"A PROCESS FOR THE PREPARATION OF DABIGATRAN ETEXILATE AND PHARMACEUTICALLY ACCEPTABLE SALTS THEREOF"
FIELD OF THE INVENTION:
The present invention relates to a process for the preparation of Dabigatran etexilate and pharmaceutically acceptable salts thereof of formula (I) with high purity and good yield.
BACKGROUND OF THE INVENTION:
Direct thrombin inhibitors are a class of medication that act as anticoagulants by directly inhibiting the enzyme thrombin. Dabigatran is an oral anticoagulant from the class of direct thrombin inhibitors.
Dabigatran etexilate mesylate is chemically known as P-Alanine, N-[[2-[[[4-[[[(hexyloxy) carbonyl] amino] iminomethyl] phenyl] amino] methyl]-l-methyl-lH-benzimidazol-5-yl] carbonyl]-N-2-pyridinyl-, ethylester, methanesulfonate and is structurally represented as follows.
Its empirical formula is C34H41N7O5.CH4O3S with a molecular weight of 723.86 (mesylate salt) and 627.75 (free base). Dabigatran etexilate mesylate is commercially marketed under the trade name of PRADAXA® in United States, Europe, Australia and Canada and under the trade name of PRAZAXA® in Japan.
The active ingredient, Dabigatran etexilate of formula (I) has been disclosed in US 6,087,380 (hereinafter referred to as US '380) and WO 98/37075. The patent US '380 also discloses a process for the preparation of Dabigatran etexilate, which comprises the reaction of compound of formula (VI) with a compound of formula (V) in presence of carbonyl diimidazole (CDI) in tetrahydrofuran (THF) and further cyclization with acetic acid (AcOH) to produce the compound of formula (III) in the form of free base or acetate. The compound of formula (III) is subjected to Pinner reaction to produce the compound of formula (II).

Finally acylation of compound of formula (II) with hexylchloroformate to produce the Dabigatran etexilate of compound of formula (I), which is depicted in scheme-I given below:
5
In the first step, corresponds to the preparation of free base or acetate of the compound of formula (III) requires a purification by column chromatography, which is very difficult to apply in the industrial scale. This purification method burdens the process economy very much and has a negative impact on the yield.
) In the next step, that compound of formula (II) prepared by this method required subsequent chromatographic purification as it was an oily substance with a relatively high content of impurities. The compound of formula (II) in its hydrochloride salts was obtained in its amorphous form with a relatively high content of impurities, this compound subsequent required chromatographic purification.
In the last step, thus the obtained Dabigatran etexilate free base of formula (I) is also purified by column chromatography.
However, the US '360 process for the preparation of Dabigatran etexilate with high purity and good yield is not applicable in the industrial scale.

J. Med. Chem. 2002, 45, 1757-1766 discloses a process for the preparation of Dabigatran etexilate as same as described in US '360.
WO 2014/192030 discloses an improved process for the preparation of dabigatran etexilate and pharmaceutically acceptable acid addition salts thereof, which comprises, reaction of a diamine compound of formula (VI) with a compound of formula (V) in presence of coupling agent like carbonyldiimidazole (CDI) and an additive; further cyclization with acetic acid or p-toluene sulfonic acid (PTSA) to produce the cyano compound of formula (III) in the form of free base or acid addition salt. The cyano compound of formula (III) is subjected to Pinner reaction to produce the benzimidazole compound of formula (II) in the form of free base or acid addition salt. Finally conversion of the benzimidazole compound of formula (II) into Dabigatran etexilate or pharmaceutically acceptable acid addition salts thereof of formula (I), which is depicted in scheme-II given below:

The major disadvantage with the above process, the use of an additive along with a coupling agent making the process more cost effective and suitable only for particularly on large scale operations.
Further, the processes for the preparation of dabigatran etexilate and its methanesulfonate salt are also described in PCT Publication Nos. WO 2012/004396, WO 2013/144903, WO 2014/020555, WO 2015/128875, WO2015/132794 and WO 2016/027077.

