close

Вход

Забыли?

вход по аккаунту

?

Патент USA US3049559

код для вставки
United States Patent 0 ”
2
1
cin-active material may be recovered by acid precipitation.
The product thus obtained may be puri?ed by recrystal
3,049,550
PROCESS FOR THE PRODUCTION OF 3-AM1N0
lization from an aqueous acidic alcohol solution.
4,7-DIHYDROXYCOUMARIN HYDROCHLORIDE
This new antibiotic, novobiocin, is comprised of the ele
ments carbon, hydrogen, nitrogen, and oxygen, combined
in a substance having approximately the formula
Claude Spencer, Chatham, and Edward Walton, Scotch
Plains, N.J., assignors to Merck & Co., Inc., Rahway,
N.J., a corporation of New Jersey
No Drawing. Original application Apr. 19, 1956, Ser
No. 579,401. Divided and this application Jan. 13,
1960, Ser. No. 7,504
2 Claims.
(Cl. 260—343.2)
3,049,550
Patented, Aug. 14, 1962
according to present data. It reacts as an acidic organic
10 compound toward and is easily soluble in alkaline 're
This invention relates to antibiotic substances and meth
ods of synthesizing the same. More particularly, it is con
agents, such as aqueous solutions of alkali-metal hydrox
ides, carbonates, and bicarbonates; it has two base-binding
groups and can be precipitated from its solution in alkalies
cerned with the synthesis of desmethylnovobiocin and
by acidi?cation. It is soluble in the lower alk-anols, lower
related antibiotic substances.
This application is a division of the copending applica 15 aliphatic ketones, acetic acid, ethyl acetate, dioxane; and
it is insoluble or merely sparingly soluble in ether, benzene,
tion Serial No. 579,401, ?led April 19, 1956, now aban
chloroform, carbon tetrachloride, ethylene dichloride,
doned.
water, and hydrochloric acid.
Novobiocin (the ‘generic name for an antibiotic the
Substantially pure novobiocin has been obtained in two
proprietary name for one brand of which is “Cathomy
cin”), is produced by growing, under controlled condi 20 crystalline modi?cations: a form crystallizing as rosettes
and melting at about 152°—154° C., and another form
tions, a previously unknown species of microorganism
having the appearance of ?at needles, melting at about
which has been named Streptomyces spheroides. This
170°—172° C. Each of these crystalline forms of the anti
microorganism, which was isolated from a sample of soil
from an old sod pasture in Vermont, has been designated
biotic can be converted into a ‘so-called normalized form,
Streptomyces spheroides MA-319 in the culture collection 25 which may be an amorphous or submicrocrystalline form,
by dissolving the crystals in acetone, quickly adding to
of Merck & Co., Inc., Rahway, New Jersey. A viable
this solution a relatively large volume of petroleum ether,
culture thereof has been deposited with the Fermentation
and recovering the precipitated normalized material by
section of the Northern Utilization Research Bureau,
?ltration.
United States Department of Agriculture at Peoria, Illi
Alkaline aqueous solutions of novobiocin and mineral
nois, and added to its permanent culture collection as 30
oil suspensions of the normalized form of the antibiotic
NRRL 2449.
exhibit characteristic absorption, the former in the ultra~
Aqueous media that are suitable ‘for aerobic cultivation
violet and the latter in the infrared portions of the radia
of strains of Streptomyces spheroides to produce novobio
tion spectrum. A solution of substantially pure novo
cin are, generally stated, those suitable for the production
of other antibiotics by cultivation of other Streptomyces 35 biocin in 0.1 N aqueous sodium hydroxide exhibits a
characteristic ultraviolet absorption peak at 3070 A. This
organisms. Such media contain ‘sources of assimilable
absorption peak is indicative of a substantially pure ma
carbon, such as carbohydrate; of assimilable nitrogen, such
terial having a speci?c absorbency of 600, measured at this
as corn steep liquor, casein hydrolysate, distiller’s solubles,
wavelength, using a solution containing one gram of pure
or the like; and inorganic salts, including trace metals, re
quired for proper metabolism of the microorganism. Pref 40 novobiocin per hundred milliliters of the solution, con
tained in a cell having an absorption path of one cen
erably, the medium is maintained at ‘a temperature of
timeter. A solution of pure novobiocin in 0.1 N aqueous
24°-28° C. during the period, usually about one to seven
methanolic hydrochloric acid exhibits a characteristic
days, in which the microorganism is cultivated, and aera
ultraviolet absorption peak at 3240 A. with
tion is provided for optimum growth of the organism and
production of novobiocin. Fermented broths produced 45
EH”... 390
in this manner have an activity of about 150-2000 novo
A
mineral
oil
suspension
of substantially pure normalized
biocin units per milliliter, and the fermentation broth solids
novobiocin exhibits characteristic infrared ‘absorption
have an activity of the order of about 2.25 novobiocin
peaks at the following wavelengths, expressed in microns:
units per milligram of solids. The antibioactive material
can be puri?ed and recovered in purer form by any of 50 5.8-6.0 (broad), 6.10, 6.21, 6.30, 6.49, 6.63, 7.4-7.6
(broad shoulder), 7.78, 7.96, 8.27 (Weak), 8.60 (shoul
several procedures.
der), 8.7 (shoulder), 9.13, 9.40, 10.0~10.1 (broad), 10.28,
For example, the whole broth may be ?ltered at the hy
10.60 (broad), 12.0-12.30 (broad), 12.60-12.75 (broad),
drogen-ion concentration of harvest, usually about pH
13.07, and 13.39.
