Note: Descriptions are shown in the official language in which they were submitted.
~310Z GG172/162
This invention relates to new 7-methoxy thienyl- and uryl-
ureido-cephalosporins o the general formula
NHCNH2 OCH3
Y - ~ CH-Il-NH ~ ~ ~ (V)
O O N ~ C~I2R
C02~[
wherein R is H, heterocyclic thio, e.g. pyridyl-l-oxo-2-thio,
3-methyl-1,2,4-thiadiazolylthio, 1-methyl-tetrazolyl-5-thio,
2-methyl-1,3,4-thiadiazolyl~5-thio; pyridinium, carbamoyloxy
or acetoxyS Y i9 hydrogen, Cl, Br, F, nitro or methylsulfonyl;
Z is O or S; and pharmaceutically acceptable carboxylate salts,
lower alkyl esters, haloloweralkyl esters or acyloxymethyl
esters thereof. The compounds of the present invention are
effective antibacterial agents and are useful in the treatment
of many gram negative and gram positive infections. These
compounds are useful as disinfectants and also as nutritional
supplements in animal feeds. The 7-methoxy group occupies the
~-configuration.
The compounds of the present invention may be prepared
from a heterocyclic aldehyde o the formula
Y ~ CHO (I)
Z
.~
106310Z GG172/162
The heterocyclic aldehyde (I) is first converted to an :~
amino acid by known methods, for example, the well known
Strecker amino acid synthesis, Ann 75, 27 (1850); 91, 349
(1854). This method involves the synthesis of a-amino acids ;~
by the simultaneous reaction of aldehydes with ammonia and
hydrogen cyanide followed by hydrolysis of the resulting amino
nitriles. As applied to the heterocyclic aldehyde I of the
present invention the reaction is as follows:
NH2 NH
Y~ CHO ~ L~--~1HC~ ~ Y~l HCO2H
In addition, various well known modifications of the Strecker
synthesi~ may be employed. Fo.r example the Erlenmeyer
modification:
. ~H2 1~ HCN IH2
Y~CHO + IIH3 -~ Y~3 CHOH 2 ) H20/H+ Y~CHCO2H
the Tiemann modification:
OH NH
Y ~ CHO + HCN ---~ Y~1HCN -~ ~ Y~ 1HCN
H20/H .,
or OH ~
\ ~ INH2 '
Y~ CHC02H
--2
.
,
:: . . , ~: . ~ . .. .
10~3iOZ GG172/162
the Zel~nsky-Stadnikoff modification: ~
~H2
Y ~ CHO + NH4Cl -~ KCN ~ Y ~ CHCN
! H O/H+
lH2
Y~ HC02H
. ~.
and the Bucherer modification:
1) HCN ¦ (NH ~ CO
Y- ~ CHO ~ Y ~ CHCN 4 2 3> F ~=
CO-NH
H20/H
NH2
Y ~ CHCO2H
The amino acid o the formula
NH
2 (II)
Y~CH-C02H
produced by any suitable method, for example, those described
above, is then raacted with an alkali metal cyanate or an
alkaline earth metal cyanate to form an a-ureido compound of
the formula l (III) ~: -
NH--C--
y~CH~C02H
: _3_
' ~ ' , .' ' ' ' ' . .
, .
. .
lQ~3102 GG172/162
This reaction takes place by treating an aqueous suspension
of the ~-amino acid with the alkali or alkaline earth metal
cyanate. Acidification with hydrochloric acid precipitates
the ~-ureido acid in good yield. A solution of th~ a-ureido
acid in an organic solvent containing a tri-(lower)alkyl amine
is converted to a mixed carbonic or other anhydride by treating
with an anhydride forming reagent, e.g., a lower alkyl chloro-
formate, an aryl chloroformate, or an acyl halide, at reduced
temperatures of from about 0C to about -20C. The reaction
takes place in a solvent such as methylene chloride, chloroform,
dioxane, dimethoxyethane, benzene, aqueous acetone or dlmethoxy-
ethane, acetonitrile, or dimethylformamide.
Reaction at reduced temperatures o the mixed anhydride
with a protected ester, e~g., benzhydryl, trichloroethyl, or
tertiarybutyl, o a compound o the formula
OCH
3f ~ : ::
20o// ~ H2R (VII)
COOH
wherein R is as previously defined yields the compounds of the
present invention after removal of the ester group. `
In general, a compound of formula V may be obtained by
converting a compound of formula III to an activated ester or
by reacting a compound of formula III with a carboxyl group
activating agent, such as, or example, dicyclohexylcarbodiimide
or bisimidazole carbonyl, and then coupling the activated form
o the compound o formula III with a protected ester of the
,~ . :,-~ ' .
1~3~02 GGl72/162
compound of Formula VII. In some cases, as will be obvious
to those skilled in the art, the carboxyl group may be
activated by conversion to an acid halide, e,g., the chloride,
or to an azide prior to coupling with a protected ester of the
Formula VII compound. A more detail~d discussion of carboxyl
activating groups may be obtained by reference to standard
works on peptide synthesis, for example, Bodanszky et al.,
"Peptide Synthesis", Interscience, 1966.
Alternatively, the ~-amino acid (II) produced, for example
by subjecting the heterocyclic aldehyde to the Strecker
reaction or a modification thereof, may be used to acylate a
protected ester of the Formula VII compound. The resulting
~-amino acid derivative of the Formula VII compound is then
reacted with an alkali metal cyanate or an alkaline earth metal
cyanate in the presence of water to form the compounds (V) of
the present invention.
