Note: Descriptions are shown in the official language in which they were submitted.
~ 8
BRIEF SUMMARY OF THE INVENTION
This invention relates to novel compositions of
matter useful as anti-inflammatory agents and as inhibitors
of the progressive joint deterioration characteristic of
arthritic disease. More particularly, it relates to thera-
peutic compositions containing certain cis-2-benzoyl-3-
hydroxy-2-alkenonitriles or cationic salts thereof which melio-
rate inflammation and inhibit arthritic joint deterioration
in mammals. The invention includes the new compositions of
matter and the methods of meliorating inflammation and of
inhibiting joint deteriora~ion in mammals therewith. The
active ingredients of the novel compositions of this inven-
tion may be represented by the following structural formula:
~ C~ ~CN
1~
Mo~C~R
wherein R is alkyl having from one to four carbon atoms and
M is hydrogen or a pharmaceutically acceptable cation. The
useful pharmaceutically acceptable salts of the compounds of
the above structural formula wherein M is hydrogen are those
with pharmacologically acceptable metal cations, ammonium,
amine cations or quaternary ammonium cations. Preferred
metal cations are those derived from the alkali metals, e.g.
lithium, sodium and potassium, and from the alkaline earth
metals, e.g. magnesium and calcium, although cationic forms
of other metals, e.g. aluminum, zinc, iron and in particular
-- 1 --
1~184~8
copper, are within the scope of the invention.
Pharmacologically acceptable amine cations and those
derived from primary, secondary or tertiary amines such as
mono-, di- or trimethylamine, ethylamine, dibutylamine, tri-
isopropylamine, N-methylhexylamine, decylamine, dodecylamine,
allylamine, crotylamine, cyclopentylamine, dicyclohexylamine,
mono- or dibenzylamine, ~- or ~-phenylethylamine, ethylenedi-
amine, diethylenetriamine, and aryliphatic amines containing
up to and including 18 carbon atoms, as well as heterocyclic
amines, e.g. piperidine, morpholine, pyrrolidine, piperazine
and lower alkyl derivative thereof, e.g. l-methylpiperidine,
4-ethylmorpholine, l-isopropylpyrrolidine, 2-methylpyrroli-
dine, 1,4-dimethylpiperazine, 2-methylpiperidine, and the
like, as well as amines containing water-solubilizing or hy-
drophilic groups, e.g. mono-, di-, or triethanolamine, ethyl-
diethanolamine, N-butylethanolamine, 2-amino-1-butanol, 2-
-amino-2-ethyl-1,3-propanediol, 2-amino-2-methyl-1-propanol,
tris(hydroxy-methyl)aminomethane, N-phenylethanolamine, N-
-(p-tert-amylphenyl)diethanolamine, galactamine, N-methylgluc-
amine, N-methylglucosamine, ephedrine, phenylephrine, epi-
nephrine, procaine, and the like.
Examples of suitable pharmacologically acceptable
quaternary ammonium cations are tetramethylammonium, tetra-
ethylammonium, benzyltrimethylammonium, phenyltriethylammonium
and the like.
The cis-2-benzoyl-3-hydroxy-2-alkenonitriles of the
present invention may exist in other tautomeric forms as fol-
lows:
4'~8
o _ ._
C6H5-C f N 8 M \ /
C ~ ~ C6115- C- C- CN \ C
C \ O=C-R / C \
MO R R-IC CN
In another aspect, the present invention provides a process of
forming a cis-2-benzoyl-3-hydroxy-2-alkenonitrile having the following
general formula
H c f CN
MO-C-R
wherein R is an alkyl group having from 2 to 4 carbon atoms and M is hydrogen
or a pharmacologically acceptable cation and the tautomers thereof, which
comprises:
a) acylation of a benzoylacetonitrile anion (I) with an acyl
halide (II) as follows:
OM
C6H5 -C=CHCN + R.COX ~ C6H5CO-C-CN
(I) (II) /C\ (III)
HO R
whcrein M and R as hereinbefore defined, and X is a halogen;
b) condensation of benzoylacetonitrile with a trialkyl artho-
cstcr as follows:
CGH5co-cH2cN 3` C6H5CO-C-CN > C6H5CO-!CI-CN
/ C / C \ (III)
R' R HO R
(IV)
whcrcirl 1~ is a hcrcirlbci~orc dcfincd and R' is alkoxy, rmd then hyclrolyzing
ttlC compoulld (~[V) unde-r acidic conditions to provide (II[); or
- 3 -
c) addition of a cyanoenolate (V) to benzoyl chloride as
follows:
ONa
C H COCl -~ R-l=CIICN ~ C61-15CO-6-CN
(V) /C (III)
HO
wherein R is as hereinbefore defined;
d) condensation of an acylacetophenone (VI) with N,N-dimethyl
formamide dimethyl acetal to give a compound of general formula (VII) as
follows:
C6H5CO-CH2COR C6H5CO-C COR
H N(CH3)2
(VI) (VII)
wherein R is as hereinbefore defined and then treating the compound VII
1,~ with hydroxylamine to give a mixture of products of formulae VIII and IX
which are then reacted with a strong basa to give a cis-2-benzoyl-3-hydroxy-
2-alkenonitrile of general formula III as follows:
_ _ O
/ \ ,0 ' 11
R-C N + C6H5-1C N 6 5 ll
C - Cll IC - Cl-l HO - C - R
_ COC6~15 COR l
(VII[) (IX) (III)
whcrein R :is as hereinbefore defined.