7.0-7 .0; the ?ltrate may be extracted at a hydrogen-ion
Novobiocin units are related to the microbiological
concentration within the acid range below about pH 7.0 55
activity of substantially pure crystalline novobiocin; the
with a substantially neutral, merely slightly polar, Water
immiscible, liquid organic solvent soluble in cold concen- _
microbiological activity of substantially pure crystalline
trated sulfuric acid and in cold syrupy orthophosphoric
novobiocin has been arbitrarily taken as 5,000 units per
milligram, as determined by standard cup-plate diffusion
acid; and the organic extract may be extracted with an
aqueous alkaline buffer solution at a hydrogen-ion concen 60 methods, using Bacillus subtilis .A'ITC 12,432 as the test
organism.
tration of at least pH 8.5 to obtain a solution containing
Novobiocin is optically active, [a]D25=——27° (C., 1
a substantial concentration of novobiocin salt. The two
extraction steps may be repeated in succession, to obtain
an even more concentrated solution from which novobio
in 1 N sodium hydroxide) and [u]D25=——4-4° (0., 1 in
pyridine).
3,049,550
3
4
Novobiocin is active in inhibiting growth of gram-positive microorganisms primarily, although it also exhibits
some activity against gram-negative microorganism. It
inhibits growth of the following organisms, inter alia:
The foregoing described reactions can be shown struc
turally as follows:
H0
M. pyogenes var. albus
/0\_0
5 IX'R +
M. pyogenes var. aureus
NHCO—
Diplococcus pneumoniae
I
Corynebacterium diphtheriae type gravis
0H
CH’
L
OR I H
Corynebacterium diphtheriae type intermedius
C—GHa
Corynebacterium diphtheriae type mitis
10
(‘3H3
Corynebacterium xerose
VLR
Corynebacterium renale
Neisseria
meningitidis
Sarcina lutea
(VD)
J)_(3H0
I
.
.
15
M. pyogenes var. aureus resistant to aureomycm
M. pyogenes var. aureus resistant to streptomycin-strepto-
Mthrwm
.
o
py genes v ar .
'
au
O
CHOH
.
NHCO
CH3
CH0 CONE:
. m
CHOCH;
re u s res1stant to penici
n
$11
C_CH3
Novobiocin salts ‘also have antibiotic activity. For 20
example, the sodium salt of novobiocin, when tested by "
on on
C_OHB
OH,
H3
X
the agar streak dilution assay, was found to inhibit the
growth of various strains of M. pyogenes var. aureus, M.
Hopyogenes var. albus, Neisseria meningitidis (No. 274),
IX-R +
and Sarcina lutea (VD) at concentrations below 0.5 mcg. 25
0
0
g
on,
NHOO-
per m1. Other microorganisms are also a?ected by novo- ’
'
biocin to its salts in varying degrees.
It is now discovered, according to the present invention,
OH
03 OH,
CH CH
that S-desmethyl novobiocin and related substances can
3_
be prepared by reacting 2-acyloxy-3-carbamyl-4~methyl- 30
H3
novobiosyl chloride (IX-R) with a 3-acylamido-4,7-di- 'T 3
VII-R
.
hydroxy-coumarin (V-R) in the presence of silver oxide,
0
and then hydrolyzing the resulting reaction product. This
‘
non
NHCO
35
c1101
HO
-
CH0
reaction can be shown as follows:
l____
CHOR
NHR
cnoo ONE:
CH:
CHOCONH:
“
0
‘
HOOHs
5H
'
on
c-orn
(
v
v
‘
40
rn
ore-on
CH]
CH:
CH0 CH3
cno—
/
0
o
0
I‘JJHOH
VIII-R
o oHoooNH,
I
0110011,
CH
__
CH2:
.
0
OH
50
CH0
0
([7]CHOH
a
.
CH
CH0 0 ONE,
s
XIII-R
.
5
wherein R represents an acyl radical.
CH’
CH0 CHa
5
_
Thus, when 3-(3-['y,'y-dimethyla1lyl]-4—acyloxybenzami-
6H
\0-
CH;
CH:
H3
vdo)-4,7-dihydroxycourn-arin (VI-R) is reacted with 2-
'
acyloxy-3-carbamyl-4-methylnovobiosy1 chloride in the
presence of silver oxide and the condensation product is
hydrolyzed, 8-desmethyl-novobiocin (X) is produced. _
XII
0
LX~R +
—->
NHCOCHa
Similarly, when 3-(3-[3-methylbutyl]-4—acyloxybenzami
'do)-4,7-dihydroxycoumarin (VII-R) is reacted with 2
acyloxy-3-carbamyl-4~methylnovobiosyl chloride and the
VA’H
'intermediate acylated derivative is hydrolyzed, 8-desmeth- 65
yl-d-ihydronovobiocin (XI) is obtained.
In this way
0 _ 0
other’ compounds related to desmethylnovobiocin and
CH0
.desmethyldihydronovobiocin and having antibiotic ‘activ-
JJHOH
,ity can be produced. Thus, when 3-(2,2-dimethylchro-
'
man - 6 - carboxa-rmdo) - 4,7 -d1hydroxycoumar1n (VIH- 70
‘Room
Y R) and 3-acetamido-4,7-dihydroxycoumarin (V) are re
acted with 24acyloxy-3 -carbamyl-4-methylnovobiosyl chloride and the resulting condensation products are hydrolyzed, 8-desmethylcyclonovobiocin (X11) and 3,-ace—
tamino - 7 - (3 - carbamyl - 4 - methylnovobiosyloxy)-
NHCOOH
0 é?ocoNHz
_
OH
(ll-CH:
CH3
v
XIILC
.7
_
I
.
