 4.23 (lH), 4.45 (lH), 5.01
~ (lH), about 5.75 (lH), 5.84 (lH), 6.43 (lH), 7.37
;1 ~5H), 7.81 ~lH), 8.43 ~lH~, and 9.42 (lH).
Example_23
. . .
To 40 ml. of water were added 0.63 g. of 7-LD-~-(4-hydroxy-
nicotinoylamido)~ phenylacetamido]cephalosporanic acicl, 0.25 g.
36
' ` "~ ;'
of 2-mercapto-4-methylthiazole-5-acetic acid, and 0.3 g~ of
sodium bicarbonate and the resultant mixture was stirred for one
hour at about 55 C. The p~ of the reaction mixture thus obtained
was adjusted to about 2 with 1 N hydrochloric acid under ice-
cooling and the precipitates formed were recovered by filtration,washed with water, and dried to provide 0.55 g. of 7-tD~-(4-
hydroxynicotinoylamido)-~-phenylacetamido]-3-(5-carboxymethyl-4-
methylthiazol -2-yl)thiomethyl-d -cephem-4-carboxylic acid. ~-
Infrared absorption spectrum:
~KBr cm -1 1772 (~-lactam)
Nuclear magnetic resonance spectra (D6-DMSO):
ppm): 2.20 (3H), 3.53 (2H), 3.72 (2H), 4.02 (lH), 4.46
~lH), 4.97 (lH), 5.71 (lH), 5.82 (lH), 6.42 (lH),
7.33 (5I~), 7.78 (lH)~ 8.40 (lH), 9.39 (lH), 11.15
(lH), and 12.02 (lH).
Example 24
: . ~. .. :
~ To 50 ml~ o~ water were added 0.8 g. of sodium 7- ~D-~-(4- ~
_
hydroxynicotinoylamido)-~-phenylacetamido cephalosporanate, ;`
0.386 g. of (2-mercapto-4-methylthia~ol -5-yl)thioacetic acid,
and 0.295 g. o~ sodium bicarbonate and the resultant mixture was
stirred for 20 minutes at about 55 C. Then, the pH of the
reaction mixture thus obtained was adjusted to about 2 with 1 N
, hydrochloric acid under ice-cooling and the precipitates formed
......
were recovered by fil~ration, washed with water, and dried to
provide 0.4 g. of 7- LD-~-(4-hydroxynicotinoylamido)-q-phenylace-
tamido]-3-(5-carboxymethylthio-4-methylthiazol -2-yl)thiomethyl-
~3-cephem-4-carboxylic acid.
~ :: . :
!: : Infrared absorption spectrum: ~
;. i .
~max cm- 1768 (~-lactam)~ 'J '~.';;''
Nuclear magnetic resonance spectra (D6-DMS0
37
''- ':.''. :'.
~(ppm): 2.35 (3H), 3.50 (2H), 3.55 (2H~, 4.07 (lH), 4.52
(lH~, 4.99 (lH), 5.74 (lH), 5.84 (lH), 6.43 (lH),
7.34 (5H), 7.80 ~lH), 8.43 (lH), 9.41 (lH), 11.29
- .:
( 1H), and 12.16 (lH).
Example 25
.
To 50 ml. of water were added 0.8 g. ol- sodium 7-tD-~-(4- -
hydroxynicotinoylamido)-~-phenylacetamido~cephalosporanate,
0.284 g. of 2-mercapto-1,3,4-thiadiazole-5-acetic acid, and
,.~,
0.271 g~ of sodium bicarbonate and th~ resultant mixture was
stirred for 21 hours at 55 C. The pH of the reaction mixture
thus obtained was adjusted to about 2 with 1 N hydrochloric acid
. .
under ice-cooling and the precipitates formed were recovered by
filtration, washed with water, and dried to provide 0.45 g. of
7-~D-~-(4-hydroxynicotinoylamido)-o~-phenylacetamido3-3-(5-
carboxymethyl-1,3,4-thiadiazol -2-yl)thiomethyl-~ -cephem-4-
carboxylic acid. ` -~
Infrared absorption spectrum: ~ -
ma cm. : 1774 (~-lactam). -
Nuclear magnetic resonance spectra (D6-DMS0):
'~ 20 ~(ppm): 3.58 (2H), 3.17 (2H), 4.19 (1~), 4.53 (lEI)~ 5.00
(lH), 5.71 (lH), 5.82 (lH), 6.42 (lH), 7.35 (5H),
l 7.79 (lH), 8.42 (lH), 9.41 (lH), 11.19 (lH), and ~
.... . .
12.04 (lH).
