Concentrating to dryness

N-Acetyl-(R)-phanylalanlna (6). The rhodium catalyst was obtained by adding (-) dlop 5 (from diethyl tartrate) to a benzene solution of [RhCi(cyclooctene)2]2 under Ar, and stirring for tS mn A solution of the Rh catalyst (1 mM in EtOH PhH 4 1) was introduced under Hj to a solution of a-N acetylamino- phenytacrylic acid 4 (molar ratio Rh 4 1.540) The solvent was evaporated, the residue dissolved In 0 5 N NaOH, the catalyst was filtered and the solution acidified and concentrated to dryness to give 6 (81% ee) in 90 95% yield... 

S-Chloro-2-phenyllndole (2). A stirred solution ol N-(4-chioro-2-methylphenyl)-benzamkle 1 (12 25 g, 50 mmol) m THF (100 ml.) was maintained under N2 at 0°C and treated with n-txityllilhlum in hexane (0.1-0 15 mol). The stirred mature was kept at room (emperalure lor 15 h, treated with 2N HCI (60 mL) under ice cooling and extracted with PhH. The combined organic layers were concentrated to dryness to aftord 10 6 g ol 2 (94%). mp 195-196 C (trom Et20-PhH). 

Hydrolysis of 6p,l9-Oxido-3a,5a-cyclo steroids.A solution of 0.1 g of 3a,5a-cyclo-6)5,19-oxidocholestane in 20 ml of acetone is treated with 1.0 ml of water and 0.5 ml of 1 N sulfuric acid. After standing for 2 hr at room temperature, the solution is neutralized with saturated sodium bicarbonate solution and extracted with ether. The combined ether extracts are washed with water and concentrated to dryness under reduced pressure. Recrystallization of the crude product from methanol yields 85 mg choIest-5-ene-3, 19-diol mp 147-149° [aj —30°. 

P-Hydroxy-A-norpregn-3(5)-en-2-one (7) A solution of the hydroxy-methylene steroid (5) (24.8 g) dissolved in 240 ml of acetic acid and 240 ml of ethyl acetate is ozonized at — 10° with one molar equivalent of ozone. The resulting solution is diluted with 240 ml. of water and 60 ml of 30 % hydrogen peroxide and allowed to stand overnight. The solution is diluted with 1.5 liters of water and extracted with 3 x 700 ml portions of ethyl acetate. The combined extracts are washed with water, saturated sodium chloride solution, dried over sodium sulfate and concentrated to dryness under vacuum, leaving 23.4 g of a colorless amorphous residue of crude diacid. This material shows a maximum in the ultraviolet spectrum at 224 mp (s 6,400) indicating a 53 % yield of unsaturated acid (6). It is used without further purification. 

Twenty grams of the resulting oily substances were mixed with 20 grams of silicic acid (Mal-linckrodt Chemical Co.),applied to a column 40 cm in length and 4.5 cm in diameter filled with silicic acid,and eluted with a be nzene-ecetone-methanol mixture. The initial eluate which eluted with a 1 1 0 mixture was discarded and the active fractions eluted with 1 3 0 and 1 3 0.3 mixtures were collected and concentrated to dryness in vacuo. 11 g of this crude substance was then dissolved in a small amount of ethyl acetate and applied to the same silicic acid column as above. After discarding the initial eluates by the 1 1 and 2 1 benzene-acetone mixtures, aclacinomycin 8 fractions were first eluted with the above mixtures of 1 3 and 1 5 ratio, and aclacinomycin A fractions were then eluted with the 1 5 0.5 and 1 5 1 benzene-acetone-methanol mixtures. The eluates were dried over anhydrous sodium sulfate and concentrated to dryness in vacuo. 4,8 g of crude aclacinomycin A and 3.5 g of aclacinomycin 8 were obtained as yellow powder. 

Chloro-1 -methyl-5-phenyl-s-trizolo[4,3-a]quinoline A stirred mixture of 6triethyl-orthoacetate (0.925 g,0.0057 mol) and xylene (100 ml) was refluxed, under nitrogen, for 2 hours 40 minutes. During this period the ethanol formed in the reaction was removed by distillation through a short,glass helix-packed column. The mixture was concentrated to dryness In vacuo and the residue was crystallized from methanol-ethyl acetate to give 1.28 g of 7-chloro-1-methyl-5-phenyl-s-triazolo[4,3-a]-quinoline (83.9% yield). The analytical sample was crystallized from methylene chloride methanol and had a melting point 252.5°-253.5°C. 