In the case of the products of Formula V wherein R is
heterocyclic thio, the heterocyclic thio group may be inserted
as the last step by displacement of the acetoxy group of the
corresponding product of Formula V wherein R is acetoxy with
the desired heterocyclic thio group by reaction with the desired
mercaptoheterocycle according to conventional methods. The
insertion can also be effected at an earlier stage by reacting
a compound of Formula VII wherein R is acetoxy with the desired
mercaptoheterocycle, see also Abraham and Newton (1950) Ciba
Foundation Symposium, Amino Acids, Peptides, Antimetabolic
Activity, p. 205; U. S. Patent 3,225,038; and Belgian patents
641 r 338 and 652,148.
, , ~.- ~ .,
GG172/162
~L~63~0Z
The a-amino acid (II) or the ~-ureido acid (III) obtained
occurs as a racemate of D and L optical isomers. It is gen-
erally possible to resolve these compounds by using optically
pure bases (or acids) using methods known to the art, for
example, as described by L. Velluz, "Substances Naturelle de
Synthesese," 9, pp. 119-174 (1954), or E.L. Eliel, "Stereo-
chemistry of Carbon Compounds," Chapter 4, McGraw-Hill Book Co.,
N.Y. (1962). The resulting pure D or L acid may then be
coupled to a protected ester of the Formula VII moiety. ; ~ ?
10 Generally the coupling of the D form leads to the more active ~ ;
product.
When preparing compounds of the present invention according
to the reaction sequence wherein a protected ester of the com-
pound of Formula VII i9 acylated with an a-amino aaid of
Formula II, it may be desirable to protect the a-amino group
during the acylation reaction by means of an amino protecting
group. Such amino protecting groups are well known in the art
and are described, for example, by Bodanszky et al., "Peptide
Synthesis," supra. ~
Specific examples of methods for acylating the cephalo- ~ ;
sporin of Formula VII are similar to those described for the
acylation of 7-ACA or 7-ADCA, for example, in Netherlands
Patent 6,812,382, Belgian Patent 675,298, as well as in the ~;
following articles:
Spencer et al., J. Med. Chem., 9, 746 tl966);
Ryan et al., ibid, 1~, 310 tl969).
A general reaction scheme for preparing compounds of the
. ~;, .. .
present invention is shown below wherein for ease of represen ~-
tation A is the residue of the cephalosporin moiety of
Formula VII.
- . .::. . . . . .
~0~3~z GGl72/162
1) HCN ~ 1 2
HO 3) H2O/H y_ ~ CH-cO2H
(I) / (II)
/ ~alkali or alkaline earth
cyanate/H20
/ \ ' 1l'
~ / NHC-NH (III)
Y~ CH-C02H
NH2
Y- ~ ~ CH-C-A (VI) ~:
~Z O 1) tri-(lower)alkyl
\ 2) anhydride forming
\ reagent :~
~ / NH--C-NH2
\ Y ~ CH-C-OCR (IV) : :
alkali or \
alkaline earth \ . .
cyanate/H20 \ .,
AH '~
\ ~ NH-Il-NH
y ~ CH-~-A (V)
..
7- ? '
: ' . . ' ' ' ' ' ' ' '
'', ,. ' ~ ' , ` ; : .
GG172/162
1~3~
The carboxylate salts of the compounds (V) of the present
invention are formed by reacting the carbo~yl group of the ceph-
alosporanic acid moiety with a salt formin~ ion, e.g., an alkali
metal such as sodium or potassium, or an a]Lkaline earth metal
such as magnesium or calcium, or a metal of group IIIA such as
aluminum, or the radical of an organic base such as dibenzyl-
amine, N,N-dibenzylethylenediamine or other organic bases known
to form salts with cephalosporanic acids.
The lower alkyl esters may be obtained by esterifying the
carboxyl group of the cephalosporanic acid moiety with a str~aight
or branched chain lower alkyl halide of from 1 to 3 carbon
atoms, e.g., methyl chloride, ethyl bromide and the like, or
with a diazoalkane of from 1 to 3 carbon atoms, e.g., diazo-
methane, diazoethane, and the like. The resulting ester group
is then formed by a radical such as methyl, ethyl, propyl, or
isopropyl.
The acyloxymethyl esters may be obtained according to
known methods, for example, a method adapted from that of `
Daehne et al., J. Med. Chem~ 13, 607 (1970), by reacting the
carboxyl group or a metal ~alt thereof of the cephalosporanic
acid moiety with a halide of the formula
O . ~'
Il ~.. ..
X-CH2-O-C-R'
! .
wherein R' may be lower alkyl of up to 5 carbon atoms, phenyl, -
benzyl or phenethyl, and X is chlorine or bromine. Thus, suit~
able compounds include acetoxymethyl chloride, propionyloxy~
methyl chloride, butyryloxymethy1 chloride, pivaloyloxymethyl
chloride, valeryloxymethyl chloride, benzoyloxymethyl chloride,
or phenacetoxymethyl chloride, and the like.
. ~ . . .
.
~31~2 GG172/162
It will be appreciated that certain of the compounds of
this invention exist in various states of solvation as well as
in different optically active forms. The various forms as
well as their mixtures are within the scope of this invention.