,~ Example_26-a
In 150 ml. of methylene chloride was suspended 7.5 g. of
~ ~ cep~aloglycin and after adding 5.0 g. Oe triethylamine and 5.0 g
;1 ~ of magnesium sul:Eate to the suspension, the resultant mixture ;~
was stirred ~or 30 minutes at room temperature. After the reac~
tion was over, the reaction mixture was riltered and while cooling ~-
' 30 the :eiltrate to -30 C., 380 g. of 4-methoxynicotinic acid chloride
38
,~,, .. .. :
was added all at once swoop to the filtrate followed by stirring
for 2 hours at temperatures from -20 C. to -30 C. The reaction
mixture was allowed to stand overnight at -~ C. and then stirred
for 2 hours at room temperature~ AEter the reaction was over,
5 the solvent was distilled off a-t room temperature under reduced ~;~
pressure from the reaction mixture, the residue formed was
dissolved in 300 ml. of water. A small amount of insoluble matter
present in the solution was filtered off and after adjusting the
pH of the filtrate by adding an aqueous 5% hydrochloric acid
solution, the filtrate was stirred for 30 minutes under ice-
cooling~ The precipitates thus formed were recovered by filtra-
tion, washed with water, an aqueous 1% hydrochloric acid solu-
tion, and then water sufficient]y, and dried over phosphorus
pentoxide under reduced pressure to provide 6.3 g. of the powder
15 of 7-LD-d~(4-methoxynicotinoylamido)-~-phenylacetamido~- i
cephalosporanic acid.
Infrared absorption spectra: ;
m ~cm. : 1774 t~-lactam), 1660 (acid amLde), 1728 ; ~ ;
(acetate) '
Nuclear magnetic resonance spectra (D6-DMSO):
~(ppm): 3.74 (3H, -OMe), 6.42, 7.76, 8.42 (lH, lH, l~I, the
proton of pyridine nucleus), 2.00 (3~, -CH2OCOCH3),
Example 26-b
~ In 44 ml. of water were suspended 0.70 g. Of 7 LD-~- ( 4 -;
methoxynicotinoylamido)-d-phenylacetamido~cephalosporanic acid
and 0.26 g. of 5-carboxymethylthio-2-mercapto-1~3,4-thiadiazole
and after Eurther adding 0.37 g. of sodium bicarbonate to the
suspension, the resultant mixture was stirred for 22 hours at
, ~ . . :,
55 C. After the reaction was over~ a small amount of insoluble
, - - ~ :
i 30 matters were filtered off from the reaction mixture and then the ;
~ 39
.. .. .
. .
pH of the filtrate was adjusted to 1 by adding an aqueous 5%
hydrochloric acid solution. The precipitates thus formed were
recovered by filtra-tion, washed well with water and then ether,
and dried over phosphorus pentoxide under a reduced pressure to
provide 0.76 g. of the white powder of 7-~D-~-(4-methoxynicoti-
noylamido)-~-phenylacetamido~-3-(5-carboxymethylthio-1,3,4-
thiadiazole -2-yl)thiomethyl-~ -cephem-4-carboxylic acid.
Infrared absorption spectra:
cm. : 1770 (~-lactam), 1660 (acid amide).
max
Nuclear magnetic resonance spectra (D6-DMSO):
ppm): 3.76 (3~, -OMe), 3~56 (2H, ~ ), and 4.14
(2H, -SCH2COOH).
Example 27-a
To 18 ml. of dichloromethane were added 300 mg. of powder
15 of 4-hydroxy-5-methylnicotinic acid and 230 mg. of triethylamine
and the resultant mixture was stirred overnight at room tempera- ~;
ture. To the resultant suspension was added 241 mg. of thionyl
1 chloride under ice-cooling, then stirred for 30 minutes under
i ice cooling and further 1 hour at room temperature. The solvent
1 20 was distilled off from the reaction mixture under a reduced
pressure. A mixture of 500mg. of cephaloglycin and 280 mg. of ~ ;
trlethylamine, and 18 ml. of dichloromethane was cooled to -20 C ~;
and was added to the white powdery residue thus formed followed by
stirring for 2 hours at temperatures from -10 C. to -20 C. The
25~ mlxture was allowed to stand overnight at the same temperature as
above and then stirred for 2 hours at room temperature. The
~a~ solvent was distilled off under a reduced pressure from the reac-
tion mixture and then about 6 ml. of water was added to the
residue formed. Insoluble precipitates (i.e., unchan~ed 4-
j ~ ..
hydroxy-5-methylnicotinic acid) were filtered off from the reaction
- ~
mixture and the pH of the filtrate was adjusted to about 2 wlth
l N hydrochloric acid under ice-cooling. The white precipitates
thus ~ormed were extracted with a 1 : l mixture of n-butanol and
ethyl acetate and the extract was washed wit:h water and -then an
aqueous saturated sodium chloride solution al~d dried with magne-
sium sulfa-te. After filtering off magnesiurn sulfate, a slightly
excessive amount of a butanol solution of sodium 2-methylhexano-
ate and the precipitates formed were recovered by filtration, ~;
i washed with ethyl acetate and ether, and dried to provide 400 mg.
of 7-LD-~- (4-hydroxy-3-methylnicotinoyla~ido)-~-phenylacetamido~-
cephalosporanic acid. ~
i Nuclear magnetic resonance spectra (D6-DMS0): ~ ;
~(ppm): l.91 (3H), l.99 (3H), about 3.26 (2H), 4.70 (2H),
4.98 (lH), 5.73 (lH), 6.40 (lH), 7.24 ~5H~, 7.82 ; ;
~ 15 (lH), 8.28 (lH), and 9.34 (lH3.