4 - hydroxycoumarin (XIII-C) respectively are produced. 75' wherein R is an acyl substituent.
3,049,550
5
5
amino-4,7-dihydroxycoumarin.
In carrying out this process for the preparation of
novobiocin and the compounds related thereto pursuant
to the present invention, the 3-acylamido-4,7-dihydroxy
This reaction can'be
shown structurally as follows
coumarin is dissolved in a suitable solvent, such as di
oxane, ‘and silver oxide is added thereto. To the result
HO
/
0
—o
no-
O
~0
+ Acylating ——->
ing suspension is then added the 2-acyloxy-3-carbamyl-4
agent
NHg
NHR
methylnovobiosyl chloride and the reaction mixture is
I
stirred for sufficient time to complete the formation of
OH
OH
the condensation product. Usually, it is found that stir
V
V—R
ring at room temperature for 5-10 hours is su?icient to 10 wherein R represents an acyl substituent.
complete the reaction.
Thus, the 3-amino-4,7-'dihydroxycoumarin can be
In general, it is found desirable to carry out the re
acylated by reaction with an acid anhydride in the
action in the absence of light and under substantially an
presence of pyridine, for example, acetic anhydride in
hydrous conditions in order to obtain optimum yields of
the presence of pyridine, to produce the corresponding
the desired reaction product. Also, it is found that the
acylated derivative. Alternatively, the 3-amino-4,7-di=hy
reaction proceeds more rapidly if a small amount of
droxycoumarin can be acylated by reaction with an acid
iodine is added to the reaction mixture, although the
chloride. For example, when 3-amino-4,7-dihydroxycou
addition of iodine is not essential to the condensation
reaction.
marinhydrochloride (III) is reacted with 3-('y,'y-dimethyl
allyl)-4~acyloxybenzoyl chloride, 3-(3-methylbutyl) -4
After completion of the condensation reaction, the acyl
substituent of the novobiosyl moiety is readily removed
by hydrolysis with alkali. Thus, 1after ?ltering the re
acyloxybenzoyl chloride or 2,2-d-imethylchroman-6-car
bonyl chloride in the presence of sodium acetate, 3-(3
action mixture to remove insoluble by-products and fol
lowing reduction of the excess iodine with sodium bisul
droxycoumarin (Vi), 3-(3-[3-methylbutyl]§-4-acyloxy
[7,7 - dimethylallyl] - 4 - acyloxybenzamido) - 4,7 - dihy
benzamido)-4,7-dihydroxycoumarin (VII), and 3-(2,2-di
?te, the ?ltrate is concentrated to a small volume at re
duced pressure and treated with sodium hydroxide to a
methylchroman - 6 - carboxamido) - 4,7 - dihydroxycou
mar-in (VIII) respectively ‘are produced. These reactions
pH of about 7 which hydrolyzes off the acyl substituent
of the novobiosyl moiety. The S-desmethyl novobiocin
or 8-desmethyl novobiocin-like compound is conveniently
recovered from the neutralized solution by lyophilization. 30
The product so obtained can be further puri?ed by re
crystallization from suitable solvents or solvent mixtures.
can be shown structurally as follows:
0100
111+
CH2
I
(‘E
o
/ \
The 2-acyloxy-3-carbamyl-4-methylnovobiosyl chloride
on;
(IX—R) used in the condensation reactions described
above can be prepared by acylating 3-carbamyl-4-methyl 35
novobiose (X) to form the diacyl derivative, 1,2-di
acyloxy-3-carbamyl-4-methylnovobiose (XLR), and then
w 4 To
\/—NHCO—
A)
reacting this di'acylated compound with hydrogen chloride
These re
actions can be shown structurally as follows:
CH3
0
under anhydrous conditions to produce 2-acyloxy-3-car
bamyl-4-methylnovohiosyl chloride (IX-R) .
_->
—OR I!
11
40
on:
on
11
/C\
CH3 CH3
OR
v1
0100
OHOH
45
CH2
111+
(13112
—>
on é
CHOH
Céa \ CH3
—)
CHOCONHz
0
(‘3H0 CH3
HO—
OHy-s?
—0
NHCO
X
(g H
CH O R
HO R
OR
f’? CH3
C H 01
.
(‘2112
CH2
l
CH3
CH 0 R
v11
————>
CHOCONHQ
HOOONH-z
(‘3H 0 C H3
CH3—C|1
0100
0 H O C H3
III-l-
CH3—
CH3
XI-R
/CH3 ———)
\O/—CHa
CH3
0
IX—R
HO-—-
/ To
65
NHGO~
wherein R represents an acyl radical.
The 3-carbamyl-4-methylnovobiose (X) used as the
starting material in the above process can be prepared
by processes which are more fully described in the co
6
OH;
H
\O/_cn3
VIII
pending application of Clifford Shunk, Serial No. 579,142, 70 wherein R respresents an acyl group.
?led April 19, 1956.
These reactions are conveniently carried out by suspend
Pursuant to another embodiment of the present inven
tion, it is now found that the 3-acylamido-4,7-dihydroxy
coumarin (V-R) used as a starting material in the proc
esses described above can be prepared by acylating 3
ing the coumarin compound in water adding sodium ace
tate thereto, and then adding ‘the acyl chloride in small
amounts while agitating the reaction mixture. After com
75 pletion of the reaction, the desired product is obtained by
3,049,550
by crystallization from suitable solvents.