ETAILED DESCRIPTION OF THE INVENTION
The cis-2-benzoyl-3-hydroxy-2-alkenonitriles and cationic salts
-
thercof of the present invention have been found to be higllly useful for
mcliorilting, inEIalnmati.on and inhibiting joint deterioratioll in man~nals when
a~ inistcrcd in an10urlts ranging from about one milligralll to about 250 mg.
2() per ki,logralll of body weight per day. A preferred dosagc rcgimen Eor optimum
- 3a -
<,, ,
i~l84413
results would be from about 5 mg. to about 100 mg. per kilogram of body
weight per day, and such dosage units are employed that a total of from
about 0.35 gram to about 7.0 grams of the active ingredient for a subject
of about 70 kg. of body weight are administered in a 24 hour period. This
dosage regimen may be adjusted to provide the optimum therapeutic response.
For example, several divided doses may be administered daily or the dose
may be proportionally reduced as indicated by the exigencies of the thera-
peu~ic situation. A decided practical advantage of this invention is that
the active ingredient may be administered in any convenient manner such as
by the oral, intravenous, intramuscular, topical or subcutaneous routes.
Compositions according to the present invention having the
desired clarity, stability and adaptability for parenteral use are obtained
by dissolving from 0.10% to
. .,.~s '
10.0% by weight of active compound in a vehicle consisting
of a polyhydric aliphatic alcohol or mixtures thereof. Es-
pecially satisfactory are glycerin, propylene glycol, and
polyethylene glycols. The polyethylene glycols consist of
a mixture of non-volatile, normally liquid, polyethylene
glycols which are soluble in both water and organic liquids
and which have molecular weights of from about 200 to 1500.
Although the amount of active compound dissolved in the above
vehicle may vary from 0.10 to 10.0% by weight, it is pre-
ferred that the amount of active compound employed be from
about 3.0 to about 9.0% by weight. Although various mixtures
of the aforementioned non-volatile polyethylene glycols may
be employed, it is preferred to use a mixture having an aver-
age molecular weight of from about 200 to about 400~
In addition to the active compound, the parenteral
solutions may also contain various preservatives which may
be used to prevent bacterial and fungal contamination. The
preservatives which may be used for these purposes are, for
example, myristyl-gamma-picolinium chloride, phenyl mercuric
nitrate, benzalkonium chloride, phenethyl alcohol, p-chloro-
phenyl-~-glycerol ether, methyl and propyl parabens, and
thimerosal. As a practical matter it is also convenient to
employ antioxidants. Suitable antioxidants include, for ex-
ample, sodium bisulfite, sodium metabisulfite, and sodium
formaldehyde sulfoxylate. Generally, from about 0~05 to
about 0.2% concentrations of antioxidant are employed.
For intramuscular injection, the preferred con-
centration of active compound is 0.25 to 0.50 mg./ml. of the
finished compositions. The cls-2-benzoyl-3-hydroxy-2-alkeno-
nitriles are equally adapted to intravenous administration
-- 4 --
~ ~Lh~34'i8
when diluted with water or diluents employed in intravenous
therapy such as isotonic glucose in appropriate quantities~
For intravenous use, initial concentrations down to about
0.05 to 0.25 mg./ml. of active compound are satisfactory.
The active compounds of the present invention may
be orally administered, for example, with an inert diluent
or with an assimilable edible carrier, or they may be enclosed
in hard or soft shell gelatin capsules, or they may be com-
pressed into tablets, or they may be incorporated directly
with the food of the diet. For oral therapeutic administra-
tion, the active compounds may be incorporated with excipients
and used in the form of tablets, troches, capsules, elixirs,
suspensions, syrups, wafers, and the like. Such compositions
and preparations should contain at least 0.1% of active com-
pound. The percentage of the compositions and preparations
may, of course, be varied and may conveniently be between
about 2% to about 60% of the weight of the unit. The amount
of active ingredient in such therapeutically useful composi-
tions is such that a suitable dosage will be obtained. Pre-
ferred compositions or preparations according to the present
invention are prepared so that an oral dosage unit form con-
tains between about 50 and 250 milligrams of active compound.
The tablets, troches, pills, capsules and the like
may also contain the following: a binder such as gum traga-
canth, acacia, corn starch or gelatin; excipients such as
dicalcium phosphate; a disintegrating agent such as corn
starch, potato starch, alginic acid and the like; a lubri-
cant 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 fla-
4'~
voring. When the dosage unit form is a capsule, it may con-
tain, 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 cap-
sules may be coated with shellac, sugar or both~ A syrup
or elixir may contain the active compound, sucrose as a
sweetening agent, methyl and propylparabens as preservatives,
a dye and 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.