!~ Example 27-b ~;
, Z :: . .: :
~ To 30 ml. of water were added 400 mg. of sodium 7 ~D-ot-(4-
i . .. :
hydroxy-3-methylpyridin -5-ylcarboxamido)-~-phenylacetamido~
~ cephalosporanate, 193 mg. of 2-carboxymethylthio-5-mercapto-1,3,- ~ -
,,,,,,!~Z 20 4-thiadiazole, and 172 mg. of sodium bicarbonate and the resultant
mixture was stirred for 22 hours at 50-53 C. Then, by treating
,~ the reaction mlxture as in Example 21, 300 mg. of 7_CD_~_(4_ -
hydroxy-3-methylpyridin -5-ylcarboxamido)-~-phenylacetamZido~ 3
~ 5-carboxymethylthio-1,3,4-thiadiazol -2-Y1)thiOmethY1-a3-CePhem- :~
;
J 25 4~carboxyIic acid.
Infrared absorption spectrum:
. :i ,~ ,
r cm.~J' 1778 (~-lactam).
Z Nuclear ma~netic resonance spectra (D6-DMSO):
(ppm): 1.94 (3~l) 3.55 (2~), 4.11 (2H) about. 4.25 (2H),
5.00 (1H) 5.74 (2H), 7.34 (5H) 7.73 ~lH~, 8.34
, . . . ~ ; . .
~ ~ 41 ;. ;
. . . ~.. ~.
~4~3
~lH), and 9.38 (lH)~
Example 28-a
In 20 ml. of methylene chloride was suspended 0.50 g. of
6-hydroxy-4-me-thoxynicotinic acid and after adding 0.33 g. of
triethylamine to the suspension, the mixture was stirred for 10
minutes. Then, after cooling the mixture to -30 C., a solution
of 0.40 g. of thionyl chloride in 6 ml. of methylene chloride
was added dropwise to the mixture. Thereafter, the resultant
mixture was stirred for one hour at -25 C. Separately, 1.0 g.
of cephaloglycin was suspended in 20 ml. of methylene chloride
and after adding thereto 0.65 g. of triethylamine and then 1.0 g.
of magnesium sulfate, the mixture was stirred for 20 minutes at
, room temperature. After the reaction was over, insoluble matters
j~ were filtered off from the reaction mixture, and after cooling
15 the filtrate to -45 C., the methylene chloride solution of the ~;
acid chloride cooled prepared above was added all at once to the
ll filtrate.
Then, after stirriny the mixture for 3 hours at temperatures
from -20 C. to -30 C. and then for 1.5 hours at room temperature,
the solvent was distilled o~f under reduced pressure and 30 ml.
of water was added to the residue thus formed. ~he pH of the
mixture was adjusted to 8 by adding an aqueous 5% sodium bicar-
;,: .. :
i~ bonate solution. Thereafter, the p~ of the system was further
$~ adjusted to 2 by adding thereto an aqueous 5% hydrochloric acid
.j~ :
solution and the precipitates formed were recovered by filtration~washed sufficiently with an aqueous 1% hydrochloric acid solution
:. ~
!~ and then water~ and dried over phosphorus pentoxide under reduced ~;-
.~
pressure to provide 0.80 g. of the white powder of 7-~D~ 4~
methoxy-6-hydroxynicotinoylamido)-~-phenylacetamido~cephalosporanic
~ ~ 30 acid.
42
,:
Infrared absorption spectra:
cm. : 1778 ~-lactam), 1662 (acid amide)~
Nuclear magnetic resonance spec-tra (D6-DMS0):
~(ppm): 2.00 (3H), 3.40 (2H, 3H), 4.82 (2H), 5.04 (1~),
- 5 5~70 (lH), 5.78 (lH, lH), 7.36 (5H), 8.42 (lH),
8.98 (lH), and 9.44 (lH).
, . ~ . .
Exa~ple 28-b ~; ~
In 43 ml. oE water were suspended 0.69 g.of 7- LD~-(6- ~
hydroxy-4-methoxynicotinoylamido)-O~phenylacetamido~Gephalosporanic - ~ ;
acid and 0~28 g. of 2-carboxymethylthio-5-mercapto-1,3,4-thia-
'. ~
diazole and after adding 0.37 g. of sodium bicarbonate to the ~; --
: :-.
suspension, the resultant mixture was stirred for 23 hours at
55 C. Ater the reaction was over, a small amount oE insoluble
?j;!, :.: !
~ matters ormed were filtered off from the reaction mixture and
.,~ ... .
the pH of the filtrate was adjusted to 2 by adding an aqueous 5%
¦ hydrochloric acid solution. The precipitates formed were
; recovered by -filtration, washed with water and then ether, and
dried over phosphorus pentoxide under reduced pressure to provide i;
-.- . ~ . . .
0.51 g. of the crystalline powder of 7-LD-~-(6-hydroxy-4-methoxy- ;
n.icotinoylamido)-c~~phenylacetam:Lclo~-3-(5-carboxymethylthio-1,3,4- ~ ;
~ 3 - -
i~ thiadiazol -2-yl)thiomethyl-~ -cephem-4-carboxylic acid.