'
In accordance with another embodiment of this inven
tion, it is now discovered that 3-amino—4,7-dihydroxy-cou
uct with oxalyl chloride at lower temperatures in a suit
able solvent medium such as benzene. After completion
of the reaction the precipitated sodium chloride is re
moved and the ?ltrate concentrated at reduced pressure
marin hydrochloride (111) can be prepared starting with
4,7-dihydroxycoumarin.
Thus, 4,7-dihydroxy-coumarin
(I) is treated with nitrous acid to produce 2,4-dioxo-7’hy
droXy-3-oximinochroman (II), and this product’ is re
10 to obtain the acid chloride.
3-amino-4,7-dihydroXy-coumarin
———->
no-
/o ~o
--—->
HO
obtained in ‘a manner similar to that described above for
the preparation of 3-('y,'y-dimethylallyl) -4-acyloxyben
zoyl chloride using 3-(3-methylbutyl)-4¢hydroxybenzoic
hydrochloride
(III). These reactions can be shown structurally as fol
lows:
15 acid as the starting material.
chloride to produce 3-(3-methylbutyl)-4-acyloxybenzoyl
NOH
I
H
'
n
o
no
chloride.
20
O
,r
—0
-—>
.HCl
Thus, the starting material
is ?rst acylated to form the corresponding acylated de
rivative and this acylated derivative is reacted with thionyl
/OT0
OH
‘
The 3-(3-[3-methylbutyl] -4-acyloxybenzoy1 chloride is
duced with hydrogen in the presence of a noble metal cat—
alyst, such as palladium on charcoal, to produce the de
sired
8
action mixture with water, acidifying the resulting solution
and recovering the precipitated product by ?ltration.
The second step in the process of preparing 3-('y,'y-di
methylallyl)-4-acetoxybenzoyl chloride is readily accom
plished by treating the sodium salt of the acylated prod
vacidifying the aqueous solution to a pH of about 2. At
this pH the product is insoluble and can be recovered by
?ltration. If desired, the product can be further puri?ed
.
'
.
'
The 2,2-dimethylchroman-64carbonyl chloride utilized
in the foregoing described process can be readily prepared
‘by reacting 2,2-dimethylchroman-6-carboxylate with
thionyl chloride at room temperature. After completion
of the reaction, the mixture is concentrated under reduced
25 pressure to obtain the desired product in solid form.
The 3-acyla.mino-4,7-dihydroxycoumarin which is used
as a starting material in the foregoing described reactions
on
III
is readily prepared by reacting 3-amino-4,7-dihydroxycou
marin with an acylating agent to produce the diacylated
product which on selective hydrolysis is converted to the
The step of converting the 4,7-dihydroxycoumarin to
2,4-dioxo-7-hydroxy-3-oximinochroman (II) is readily
3-acylamino-4,7-dihydroxycouma1in (V).
These reac
tions can be shown as follows:
carried out by suspending the starting material in water,
and adding ‘a solution of sodium nitrite. To insure com
pletion of the reaction, the mixture is shaken frequently
for 3 hours during which time the coumarin compound
gradually dissolves. After [all the coumarin compound is
dissolved, the reaction mixture is acidi?ed with acetic acid
HO—
O‘ —o
——>
NH;
611
and, cooled‘causing the oximino compound to precipitate
III
in crystalline form and it can be readily recovered by
?ltration.
The ?nal step in the process of preparing 3-amino-4,7
RO-
dihydroxycoumarin hydrochloride is conveniently carried
out by dissolving the 2,4-dioxo-7-hydroxy-3-oximino
0
0
To
chroman in a suitable solvent such as ethanol, adding a
\/—NHR
(5H
noble metal catalyst, vand at least one equivalent of hydro
IV
—)
HO
0
NHR
OH
V
chloric acid, and hydrogenating the resulting reaction
wherein R represents an acyl group.
mixture. The hydrogenation is continued until the re
In thus preparing Compound V, it is preferred to uti
quired amount of hydrogen has been absorbed. In carry
lize lower aliphatic acid esters for this purpose. Thus,
ing out this reduction, it is preferred to use a palladium 50 the diacylated derivative is conveniently obtained by react
catalyst suspended on charcoal, although other forms of
ing the starting material in the presence of pyridine with
noble metals or suspended noble metals on inert carriers
an anhydride of a lower aliphatic acid having from 2-8
can be used for this purpose. When the reaction is car
carbon atoms. The diacylated derivative so obtained is
ricd out in this manner the product is readily recovered by
selectively hydrolyzed by reaction with a solution of an
?ltering off the catalyst and evaporating the resulting solu
alkali such as sodium hydroxide.
tion to dryness.
1In the reactions described above, 3-amino-4,7-dihy
The 3-('y,'y-dimethylallyl)-4-acyloxybenzoyl chloride
used ‘as a starting material in the above-described process
droxycoumarin and its N-acylated derivatives have been
described and written structurally as coumarin com
can be prepared by acylating 3-('y,'y-dimethylallyl)-4-hy
pounds. However, these compounds may exist and react
droxybenzoic acid and treating the sodium salt of the acyl 60 as chromone compounds in which the positions of the
4-hydroxy ‘group and the keto group are reversed. Thus,
ated product with oxalyl chloride.
these compounds may be shown structurally as follows:
In carrying out the acylation of 3-('y,'y-dimethylallyl)
4-hydroxybenzoic acid any of the several methods for ef
fecting the acylation of phenolic compounds can be used.