Hence, there is consequently a need for a process for the preparation of Dabigatran etexilate which does not involves the problems described above. So, the inventors of the present invention have developed a process for preparing Dabigatran etexilate towards as high purity and good yield.
SUMMARY OF THE INVENTION
The present invention is relates to a process for the preparation of Dabigatran etexilate and pharmaceutically acceptable salts thereof of formula (I) with high purity and good yield.
In one aspect of the present invention provides a process for the preparation of dabigatran etexilate of formula (I), or a pharmaceutically acceptable salt thereof,

by comprising the steps of;
a) reacting the compound of formula (VI) with a compound of formula (V)in presence of a
coupling agent to produce the compound of formula (IV),

b) cyclizing the product of step a) with acetic acid and followed by addition of p-toluene
sulfonic acid to produce the p-toluene sulfonic acid (PTSA) salt of the compound of
formula (IIIA),


c) reacting the product of step b) with an acid and followed by a base to in presence of suitable solvent to produce the p-toluene sulfonic acid (PTSA) salt of the compound of formula IIA,
d) reacting the product of step c) with n-hexyl chloroformate in presence of base and suitable solvent to produce dabigatran etexilate of formula (I); and
e) converting the dabigatran etexilate of formula (I) into a pharmaceutically acceptable salt thereof.
DETAILED DESCRIPTION OF THE INVENTION
The main objective of the present invention relates to a process for the preparation of Dabigatran etexilate and pharmaceutically acceptable salts thereof of formula (I) with high purity, cost effective and good yield.
In one embodiment of the present invention relates to a process for the preparation of dabigatran etexilate of formula (I), or a pharmaceutically acceptable salt thereof,

by comprising the steps of;

a) reacting the compound of formula (VI) with a compound of formula (V)in presence of a
coupling agent to produce the compound of formula (IV),

b) cyclizing the product of step a) with acetic acid and followed by addition of p-toluene sulfonic acid to produce the p-toluene sulfonic acid (PTSA) salt of the compound of formula (IIIA),
c) reacting the product of step b) with an acid and followed by a base to in presence of suitable solvent to produce the p-toluene sulfonic acid (PTSA) salt of the compound of formula IIA,
d) reacting the product of step c) with n-hexyl chloroformate in presence of base and suitable solvent to produce dabigatran etexilate of formula (I); and
e) converting the dabigatran etexilate of formula (I) into a pharmaceutically acceptable salt thereof.

According to the embodiment of the present invention, theethyl-3-(3-amino-(4-methylamino)-N-(pyridine-2-yl) benzamidopropionate of formula (VI) is reacted with 2-((4-cyanophenyl) amino) acetic acid in presence of coupling agent and dimethyl formamide (DMF) to produce ethyl 3-(3-(2-((4-cyanophenyl) amino) acetamide)-4-(methylamino)-N-(pyridin-2-yl) benzamido) propanoate of formula (IV), wherein the coupling agent is selected from carbonyldiimidazole (CDI), carbonyl-di (1, 2, 4-triazole), dicyclohexylcarbodiimide (DCC) and l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC).The reaction mass carried out at temperature raised to 35-40° C, maintained it for 10-12 hrs to complete the reaction. After completion of the reaction, the mixture was allowed to cool for 20-25° C.
To the resulted mixture was added purified water to separate the two layers; further added dichloromethane to the aqueous layer, stir it for 1 Omin and settle for 20min to separate two layers. Combine both dichloromethane layers and distilled out solvent at below 45° C under pressure.
According to the embodiment of the present invention, theethyl3-(3-(2-((4-cyanophenyl) amino) acetamide)-4-(methylamino)-N-(pyridin-2-yl) benzamido) propanoate of formula (IV) is cyclized with acetic acid to produce ethyl 3-(2-(((4-cyanophenyl) amino) methyl)-1-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido) propanoate of formula (III). The reaction mass carried out at temperature raised to 70-75° C, maintained it for 6-8 hrs to complete the reaction. After completion of the reaction, the mixture was allowed to cool for 10-15° C. Add dichloromethane and purified water to the reaction mixture to separate the two layers.
Further added purified water and ammonia solution into aqueous layer, adjust the reaction mass at pH 8.0-9.0, stir it for lOmin and settle for lOmin to separate two layers. Combine both dichloromethane layers and distilled out solvent completely under pressure at below 45° C.
According to the embodiment of the present invention, the addition of p-toluene sulfonic acid (PTSA) in presence of acetone to ethyl 3-(2-(((4-cyanophenyl) amino) methyl)- 1-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido) propanoate of formula (Ill)to produce