Infrared absorption spectra:
3~ KBx cm. : 177~3 (~-lactam) and 1664 (acid amide).
j~ ~ Nuclear magnetic resonance spectra (D6-DMS0)_ ;;
~(ppm~: 3.44 t2E, 3~), 4.14 (2E), 4.96 ~2H), 5.04 (lH),
~ 5.66 (lH), 5.76 (lH, lM~, 7.36 (5E), 8.40 (lH~
i~ 9.00 ~lH~, and g.46 (lH).
~ Example 29-a
.j . .
.,::
To 30 ml~ o~ methylene chloride were added 1 g. of cephalo-
glycin and 0.62 g. o~ triethylamine with stirring and a~ter
;- . ;
~ 43
:: . :.
, .. . . .
Eurther adding 0.53 g. of 4-hydroxy-6-methylnicotinic acid
chloride to the mixture at -10 C., the resultant mixture was
; stirred for 4 hours at temperatures of from -10 C. to 0 C.
Thereafter, the reaction mixture was allowecl to stand overnight
5 at -20 C. and after stirring the mixture for 3 hours at 10-15 C.,
the solvent was distilled off ~rom the reaction mixture under
- reduced pressure. Then, 30 ml. of water was added to the residue
to dissolve the residue and after adjusting the pH of the solu-
tion to 1-2 by adding 1 N hydrochloric acid under ice-cooling, -~
the product was extracted with a mixture of n-butanol and ethyl
acetate in 1 : 1 by volume ratio. The extract was washed with
water and then an aqueous saturated sodium chloride solution,
., . ~
and dried by anhydrous magnesium sulfate. Then, magnesium sulfate
was ~iltered of from the system and the filtrate was mixed w.ith
15 a slight excess of a n-butanol solution of sodium 2-methylhexano- ~;
ate with stirring. The precipitates thus formed were recovered
by filtration and washed with ether to provide 0.4 g. o~ sodium -
7-LD-d- (4-hydroxy-6-methylnicotinoylamido~-~-phenylacetamido]-
cephalosporanate.
Nuclear magnetic resonance spectra (D6-DMS0): ;
~, (ddm): 1.91 (3H), 2.28 (3H), 3.14 tlH), 3.40 (lH), 4.71 ~ ~
-i~ (2H), 5.00 (lH), 5.76 (lH), 6.33 (lH), about 6.43 ~;;
(lH~, 7~23 (5H), 8.21 llH), and 9.32 (lH).
Example 2~-b
To 20 ml. of water were added 250 mg. of 7 LD-~- (4-hydroxy-
6-~ethylnicotinoylamido)-d-phenylacetamido~cephalosporanic acid,
, :. . . .
120 mg. or 2-mercapto-5-carboxymethylt~io-1~3,4-thiadiazole, and
102 mg. of soalum bicarbonate and the resultant mixture was
heated for 24 hours to 50-53 C. with stirring. Then, the pH of
~ ,; . ,
the reaction mixture was adjusted to 1-2 with 1 N hyclrochloric
~ ',
s;~
acid under ice-cooling and the precipitatesZ formeZr~l were recovered
by f iltration, washed with water and then e-ther, and dried to
provide about 170 mg. of 7-~D-o~-(4-hydroxy-6-methylnicotinoyl-
amido)-d-phenylacetamido~-3~(5-carboxymethylthio-1,3,4-thiadiazol-
5 2-yl) thiomethyl-a3-cephem-4-carboxylic acid.
Infrared absorption spectrum:
`~ ma cm . : 1776 ( ~3- lactam) .
Nuclear magnetic resonance spectra (D6-DMS0):
Z,r(ppm) 2.26 (3H), 3.59 (2H), 4.15 (2H), 4.19 (lH), 4~50
10 (lH), 5.03 (lH), 5.79 (2H), 6.30 (lH), 7.39 (5H),
8.32 (lH) and 9.47 (lH).
Example 30
In 47 ml. O.e water were suspended 0.75 g. of sodium 7- tD~-
(4-hydroxynicotinoylaZmido) ~-phenylacetamido] cephalosporanate and ~;
0.29 g. of 1-amino-2-carboxymethylthio-5-mercapto-1,3,4-tria~ole
and after adding further 0.29 g . of sodium bicarbonate to the
suspension, the resultant mixture was stirred for 22 hours at
55 C. After the reaction was over, a small amount of insoluble
matters in the mixture were filterèd off and then the pH oE the
filtrate was adjusted to 2 by aclding an aqueous 5% hydrochloric
acid solution to the mixture. The precipitates thus formed were
recovered by filtration, washed sufficiently with water and then
ether, and dried over phosphorus pentoxide under reduced pressure
to provide 0.62 g. of the crystalline powder of 7-CD~ 4-hydroxy-
nicotinoylaZmido3 -c~-phenylacetamido~-3- (1-amino-5-carboxymethyl-
thio-1,3,4-triazol -2-yl~thiomethyl-~ -cephem~4-carboxylic acidu ~ ;
Z Infrared absorption spectra:
Z ~ax cm. : 1772 (~i-lactam~ and 1660 (acid amiZde).