It is ‘found that the acyl derivatives of lower aliphatic car 65
boxylic acids having vfrom 2-8 carbon atoms are particu
larly advantageous in the described process and are there
0
HO
TOE
NHR1
l
0
fore preferred intermediate products. Such acylated de
rivatives are readily prepared by reaction with the acid
wherein R1 represents hydrogen or an acyl group.
anhydride in the presence of pyridine. For example, the 70 The novel novobiocin-like compounds of the present
.acetoxy derivative is conveniently obtained by mixing the
invention, namely, 8-desmethylnovobiocin, S-desmethyl
.hydroxybenzoic acid with acetic anhydride in the presence
dihydronovobiocin, S-desmethylcyclonovobiocin and 7
of dry pyridine and allowing the resulting reaction mix
v(3 - carbamyl-4-methylnovobiosyloxy)-3-acetamino-4-hy
ture to stand overnight. The acetylated product can be
droxycoumarin, are useful antimicrobial agents. For ex
recovered by conventional means such as treating the re 75 ample, they can be utilized to remove susceptible micro
3,049,550
10
organisms from pharmaceutical equipment and the like,
EXAMPLE 5
or to separate certain microorganisms from solutions con
Preparation of 3-Carbamyl-4-Methylnovobiose
taining mixtures of several microorganisms. In addition,
these new novobiocin-like compounds are also effective
in the treatment and control of plant diseases.
To a solution of 20 g. of novobiocin in one liter of
methyl alcohol is added 10 ml. of hydrochloric acid (sp.
gr. 1.19). The resulting solution is re?uxed for about
1% hours, after which it is diluted with an equal volume
of water and cooled. The resulting precipitate is collected
and the ?ltrate is concentrated to about 300 ml. under
reduced pressure. After ?ltering the concentrated solu
tion it is made alkaline with sodium bicarbonate and
The following examples are presented as illustrative
embodiments of the present invention.
EXAMPLE 1
Preparation of 8-Desmethylnovobiocin (X)
A solution of 2.1 g. of 4,7-dihydroxy-3-(3'-[~y,'y-dimeth
lyophylized.
ylallyl] -4'-acetoxybenzoylamino) -coumarin (VI) in 50
The residual solids are extracted portionwise with about
ml. of puri?ed anhydrous dioxane is stirred while about
300 ml. of acetone and the combined acetone extracts are
4 g. of anhydrous calcium sulfate and 4.6 g. of silver ox
concentrated to about 25 ml. Methyl 3-carbamyl-4-meth
ide are added. The mixture is protected from light and
ylnovobioside crystallizes from the resulting solution, and
stirred for one hour. A small amount of iodine is added
followed by a solution of 2.9 g. of 2-acetyl-3-canbamyl-4
is collected and washed with acetone. After recrystalliza
tion from hot acetone the product is found to melt at 190
192° C.
One and one-half grams of methyl 3-carbamyl-4-meth
yl-novobioside is heated on the steam ‘bath in 150 ml. of
0.1 N hydrochloric acid about forty-?ve minutes until the
methylnovobiosylchloride in 10 ml. of pure anhydrous
dioxane. The reaction mixture is stirred for 5-10 hours.
The mixture is ?ltered to remove insoluble byproducts
and following reduction of the excess iodine with sodium
bisul?te, the ?ltrate is concentrated under reduced pres
rotation of the solution reaches a constant value. The ob
sure. The residue is hydrolyzed with dilute sodium hy
served rotation of the solution at the end of this time is
droxide solution at room temperature. The sodium hy
droxide is added until the pH of the solution becomes con 25 +0.50“ in a 1 cm. tube (D line of sodium used). The
resulting solution is lyophilized to produce 3-carbamyl-4
stant at about 7. The solution is lyophilized and the resi
methyinovobiose as a glass [a]D22=+46° (0., 1 in metha
due is leached with Warm methanol which dissolves the
nol).
mono sodium salt of S-desmethylnovobiocin. The sodi
EXAMPLE 6
um salt can be crystallized by adding ‘benzene or the free
acid can Ibe precipitated by adding acetic acid followed by 30
water.
Preparation 0]‘ 2-Acety[-3-Carbamyl—4-Methyln0v0bi0syl
Chloride From 3-Carbamyl-4-Methylnovobi0se
EXAMPLE 2
Preparation of 8-Desmethyldihydronovdbiocin (XI)
A. Preparation of 1,2-diacetyl-3-carbamyl-4-methyl
novobiose.—A 2.35 g. (0.01 mole) portion of 3-carbamyl
4-methyl-novobiose is added to 15 ml. of acetic anhydn'de
A solution of 2.1 g. of 3-(3-[3-rnethyl-butyl] ~4-acetoxy 35 and 12 g. of sodium acetate and the mixture is warmed
'benzoylamino)-4,7-dihydroxycoumarin (VII) in 50 ml.
at 100° C. for about an hour. The cooled reaction mix
of puri?ed anhydrous dioxane is stirred while about 4 g.
ture is diluted with 30 ml. of ice and water and neutralized
of anhydrous calcium sulfate and 4.6 g. of silver oxide
with sodium vbicarbonate. The product is extracted into
are added. The mixture is protected from light and 40 chloroform which is Washed with Water and dried. Re
stirred for one hour. A small amount of iodine is added
followed {by a solution of 2.9 g. of 2-acetyl-3-carbamyl-4
methylnovobiosyl chloride in 10 ml. of pure anhydrous
dioxane. The reaction mixture is stirred for 5-10 hours.
moval of the chloroform gives 1,2-diacetyl-3-carbamyl-4
methylnovobiose (XI) .