ethyl 3-(2-(((4-cyanophenyl) amino) methyl)-! -methyl-N-(pyridin-2-yl)-l H-
benzo[d]imidazole-5-carboxamido) propanoate p-toluene sulfonic acid of formula (IIIA).
The reaction mixture heated up to 50-55° C, stir it for 60min and allowed to cool to 25-30° C, stir it for 30min; further cool the reaction mixture to 0-5° C, stir it for 30min. The obtained compound was filtered, washed with acetone and load the wet material into the tray drier, dry the material at 55-60° C for 6-8 hrs to afford ethyl 3-(2-(((4-cyanophenyl) amino) methyl)-l-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido) propanoate p-toluene sulfonic acid of formula (IIIA).
According to the embodiment of the present invention, the ethyl 3-(2-(((4-cyanophenyl)
amino) methyl)-l-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido)
propanoate p-toluene sulfonic acid of formula (IIIA) is reacted with an acid and followed by a
base to in presence of suitable solvent to produce ethyl 3-(2-(((4-carbamimidoylphenyl)
amino) methyl)-l-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido)
propanoate p-toluene sulfonic acid of formula (IIA), wherein the acid is selected from hydrochloric acid or hydrobromic acid; the base is selected from ammonia or ammonium carbonate.
When analytical HPLC revealed completion of the reaction, temperature raised to 50-55°C and solvent distilled out completely under reduced pressure. Added purified water, stir the reaction mixture at 25-30° C for 60-90min. The obtained compound was filtered, washed with purified water and then recrystallized in the mixture of ethyl acetate and ethanol to afford ethyl 3-(2-(((4-carbamimidoylphenyl) amino) methyl)-l-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido) propanoate p-toluene sulfonic acid of formula (IIA).
According to the embodiment of the present invention, the ethyl 3-(2-(((4-carbamimidoylphenyl) amino) methyl)-l-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido) propanoate p-toluene sulfonic acid of formula (IIA) is reacted with n-hexyl chloroformate in presence of base and suitable solvent to produce (E)-ethyl 3-(2-((4-(N-ethyl 3-(2-(((4-(N-((hexyloxy) carbonyl) carbamimidoyl) phenyl) amino) methyl)-1-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido) propanoate of formula (I), wherein the base is selected from potassium carbonate.

When analytical HPLC revealed completion of the reaction to obtain the compound. The obtained compound was filtered, washed with purified water, recrystallized in the mixture of acetone and acetonitrile and further recrystallized in ethyl acetate to afford (E)-ethyl 3-(2-((4-(N-ethyl 3-(2-(((4-(N-((hexyloxy) carbonyl) carbamimidoyl) phenyl) amino) methyl)-1-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido) propanoate of formula (I) i.e. Dabigatran etexilate.
According to the embodiment of the present invention, the Dabigatran etexilate of formula (I) is further converted to Dabigatran etexilate mesylate by treating Dabigatran etexilate with methane sulfonic acid in presence of suitable solvent.
The suitable solvent is selected from alcohols or ketones; wherein alcohol solvent is selected from methanol, ethanol, isopropyl alcohol, butanol and isobutanol; ketone solvent is selected from acetone, butanone, pentanone and acetophenone.
Advantages of the present invention:
1. The present invention has developed a process for the preparing Dabigatran etexilate pharmaceutically acceptable salts thereof of formula (I) with high purity and good yield.
2. The present invention is also provides simple, scalable, operation friendly and industrially applicable process.
3. The present invention PTSA salt of formula (IIIA) is giving good conversion and in low time period compare free base of formula (III) conversion.
4. The present invention gives re crystallization of formula (I) in acetone and acetonitrile mixture to remove amide impurity.
5. The present invention gives stable pure form-I by adding acetone and methane sulphonic acid solution to free base of dabigatran at 35-40° C.