Nuclear magnetic resonance spectra (D6-DMS0~.
30 J~ppm); 3.58 (2H), 3.98 (2H), 5.04 (2H), 5.40 (2H), 5.70 ;-
. .. . , , , , , ,, ,,,, , , ,:
t1H), S.78 (lH), 5.82 (lH), 6.46 (lH), 7.36 (5H),
7.78 (lH), 8.46 (lH), 9.40 (lH), and 11.16 (lH).
Example 31-a
To 10 ml. of chloroform were added 500 mg. of 7~(D~-amino~
~-p-hydroxyphenylacetamido) cephalosporanic acid trifluoroacetate
and 190 mg~ of triethylamine and after cool:ing the resultant
mixture to -20 C., 163 mg. of 4-thiopyrone-3-carboxylic acid
chloride was added to the mixture with stirring. rrhen, aftPr
stirring the mixture for one hour at the same temperature as
10 above and for 2 hours at room temperature, the solvent was dis- `
.
' tilled off from the reaction mixture under reduced pressure. To
. -
'j the residue thus formed was added 10 ml. of water and after
l acidifying the solution with 1 N hydrochloric acid, the product
;1 was extracted twice with 25 ml. of a 1 : 1 solution of n-butanol
15 and ethyl acetate. The extract was washed with water and then -
dried and the solvent was distilled off under reduced pressure.
i~ Thereafter, ether was added to the residue thus ~ormed and the
~,' precipitates formed were recovered by filtration to provide 380
mg. of 7_LD_~ (4-thiopyrone-3-carboxamido)-~-p-hydroxyphenyl-
~ s ~
acetamido]cephalosporanic acid.
` Infrared a~sorption spectrum: :
: ~m cm. : 1780 (~-lactam).
Nuclear magnetic resonance spectra (D6-DMSO):
ppm): 2.00 (3H), 3.40 ~2H), 4.80 (2H), 5.02 (lH), 5.68
(2H), 6~70 (2H), 7.20 (2E), 7.22 (lH), 8.40 (lH),
and 9.30 (1~
1: :
Example 31-b
To 10 ml. of water were added 150 mg, of 7-~D~ (4-thio-
pyrone-3-carboxamido)-~-p-hydroxyphenylacetamido~cephalo~poranic
: . . .
30 acid, 61.5 mg. of 2-carboxymethylthio-5-mercapto-1,3,4--thiadia- `~
~6 ` ~ ~
'' `" '.,.,~ :`' '.:
.
. .. ... . . .... ,.. ,. ,., . .. ~ . .. , ... , , . " ~ . . " , . , . . ., , " .. . ... .
~ 3
zole, and 81.0 mg. of sodium bicarbonate and the mixture was
stirred for 20 hours at 50-55 C.
After the reaction was over, the reaction mixture thus
obtained was acidified by adding 1 N hydrochloric acid under ice-
cooling and the precipi-tates formed were recovered by filtration
to provide 120 mg. of 7 CD ~ (4-thiopyrone~3-carboxamido3-~-p-
hydroxyphenylacetamido~-3- ~5~carboxymethylthio-1,3,4-tl~iadiazol
-2-yl~thiomethyl-~3-cephem-4-carboxylic acid.
Infrared absorption spectrum: ;
~m cm. : 1770 ~-lactam).
Nuclear magnetic resonance spectra (D6-DMSO):
~ (ppm): 3.56 (2~), 4.12 (2~I), 4.30 (2H), .00 (lE[), 5.64
~ (2H), 6.70 (2H), 7.10 (lH), 7.20 (2H), 8.36 (lH),
I and 9.30 (lH).
! 15 Exa ple 32
- ~. ,, , - , .
To 12 ml. of water were added 170 mg. of 7-~D-~-(4-thio~
pyrone-3-carboxamido)-~-p-hydroxyphenylacetamido~cepha~osporanic
acid, 87 mg. of 2-carboxymethyl-5-mercapto-1,3,~-thiadizole, and
,, ~ .
92 mg. of sodium bicarbonate and the ~ixture was stirred for 15 '
hours at 50-55 C. Then, by treating the mixture as in Example
, 31-b, 120 mg. O~ 7-LD-~-(4-thiopyrone-3-carboxamido)-~-p-hydroxy-
! phenylacetamido~-3-t5-carboxymethyl-1,3,4-thiadiazol -2-yl~thio-
i,~ methyI-~ -cephem-4-carboxylic acid was obtained.
,~ Infrared absorption spectru~:
, 25 ~mKBr cm. 1 1775 (~-lactam).
Nuclear magnetic resonance spectra (D6-DMSO):
ppm)1 3.58 (2H), 4.16 (2H), 4.36 (2H), 5,00 (lH), 5.64
~2H~, 6.70 (2H),7~20 (2H), 7.22 (lH), 8.38 (lH), ;;
and 9.10 (lH).
.................................................................................. : ,; :- .
Example 33
~7
'..