B. Preparation of Z-acetyl-3—earbamyl-4-methylnov0~
biosyl chloride-A solution of 1.6 g. (0.005 mole) of 1,2
The product is isolated in the same manner as described
45 diacetyl-3-carbamyl-4-methylnovobiose in 40 ml. of anhy
above in Example No. 1.
drous ether is saturated (at about 0° C.) with anhydrous
hydrogen chloride. The reaction mixture is kept at ice
EXAMPLE 3
Preparation of 8-Desmethylcyclonovobiocin (XII)
A solution of 2.1 g. of 4,7-dihydroxy-3~(2,2-dimethyl~
chroman-6-carboxamido)-coumarin (VIII) in 50 n11. of
puri?ed anhydrous dioxane is stirred while about 4 g. of
anhydrous calcium sulfate and 4.6 g. of silver oxide are
added. The mixture is protected from light and stirred
temperature for an extended period of time. The ether
and excess hydrogen chloride are removed at reduced pres
The last traces are removed by dissolving the
50 sure.
residue in benzene and removing it at reduced pressure to
yield 2-acetyl-3~carbamyl-4-methylnovobiosyl chloride.
EXAMPLE 7
for one hour. A small amount of iodine is added followed
Preparation of 4, 7-D ihydr0xy-3~ (3 '- ['y,'y-D imethylallyl] -
by a solution of 2.9 g. of 2-acetyl-3-carbamyl-4-methy1
novobiosyl chloride in 10 ml. of pure anhydrous dioxane.
The reaction mixture is stirred for 5-10 hours. The prod
4 '-A cetoxybenzoy lam ino) -C0umarin (VI)
uct is isolated in the same manner as described above in
Example No. 1.
EXAMPLE 4
Preparation of 7-(S-Carbamyl-4-Metlzylnovobiosyloxy)
3-Acetamino-4-Hydroxycoumarin
A solution of 1.2 g. of 3-acetamino-4,7-dihydroxycou
marin (V) in 50 ml. of puri?ed anhydrous dioxane is
A suspension of 11.5 g. of 3-amino-4,7-dihydroxycou
marin hydrochloride (III) in 100 ml. of water containing
60 16 g. of sodium acetate is treated portionwise with 13.4 g.
of 3-('y,'y-dimethylallyl)-4-acetoxybenzoyl chloride with
shaking. The mixture is shaken for about one hour after
the ?nal addition of acid chloride. The mixture is acidi
?ed to about pH 2 with hydrochloric acid. The product is
removed and recrystallized.
-
EXAMPLE 8'
Preparation of 3- (3- [3-Methylbntyl] -4-A cetaxy benzoyl
stirred while 4 g. of anhydrous calcium sulfate and 4.6 g.
amino) -4,7-D ihydroxycoumarin ( VII)
of silver oxide are added. The mixture is protected from
light and stirred for one hour. A small amount of io 70
A suspension of 11.5 g. of 3-amino-4,7-dihydroxycou
dine is added followed by a solution of 2.9 g. of 2-acetyl
3-carbamyl-4~methylnovobiosyl chloride in 10 ml. of pure
anhydrous dioxane. The reaction mixture is stirred for
marin hydrochloride (III) in 100 ml. of water containing
16 g. of sodium acetate is treated with 13.4 g. of 3-(3
methylbutyl)-4-acetoxybenzoyl chloride with shaking.
5-10 hours. The product is isolated in the same manner
The mixture is shaken for about one hour after the ?nal
as described above in Example No. 1.
75 addition of acid chloride. The mixture is acidi?ed to
3,049,550 '
,
12
11
?ltration ‘to remove sodium bromide, the toluene solution
about pH 2 with hydrochloric acid. The product is re
moved and recrystallized.
EXAMPLE 9
is concentrated under reduced pressure to 100 ml. and
~ extracted three times with 100 ml. portions of 2.5 N so
dium hydroxide. The alkaline extracts are combined,
cooled to 0° C. and carefully acidi?ed to pH 6 with 2 N
sulfuric acid. The heavy oil which separates is extracted
with ether, dried over magnesium sulfate and concentrated
in vacuo; weight 42 g. This oil is dissolved in cyclohex
Preparation of 4,7—Dihydroxy-3-(2,2-Dimethylchroman-o
Carboxamido) -C0umarin (VIII )
A suspension of 11.5 g. of 3-amino-4,7-dihydroxycou
.marin hydrochloride (III) in 100 ml. of water containing
16 g. of sodium acetate is treated portionwise with 11.2 g.
ane and extracted with a saturated solution of sodium car
10 bonate to remove some unreacted ethyl p-hydroxybenzo
of 2,2-dimethylchroman-6-carbonyl chloride with shaking.
ate. The cyclohexane layer is dried over magnesium’sule
fate and concentrated in vacuo. The residue is dissolved
The mixture is shaken for about one hour after the ?nal
addition of acid chloride. The mixture is acidi?ed to about
pH 2 with hydrochloric acid. The product is removed and
in a boiling mixture of equal parts of cyclohexane and
petroleum ether (B.P. 30-60“). After cooling at room
recrystallized.
temperature for a few hours and 3° overnight, the color
EXAMPLE 10
less prisms are collected on a ?lter; M.P. 62-66°.