The process details of the invention are provided in the examples given below, which are provided by way of illustration only and therefore should not be construed to limit the scope of the invention.
EXPERIMENTAL PROCEDURE:
Example-1:
Synthesis of Ethyl 3-(2-(((4-cyanophenyl) amino) methvl)-l-methvl-N-(pvridin-2-yl)-lH-benzofdlimidazole-5-carboxamido) propaiurate p-toluene sulfonic acid (MA)
2-((4-cyanophenyl) amino) acetic acid (64.0gm 0.368 mol) and carbonyl di imidazole (65.0gm 0.400 mol) were added to a suspension of Dichloromethane (800ml), N, N Dimethyl formamide (100ml) and heated to 35-40°C. The reaction mixture was stir for 2-3 hrs at 35-40°C,allow to cool to 15-20°C, followed by portion wise addition of ethyl-3-(3-amino-(4-methylamino)-N-(pyridine-2-yl) benzamidopropionate(100.0gm 0.291 mol) at below 20°C. The reaction mixture temperature was raised to 35-40°C and maintain temperature for 10-12 hrs. After completion of the reaction, cool to 20-25°C and added purified water (500ml) into the flask at 20-25°C to separate two layers. The obtained dichloromethane layers was distilled out solvent at below 45°C under reduced pressure, cool the mixture to 25-30°C, added Ethylacetate (300ml) and acetic acid (250ml) into the flask at 25-30°C, the reaction mixture temperature was raised to 70-75°C for 6-8 hrs to complete the reaction. After completion of the reaction cool the mixture to 10-15°C, add dichloromethane (600ml) and purified water (500ml) to the reaction mixture atlO-15°C, stir it for lOmin and settle for 20min, separate two layers. The obtain dichloromethane layers was distilled out solvent completely under reduced pressure at below 45°C, added acetone (500ml) and p-toluene sulphonic acid (55.0gm 0.291 mol) and allow to stir for 60min at 50-55°C. The reaction was cool to 25-30°C and further cooled to 0-5°C to obtain the solid. The resultant solid was filtered, washed with acetone (50ml) and dried for 6-8 hrs at 55-60°C to get pure Ethyl 3-(2-(((4-cyanophenyl) amino) methyl)-1 -methyl-N-(pyridin-2-yl)-1 H-benzo[d]imidazole-5-carboxamido) propanoate p-toluene sulfonic acid. Yield: 120 gm

Example-2:
Synthesis of Ethyl 3-(2-(((4-cyanophenyl) amino) methyl)-l-methvl-N-(pvridin-2-yl)-lH-bcnzo|dliinida7,olc-5-carboxamido) propanoate p-toluene sulfonic acid (111A)
2-((4-cyanophenyl) amino) acetic acid (64.0gm 0.3638 mol) and carbonyl diimidazole (65.0gm0.400 mol) were added to a suspension of Ethylacetate (800ml), N, N Dimethyl formamide (100ml) and heated to 35-40°C. The reaction mixture was stir for 2-3 hrs at 35-40°C, allow to cool to 15-20°C, followed by portion wise addition of ethyl-3-(3-amino-(4-methylamino)-N-(pyridine-2-yl) benzamido propionate (100.Ogm 0.291 mol) at below 20°C. The reaction mixture temperature was raised to 35-40°C and maintain the temperature for 1-2 hrs. After completion of the reaction, cool to 20-25°C, added acetic acid (250ml) into the flask at 25-30°C, the reaction mixture temperature was raised to 70-75°C for 6-8 hrs to complete the reaction.
After completion of the reaction cool the mixture to 10-15°C, add dichloromethane (600ml)
and purified water (500ml) to the reaction mixture atlO-15°C, stir it for lOmin and settle for
20min, separate two layers. The obtain dichloromethane layers was distilled out solvent
completely under reduced pressure at below 45°C, added acetone (500ml) and p-toluene
sulphonic acid (55.Ogm 0.291 mol) and allow to stir for 60min at 50-55°C. The reaction was
cool to 25-30°C and further cool to 0-5°C to obtain the solid. The resultant solid was filtered,
washed with acetone (50ml) and dried for 6-8 hrs to get pure Ethyl 3-(2-(((4-cyanophenyl)
amino) methyl)-1-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido)
propanoate p-toluene sulfonic acid. Yield: 120 gm
Example-3:
Synthesis of Ethyl 3-(2-(((4-carbaniimidovlnhenvl) amino) mctlivD-l-inethvl-IN-fpyridin-
2-yl)-lH-benzo|dliinidazole-5-carboxamido) propanoate p-toluene sulfonic acid (HA)
Ethanolic hydrochloride solution (500ml) was added to a mixture of compound of Ethyl 3-(2-(4-cyanophenyl) amino) methyl)-l-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido) propanoate p-toluene sulfonic acid (lOOgm 0.152 mol), the temperature of the reaction mixture was raised to 25-30°C and maintained for 15 hrs to complete the reaction. When analytical HPLC revealed completion of the reaction, nitrogen gas was purged for 2hrs,