To 10 ml. of water were added 150 mg. of 7- ~D~ (4-hydroxy-
nicotinoylamido)-~-p~hydroxyphenylacetamido~cephalosporanic acid,
63 mg. of 2-carboxymethylthio-5-mercapto-1,3,4-thiadiazole, and
83.3 mg. of sodium bicarbonat0 and the mixture was stirred for 20
hours at 50-52C. After ~he reaction was over, the reaction mix-
ture was acidified by adding 1 M hydrochloric acid under ice-
cooling and the precipitates formed were recovered by filtration
to provide 60 mg. of 7 [D-~-(4-hydxoxynicotinoylamido)-~p- ~:
hydroxyphenylacetamido~-3- ~-carboxymethylthio-1,3,4-~3-thiadiazol-
2-yl~thiomethyl-cephem-4-carboxylic acid.
Infrared absorption spec-trum: ~ .
m cm. : 1770 (~-lactam).
Nuclear magnetic resonance spectra (D6-DMSO)~
~(ppm): 3.52 (2H), 4.08 (2H), 4.28 (2H), 4.64 (2H), 4.97
(lH), 6.36 (lH), 6.68 (2Hl, 7.20 (2H), 7.74 (lH),
' 8.39 (lH).
Example 34 ; .
. . .
In 23 ml. of water were suspended 0.36 g. of sodium 7-[D-~ :
(4~hydroxynicotinoylamido)-~-phenylacetamido~cephalosporanate
20 and 0.17 g. of 5-carboxymethylthio-~-mercap-to-[1,2, ~ triazolo-
' C3,4-b~ ~t3,4~-thiadiazole and after adding 0.15 g. of sodium
! bicarbonate to the suspension, the resultant mixture was stirred
for 26 hours at 55 C. After the reaction was over, insoluble ~
matters formed were filtered off from the reaction mixture and ~
25 then the pH of the filtrate was adjusted to 2 by adding aqueous . .
5% hydrochloric acid solution. Then, the pxecipitates thus formed
-
'7 were recovered by $iltration, washed sufficiently with water and ~
' ~ then ether, and dried over phosphorus pentoxide to provide 0~27 gO .~ .
,~ of the crystallins powder of 7-~D-~-(4-hydroxynicotinoylamido)~
`~ 30 phenylacetamido~-3-(5-carboxymethylth:io- [1,2,4~--triazolo¦3,4-b3-
.
48 ~`~
.
~1,3,4I-thiadiazol -2-yl)thiomethyl-~ -cephem-4-carbo~ylic acid.
Infrared absorption spec-tra:
cm. : 1776 (~-lactam) and 1660 ~acid amide).
Nuclear magnetlc resonance spectra (D6 DMS0):
~(ppm): 3.56 (2H), 4.20 (2H), 4.96 ~2H), 5.60 (lH), 5.76
(lH), 5.84 (lH), 6.42 (lH), 7.36 (5H), 7.80 (lH3,
9.42 (lH), 9.40 (lH), and 11.14 (lH).
Example 35
In 150 ml~ of water were dissolved 2.4 g. of sodium 7-D-
C~ -~4-hydroxynicotinoylamido)-~-phenylacetamido~cephalosporanate
and 1.1 g. of 2-mercapto-5-(3-carboxypropionylamino)-1,3,4-thia-
diazol, and after adling 9 ml. of 10~/o aqueous sodium bicarbona-te
`~'! solution to the solution, the mixture was stirred for 22 hours ~ `
j while heating to 55 C. After the reaction was over, insoluble
, ' ,
` 15 matters were filtered off from the reaction mixture and the pH - ~ -
3 of the filtrate was adjusted to 2 with 2N hydrochloric acid. The
precipitates thus formed were recovered by filtration, washed
~ well with water, twice each with 45 ml. of a methanol-ether
'~ mixture in a 1:5 volume ratio and then ether respectively and
dried over phosphorus pentoxide under reduced pressure to give
1.5 g. of the light yellow powder oE 7-D-~ -(4-hydroxynicotinoyl-
! amido)-~-phenylacetamido~-3-~5-(3-carboxypropionylamino)-1,3,4-
,
,~ thiadiazol~2-y ~ thiomethyl-~ -cephem-4-carboxylic acid.
Melting point 218-221C (decomp.)
. ` .
Infrared absorption spectra:
~ ma cm~ : 1772 (~-lactam), 1660
;' Nuclear magnetic resonance. spectrum (D2O-NaOD):
~: 2.54 (4H, -NHCOCH2CH2COONa)
', ---- , ,, :.
-i Example 36 -
: ! , .
In 50 ml. of water were dissolved 0.8 g. of sodiurn 7-D-~ - -
.. .. ~ .. .. . .. , . , ., , , , " , . . . .. .. . .. . . . ..