Preparation of 3-Acetaminc-4,7-Dihydr0xyc0umarin
Re
crystallization from a mixture of cyclohexane and petro
leum ether (B.P. 30-60° C.) gives 18.9 g. of colorless
prisms; M.P. 66—69° C.
A solution of 115 mg. of 3-amino-4,7-dihydroxycou
marin hydrochloride (III) in 4 ml. of pyridine is cooled
in an ice-bath and 0.3 ml. of acetic anhydride is added. 20
The solution is allowed to warm to room temperature and
B. Preparation of 3-('y,'y-dimethylallyl)-4-hydr0xyben
zoic acid.—A solution of 7 g. of ethyl 3-('y,'y-dimethyl
allyl)-4-hydroxybenzoate in 30 ml. of 4 N sodium hy
is kept overnight. The mixture (containing some crystal
droxide is heated on the steam cone for four hours. After
line material) is poured into 20 ml. of ice and water, acid
cooling in an ice bath, the resulting solution of the sodium
i?ed to pH 2 with concentrated hydrochloric acid and
extracted with four 25
portions of chloroform. The 25 salt of 3-('y,'y-dimethylallyl)-4-hydroxybenzoic acid is
acidi?ed with dilute hydrochloric acid and extracted with
combined chloroform extracts are dried over magnesium
ether. The combined ether extracts are washed with
sulfate and evaporated to dryness under reduced pressure.
water, dried over magnesium sulfate, ?ltered, and con—
The solid 3-acetamino-7-acetoxy-4#hydroxycoumarin is
crystallized from ethyl acetate.
centrated in vacuo. The residue is dissolved in hot ben
-
One gram of 3-acetamino-7-acetoxy-4-hydroxycoumarin 30 zene and crystallized by slowly adding cyclohexane; M.P.
80-84". Recrystallization from a mixture of benzene and
(IV) is dissolved in -10 ml. of 10% sodium hydroxide solu
cyclohexane gives colorless prisms; M.P. 87—89°. Several
tion and kept at room temperature for about twenty min
recrystallizations from benzene give prisms melting at 94
utes. The solution is then cooled and acidi?ed to pH 2
95° C. The 3-('y,'y-dimethylallyl) -4-hydroxybenzoic acid
with concentrated hydrochloric acid. The solid product is
collected on a ?lter, washed with a small amount of cold 35 can be puri?ed by dissolving it in a solution of sodium
carbonate and extracting with ether. After acidifying the
Water and dried.
ice cold sodium carbonate solution with hydrochloric acid,
EXAMPLE 11
Preparation of 3-Amin0-4,7-Dihydr0xycoumarin
Hydrochloride
the product is collected on a ?lter and Washed with cold
water. Recrystallization from a mixture of methanol and _
40 water gives colorless needles; M.P. 101-103° C.
To a suspension of 0.50 g. of 4,7-dihydroxycoumarin
[Sonn, Ben, 50, 1299 (1917)], in 200 ml. of water is added
EXAMPLE 13
Preparation of. 3-('y,'y-Dimethylallyl)-4-Acetoxybenzoyl
a solution of 0.20 g. of sodium nitrite in 5 ml. of Water.
‘The mixture is kept at room temperature and shaken fre
Chloride From 3-('y,'y-Dimethylallyl) -4-Acetoxybenz0ic
quently. After all the dihydroxycoumarin has dissolved 45
the solution is acidi?ed with acetic acid. The oxirnino
compound 2,4-dioxo-7-hydroxy-3-oximinochroman (II),
is collected on a ?lter, washed with a small amount of
A solution of 97 mg. of 2,4-dioxo-7-hydroxy-3-oximino
.
zoic acid.—A solution of 20.6 g. (0.1 mole) of 3-('y,'y—di
methylallyl) -4~hydroxybenzoic acid (VI) in 100 ml. of dry
separates in small yellow plates and after thorough cooling
cold water and dried.
Acid
A. Preparation of 3-('y,'y-dimethylallyl)-4-acet0xyben
pyridine is treated with 12.8 g. (0.125 mole) of acetic an
50 hydride and kept at room temperature overnight. The
reaction mixture is diluted with 400 ml. of ice and water '
chroman (II) in 20 ml. of absolute ethanol is added to a
and acidi?ed, with concentrated hydrochloric acid. The
suspension of 500 mg. of prereduced palladium on acti
precipitated product is removed by ?ltration. The crude
vated charcoal (10%) in 20 ml. of absolute ethanol con
product is recrystallized from petroleum ether (B.P. 85
taining 0.58 ml. of 2.5 N hydrochloric acid. The mixture 55 100“) to yield puri?ed 3-('y,'y-dimethylallyl)-4-acetoxy
is shaken with hydrogen at room temperature and atmos
benzoic acid.
pheric pressure until two molar equivalents of hydrogen
B. Preparation of 3-('y,'y-dimethylallyl)-4-acet0xyben
have been absorbed and the reaction stops. The catalyst
zoyl chl0ride.—-A suspension of 24.8 g. (0.1 mole) of 3
is removed by ?ltration and washed with a little ethanol.