The compounds of this invention have a broad spectrum of
antibacterial activity against both gram positive and gram
negative organisms such as Staph~lococcus aureus, Salmonella
schottmuelleri, Proteus vul~aris~ Escherichia coli and
Streptococcus pyogenes. They may be used an antibacterial
agents in a prophylactic manner, e.g., in cleaning or dis-
infecting compositions, or otherwise to combat infections due
to organisms such as those named above, and in general may be
utilized in a manner similar to cephalosporin C, cephalothin
and other cephalosporins. For example, a compound of formula V
or a physiologically acceptable salt or ester thereof may be
used in various animal species in an amount of about 1 to 200
mg./kg. daily, orally or parenterally, in single or two to four
~,
divided doses to treat infections of bacterial origin.
Up to about 600 mg of a compound of formula I or a
pharmaceutically acceptable salt or ester thereof may be incor-
porated in an oral dosage form such as tablets, capsules or
eli~irs or in an injectable form in a sterile aqueous vehicle
prepared according to conventional pharmaceutical practice.
In cleaning or disinfecting compositions, e.g., in barns
or dairy equipment, a concentration of about 0.01 to 1~ by
weight of such compounds admixed with, suspended or dissolved
in conventional inert dry or aqueous carriers for application
by washing or spraying may be used.
They are also useful as nutritional supplem~nts in animal
feeds.
.:
:~ :
GG172/162
~63iOZ ~ .
The compounds of the present invention in the described
dosages may be administered orally; however r other routes such
as intraperitoneally, subcutaneously, intramuscularly or intra-
venously may be employed.
The active compounds of the present invention are orally
administered, for example, with an inert diluent or with an
assimilable edible carrier, or they may be enclosed in hard or
soft gelatin capsules, or they may be compressed into tablets,
or they may be incorporate~ directly with the food of the diet.
For oral therapeutic administration, the active compounds Of n
this invention may be incorporated with excipients and used
in the form of tablets, troches r capsules, elixirs, suspensions,
syrups, waers, chewing gum, and the like. The amount of active
compound in such therapeutically useful compositions or pre- ;
parations is such that a suitable dosage will be obtained.
The tablets, troches, pills, capsules and the like may -~
also contain the following: a binder such as gum tragacanth,
acacia, corn starch or gelatin; an excipient such as dicalcium ;
phosphate; a disintegrating agent such as corn starch, potato
starch, alginic acid and the like; a lubricant such as magnesium `
stearate; and a sweetening agent such as sucrose, lactose or
saccharin may be added or a flavoring agent such as peppermint,
oil of wintergreen, or cherry flavoring. When the dosage unit
form is a capsule, it may contain in addition to materials of
the above type a liquid carrier such as a fatty oil. Various
other materials may be present as coatings or to otherwise
modify the physical form of the dosage unit, for instance,
tablets, pills or capsules may be coated with shellac, sugar,
or both. A syrup or elixir may contain the active compounds,
sucrose as a sweetening agent, methyl and propyl parabens as
--10--
. , ~ . . . . .
GG172/162
1~3~0Z
preservatives, a dye and a flavoring such as cherry or orange
flavor. Of course, any material used in preparing any dosage
unit form should be pharmaceutically pure and substantially
non-toxic in the amounts employed.
In the compound of Formula VII wherein R is as previously
defined
OCH
H2~ ~
~ ~ C 2
I `'
COO~ , ~. ...
the 7-methoxy group has the a-configuration. :~
'.
-. , ~ ; . .
1~6310Z GG172/162
The following exampl~s illustrate the present invention
without, however, limiting the same thereto.
Example 1
7-[2-Ureido-2-(2-thien~l)acetamidol-7-methox~cephalosporanic acid
Method A
2-Thiophene carboxaldehyde is reacted with ammonium cyanide
prepared in situ from ammonium chloride and sodium cyanide. The
resulting amino nitrile is hydrolyzed to yield thienyl-2-(2-
amino)acetic acid. A suspension of the latter compound (0.10 ~,
moles in 150 ml of water is treated with 8.1 g of potassium
cyanate. The resulting mixture is heated to about 80C to gi~e
a clear solution which is then allowed to stand at roorn
temperature for about 24 hours, Acidification to pH 3.5 with
hydrochloric acid preclpitates the a-ureido compound~ A solution
containing 0.10 moles of the a-ureido acid in 100 ml of
acetone containing 10.1 g. of triethylamine at a temperature of
from about 0C ~o about -20C is converted to a mixed carbonic
anhydride by treating with 10.~ g of ethyl chloroformate for
about 30 minutes. A cold (about -10C) solution of 0~10 mole of
7-amino-7-methoxy-cephalosporanic acid, benzhydryl ester [Cama
et al., J.A.C.S., 94, 1408 (1972)] in 400 ml o 1:1 acetone
containing 10.1 g of triethylamine is added to the solution
of mixed anhydride and the reaction mixture stirred vigorou~ly
at about 0C for approximately 30-45 minutes. The volume of
the solution is reduced by evaporating the bulk of the acetone
at reduced pressure at room temperature or below.
One liter of ethyl acetate is added and the solution
washed with 2 X 200 ml of ice cold 5~ aqueous sodium bicarbonate
100 ml of water, 2 X 200 ml of 0.5 molar hydrochloric acid, and
100 ml again of water. The ethyl acetate solution is dried
:: ~ ' '::,: .