~43~
(4-hydroxynicotinoylamido)-~-phenylacetamidoJcephalosporanate
and 0.41 g. of 2-mercapto-5-o-carboxybenzoylamino-1,3,4-thiadiazol
and after adding 3 ml~ of l~/o a~ueous sodium bicarbonate solution .
to the solution, the mixture was stirred for 21 hours while
heating to 55 C~
After -the reaction was over, insoluble matters were filtered
of~ from the reac-tion mixture and the pH of the filtrate was
adjusted to 2 with 2 N hydrochloric acid. The precipitates thus .
formed were recovered by filtration, washed well with water~
twice each with 15 ml. of a methanol-ether mixture in a 1:5
volume ratio and then ether respectively and dried over phosphorus
pentoxide under reduced pressure to give 0.54 g. of the light
yellow of 7-D- ~ -(4-hydroxynicotinoylamido)-~-phenylacetamido~-3-
(5-o-carboxybenzoylamino-1,3,4-thiadiazol-2-yl)thiomethyl.-
~
15 cephem-4-carboxylic acid. ~ -
Melting point 207 - 211C
, Infrared absorption spectrum: .. ..
~i ~max cm. 1770 (~-lactam), 1658
Example 37
In 50 ml. oE water were dissolved 0.8 g. o~ sodium 7-D-~
(4-hydroxynicotinoylamido)-~-phenylacetamido3cephalosporanate ..
.: .
71 and 0.27 g. of 2-mercapto-5-carboxymethyl-1,3,4-triazole, and .~ .. .
,';~ after adding 3 ml. of loo/O aqueous sodium bicarbonate solution ..
1 : to the solution, the mixture was stirred for 22 hours while
!~ 25 heating to 55 C. `.... -.
By treating the reaction mixture thus ob-tained as in Example ~ .-
36, 0.28 g. of the li~ht yellow powder of 7-D-r~ (4-hydroxy~
nicvtinoylamido~ -phenylacetamido~-3-(5~carboxylmethyl-1,3,4- ..
~ triazol-2-yl)thiomethyl-d -cephem-4-carhoxylic acid.
: 30 Melting point 210 - 213 C .
50 -~ :
~: :
- Nuclear magnetic resonance spec-trum (DMSO): 3.32 (2~,-CH2-
COOH)
Infrared absorption spectra: -
~ max cm. : 1775 (~3-lactam~, 1658
Example 33
To 20 ml. of water were added 300 mg . of sodium 7- LD-C~- (4- :
. .
hydroxynicotinoylamido~-~-phenylacetamido~cephalosporanate, 140 : -
mg. of 4-(5-mercapto-1,3,ar-thiadiazol -2-yl'?butyric acid, and 120
!'`: :,...:.;, ':,...
mg. of sodium bicarbonate and tha resultant mi~ture was stirred
10 for 24 hours at 50-52 C. Then, the pH of the reaction mixture
was adjusted to about 2 with 1 N hydrochloric acid and the white
precipiates thus Eormed were recovered and washed with water and
:-, !. ' '
then ether and dried to provide 200 mg. oE 7-CD-~-(4-hYdrOXY_
', n ico t inoy lamido ) -o~-pheny lace tamido l- 3- L5 (3-carboxypropy l) - 1,3,4-
:J' 15 thiadiazol -2-yllthiomethyl-~ -cephem-4-carboxylic acid. -
l : .....
, Infrared absorption spectrum:
m cm. : 1780 (~-lactam).
Nuclear magnetic resonance spec-tra ( D6-DMSO):
~ppm: 1.91 (2H), 2.32 (2H), 3.06 (2H), 3.58 (2E), 4.17
(lH), 4.51 (lH), 5~01 (lH), 5.01 (lH), about 5.76
(lH), 5.84 (lH), 6.44 (lH), 7.38 (5H), 7.82 (lH),
1 8.45 ( lH), and 9.47 (1~) .
.~ ~; Example 39
~,}~ ~o 20ml. of water were added 300 mg, of 7- ~ (4-hydroxy-
25 6-methylnicotinoylamido)-clrphenylaceti~nido~cephalosporanic acid,
?~ ', ' '
142 mg. o~ 4-(5-mercapto-1,3,4-thiia;zol -2-yl~butyric acid, and
169 mg. of sodium bicarbonate and the mixture was stirred Eor 24
hours at 50~52 C. ~hen, the pH o E the reaction mixture was adjus-
- ~ ~ ted to about 2 with 1 N hydrochloric acid under ice-cool:ing and
30 the precipitates formed were recovered by filtration, ~ashed with
~ 51 ~;
,:, ~ ~,. ~ .
- .
4~83
~ Y D15~5~
water and then ether, and dried to provide 200 mg. of
7-[D-a-(4-hydroxy-6~methylnicotinoylamido)-a-phenylacetamido]-
3-[5-(3-carboxypropyl)-1,3,4-thiadiazol-2-yl]thiomethyl-
-cephem-4-carboxylic acid.
Infrared absorption spectrum:
~ KB cmv : 1778 (~-lactam).
Nuclear magnetic resonancespectra (D6-DMS0):
ppm: l.91 (2H), 2.25 (3H), 2.32 (2H), 3.06 (2H), 3.58 (2H~,
4.18 (lH), 4.51 (lH), 5.00 (lH), about 5.75 (lH),
;~ lO 5.83 (lH), 6.29 (lH), 7.38 (5H), 8.32 (lH3, and
~9__OS(A Y ~
Example 40
~, a)In 30 ml. o methylene chloride were added S00 mq. o
4-methoxy-6-methyl-nicotinic acid and 360 mg. of triethylamine
and the mixture was cooled to -20 C with stirring. Then, ~
~! 280 mg. of ethyl chlorocarbonate was added to the mixture and -
the resultant mixture was stirred to conduct a reaction ~or
.`1 :
2.5 hours at the same temperature.