The benzyloxycarbonyl-L-proline thus obtained (180 g) is dissolved in a mixture of dichloro-methane (300 ml), liquid isobutylene (800 ml) and concentrated sulfuric acid (7.2 ml). The solution is shaken in a pressure bottle for 72 hours. The pressure is released, the Isobutylene is allowed to evaporate and the solution is washed with 5% sodium carbonate, water, dried over magnesium sulfate and concentrated to dryness in vacuo, to obtain benzyloxycarbonyl-L-proline tert-butyl ester, yield 205 g. 

The next step yields 1-(3-acetylthio-2-methylpropanoyl)-L-proline tert-butyl ester. L-proline tert-butyl ester (5.1 g) is dissolved in dichloromethane (40 ml) and the solution stirred and chilled in an ice bath. Dicyclohexylcarbodiimide (15 ml) is added followed immediately by a solution of 3-acetylthio-2-methylpropanoic acid (4.9 g) in dichloromethane (5 ml). After 15 minutes stirring in the ice bath and 16 hours at room temperature, the precipitate is filtered off and the filtrate is concentrated to dryness in vacuo. The residue is dissolved in ethyl acetate and washed neutral. The organic phase is dried over magnesium suifateand concentrated to dryness in vacuo. The residue 1-(3-acetylthio-2-methylpropanoyl)-L-proline tert-butyl ester is purified by column chromatography (silica gel-chloroform), yield 7.9 g. 

Then, 1-(3-acetylthio-2-methylpropanoyl)-L-proline is produced. The 1-(3-acetylthio-3-methyl-propanoyl)-L-proline tert-butyl ester (7.8 g) is dissolved in a mixture of anisole (55 ml) and trifluoroacetic acid (110 ml). After one hour storage at room temperature the solvent Is removed in vacuo and the residue is precipitated several times from ether-hexane. The residue (6.8 g) is dissolved in acetonitrile (40 ml) and dicyclohexylamine (4.5 ml) is added. The crystalline salt is boiled with fresh acetonitrile (100 ml), chilled to room temperature and filtered, yield 3 g, MP 187°C to 188°C. This material is recrystallized from isopropanol [ttlo -67° (C 1.4, EtOH). The crystalline dicyclohexylamine salt is suspended in a mixture of 5% aqueous potassium bisulfate and ethyl acetate. The organic phase is washed with water and concentrated to dryness. The residue is crystallized from ethyl acetate-hexane to yield the 1-(3-acetylthio-2-D-methylpropanoyl-L-proline, MP83°Cto 85°C. 

The filtrate is concentrated to dryness in vacuo and the residue is taken up in 25 cc of ethanol. The residual ammonium bromide Is removed by filtration and to the filtrate... 

A stirred mixture of 11 g of 6-chloro-a-methylcarbazole-2-acetic acid ethyl ester, 100 ml ethanol and 100 ml of 3N sodium hydroxide was heated (N2 atmosphere). After 2 hours at reflux, the reaction mixture was concentrated to dryness under reduced pressure. Water (300 ml) and ice (200 g) were added to the residue and concentrated hydrochloric acid was added until the mixture was strongly acid. The acidic mixture was extracted with ether (3 X 200 ml). The ether extracts were combined, washed by extraction with water (3 x 100 ml) and dried over anhydrous magnesium sulfate. Following filtration of the desiccant and evaporation of the solvent, a yield of 9.8 g (98.2%) was obtained. Crystallization from CHCI3 yielded 6.2 g (62.0%) of 6-chloro-0 -methylcarbazole-2-acetic acid, MP 197°-198°C. A second crop of 1.6g,MP 195°-199°C was obtained from the mother liquors. 