10~3~0Z GG172/162
(Na2SO4) and evaporated to deposit the benzhydryl ester of
the title compound.
The free acid is obtained by dissolving the ester (1 g)
and anisole (500 mg) in 20 ml of ice cold trifluoroacetic acid
and keeping it at 0-5C for 30 minutes. The acid solvent is
evaporated at reduced pressure. The residue is treated with
50 ml of water and the pH adjusted to 7.5 to dissolve the ;
product. The solution is washed with ethyl acetate.
(Lyophilization of the aqueous solution deposits the sodium ~;
10 salt of the title compound). The aqueous layer is acidified "
to precipitate the title compound.
Method B
A solution containing 0.1 mole of 2-(p-methoxybenzyloxy-
carbonylamino)-2-thiophene acetic acid in 100 ml of acetone
containing 10.1 g o triethylamine at a temperature of -10C
is converted to a mixed carbonic anhydride by treating with
10.8 g of ethyl chloroformate for about 30 minutes. ~ cold
(about -10C.) solution of 0.1 mole of 7-amino-7-meth~xy
cephalosporanic acid benzhydryl ester in 400 ml of 1:1 acetone
containing 10.1 g of triethylamine is added to the solution
of mixed anhydride and the reaction mixture stirred vigorously
at about 0C for approximately 30-45 minutes. The volume of
the solution is reduced by evaporating the bulk of the acetone
at reduced pressure at room temperature.
One liter of ethyl acetate is added and the solution -
washed with 2 X 200 ml of ice cold 5% aqueous sodium bicarbonate,
100 ml of water, 2 X 200 ml of 0.5 molar hydrochloric acid,
and 100 ml again of water. The ethyl acetate solution is dried
(Na2SO4) and evaporated to yield the 7-[2~p-methoxybenzyloxy-
-13-
,
1063102 GG172/162
~, '
carbonylamino)-2-thiophene acetamido]-7-methoxy cephalosporanic
acid benzhydryl ester.
The free amino acid i~ obtained by dissolving the ester ;~
(11 g.) and anisole (30 ml) in 150 ml of ice cold trifluoro- ;
acetic acid and keeping it at -5C for 20 minutes. The acid
solvent is evaporated at reduced pressure. The residue is
triturated with ether and dried
.
The 7-thienylglycyl-7-methoxy-cephalosporanic acid (5 g.) ~
and potassium cyanate (1.4 g.) are dissolved in water (35 ml.). ~ - -
The mixture is stirred for three hours, filtered and the
filtrate adjusted to pH 1.5 to precipitate the compound of the
title.
Ex mple 2
7-[7-Ureido-2-(2-thienyl)aaetamido]-7-methoxy-3-desaaetoxy-
cephalosporanic acid
Following the procedure of example 1 but substituting ~
0.1 mole of 7-amino-7-methoxy-3-desacetoxycephalosporanic acid ~ ;
benzhydryl ester for 7-amino-7-methoxycephalosporanic acid
benzhydryl ester, the title compound is obtained.
,
Example 3
Pivaloyloxymethyl ester of 7-[2-ureido-2-(2-thienyl)-acetamldo]-
7-methoxy-3-desacetoxy cephalosporanic acid
Chloromethylpivalate (20 mmole) is combined with 10 mmole
of the product of example 2, 0.4 ml o a 5% aqueous sodium ;
iodide solution, and 170 ml of acetone. Triethyl amine, 2.0 gm.
(20 mmole), is added and the mixture stirred for 10 hours, then
refluxed for one hour. The reaction mixture is cooled and con-
centrated ln vacuoO The residue is partitioned between ethyl
acetate, and 5% aqueous sodium bicarbonate. The organic layer
is dried over sodium sulfa~e and evaporated to give the crude
-14-
GG172/162
~33L~2
product which is obtained as a powder upon trituration wi~h
ether.
Example 4
Pivaloyloxymethyl ester of 7-[?-ureido-2-(2-thienyl)-acetamido~-
7~methox ce halos oranic acid
Y P P
Following the procedure of example 3 but substituting
10 mmole of the product of example 1 for the produat of example
3, the title compound is obtained.
Exam~le 5
7-[2~Ureido-2-(2-thienyl)acetamido]-7-methoxy-3-desacetoxy
cephalosporanic acid, methyl ester
A 0.1 molar solution of the product of example 2 in
dimethoxyethane i9 treated with an excess o ethereal diazo- ,~
methane. Evaporation o~ the solvent at reduced pressure deposits
the title compound.
Example 6
Aceto~methyl ester of 7-[2-ureido-2-(2-thienyl)acetamido]-
7-methoxy~cephalosporanic acid
,: . .
Following the procedure of example 4 but substituting
chloromethylacetate for chloromethylpivalate, the title compound
is obtained.
Example 7
7-~2-Ureido-2~(2-thienyl)acetamido~-7-methoxycephalosporanic
acid, potassium salt
.
One millimole of the product of example 1 is dissolved
in 10 ml of a 0.01 N aqueous KOH solution. Lyophilization
of the solution yields the title compound.
-15-
... .