After the reaction was over, the solution which was
l 20 prepared by dissolving lg. o cephaloglycin and 27S mg. of
} ~ triethylamine in 20 ml. o methylene chloride was added to the
~ reaction mixture~ and then the resultant mixture was stirred
1 ~
~ for 2 hours at th~ temperature o _20U C. The reaction
, :,
mixture was allowed to stand overni~ht at the same temperature
and then stirred ~or one hour at the temperature o~ 0 C.
f~er~tn~ reaction was Gve~ the solvent of the
rsaction~mixture was distillea of~ under reduced pressure and
ths residue thus obtained was dissolved by addin~ about 40 ml.
of~water. The pH of the solution was adjusted with 1 N
.~ ~ ' - .
~ ` '
~ f~
hydrochloric acid to 2 under ice-cooling, whereby precipitates
were formed. The precipitates thus formed were recovered by
. ........ ..
filtration, washed with water and dried over, whereby 850 mg. `
. .~
of 7- [D-~- (4-methoxy-i~-methyl-nicotinoylamido)-a-phenylacetamido]-
cephalosporanic acid was obtained. ~ -
Infrared absorption spectra
~ KBr cm- 1 1778 (~-lactam)-
Nuclear magnetic resonance spectra (D6-dimethylsulfoxide)
p.p.m.: 2.01 (3~), 2.32 ~3~), 3.45 (2H), 3.72 (3~),
4.67 (lH), 4.99 (lH), 5.03 (lH), 5.81 (2H),
6.42 (lH), 7.38 (5H), 8.50 (lH), 9.50 ~lH)
b) in 23 ml. of water were added 300 mg. 7-[D~-(4-
j methoxy-6-methylnicotinoylamido)-a-phenylacetamido]-cephalo- - i
~ sporanic acid, 140 mg. of 2-carboxymethylthio-5-mercapto-1,3,
Jl 4-thiadiazole and 1~3 mg. o~ sodium bicarbonate, and the
f
resultant mixture was conducted to react ~or 24 hours at the :
f temperature of 50 C.
3i~ After the reaction was over, the pH of the reaction `
mixture was adjusted to about 2 with 1 ~ hydrochloric acid
20 under ice-cooling, whereby precipitates were formed. The
precipitates thus formed were recovered by filtration~ washed `~:
with water~nd then with ether and dried over~ whereby 250 mg. ~ ;
of 7-[D-~-(4-methoxy-6-methylnicotinoylamido)-a-phenylacetamido]-
'~: :., ':,
3-(5-carboxymethylbhio-1,3,4,thladiazole-2-yl)-thiomethyl- ~ -
Q -cephem-4-carboxylic acid was obtained. :;
Infrared absorption spectra i`
maX cm- : 1778 (~-lactam) ~;
,1 ~ h~
~ Nuclear magnetic resonance spectra (D6-dimethylsulfoxide)
; ~ p.p.m.: 2.54 (3H), 3.60 (2H), 3.74 (3H), 4.18 (2H),
4.21 (lH), 4.52 (lH), 5.05 (lH), 5.83 (2H),
- 6.45 (lH), 7.43 (5H), 8.52 (lH), 9.53 (lH)
Example 41
In 20 ml. of water were added 300 mg. of 7-[D-a-
(4-hydroxy-6-methylnicotinoylamido)_a-phenylacetamido]-cephalo-
sporanic acid, 127 mg. of 2-carboxymethyl-5-mercapto-1,3,4-
thiadlazole and 174 mg. of sodium bicarbonate, and the resultant
10 mixture was conducted to react for 22 hours at the temperature - -
of 50 C.
.. . .
Ater the reaction was over, the insoluble matter in
the reaction mixture was filtered of~ and the pH of the fil-
trate was adjusted to ~bout 2 with 1 N hydrochloric acid under
ice-cooling and then the product was extracted with a mixture
, of n-butanol and ethyl acetate (1 : 1 in volume ratio). The ~;~
extract was washed with saturated aqueous solution of sodium ~;
;, ~ . .
1~ chloride and then dried over anhydrous magnesium sulfate.
: ~ .
Then, slightly excessive amount of sodium 2-methylhexanate was
added to the solution, whereby precipitates were formed. The
,5 ~ ' . '; .
precipitates thus formed were recovered by filtration, washed
with~water and then with ether and dried over, whereby 250 mg.
of sodiUm salt of 7-[D-a-(4-hydroxy-6-methylnicotinoylamido)-
a-phenylacetamido]-3-(5-carboxymethyl-1,3,4-thiadiazole-2-yl)- ;
thiomethyl-~ -cephem-~-carboxylic acid was obtained.
Infrared absorption spectra
KBr cm -1 1765 (~-lactam)
i ! :
~ - 54 ~ ; ~