The following test shows the activity of the ClS-
-2-benzoyl-3-hydroxy-2-alkenonitriles against chronic inflam-
mation in adjuvant induced arthritis which is accompanied by
joint destruction. Groups of three Royal Hart, Wistar strain
rats weighing 200+10 grams each were injected intradermally in
the right hind paw with Freund's adjuvant (dried human tuber-
cle bacilli in a mineral oil vehicle) at a dose of 2 mg./kg.
of body weight. Test compound was administered orally in
a 1.5% starch vehicle at various doses once daily on days
0 through 13 post challenge. Control rats were treated in
a similar manner, but given only starch vehicle. On the 14th
and 21st day post challenge the diameter of the injected paw (pr
(primary lesion) was measured by micrometer caliper. The
volume of inflamed paws were estimated from these measurements
and the results are expressed as percent inhibition of swell-
ing as compared to controls. At the same time, the other
inflamed sites, such as ears, paws and tail (seconday lesions)
were observed and each rat was graded as to degree of inflam-
-- 6 --
3448
mation and swelling present. The grading is based on a scale
of 0 to 24, where 0 represents a complete absence of induced
arthritic nodules and 24 represents the maximum degree of in-
flammation. The mean grade for each treated group is calcu-
lated and the effects of each compound are expressed as per-
cent inhibition of the control grade. Table I records the
results of tests conducted with a typical compound of the pre-
sent invention and known anti-inflammatory agents. The c~s-
-2-benzoyl-3-hydroxycrotononitrile appears to suppress the
progression of the arthritis and associated joint deteriora-
tion.
-- 7 --
:
4'~8
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34~8
Adjuvant induced experimental polyarthritis is a
specific systemic desease of the rat which shares interesting
similarities with rheumatoid arthritis. Specifically, the his-
tology of the two diseases bears a remarkable resemblence as
shown by C. M. Pearson et al., Am. J. Path. 42, 73 (1963).
E. M. Glenn, Am. J. Vet. Res. 27 (116), 339 (1966) has classi-
fied adjuvant induced polyarthritis as a crippling and perm-
anent deformity resulting from diffuse connective tissue in-
volvement around certain susceptible joints in the rat. Zahiri
et al, Can. Med. Ass. J. 101, 269 (1969) have shown that the
fusiform swelling of the distal joints is associated with edema,
congestion and synovitis including pannus formation, all of
which precede the ultimate destruction of bone and cartilage.
Furthermore, Zahiri et al. indicate that the cartilage destruc-
tion in the joint is due to an invasive pannus which originates
in the marginal synovium and extends across the articular sur-
face to erode it. When non-steroidal, anti-inflammatory agents
such as indomethacin inhibit arthritic paw swelling, which is -
composed of inflammatory cell infiltrates, they have also been
shown to prevent joint and bone deterioration [see S. Wong et
al., J. Pharm. & Exp. Ther. 185, 127 (1973) and G. R. Bobalick
et al., Agents and Actions 4, 364 (1974)]. The most pertinent
reference showing the relationship between arthritis and joint
; deterioration is an X-ray analysis of adjuvant arthritis in
the rat by Blackham et al., Agents and Actions 7, 145 (1977).
In a similar manner, inhibition of the progress of arthritis
in paws of rats treated with the compounds of this invention
~ also lessens associated joint deterioration.
-- 10 --
4~
Another method of determining a drug effect on con-
ditions which result in inflammation is by measuring the ef-
fect on ultraviolet induced erythema in guinea pigs. Albino
guinea pigs were depilitated on their flanks, the evening be-
fore testing, with a standard mixture of barium sulfide and
gum acacia. On the morning of the test, groups of four guinea
pigs were dosed by gavage one hour prior (-l hour) to ultra-
violet exposure. At 0 hour they were restrained in a plastic
container which allows exposure of 3 circular spots. They
were then exposed to ultraviolet irradiation from a "Hanovia"
Kromayer lamp, model 10, for 60 seconds. At one and four hours,
the degree of erythema for each of the three sites was assessed
according to the following scoring system: 0 = no erythema,
0.5 = incomplete circle or faint erythema, and 1.0 = complete
circle of distinct erythema. Thus, the maximum score for each
animal was 3Ø The following Table II summarizes the results
of this test with a typical compound of the present invention
and other drugs known to have a beneficial anti-inflammatory
effect in suppressing ultra-violet induced erythema in warm
blooded animals.
-- 11 --
~L18~48
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-- 12 --
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34~8
In determining the acute anti-inflammatory acti-
vity of the cls-2-benzoyl-3-hydroxy-2-alkenonitriles of the
present invention, Royal Hart, Wistar strain rats, ranging in
weight from 80 to 90 grams were used. The rats were fasted
overnight prior to dosing but had free access to water. The
test compounds were administered in aqueous suspension, by
gavage, in a volume of 1.7 ml. per 50 grams of rat [corres-
ponds to hydration volume used by Winter, et al., Proc. Soc.