,
:
GG172/162
i~363~0Z
Example 8
7-[2-Ureido-2-(2~thienyl)acetamido]~7-methoxy-3-desacetoxy
cephalosporanic acid, calcium salt
. ~
One millimole o the product of example 2 is dissolved
in 10 ml of a 0.05 N aqueous Ca(OH~2 solution. Lyophilization ~ ~
of the solution yields the title compound. - i
Example 9
:. :
7-[2~Ureido-2-(2-thienyl)acetamido]-7-methoxy-3-[2-(5-methyl-
1,3,4~thiadiazolyl~thiomethyl-~3-cephem-4-carboxylic acid
A solution (0.026 mole) of the product of example 1, NaHCO3
(2.1 g) and 3.8 g 2-mercapto-1,3,4-thiadiazole in 200 ml of pH -
6.4 phosphate buffer is stirred for 5.5 hours at 60C. The
reaction is cooled to room temperature, acidified to pH 3 and
extracted with ethyl acetate. The ethyl acetate layer is
worked with saturated NaCl solution, dried (Na2SO4), and
evaporated at reduced pressure to deposit the product.
Exam~le 10
7-[2-Ureido-2-(2-thienyl)acetamido~-7-methoxyce~haloSporaniC
acid, N,N'-dibenzylethylenediamine salt
A solution of 0.010 mole of the product of example 1 in
25 ml of ethanol is added to a solution of 1.20 g of N,N-
dibenzylethylenediamine in 25 ml of ethanol at room temperature.
- After 15 minutes stirring the solvent is evaporated to deposit
the title compound. ~-
Example 11
7-[2-Ureido-2-(2-th:ienyl)acetamido]-7-methoxycephalosporanic
acid
2-Thiophene carboxaldehyde is reacted with ammonium cyanide
preparad ln situ from ammonium chloride and sodium cyanideO The
resulting amino nitrile is hydrolyzed to yield thienyl-2-(2-
-16-
; : . ~ . - ~ .
.
.: , .i . . .. ..
GG172/162
1~63~ \2
amino)acetic acid. A suspension of the latter compound (0.10
moles) in 150 ml of water is treated with 8.1 g of potassium
cyanate. The resulting mixture is heated to about 80~C to give
a clear solution which is then allowed to stand at room tem-
perature for about 24 hours. Acidification to pH 3.5 with
hydrochloric acid precipitates the ~-ureido compound. A
solution containing 0.10 mol~s of the ~-ureido acid in 100 ml.
of acetone containing 10.1 g of triethylamine at a temperature
of from about 0C to about -20C is converted to a mixed
carbonic anhydride by treating with 10.8 g of ethyl chloro-
formate for about 30 minutes. A cold (about -10C.) solution ;
of 0.10 mole of 7-amino-7-methoxy-trichloroethyl ester
(prepared as described in Netherlands Patent 7r204,982) in
400 ml. of 1:1 acetone containing 10~1 g of triethylamine is
added to the solution of mixed anhydride and the reaction
mixture stirred vigorously at about 0C for approximately ~;
30-45 minutes. The volume of the solution is reduced by
evaporating the bulk of the acetone at reduced pressure at
room temperature or below.
One liter of ethyl acetate is added and the solution
washed with 2 X 200 ml. of ice cold 5% a~ueous sodium bicar-
bonate, 100 ml of water, 2 X 200 ml of 0.5 molar hydrochloric
acid, and 100 ml again of water. The ethyl acetate solution -~
dried (Na2SO4) and evaporated to deposit the trichloroethyl ;
ester of the title compound. ;
The free acid is obtained by dissolving the ester (1 g.)
in 30 ml. of cold 90% acetic acid and adding 1 g of zinc to
:,. .
the vigorously stirred solution. After one hour the acid ~ ~
solvent is evaporated at reduced pressure. The residue is ~ ~ -
30 treated with 50 ml of water and the pH adjusted to 7.5 to `-
-17-
.: .
;. ,. : : . . ~
~G172/162
~C~63~0Z ~
dissolve the product. The solution is washed with ethyl
acetate. (Lyophilization of the aqueous solution deposits
the sodium salt of the title compound). The aqueous layer ;~
is acidified to precipitate the title compound.
Example 12
7-~2-Ureido-2 (2-thienyl)acetamidol-7-methoxy-3-carbamoyloxy-
.
methyl-~3-cephem-4-carboxylic acid
2-Thiophene carboxaldehyde is reacted with ammonium
cyanide prepared in situ from ammonium chloride and sodium
cyanide. The resulting amino nitrile is hydrolyzed to yield
thienyl-2-~2-amino)acetic acid. A suspension of the latter
compound (0.10 moles) in 150 ml of water is treated with 8.1 g.
of potassium c~anate. The resulting mixture is heated to
about 80C. to give a clear solution which is then allowed to
stand at room temperature for about 24 hours. Acidification
to pH 3.5 with hydrochloric acid precipitates the ~-ureido
compound. A solution containing 0.10 moles of the ~-ureido
acid in 100 ml of a~etone containing 10.1 g of triethylamine
at a temperature of from about 0C to about -20C is converted ~`
to a mixed carbonic anhydride by treating with 10.8 g of ethyl
chloroformate for about ~0 minutes. A cold (about -10C)
solution of 0.10 mole of 7-amino-7-methoxy-3-carbamoyloxymethyl-
~3-cephemr4-carboxylic acid, benzhydryl ester in 400 ml of 1:1 ,
acetone containing 10.1 g of triethylamina is added to the
solution of mixed anhydride and the reaction mi~ture stirred
vigorously at about 0C for approximately 30-45 minutes. The
volume of the solution is reduced by evaporating the bulk of
the aceton~ at reduced pressure at room temperature or below.