further cool to 0-5°C. Added aqueous ammonia solution to adjust the reaction mixture pH to 8.0-9.0, temperature was raised to 25-30°C and maintain for 8- 10 hrs. When analytical HPLC revealed completion of the reaction, temperature raised to 50-55°C and the solvent was completely distilled out under reduced pressure. Added purified water, stir the reaction mixture at 25-30°Cfor 60-90min to obtain the solid. The obtained solid was filtered, washed with purified water and then recrystallized in the mixture of ethyl acetate and ethanol to get pure Ethyl 3-(2-(((4-carbamimidoylphenyl)amino)methyl)-l-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido)propanoate p-toluene sulfonic acid. Yield: 80.0 gm
Example-4:
Synthesis of Ethyl 3-(2-(((4-carbaniiniidoylpheiiyl) amino) methvl)-l-nicthvl-N-(pyridiii-2-vl)-lH-bcnzo|d|iinidazole-5-carboxai]iido) propanoate p-toluene sulfonic acid (HA)
Ethanolic hydrochloride solution (500ml) was added to a mixture of compound of Ethyl 3-(2-(4-cyanophenyl) amino) methyl)-1 -methyl-N-(pyridin-2-yl)-1 H-benzo[d]imidazole-5-carboxamido) propanoate p-toluene sulfonic acid (lOOgm 0.152 mol), the temperature of the reaction mixture was raised to 25-30°C and maintained for 15 hrs to complete the reaction. When analytical HPLC revealed completion of the reaction, nitrogen gas was purged for 2hrs, further cool to 0-5°C. Added ammonium carbonate to adjust the reaction mixture pH to 8.0-9.0, temperature was raised to 25-30°C and maintain for 8- 10 hrs. When analytical HPLC revealed completion of the reaction, temperature raised to 50-55°C and the solvent was completely distilled out under reduced pressure. Added purified water, stir the reaction mixture at 25-30°C for 60-90min to obtain the solid. The obtained solid was filtered, washed with purified water and then recrystallized in the mixture of ethyl acetate and ethanol to get pure Ethyl 3-(2-(((4-carbamimidoylphenyl)amino)methyl)-l-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido) propanoate p-toluene sulfonic acid.
Yield: 80.0 gm