Exp. Biol. & Med., 111, 544-547 (1962)]. The phlogistic agent
used was carrageenin prepared as a sterile 1% suspension in
0.9~ aqueous sodium chloride for routine testing. A volume
of 0.05 ml. was injected through a 26 gauge needle into the
plantar tissue of the right hind paw. Measurements were made
5 hours after drug administration (4 hours after carrageenin
challenge). Volumes of both the normal and carrageenin in-
- flammed feet were determined. The difference between the two
measurements is considered to be the increased edema due to
the carrageenin administration. Results are expressed as a
CtT efficacy ratio (edema of control animals/edema of treated
animals) and a C/T ratio of greater than 1.41 is considered as
active. Table III records the results of this test at the in-
dicated dose level of a typical compound of the present inven-
tion and demonstrates the anti-inflammatory effect of this
compound in comparison with known anti-inflammatory agents.
- 14 -
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-- 15 --
This invention will be described in greater detail
in conjunction with the following specific examples.
EXAMP;LE 1
Benzoylacetonitrile, thallium (I) salt
A suspension of 10 g. of benzoylacetonitrile in
200 ml. of dry diethyl ether is stirred at room temperature
while 17.2 g. of neat thallium (I) ethoxide is slowly added.
The resulting reaction mixture is stirred for one hour at
room temperature and then filtered. The collected precipitate
is washed with diethyl ether and air dried whereby there is
obtained 23.7 g. (99% yield) of the thallium (I) salt of ben-
zoylacetonitrile which may be stored indefinitely at room
temperature.
EXAMPLE 2
Cis-2-benzoyl-3-hydroxycrotononitrile
-
To a suspension of 2.2 g. of benzoylacetonitrile,
thallium (I) salt in 20 ml. of tetrahydrofuran is added 2 ml.
of acetyl fluoride with stirring at room temperature. After
one hour, another 2 ml. of acetyl fluoride is added, followed
by 2 ml. more after twleve hours. Twelve hours later, the
precipitated thallium (I) fluoride is removed by filtration
and the filtrate is evaporated to dryness. The off-white
residue is recrystallized from ethanol to provide the c15-2-
-benzoyl-3-hydroxycrotononitrile which has been described by
Musante, Gazz. Chim. Ital. 69, 523 (1939).
EXAMPLE 3
Cis-2-benzoyl-3-hydroxy-2-pentenonitrile
To a suspension of 5 g. of benzoylacetonitrile,
thallium (I) salt in 50 ml. of diethyl ether is added a solu-
tion of 5 g. of propionyl fluoride in 10 ml. of diethyl ether
- 16 -
~i34~8
with stirring at room temperature. After 12 hours, the re-
action mixture is filtered and the filtrate evaporated and
then pumped dry on a vacuum pump for one hour. The residue
is then treated with chloroform and filtered and the solid
collected provides the title compound.
XAMPLE 4
Cis-2-benzoyl-3-hydroxy-2-hexenonitrile
The procedure of Example 2 is repeated substituting
an equimolecular amount of butyryl fluoride for the acetyl
fluoride employed in that example. There is thus obtained
the cis-2-benzoyl-3-hydroxy-2-hexenonitrile after purification
by column chromatography.
EXAMPLE 5
Cis-2-benzoyl-3-hydroxy-4-methyl-2-pentenonitrile
By replacing the acetyl fluoride employed in Exam-
ple 2 with an equimolar amount of _ butyryl fluoride, there
is obtained the corresponding cis-2-benzoyl-3-hydroxy-4-meth-
yl-2-pentenonitrile after purification by column chromatography.
EXAMPLE 6
Cis-2-benzoyl-3-hydroxy-2-heptenonitrile
The general procedure of Example 2 is repeated but
replacing the acetyl fluoride employed in that example with
an equivalent amount of valeryl fluoride whereby there is
obtained the cls-2-benzoyl-3-hydroxy-2-heptenonitrile
after purification by column chromatography.
EXAMPLE 7
2-Methylbutyryl fluoride
A sample of 2-methylbutyryl chloride in dimethyl-
formamide is treated with a threefold excess of sodium fluo-
ride. The temperature is raised and the pure acid fluoride
is collected by distillation from the reaction mixture.
34 ~8
EXAMPLE 8
Cis-2-benzoyl-3-hydroxy-4-methyl-2-hexenonitrile
Following the general procedure of Example 3, ben-
zoylacetonitrile, thallium (I) salt is treated with 2-methyl-
butyryl fluoride to give the cis-2-benzoyl-3-hydroxy-4-meth-
yl-2-hexenonitrile after purification by column chromatography.
EXAMPLE 9
Cis-2-benzoyl-3-hydroxy-5-methyl-2-hexenonitrile
_
Treatment of be~nzoylacetonitrile, thallium (I) salt
with lsovaleryl fluoride by the procedure described in Exam-
ple 3 is productive of the cis-2-benzoyl-3-hydroxy-5-methyl-
-2-hexenonitrile after purification by column chromatography.