One liter of ethyl aceta~e is added and the solution
30 washed with 2 X 200 ml of ice cold 5% aqueous sodium bicarbonate,
-18-
. . , :
GG172/162
~3~(~Z
100 ml of water, 2 X 200 ml of 0.5 molar hydrochloric acid, and
100 ml again of water. The ethyl acetate solution is dried (Na2SO4)
and evaporated to deposit the benzhydryl ester of the title compound.
The free acid is obtained by dissolving the ester (1 g) and
anisole (500 mg~ in 20 ml of ice cold tri~luoroacetic acid and
keeping it at 0-5C for 30 minutes. The acid solvent is evaporated
at reduced pressure. The residue is treated with 50 ml of water
and the pH adjusted to 7.5 to dissolve the product. The solution
is washed with ethyl acetate. (Lyophilization of the aqueous
solution deposits the sodium salt of the title compound). The
aqueous layer is acidified to precipitate the title compound.
Examples 13-15
Following the procedure of example 9 but s~bstituting for
2-mercapto-1,3,4-thiadiazole, the heterocyclicthio compound
listed below in column I, there is obtained respectively the
~inal compound of column II.
I II
13. Pyridinyl-l-oxo-2-thiol 7-E2-Ureido-2-(2-thienyl)-
acetamido]-7-methoxy-3-~2-
(l-oxopyridinyl)thiomethyl]-
~3-cephem-4-carboxylic acid.
14. 3-methyl-1,2,4-thiadia- 7-[2-Ureido-2-(2-thienyl)-
zolyl-5-thiol acetamidol-7-methoxy-3-[5-(3-
methyl-1,2,4-thiadiazolyl)-
thiomethyl]-~3-cephem-4-car-
boxylic acid
15. 1-methyltetrazolyl-5- 7-[2-Ureido-2-(2-thienyl)-
thiol acetamido]-7-methoxy-3-(5-(1- ~-
methyltetrazolyl)-thiomethyl]-
~3-cephem-4-carboxylic acid
-19--
: ' ' '' .'' '
~0~3102 GG172/162
Example 16
7-[2-Ureido-2-(2~thienyl)acetam_do-7-methoxy-3-[1-pyridinium-
methyl] ~3-cephem-4-carboxylic acid
.
.
A solution of 0.046 mole of the product of example 1, -
sodium salt, 10 g of potassium thiocyanate and 10 ml of pyridine ~;
in 50 ml of water is adjusted to pH 6.5 with 85% phosphoric
acid and warmed to 60 for 6 hours~ The solution is cooled to
room temperature and extracted with a 25% solution of Amberlite
LA-l (acetate form~ in methyl isobutylketone (6 X 100 ml). After ;
standing overnight at 0 to 5C the title compound separates.
Example 17
':
7-[2-Ureido-2-(2-thienyl)acetamido-7-methoxy-3-(5-(1-methyl-
tetrazolyl)thiomethyl]-~3-cephem-4-carboxylic acid
Following the procedure of example 1, but substituting
for 7-amino-7-methoxy-cephalosporanic benzhydryl ester, the
7-amino-7-methoxy-3-[5(1-methyltetrazolyl)-thiomethyl]-~3-
cephem-4-diphenylmethyl carboxylate (Belgium Patent 817836),
the title compound (m.p. 170-175) is obtained.
Example_18
7-[2-Ureido-2-(2-furyl)acetamido]-7-methox~cephalosporanic acid
Furfural is reacted with ammonium cyanide prepared in situ
from ammonium chloride and sodium cyanide. The resulting amino
nitrile is hydrolyzed to yield furyl-2-(2-amino)acetic acid. A
suspension of the latter compound (0.10 moles) in 150 ml of water
is treated with 8.1 g of potassium cyanate. The resulting mix-
ture is heated to about 80C to give a clear solution which is
then allowed to stand at room temperature for about 24 hours.
Acidification to pH 3,5 with hydrochloric acid precipitates the
~-ureido compound. A solution containing 0.10 moles of the ~-
ureido acid in 100 ml of acetone containing 10.1 g of triethyl-
-20-
.
. . . .. - . .. . ..
:
GG172/162
i31~f~
amine at a temperatuxe of from about 0C to about -20C is
converted -to a mixed carbonic anhydride by treating with 10.8 g
of ethyl chloroformate for about 30 minutes. A cold tabout -10C)
solution of 0.10 mole of 7-amino-7-methoxy-cephalosporanic acid,
benzhydryl ester [Cama et al., J.A.C.S., 94, 1408 (1972)] in
400 ml of 1:1 acetone-wa'cer containing 10.1 g of triethylamine
is added to the solution of mixed anhydride and the reaction
mixture stirred vigorously at about 0C for approximately 30-45
minutes. The volume of the solution is reduced by evaporating
the bulk of the acetone at reduced pressure at room temperature
or below.
One liter o-f ethyl acetate is added and the solution washed
with 2 X 200 ml of ice cold 5% aqueous sodium bicarbonate,
100 ml of water, 2 X 200 ml of 0.5 molar hydrochloric acid,
and 100 ml again of water. The ethyl acetate solution is dried
(Na2SO4) and evaporated to deposit the benzhydryl ester vf the
title compound.