Example-5:
Synthesis of(E)-ethyl 3-(2-((4-(N-ethyl 3-(2-(((4-(N-(Oiexvloxy) carbonyl)
carbamimidoyl) phenyl) amino) methvl)-l-methyl-N-(pyridin-2-yl)-lH-
benzo[dlimidazole-5-carboxamido) propanoate (I)
Ethyl 3-(2-(((4-carbamimidoylphenyl)amino)methyl)-l-methyl-N-(pyridin-2-yl)-lH-
benzo[d]imidazole-5-carboxamido)propanoate p-toluene sulfonic acid (100.0 gm 0.148 mol) was added into acetone (600ml) and purified water(300ml) at 25-30°C. The reaction mixture was allow to cool at 10-15°C, followed by addition of potassium carbonate (69.0gm 0.5 mol) at same temperature and stir it for 15 min, further added hexylchloroformate solution (37.0 gm 0.224 mol) slowly over a period of 30minutes at 10-15°C to obtain the solid compound. (When analytical HPLC revealed completion of the reaction). The obtain solid was filtered and washed with purified water (100.0ml), additionally the wet solid was recrystallized in the mixture of acetone(500ml) and acetonitrile(500ml) to get compound (I). The resultant solid was further re-purified by recrystallization with ethylacetate (600ml) to get high pure (E)-ethyl3-(2-((4-(N-ethyl 3-(2-(((4-(N ((hexyloxy) carbonyl) carbamimidoyl) phenyl) amino) methyl)-l-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido) propanoate.
Yield: 80.0 gm
Exainplc-6:
Synthesis of (E)-ethyl 3-(2-((4-(N-ethyl 3-(2-(((4-(N-((hexyloxy) carbonylcarbamimidoyl)
phenyl) amino) methyl)-l-methyl-N-(pyridin-2-vl)-lH-benzo[dlimidazole-5-
carboxamido) propanoate methane sulphonate
(E)-ethyl 3-(2-((4-(N-ethyl-3-(2-(((4-(N ((hexyloxy) carbonyl) carbamimidoyl) phenyl)
amino) methyl)-l-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido)
propanoate (lOOgrh) was suspended in acetone (1000ml 0.159 mol) and allow to stir at 40-45°C for 30 min to get dissolved. The reaction mixture was further cool to 30-35°C, followed by slow addition of methane sulphonic acid solution in acetone (methane sulphonic acid was

added into acetone (200ml) at 0-5°C over a period of 60min) and stir for 15min to get a precipitate solid . The resultant solid was filtered, washed with acetone and dried at 50-60°C for 3-5hrs to get pure (E)-ethyl 3-(2-((4-(N-ethyl-3-(2-(((4-(N ((hexyloxy) carbonyl) carbamimidoyl) phenyl) amino) methyl)-l-methyl-N-(pyridin-2-yl)-lH-benzo[d]imidazole-5-carboxamido) propanoate methane sulphonate.
Yield: 110 gm

WE CLAIM:
1. A process for the preparation of dabigatran etexilate of formula (I), or a pharmaceutically acceptable salt thereof,

by comprising the steps of;
a) reacting the compound of formula (VI) with a compound of formula (V) in
presence of a coupling agent to produce the compound of formula (IV),

b) cyclizing the product of step a) with acetic acid and followed by addition of p-
toluene sulfonic acid to produce the p-toluene sulfonic acid (PTSA) salt of the
compound of formula (IIIA),

c) reacting the product of step b) with an acid and followed by a base to in presence
of suitable solvent to produce the p-toluene sulfonic acid (PTSA) salt of the
compound of formula IIA,


d) reacting the product of step c) with n-hexyl chloroformate in presence of base and suitable solvent to produce dabigatran etexilate of formula (I); and
e) converting the dabigatran etexilate of formula (I) into a pharmaceutically acceptable salt thereof.

2. The process as claimed in claim 1, wherein the coupling agent is selected from carbonyldiimidazole (CDI), carbonyl-di (1, 2, 4-triazole), dicyclohexylcarbodiimide (DCC) and l-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC).
3. The process as claimed in claim 1, wherein the acid is selected from hydrochloric acid or hydrobromic acid; the base is selected from ammonia, ammonium carbonate and potassium carbonate.
4. The process as claimed in claim 1, wherein the suitable solvent is selected from alcohols or ketones.
5. The process as claimed in claim 4, wherein the alcohol solvent is selected from methanol, ethanol, isopropyl alcohol, butanol and isobutanol; ketone solvent is selected from acetone, butanone, pentanone and acetophenone.