EXAMPLE 10
Cis-2-benzoyl-3-hydroxy-4,~-dimethyl-2-pentenonitrile
In the manner described in Example 3, reaction of
benzoylacetonitrile, thallium (I) salt with pivalyl fluoride
provides the corresponding cis-2-benzoyl-3-hydroxy-4,4-di-
methyl-2-pentenonitrile after purification by column chromato-
graphy. EXAMPLE ll
Cis-~-benzoyl-3-hydroxycrotononitrile, sodium salt
__
A calculated concentration of sodium hydroxide in
water is prepared and an excess of c -2-benzoyl-3-hydroxy-
crotononitrile is added and stirred for half an hour. The
mixture is filtered and the filtrate is evaporated. The gummy
residue is dissolved in dry acetone and again evaporated, pro-
viding the title compound. -
EXAMPLE 12
Cis-2-benzoyl-3-hydroxy-2-pentenonitrile, triethylammonium
-
salt
A sample of cis-2-benzoyl-3-hydroxy-2-pentenonitrile
- 18 -
34 ~1~
is dissolved in dry diethyl ether and an equivalent amount
of triethylamine is added dropwise. The mixture is cooled
in ice and the precipitate is removed by filtration to pro-
vide the title compound.
EX~MPLE 13
Cis-2-benzoyl-3-hydroxycrotononitrile, copper (II) salt
A weighed sample of c1s-2-benzoyl-3-hydroxycrotono-
nitrile is dissolved in a dilute solution of ammonium hydrox-
ide and an equivalent quantity of cupric sulfate in water is
added. The blue solution is heated on a steam bath for 3
hours and cooled. The precipitate is removed by filtration
and washed with water to yield the title compound.
EXAMPLE 14
Preparation of 50 mg. Tablets
Per Tablet Per 10,000 Tablets
0.050 gM. Cis-2-benzoyl-3-hydroxy-
crotonitrile ~ O~500 gm.
0.080 gm. Lactose ...... ~...................... 800 gm~
0.010 gm. Corn Starch (for mix) ~ .O~. lO0 gm.
0.008 gm. Corn Starch (for paste) ~ O~ 75 gm.
0 148 gm 1475 gm.
0 002 gm Magnesium Stearate (1%) ..... ~ .. 15 gm.
0.150 gm. 1490 gm.
The cis-2-benzoyl-3-hydroxycrotonitrile, lactose
and corn starch (for mix) are blended together. The corn
starch (for paste) is suspended in 600 ml. of water and heated
with stirring to form a paste. This paste is then used to
granulate the mixed powders. Additional water is used if
necessary. The wet granules are passed through a No. 8 hand
screen and dried at 120F. The dry granules are then passed
through a No. 16 screen. The mixture is lubricated with 1%
magnesium stearate and compressed into tablets in a suitable
tableting machine.
-- 19 --
EXAMPLE 15
Prepa _tion of Oral Suspension
Ingredient Amount
Cis-2-Benzoyl-3-hydroxy-2-pentenonitrile ................. 500 mg.
Sorbitol solution (70~ N.F.) .............................. 40 ml.
Sodium benzoate ..~.............................. 150 mg.
Saccharin ........................................ 10 mg.
Red dye .......................................... 10 mg.
Cherry flavor .................................... 50 mg.
Distilled water... qs... ad ....................... 100 ml.
The sorbitol solution is added to 40 ml. of dis-
tilled water and the cis-2-benzoyl-3-hydroxy-2-pentenonitrile
is suspended therein. The saccharin, sodium benzoate, flavor
and dye are added and dissolved. The volume is adjusted to
100 ml. with distilled water. Each ml. of syrup contains
5 mg. of cis-2-benzoyl-3-hydroxy-2-pentenonitrile.
EXAMPLE 16
Preparation of Parenteral Solution
In a solution of 700 ml. of propylene glycol and
200 ml. of water for injection is suspended 20.0 grams of
cis-2-benzoyl-3-hydroxy-2-hexenonitrile with stirring. After
suspension is complete, the pH is adjusted to 5.5 with hydro-
chloric acid and the volume is made up to 1000 ml. with water
for injection. The formulation is sterilized, filled into
5.0 ml. ampoules each containing 2.0 ml. (representing 40 mg.
of drug) and sealed under nitrogen.
EXAMPLE 17
Preparation of Topical Cream
Ingred ent Amount
Cis-2-benzoyl-3-hydroxy-4-methyl-
-2-pentenonitrile........................... 1.0%
Ethoxylated stearyl alcohol.................. 10.0%
Benzyl alcohol................................ 0.9%
Isopropyl palmitate........................... 5.0%
~lycerin...................................... 5.0~
Sorbitol solution (USP)....................... 5.0%
Lactic acid... qs... topH 4.0-5.0
Water......... qs... ad................................. 100.0
- 20 -
4 ~8
The ethoxylated stearyl alcohol and isopropyl pal-
mitate are heated to liquifying temperature. About 95% of
the total volume of water is placed in a separate container
followed by the glycerin and sorbitol solution. This aqueous
mixture is brought to a boil and then cooled to 60-75C.