The free acid is obtained by dissolving the ester ~l g)
and anisole (500 mg) in 20 ml of ice cold trifluoroacetic acid
and keeping it at 0-5C for 30 minutes. The acid solvent is
evaporated at reduced pressure. The residue is treated with
50 ml o water and the pH adjusted to 7.5 to dissolve the
product. The solution is washed with ethyl acetate. (Lyophili-
zation of the aqueous solution deposits the sodium salt of the
title compound). The aqueous layer is acidified to precipitate
the title compound.
. .
-21-
~... .
... . . . . . . . . . . . .
,, ~
10~3~Z GG172/162
Example 19
7-[2-Ureido-2-(2-furyl~acetamido~-7-methoxy-3-desacetoxy
cephalosporanic acid
Following the procedure of example 18 but substituting
0.1 mole of 7-amino-7-methoxy-3-desacetoxycephalosporanic acid
for 7-amino-7-methoxycephalosporanic acid, the title compound
is obtained.
Example 20
Pivaloyloxymethyl ester of 7-[2-ureido-2-(2-furyl)acetamido]-7-
methoxy-3-desacetoxy cephalosporanic acid
Chloromethylpivalate (20 mmole) is combined with 10 mmole
of the product of example 19, 0.4 ml of a 5~ aqueous sodium
iodide solution, and 170 ml of acetone. Triethylamine, 2.0 gm
(20 mmole), is added and the mixture stirred for 10 hours, then
refluxed for one hour. The reaction mixture is cooled and con-
centrated ln vacuo. The residue is partitioned between ethyl
acetate, and 5% aqueous sodium bicarbonate. The organic layer
is dried over sodium sulfate and evaporated to give the crude
product which is obtained as a powder upon trituration with ether.
Exa ple 21
Pivaloyloxymethyl ester of 7-[2-ureido-2-(2-furyl)acetamido-?-
methoxy cephalosporanic acid
Following the procedure of example 20 but substituting
10 mmole of the product of example 18 for the product of
example 19, the title compound is obtained.
Example 22
- 7-[2-Ureido-2-(2-fur~l?acetamido~-7-methoxy-3-desacetoxy
cephalosporanic acid, methyl ester
A 0.1 molar solution of the product of example 19 in di-
-~0 methoxyethane is treated with an excess of ethereal diazomethane.
-22
,.~ , . . .
,,
; . . .
GG172/162
63~0Z
Evaporation of the solvent at reduced pressure deposits the
title compound.
Example 23
Acetoxymethyl eA~e r ~ 7 ~Z ~rèido-2-(2-furyl)acetamido]-7-
methox ce halos oranic acid
_ ~ P P
Following the procedure of example 21 but substituting
chloromethylacetate for chloromethylpivalate, the title compound
is obtained.
, :
7-[2-Ureido-2-(2-furyl)acetamido]-7-methoxycephalosporan:ic acid,
potassium salt
One millimole of the product of example 18 i9 dissolved in `~
10 ml of a 0.01 N aqueous KOH solution. Lyophilization of the
solution yields the title compound.
Example 25
7-[2-Ureido-2-(2-furyl)acetamido]-7-methoxy-3-desacetoxy
cephalosporanic acld, calcium salt
One millimole of the product of example 19 is dissolved in
10 ml of a 0.05 N aqueous Ca(OH)2 solution. Lyophilization of
the solution yields the title compound.
Example 26-33
:
Following the procedure of example 18 but substituting for
furfural an equivalent amount of the aldehyde indicated in
column I, there is obtained the 7-methoxy cephalosporanic acid
indicated in column II.
I II
26. 5-chloro-2-furaldehyde 7-[2~ureido-2-(5 chloro-2-
furyl)acetamido]-7-methoxy-
cephalosporanic acid
-23-
: , . . . .
Jt. I.1,, ' ~ . '
GG172/162 ~ ~
1CÇ~311~2 ~ ~
27, 5-iodo-2-furaldehyde 7-[2-ureido-2-(5-iodo-2- -
furyl)acetamido]-7-methoxy-
cephalosporanic acid
28. 5-nitro-2-furaldehyde 7-[2-ureido-2-~5-nitro-2- ~-
furyl)acetamido]-7-methoxy-
cephalosporanic acid
29. 5-phenyl-2-furaldehyde 7-[2-ureido-2-(5-phenyl-2-
furyl)acetamido]-7-methoxy-
cephalosporanic acid
30. 3-furaldehyde 7-[2-ureido-2-(3-furyl)-
acetamido]-7-methoxy-
cephalosporanlc acid
31, 5-bromo-2-furaldehyde 7-12-ureido-2-(5-bromo-2-
furyl)acetamido]-7-methoxy-
cephalosporanic acid
32. 5-methylsulfonyl-2- 7-[2-ureido-2-(5-methylsulfonyl-
furaldehyde 2-furyl)acetamido]-7-methoxy-
cephalosporanic acid
33. 4-chloro-3-furaldehyde 7-[2-ureido-(4-chloro-3-
furyl)acetamido]-7-methoxy-3-
desacetoxy-cephalosporanic acid
7-~2-Ureido-2-(2-furyl)acetamido]-7-methoxycephalosporanic
acid, N,N'-dibenzylethylenediamine salt_
:
A solution of 0.010 mole of the product of example 18 in
25 ml of ethanol is added to a solution of 1.20 g of N,N'-di- ~ ;
benzylethy~nediamine in 25 ml of ethanol at room temperature.