A mixture of 1.63 g of 5-hydroxy-3,4-dihydrocarbostyril, 2.5 g of epibromohydrin and 2 drops of piperidine was heated at a temperature of 95°C to 100°C for a period of 4 hours with stirring. The reaction mixture was then concentrated to dryness under reduced pressure and the residue was recrystallized from acetone to obtain 1.2 g of 5-(2,3-epoxy)propoxy-3,4-dihy-drocarbostyril as a colorless powder having a melting point of 172°C to 173°C,... 

B) Preparation of 7-Chloro-3-Methoxycarbony/-5-Phenyl-2-0xo-2,3-Dihydro-iH-Benzo [fl-1,4-Diazepine (4347 CB) A solution of 9.2 g (0.04 mol) of compound 4356 CB in 20 ml of methanol is added dropwise, in the course of one hour and 30 minutes, to a boiling solution of 9.2 g (0.05 mol) of the hydrochloride of methyl aminomalonate in 30 ml of methanol. When this is completed, heating under reflux is continued for 30 minutes and the product then concentrated to dryness under reduced pressure. The residue is taken up in water and ether, the ethereal layer separated, the product washed with water and dried over sodium sulfate. The solvent is evaporated under reduced pressure. The residue, which consists of the methyl ester, could not be obtained in the crystalline state. It is dissolved in 25 ml of acetic acid, heated under reflux for 15 minutes, the product evaporated to dryness and the residual oil taken up in ether. A colorless solid separates which... 

N-cyclohexyl-1-chlorophthalimlde (250 g) was dissolved in glacial acetic acid (2.5 8),concentrated hydrochloric acid (555 ml) and tin (27B g) were added and the suspension was heated on a steam bath for 16 hours. The cooled solution was filtered and concentrated to dryness in vacuo to give a white solid. This solid was dissolved in water and the precipitated oil extracted with chloroform. The chloroform solution was dried and concentrated in vacuo to give a solid which, after recrystal I izat ion, yielded 5-chloro-2-cyclohexylisoindolin-1-one (43%), MP 140°Cto 142°C. 

A solution of 1.0 g of A -3,11-diketo-20-cyano-21-acetoxy-pregnene in 10 cc of benzene is treated with 1.0 g of osmium tetroxide and 0.43 g of pyridine. After standing at room temperature for 18 hours, the resulting solution is treated successively with 50 cc of alcohol, and with 50 cc of water containing 2.5 g of sodium sulfite. The mixture is stirred for 30 hours, filtered, and the filtrate acidified with 0.5 cc of acetic acid and concentrated to small volume in vacuo. The aqueous suspension is then extracted four times with chloroform, the chloroform extracts are combined, washed with water and concentrated to dryness in vacuo. Recrystallization of the residue from acetone gives 3,11,20-triketo-17(a)-21-dihydroxy-pregnane MP 227° to 229°C. This compound is then treated with acetic anhydride and pyridine for 15 minutes at room temperature to produce 3,11,20-triketo-17(a)-hydroxy-21-acetoxy-pregnane or cortisone acetate. 

The organic phase is dried over sodium sulfate and then concentrated to dryness. 1.62 g of the thus obtained crude 1,2a-methylene-6,7a-oxido-A -pregnene-17a-ol-3,20-dione-17-acetate are dissolved in 109 cc of glacial acetic acid. This solution is then saturated at room temperature with hydrogen chloride gas and stored for 20 hours. It is then diluted with methylene chloride and washed with water until neutral. 

The final extract is faintly pale yellow in color, whereas the previous extracts are orange. The combined ether extracts are concentrated to dryness and about 3 grams of a reddish-brown residue is obtained. The residue is stirred with approximately 400 cc of petroleum ether for two to three hours, the solvent decanted and the residue treated again with approximately 400 cc of petroleum ether. A pale yellow oil constituting crude actinomycin... 

The eluate is concentrated to dryness under reduced pressure, taken up in 25 cc of hot acetone, filtered, and diluted with ether. The pigment which crystallizes as red-brick colored platelets is essentially pure but may be recrystallized if desired from hot ethyl acetate. An analysis of the product showed C = 59.01 H = 6.B1 N=13.3B. 