The cls-2-benzoyl-3-hydroxy-4-methyl-2-pentenonitrile is
added to the wax phase and the mixture is stirred until a
clear solution is obtained. The benzyl alcohol is added and
dissolved in the wax phase. The water phase is passed through
a screen into the wax phase while maintaining agitation.
Both phases are kept at about the same temperature during
transfer. The mixture is cooled while agitation is continued.
At a temperature of 50-55C. the balance of the water is
added. The pH is adjusted to 4.0-5.0 with lactic acid. The
batch is cooled with minimum agitation until the cream sets
in its final form.
EXAMPLE 18
Pre aration of Intra-articular Product
P
Ingredient Amount
Cis-2-benzoyl-3-hydroxy-2-heptenonitrile .......... 2-20 mg.
NaCl (physiological saline) ........................... 0.9%
Benzyl alcohol N.F. .................................... 0.9
Sodium carboxymethylcellulose .......................... 1-5
pH adjusted to 5.0-7.5
Water for injection ........ qs ad ...................... 100
- 21 -
EXAMPLE 19
Preparation of Injectable Depo Suspension
Ingredient % w/V
Cis-2-benzoyl-3-hydroxy-4,4-dimethyl-
-2-pentenonitrile ~ O~ O~OOOO~OO~ 0.05-5
Polysorbate 80 USP ............................. Ø2
Polyethylene glycol 4000 USP ~ O~ 3.0
Sodium chloride USP ...... ,~,................... .Ø8
Benzyl alcohol N.F. ................... ......... .Ø9
HCl to pH 6-8 ~ O~ qs
Water for injection....... ~qs ad ............... 100.0
Example 20
Cyanoacetone, sodium salt
A solution of 0.174 mole of sodium ethoxide is pre-
pared by dissolving 4.0 g. of sodium in 200 ml. of absolute
ethanol. A neat sample of 15 ml. (0.184 mole) of 5-methyl-
isoxazole is then added dropwise as a colorless precipitate
forms. When the addition is complete, the mixture is cooled
in an ice bath and then filtered. The precipitate is collected,
and washed with hexane, yielding 14.0 g. of colorless product.
Example 21
Cis-2-benzoyl-3-hydroxycrotononitrile
A mixture of 1.0 g. (9.5 mmole) of cyanoacetone,
sodium salt in 20 ml, of tetrahydrofuran is stirred at room
temperature as a solution of 0.37 ml. (3.2 mmole) of benzoyl
chloride in 5 ml. of tetrahydrofuran is added. The reaction
is heated to reflux for 2 hours and the solvent is then eva-
porated. The residue is acidified and extracted with diethyl
ether. Evaporation of the organic phase provides 0.7 g. of
a yellow oil which crystallizes on standing to provide the
title compound.
- 22 -
4~8
Example 22
l-Cyano-2-butanone, sodium salt
A solution of 0.174 moles of sodium ethoxide is pre-
pared by dissolving 4.0 g. of sodium in 200 ml. of absolute
ethanol. A neat sample of 5-ethylisoxazole (17.9 g., 0.185
mole) is then added. The reaction is stirred for one hour as
a colorless precipitate forms. The mixture is cooled in an ice
bath and filtered. The precipitate is washed with hexane and
air dried, yielding the title compound.
Example 23
Cis-2-benzo~l-3-hydroxy-2-pentenonitrile
A mixture of 3.0 g. (25 mmole) of 1-cyano-2-butanone,
sodium salt in 20 ml. of tetrahydrofuran is stirred as a sol-
ution of 0.9 g. (8.6 mmole) of benzoyl chloride in 6 ml. of
tetrahydrofuran is added. The reaction is heated to reflux
for 3 hours, then cooled and the solvent evaporated. T~e
residue is acidified and extracted with chloroform. The organ-
ic phase is extracted twice with aqueous sodium bicarbonate
which in turn is acidified and extracted again with chloro-
form. The organic phase is dried and evaporated to yield the
title compound.
4~8
Several procedures exist for the attachment of the acyl
fragment to a benzoylacetonitrile side chain. The first involves
direct acylation of the benzoylacetonitrile anion (1) with an
acyl halide (2) in an appropriate solvent. The enolate anions (1)
are prepared
OH
OM ll O
[~CN + RC ~3J~R
10(1) (2) (3)
by the treatment of the benzoylacetonitrile with the appropriate
base in an inert solvent. This enolate may be generated in situ
and acylated with (2) in the same solvent or may be isolated and
reacted in a different solvent system. When M represents sodium
where (1) has been generated by treatment of the benzoylacetonitrile
with sodium hydride, sodium amide, sodium methoxide, etc. and X
represents chlorine, yields of ( 3) are low with undesired side
products sometimes predominating. Preferably, the benzoylacetoni-
trile is dissolved in diethyl ether and one equivalent of neat
thalluim (I) ethoxide is added. The stable enolate (1), where M
is thalluim, precipitates and may be collected by filtration,
dried and stored indefinitely. Suspension of (1) in an inert
solvent such as ether, tetrahydrofuran, dioxane, etc. at room
temperature and the treatment of same with an acyl fluoride (2)
where X=F causes precipitation of thalluim (I) fluoride. This
is removed by filtration and the produc-t is extracted from the
filtrate.