(ry,'y—dimethylallyl) -4-acetoxybenzoic acid in 50 ml. of
The ?ltrate and washings are evaporated to dryness under
water is treated with 40 ml. of 2.5 N sodium hydroxide.
reduced pressure in a nitrogen atmosphere. The product
The resultant solution is lyophilized to yield a residue of
i is .crystallized from alcohol.
sodium 3_-('y,~/Pdimethylallyl)-4-acetoxybenzoate. The so
dium salt is added portionwise with stirring at ice tem
EXAMPLE 12
to a solution of 127 g. (1 mole) of oxalyl chlo
Preparation of 3-('y,ry-Dimethylallyl) -4-Hydr0xybenz0ic 65 perature
ride in 400 ml. of dry benzene. After the ?nal addition,
Acid Starting With Ethyl-p-Hydroxybenzoate
stirring is continued for about 2 hours. The precipitated
A. Preparation of ethyl 3-('y,'y-dimethylallyl) -4-hydr0xy
sodium chloride is removed and the ?ltrate is concentrated
at reduced pressure. The residue is redissolved in benzene
benzaate.-A mixture of 7 g. of sodium metal cut into
and reconcentrated to remove last traces of oxalyl chlo~
small pieces, 300 ml. of dry toluene,’ and 50 g. of ethyl
p-hydroxybenzoate is heated under re?ux and stirred for 70 ‘ride from the 3-('y,'y-dimethylallyl)-4-acetoxybenzoyl
a few hours. The mixture is cooled in an ice bath and
chloride.
EXAMPLE 14
stirred While 50 g. of 'y,'y-dimethylallyl bromide is added
dropwise during a period of two hours. The reaction
Preparation of 3- (3-Methylbutyl)-4-Hydroxybenz0ic Acid
mixture is stirred for ?fteen hours at room temperature
and then warmed to 50° C. for one-half hour. After 75
A solution of 3 g. of ethyl 3-('y,y-dimethylallyl)-4-hy
,1.
3,049,550
13
14
droxybenzoate in 50 ml. of ethanol was hydrogenated over
EXAMPLE 16
0.5 g. of platinum oxide catalyst. The theoretical amount
Preparation of 2,2-Dimethylchr0man-6-Carbonyl
of hydrogen was taken up Within one hour. After removal
of the catalyst by ?ltration, the alcohol was distilled
under reduced pressure. The ethyl 3—(3Pmethylbutyl)-4 5
hydroxybenzoate was dissolved in 20 ml. of 4 N sodium
hydroxide and heated on the steam cone for four hours.
Chloride
A mixture of 10.3 g. (0.05 mole) of 2,2-dimethyl
chrornan-6-carboxylate and 50 ml. of thionyl chloride is
kept at room temperature overnight. The mixture is
concentrated at reduced pressure. The residue is redis
solved in 50 ml. of dry ‘benzene and reconcentrated to
was extracted with ether. The ether extract was dried
over magnesium sulfate, ?ltered, and concentrated in 10 yield a residue of the desired acid chloride.
Various changes and modi?cations of the invention can
vacuo. The residue Was dissolved in hot benzene and
cyclohexane was added. After cooling the 3-(3-methy1
be made, and, to the extent that such variations incor
porate the spirit of this invention, they are intended to be
butyl)-4-hydroxybenzoic acid was collected on a ?lter.
included Within the scope of the appended claims.
Recrystallization from a mixture of chloroform and cyclo
What is claimed is:
hexane gave colorless needles; M.P. l08~9° C.
15
After vacidi?cation With hydrochloric acid, the mixture
AnaZysis.—Calcd. for C12H16O3: C, 69.20; H, 7.75.
Found: C, 68.98; H, 7.29.
EXAMPLE 15
Preparation of 3-(3-l‘t4ethylbun/l) -4-Acetoxybenz0yl
1. A process Which comprises reacting 4,7-dihydroxy
coumarin with nitrous acid to produce 2,4-dioxo-7-hy
droxy-3-oximinochroman, and reducing this compound
by reaction with hydrogen in the presence of a noble
20 metal catalyst and hydrochloric acid to produce 3-amino
4,7-dihydroxycoumarin hydrochloride.
Chloride
2. The process of claim 1 in which the noble metal
A mixture of 10.4 g. (0.005 mole) of 3-(3-methyl
catalyst is palladium on charcoal.
butyl)-4-hydroxybeuzoic acid and 50' ml. of pyridine is
treated With 10 ml. of acetic anhydride. The mixture is
References Cited in the ?le of this patent
kept at room temperature overnight. The mixture is 25
UNITED STATES PATENTS
poured into 300 ml. of ice and water. The resultant
mixture is acidi?ed to about pH 2 with concentrated
Hoeksema et al ________ __ Mar. 22, 1960
2,929,821
hydrochloric acid. The precipitated product is removed
and puri?ed by recrystallization from petroleum ether
OTHER REFERENCES
30
(B.P. 85-100“).
Hoeksema et al.: I our. Amer. Chem. Soc., vol. 77,
A mixture of 13 g. (0.05 mole) of 3-(3-methylbuty-l)~
pages 6710-11 (1955).
4-acetoxy-benzoic acid ‘and 50 ml. of thionyl chloride is
Kaczka et all; Jour. Amer. Chem. Soc, vol. 77, pages
kept at room temperature overnight. The solution is con
6404—5 (1955).
centrated at reduced pressure. The residue is dissolved
Hinman et 211.: J our. Amer. Chem. Soc., vol. 78, pages
35
in 50 m1. of dry benzene and reconcentrated at reduced
pressure to yield a residue of the desired acid chloride.
1072-4 (March 5. 1956).
Документ
Категория
Без категории
Просмотров
0
Размер файла
973 Кб
Теги
1/--страниц
Пожаловаться на содержимое документа