Ater 15 minutes stirring the solvent is evaporated to deposit
the title compound.
-24-
: ~
GG172/162
31~3Z
~ le 35
7-[2-Ureido-2-(2-furyl)acetamido]-7-methoxy-3-[2-(5-methyl-
1,3,4-thiadiazolyl-thiomethyl~-~3-cephem-4~-carbox~lic acid
A solution (0.026 mole) of the product of example 18
NaHCO3 (2.1 g) and 3.8 g 2-mercapto-1,3,4-thiadiazole in
200 ml of p~ 6.4 phosphate buffer is stirred for 5.5 hours
at 60C. The reaction is cooled to room temperature, acidiied
to pH 3 and extracted with ethyl acetate. The ethyl acetate
layer is worked with saturated NaCl solution, dried (Na~SO4),
0 and evaporated at reduced pressure to deposit the product.
Exam~le 36
7-[2-Ureldo-2-(2-furyl)acetamido]-7-methoxy-3-carbamoyloxy-
methyl-~3-cephem-4-carboxylic acid
Furfural is reacted with ammonium cyanide prepared in situ
from ammonium chloride and sodium cyanide. The resulting amino
nitrile is hydrolyzed to yield thienyl-2-(2-amino)acetic acid.
A suspension of the latter compound (0.10 moles) in 150 ml of
water is treated with 8.1 g of potassium cyanate. The resulting
mixture is heated to about 80C to give a clear solution which
is then allowed to stand at room temperature for about 24 hours.
Acidification to pH 3.5 with hydrochloric acid precipitates the
a ureido compound. A solution containing 0.10 moles o~ the ~-
ureido acid in 100 ml of acetone containing 10.1 g of triethyl-
amine at a temperature of from about 0C to about -20C is
converted to a mixed carbonic anhydride by treating with 10.8 g
of ethyl chloroformate for about 30 minutes. A co~d (about
-10C) solution of 0.10 mole of 7-amino-7-methoxy-3~carbamoyl-
oxymethyl-~3-cephem-4-carboxylic acid, benzhydryl ester in
400 ml of 1:1 acetone containing 10.1 g of triethylamine is
added to the solution of mixed anhydride and the reaction
-25-
10~3102 GG172/162
mixture stirred vigorously at about 0C for approximately
30-45 minutes. The volume of the solution is reduced by
evaporating the bulk of the acetone at reduced pressure at
room temperature or below.
One liter of ethyl acetate is added and the solution
washed with 2 X 200 ml of ice cold 5~ aqueous sodium bicarbonate,
100 ml of water, 2 X 200 ml of 0.5 molar hydrochloric acid,
and 100 ml again of water. The ethyl acetate solution is dried
(Na2SO4) and evaporated to deposit the benzhydryl ester of the
title compound.
The free acid is obtained by dissolving the ester (1 g)
and anisole (500 mg) in 20 ml of ice cold tri1uoroacetic acid
and keeping it at 0-5C for 30 minute~, The acid solvent is
evaporated at reduced pressure. The residue is treated with
50 ml o water and the pH adjusted to 7.5 to dissolve the
product. The solution is washed with ethyl acetate. (Lyophili~
zation of the aqueous solution deposits the sodium salt of the
~itle compound). The aqueous layer is acidified to precipitate
the title compound.
Examples 37-39
Following the procedures o example 35 but substituting
for 2-mercapto-1,3,4-thiadiazole, the heterocyclicthio compound
listed below in column I, there is obtained respectively the
final compound of column II.
I II
37. Pyridinyl-l oxo-2-thiol 7-[2-Ureido-2-(2-furyl)-
acetamido]-7-methoxy-3-[2-
(l-oxopyridinyl)thiomethyl]-~3-
cephem-4-carboxylic acid
-26-
~ ~ GG172/162
38. 3-methyl-1,2,4-thia- 7-12-Ureido-2-(2-furyl)~
diazolyl-5-thiol acetamido]-7-methoxy-3-[5-(3-
,, . ;,
methyl-1,2,4-thiadiazolyl)-
thiomethyl~Q3-cephem~4-
carboxylic acid
39, l-methyltetrazolyl- 7-[2-Ureido-2-(2-furyl)-
5-thiol acetamido]-7-methoxy-3-(5-(1-
methyltetrazolyl)thiomethyl-Q3-
cephem-4-carboxylic acid
Example 40
7-[2~Ureido-2-(2-furyl)acetamido-7-methoxy-3-[1-~yridlnium-
methyl]-Q3-cephem-4-carboxylic acid
A solution of 0.046 mole o~ the product of example 18
sodium salt, 10 g of potassium thiocyanate and 10 ml of
pyridine in 50 ml of water is adjusted to pH 6.5 with 85~ ~
phosphoric acid and warmed to 60 for 6 hours. The solution ~ ~ ;
is cooled to room temperature and extracted with a 25% solution
of Amberlite LA-l (acetate form) in methyl isobutylketone ;
(6 X 100 ml). After standing overnight at 0 to 5C the title
compound separates. '
.
'.'~` '
,: ,
-27
~; ''