Another approach involves addition of the acyl frayment
as a hydrolyzable portion to yield (4). This may be
o o o
CN ~ ~ ~HO I RCN
(4) ( 3)
performed by condensation of the benzoylacetonitrile with a trialkyl ortho
estcr in refluxing acetic anhydride. Evaporation Of by products and excess
acetic anhydride in vaCuo and purification of the product under anhydrous
conditions provides (4) where R/ is alkoxy. This intermediate (4), where
R/ is alkoxy, may be hydrolyzed under acidic conditions to provide the
desired product (3).
A different approach to (3) involves addition of the cyanoacyl
fragment to a benzoyl chloride. This reaction may be
CN OIH
J ~ ~ ONa > CN
,~7 R
~ / (5) (3)
/ \ NaH
\N ~
performed by gcneration Of the enolate anion (5) in situ With the 5-alkyl-
isoxazole and base such as sodium hydride or sodium amide and subsequent
addition Of the benzoylacetonitrile to affect con-
_ 25 -
,~A.
~184~3
densation to provide (3). Alternatively the ~-cyanoenolate (5)
may be prepared separately in a similar manner as above and iso-
lated. Addition of this enolate to the appropriate benzoyl
chloride in ether, tetrahydrofuran, etc. at room temperature or
at reflux provides (3).
A less desirable route to (3) involves that discribed
below. The acylacetophenone (6)
o (CH3)2N-CH (OC 3 2
R 1~ N
t6) (7) ~CH3
O OH - NH2OH~ 1
~\ R ~ j~ ~o + [~ O N
CN NaOH N/ o R
(3) (8) (9)
is condensed with N,N-dimethyl formamide dimethyl acetal at reflux
either neat or in solution with an inert solvent such as chloro-
foxm or carbon tetrachloride to provide the intermediate (7).
Treatment of this with hydroxyl amine hydrochloride in solvents
such as alcohol, dioxane or dimethyl formamide or aqueous solu-
tions of the same provides a mixture of products (8) and (9).
These are reacted as an unseparated mixture with a strong base
such as sodium hydroxide or sodium alkoxide in alcohol and provides
the desired product (3).
_26 _
11~84 ~8
xamplc 2~
Cis-3-dimethylamino-2-(p-fluorobenzoyl)-crotononitrile
To a solution of 1.6 g. of p-fluorobenzoylacetonitrile
[Pihl _ al., Reakts. Sposobnost Org. Soedin. Tartu. Gos. Vniv.,
5 (1), 27, (1968)] in 30 ml. of chloroform, cooled to -10C., is
added 1.4 g. of N,N-dimethylacetamide dimethylacetal. The reaction
mixture is stirred at -10C. for 2 hours and then evaporated in
vacuo to an oil. This oil is dissolved in 150 ml. of benzene
and filtered through magnesol. The filtrate is evaporated to 50
ml. and petroleum ether is added to effect crystallization. The
product is collected by filtration and then recrystallized from
benzene-petroleum ether with charcoal treatment giving the desired
product.
Example 25
Benzoylacetonitrile, thallium (I~ salt
A suspension of 10 g. of benzoylacetonitrile in 200
ml. of dry diethyl ether is stirred at room temperature while
17.2 g. of neat thallium (I) ethoxide is slowly added. The result-
ing reaction mixture is stirred for one hour at room temperature
and then filtered. The collected precipitate is washed with diethyl
ether and air dried whereby there is obtained 23.7 g. (99~ yield)
of the thallium (I) salt of benzoylacetonitrile which may be stored
indefinitely at room temperature.
Example 26
Cis-2-benzoyl-3-hydroxycrotononitrile
To a suspension of 2.2 g. of benzoylacetonitrile,
thallium (I) salt in 20 ml. of tetrahydrofuran is added 2 ml. of
acetyl fluoride with stirring at room temperature. After one
hour, another 2 ml. of acetyl fluoride is added, followed by 2 ml.
more after twelve hours. Twelve hours later, the precipitated
-27 -
3448
thallium (I) fluoride is removed by filtration and the filtrate
is evaporated to dryness. The off-white residue is recrystallized
from ethanol to provide the cis-2-benzoyl-3-hydroxycrotononitrile
which has ~een described by Musante, Gazz. Chim. Ital. 69, (1939).
Example 27
1-Cyano-2-butanonel sodium salt
A solution of 0.174 moles of sodium ethoxide is pre-
pared by dissolving 4.0 g. of sodium in 200 ml. of absolute ethanol.
A neat sample of 5 ethylisoxazole (17.9 g., 0.185 mole) is then
added. The reaction is stirred for one hour as a colorless pre-
cipitate forms. The mixture is cooled in an ice bath and filtered.