The mixture was refluxed gently on a steam bath for VA hours. Fifteen minutes after initiating the reaction, the reaction mixture gave a negative ferric chloride test. Most of the ethanol and acetic acid were removed by distillation in vacuo, 300 ml of water and 300 ml of ether were added to the concentrate, and the mixture was shaken. The layers were separated, the aqueous layer extracted with fresh ether, and the combined ether extracts were washed with water, dried over anhydrous sodium sulfate, filtered and evaporated to dryness in vacuo. The residue was crystalli2ed by trituration with ether, and the crystals were collected by filtration, washed with hexane and dried. The mother liquors were concentrated to dryness and dissolved in a minimum amount of acetone, whereupon a second crop was obtained. The two crops were combined, dissolved in ethyl acetate, decolori2ed with activated charcoal, and recovered by concentration. 

The product (12.1 mg) obtained in the above step was dissoived in 0.3 mi of water, to which was then added a catalytic quantity (about 5 mg) of platinum oxide. Hydrogenation was made with hydrogen gas at a pressure of 35 kg/cm for 1.5 hours. The reaction solution was filtered to remove the catalyst, and the filtrate was concentrated to dryness, giving the desired product 3, 4 -dideoxykanamycin B in the form of its monocarbonate. The yield was 11.5 mg (95%). -f 110° (c 1, water). The overall yield of 3, 4 -dideoxykanamycin B... 

At the end of the addition, an almost colorless ether layer swims on the surface of the strongly colored water layer. After removal of the ether layer, the water layer is concentrated to dryness under vacuum and a stream of an inert gas. An earthy precipitate is formed, which after recrystallization yields 100 grams of hydroquinone calcium sulfonate, which decomposes without melting above 250°C. 

A solution of 5 grams of (3-hydroxyphenyl)ethyl dimethylammonium hydroxide in about 200 cc of water was neutralized with dilute hydrochloric acid. On concentration to dryness in vacuo, (3-hydroxyphenyl)ethyl dimethylammonium chloride crystallized. The compound was recrystallized from isopropanol MP 162° to 163°C (with decomposition). 

Then, ethyl methyl(3-benzoylphenyl)cyanoacetate employed as an intermediate material is prepared as follows The sodium derivative of ethyl (3-benzoylphenyl)cyanoacetate (131 g) is dissolved in anhydrous ethanol (2 liters). Methyl iodide (236 g) is added and the mixture is heated under reflux for 22 hours, and then concentrated to dryness under reduced pressure (10 mm Hg). The residue is taken up in methylene chloride (900 cc) and water (500 cc) and acidified with 4N hydrochloric acid (10 cc). The methylene chloride solution is decanted, washed with water (400 cc) and dried over anhydrous sodium sulfate. The methylene chloride solution is filtered through a column containing alumina (1,500 g). Elution is effected with methylene chloride (6 liters), and the solvent is evaporated under reduced pressure (10 mm Hg) to give ethyl methyl(3-benzoylphenyl)cyano-acetate (48 g) in the form of an oil. 

In the final production preparation, a mixture of ethyl methyl(3-benzoylphenyl)cyano-acetate (48 g), concentrated sulfuric acid (125 cc) and water (125 cc) is heated under reflux under nitrogen for 4 hours, and water (180 cc) is then added. The reaction mixture is extracted with diethyl ether (300 cc) and the ethereal solution is extracted with N sodium hydroxide (300 cc). The alkaline solution is treated with decolorizing charcoal (2 g) and then acidified with concentrated hydrochloric acid (40 cc). An oil separates out, which is extracted with methylene chloride (450 cc), washed with water (100 cc) and dried over anhydrous sodium sulfate. The product is concentrated to dryness under reduced pressure (20 mm Hg) to give a brown oil (33.8 g). 

This oil is dissolved in benzene (100 cc) and chromatographed through silica (430 g). After elution with ethyl acetate, there is collected a fraction of 21 liters, which is concentrated to dryness under reduced pressure (20 mm Hg). The crystalline residue (32.5 g) is recrystallized from acetonitrile (100 cc) and a product (16.4 g), MP 94°C, is obtained. On recrystallization from a mixture of benzene (60 cc) and petroleum ether (200 cc), there is finally obtained 2-(3-benzoylphenyl)propionic acid (13.5 g), MP 94°C. 

---