The precipitate is washed with hexane and air dried, yielding the
title compound.
xample 28
Cis-2-benzoyl-3-hydroxy-2-pentenonitrile
A mixture of 3.0 g. (25mmole) of 1-cyano-2-butanone,
sodium salt in 20 ml. of tetrahydrofuran is stirred as a solution
of 0.9 g. (8.~ mmole) of benzoyl chloride in 6 ml. of tetrahydro-
furan is added. The reaction is heated to reflux for 3 hours, then
cooled and the solvent evaporated. The residue is acidified and
extracted with chloroform. The organic phase is extracted twice
with aqueous sodium bicarbonate which in turn is acidified and
extracted again with chloroform. The organic phase is dried and
evaporated to yield the title compound.
Example 29
-
Cyanoacetone, sodium salt
A solution of 0.174 mole of sodium ethoxide is pre-
pared by dissolving 4.0 g. of sodium in 200 ml. of absolute
ethanol. A neat sample of 15 ml. (0.184 mole) of 5-methyl-
isoxazole is then added dropwise as a colorless precipitate forms.
-28 -
1~84~8
When the addition is completc, the mixture i.s cooled in an lce
bath and then filtered. The precipitate is collected, and washed
- with hexane, yielding 14.0 g. of colorless product.
Example 30
Cis-2-benzoyl-3-hydroxycrotononitrile
A mixture of 1.0 g. (9.5 mmole) of cyanoacetone,
sodium salt in 20 ml. of tetrahydrofuran is stirred at room tem-
perature as a solution of 0.37 ml. (3.2 mmole) of benzoyl chloride
in 5 ml. of tetrahydrofuran is added. The reaction is heated to
reflux for 2 hours and the solvent is then evaporated. The residue
is acidified and extracted with diethyl ether. Evaporation of the
organic phase provides 0.7 g. of a yellow oil which crystallizes
on standing to provide the title compound.
~ e 31
Cis-2-benzoyl-3-methoxycrotononitrile
A solution containing 50 g. (0.34 mole3 of benzoylace-
tonitrile, 41 g. (0.34 mole) of trimethyl orthoacetate, and 90 g.
(0.88 mole) of acetec anhydride is heated to reflux for 5 hours.
The excess solvent and volatile by-products are then removed by
vacuum distillation and the residue is recrystallized from diethyl
ether. Alternatively, the distillation residue may be distilled
over on a Kugelrohr apparatus at 160C./0.25 mm,to provide a
vi~cous oil which may be crystallized from diethyl ether to provide
the pure title compound.
_xample 32
Cis-2-benzoyl 3-hydroxycrotononitrile
A solution of 5 g. of cis-2-benzoyl-3-methoxycroton-
onitrile in 50 ml. of ethanol is treated with 25 ml. of 3N aqueous
hydrochloric acid at room temperature for one hour. The solvent
i8 evaporated, the residue diluted with water, and the product is
-- 29 --
11184~8
extracted with chloroform. The organic phase is separated, dried,
filtered and evaporated to provide the title compound.
_xample 33
2-(Dimethylaminomethylene)-l-phenyl-butane-1,3-dione
A solution of 15 g. of benzoylacetone in 35 ml. of
N,N-dimethylformamide dimethylacetal is heated on a steam bath
and the volatile by-product (methanol) is distilled out. When
the theoretical amount of methanol has been collected, the reaction
mixture is pumped dry, then the product is collected by distillation
from the reaction pot at 150C./0.025 mm. Crystalline product
(19.5 g., 97% yield) is thus obtained upon cooling of the distillate.
Example 34
4-Acetyl-5-phenylisoxazole and 4-benzoyl-5-methylisoxazo _
To a solution of 12.8 g. of 2-(dimethylaminomethylene)-
-1-phenylbutane-1,3-dione in 130 ml. of THF is added 4.10 g. of
hydroxylamine hydrochloride, followed by 40 ml. of water. The
reaction is heated to reflux for 2 hours, then cooled and the sol-
vent is evaporated. The residue is partitioned between ether and
water and the organic layer is separated and evaporated. The
regidue i8 distilled at 100-110C./0.25 mm. to provide 9.0 g. of
an oily mixture of approximated equal amounts of 4-benzoyl-5-
-methylisoxazole and 4-acetyl-5-phenylisoxazole.
Example 35
Cis-2-benzoyl-3-hydroxycrotononitrile
A solution of 1.65 g. of a mixture of 4-acetyl-5-
-phenylisoxazole and 4-benzoyl-5-methylisoxazole in equal amounts
is dissolved in 7 ml. of methanol and treated with 5 ml. of 10~
aqueous sodium hydroxide. The solution is heated on a steam bath
for 15 minutes then cooled, acidified with lN aqueous HCl and
extracted with ehter. Evaporation of the ether provides 1.55 g.
of product. Recrystallization from isopropanol provides 1.0 g.
(66%) of cls-2-benzoyl-3-hydroxycrotononitrile, mp. 74.5-76.5C.
-30 _
